EPA R2-72-110
DECEMBER 1972
Environmental Protection Technology Series
    Oily Waste  Disposal by

    Soil  Cultivation Process
 .588.
                                 Ol
                                 O
                                 Office of Research and Monitoring

                                 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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                                          December  1972
         OILY WASTE DISPOSAL

      BY SOIL CULTIVATION PROCESS
                   By

          C. Buford Kincannon



           Project 12050 EZG


           Project Director

             Leon  H.  Myers
Robert  S.  Kerr Water Research Center
            P. 0.  Box 1198
         Ada, Oklahoma 7^820


             Prepared for

  OFFICE OF RESEARCH AND MONITORING
U.S. ENVIRONMENTAL PROTECTION AGENCY
        WASHINGTON, D.C.  20460
 For sale by the Superintendent of Documents, U.S. Government Printing Onice
             Washington, D.C., 20402 - Price $'J

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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.

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                            ABSTRACT

Disposal of oily sludges by utilizing soil microorganisms to decompose
the oil has been demonstrated at prevailing soil and climatic conditions
at Deer Park, Texas.  The oil decomposition rate was about 0.5 lbs/ft3
of soil per month without fertilizers and about 1.0 lb/ft3/month when
fertilized.  The rate of 1.0 Ib/ft3/month is about 70 bbls/acre/month
using the upper 0.5 foot of soil.  Costs of the soil disposal method,
including fertilizers, were about $7.00/bbl of oil and $3.00/bbl of
sludge containing 33 percent oil.  Major microbiological species active
in the soil were members of the genus Arthrobacter, Corynebacterium,
Flavobacterium, Nocardia, and Pseudomonas.

Differences in decomposition rate and microbial species due to hydro-
carbon type as present in crude, bunker C, and waxy raffinate oils
were minimal.

Infrared and gas chromatography examinations of oil extracted from
fertilized and unfertilized soils showed differences in organic acid
contents and boiling ranges.

Oil and fertilizer chemicals did not infiltrate vertically into the
soil at the test location under prevailing conditions.

Rainfall runoff water contained 1) up to 100 ppm extractable oils found
to be naphthenic acids and 2) up to 150 mg/1 ammonia as N when the
nutrients were excessive in the soil.

This report was submitted in fulfillment of Project Number 12050 EZG,
under the  (partial) sponsorship of the Office  of Research and Monitor-
ing, Environmental  Protection Agency.
                                iii

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                            CONTENTS
Section                                                          Page

I         Conclusions                                              1

II        Recommendations                                          3

III       Introduction                                             5

IV        Oily Sludge                                              7

V         Project Description and Objectives                       9

VI        Soil Test Plots                                         11

VII       Soil Nutrients (Fertilizer)                             15

VIII      Startup, Operation, and Appearance                      17

IX        Sampling and Testing                                    23

X         Effectiveness of the Cultivation Process                27
             Oil Decomposition Rate and Effect of Fertilizer      27
             Metals Contents of Soil                              39
             Oil and Nutrient Infiltration                        39

XI        Quantity and Cost Based Upon Conditions During
          the Project                                             41

XII       Microbial Action                                        43

XIII      Composition of Oil Extracted From the Soil              47
             Hydrocarbon Types                                    47
             Infrared Spectres copy                                47
             Gas Chromatography                                   53

XIV       Rainfall Runoff                                         59

XV        Acknowledgments                                         61

XVI       References                                              63

XVII      Appendices                                              65

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                             FIGURES




                                                                  Page




 1       EXPERIMENTAL PLOTS                                        10




 2       PREPARATION OF TEST PLOTS                                 12




 3       SOIL PLOT IDENTIFICATION MARKERS                          13




 4       SOIL CULTIVATION AND ADDITION OF OIL                      18




 5       BEFORE AND AFTER SECOND ADDITION OF OIL TO "A" PLOTS      19




 6       BEFORE AND AFTER SECOND ADDITION OF OIL TO "B" PLOTS      20




 7       BEFORE AND AFTER SECOND ADDITION OF OIL TO "C" PLOTS      21




 8       SOIL AT END OF THE DEMONSTRATION PERIOD                   22




 9       TYPICAL SAMPLING POINTS                                   25




10       OIL CONTENT OF SOIL, "A" PLOTS                            28




11       OIL CONTENT OF SOIL, "B" PLOTS                            29




12       OIL CONTENT OF SOIL, "C" PLOTS                            30




13       NITROGEN CONTENT OF SOIL, PLOT A-l                        31




14       NITROGEN CONTENT OF SOIL, PLOT A-2                        32




15       PHOSPHORUS CONTENT OF SOIL, "A" PLOTS                     33




16       TEMPERATURE-DECOMPOSITION RATE RELATIONSHIP, "A" PLOTS    35




17       TEMPERATURE-DECOMPOSITION RATE RELATIONSHIP, "B" PLOTS    36




18       TEMPERATURE-DECOMPOSITION RATE RELATIONSHIP, "C" PLOTS    37




19       MICROBIAL CONTENT OF SOIL FROM "A" PLOTS                  42




20       MICROBIAL CONTENT OF SOIL FROM "B" PLOTS                  44




21       MICROBIAL CONTENT OF SOIL FROM "C" PLOTS                  45




22       HYDROCARBON TYPE OF OIL EXTRACTED FROM "A" PLOTS          46




23       HYDROCARBON TYPE OF OIL EXTRACTED FROM "B" PLOTS          48
                                vi

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                        FIGURES (CONTD.)

                                                                  Page

24       HYDROCARBON TYPE OF OIL EXTRACTED FROM "C" PLOTS          49

25       INFRARED SPECTRA OF OILS EXTRACTED FROM "A" PLOTS         50

26       INFRARED SPECTRA OF OILS EXTRACTED FROM "B" PLOTS         51

27       INFRARED SPECTRA OF OILS EXTRACTED FROM "C" PLOTS         52

28       GAS CHROMATOGRAPH AND BOILING POINT PROFILES,
         "A" PLOTS                                                 54

29       GAS CHROMATOGRAPH AND BOILING POINT PROFILES,
         "B" PLOTS                                                 55

30       GAS CHROMATOGRAPH AND BOILING POINT PROFILES,
         "C" PLOTS                                                 56
                                VI1

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                             TABLES




No.                                                                 Page




1        Nutrients Added During the Study                            16




2        Sampling and Testing Frequency                              24




3        Oil Decomposition Rate                                      38




4        Quantity and Cost Based Upon Experimental Conditions        40
                               viii

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                            SECTION I

                           CONCLUSIONS

1.  Disposal of oily sludges (hydrocarbon) by microbial action in culti-
    vated soil has been demonstrated at prevailing soil and climatic
    conditions at Deer Park, Texas.

2.  Three simultaneous experiments with three oils, i.e., crude oil,
    bunker C fuel oil, and waxy raffinate oil, indicated decomposition
    rates for the three oils to be approximately equal and averaged
    about 0.5 pounds of oil per cubic foot per month without adding
    nitrogen and phosphorus nutrients and about 1.0 pound per cubic
    foot per month when fertilizers were added.  This is equivalent
    to about 70 barrels of oil per acre per month.

3.  Cost of the soil cultivation process based on the demonstration
    project expenses and a disposal rate of 70 barrels of oil per acre
    was $7.00 per barrel of oil.  Assuming oily sludges and waste
    materials contain 33 percent oil, the disposal cost by the soil
    cultivation process would be about $3.00 per barrel.

4.  An optimum fertilization program appears to be a) the initial addi-
    tion of chemicals, if needed based upon soil test results, to
    attain a slight excess of nitrogen, potassium, and phosphorus, and
    b) test at regular intervals, once per month, for ammonia and
    nitrate contents of the soil and add small dosages of ammonium
    nitrate as needed to maintain a positive test result (10-50 ppm)
    for ammonium and/or nitrate contents.

5.  The major species of microorganisms present are members of the
    genus Pseudomonas, Flavobacterium, Nocardia, Corynebacterium, and
    Arthrobacter.  The nature of the hydrocarbon substrate did not
    appear to influence the type of organisms present but did affect
    the number of bacteria in the soil.  Crude oil tank bottoms pro-
    duced the highest count, waxy raffinate oil produced an inter-
    mediate count, while bunker C fuel oil exhibited the lowest
    microbial population.  Temperature appeared to have no effect upon
    the microbial count and distribution.  Addition of fertilizer did
    not affect the microorganism distribution but appeared to be
    directly related with the total aerobic count.

6.  Oil decomposition rates were low when the concentration of oil in
    the soil approached the starting condition of 10 percent oil in
    the soil.  Also, the low reaction period coincided with the winter
    months and low temperature period.

7.  Both aromatic and saturated hydrocarbons were reduced with time in
    the soil for crude oil tank bottoms and bunker C fuel oil.  Only
    the saturate fraction of waxy raffinate oil appeared to be reduced

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     by soil microbial action at conditions existing during the project
     period.

 8.  Infrared and gas chromatographic analyses of the oil added to and
     extracted from soil indicated a) the absence of organic acids in
     oils added to the soil and the presence of organic acids in each
     of the extracted oils (oils from plots that were fertilized showed
     higher concentrations of organic acids),  b) the organic acid in-
     crease coincided with a decrease in total saturates, and c) the
     percent weight boiling less than 500°C generally was lower for the
     oil extracted from the soil at the finish of the project than for
     either the oil added to or the oil extracted from the soil at the
     start of'the project (the lowest percent  weight boiling up to 500 C
     was extracted from soil which had received the largest quantity of
     fertilizer materials).

 9.  Oil and fertilizer chemicals did not infiltrate vertically into the
     soil at the test location and condition.

10.  Rainfall runoff water contained 30 to 100 ppm oil.  This oil
     appeared to be essentially naphthenic acids based upon infrared
     inspection of oil fractions.  Also,  rainfall runoff water contained
     ammonia (nitrogen nutrient) approximately proportional to the
     excess ammonia content of the soil.   Phosphorus and-nitrates were
     not found in runoff water.

11.  Oil and nutrient contents of rainfall runoff water from the soil
     cultivation process can be relatively high, and this discharge
     water should receive treatment before entering public waterways.

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                           SECTION II

                         RECOMMENDATIONS

This project demonstrated that soil microorganisms decompose petroleum
oily waste and that oil and fertilizer chemicals did not penetrate the
soil at the location and conditions of the test.  However, numerous
factors are left unresolved and future experiments should gain
knowledge concerning the following uncertainties:

1.  The residual oil in the soil after the 18-month project contained
    organic acids, and those soluble in rainfall runoff water were
    primarily naphthenic acids.  Since most organic acids are water
    soluble, this suggests that naphthenic acids do not decompose in
    the soil as rapidly as aliphatic acids or that naphthenic acids
    are formed from aliphatic or aromatic materials at the soil condi-
    tions.  Speculatively, large concentrations of naphthenic acids
    may inhibit microbial growth in the soil, and water washing of
    these organic acids from the soil to a separate biotreatment
    system might improve the oil decomposition rate in the soil.

2.  Residual oil extracted from the soil was characterized by infrared
    to be polyaromatic oils, suggesting this hydrocarbon group to be
    slow reaction or nonreactive for microbial decomposition at the
    prevailing conditions.  Certain, and yet unknown, environmental
    conditions might improve the decomposition rate of the poly-
    aromatics.

3.  Accumulation and buildup of metals or salts contents with long-
    range usage perhaps affects the microorganism activity.  The
    acidity or alkalinity range, optimum for microbial action, may
    vary with the soil contents.

For application of the process, the following items must be considered:

1.  The time required for native soil bacteria to become acclimated
    oil decomposers likely depends upon the soil composition and
    temperature.

2.  Since soil profiles vary, tjhe oil penetration rate into soil needs
    to be determined at the location where the process is to be used.

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                           SECTION III

                           INTRODUCTION

Petroleum crude oil, fuels, and lubricants become waste components
when emulsified with water, solids, and/or debris and are potential
contaminants of surface and subsurface waters.  The volume of oily
waste is potentially large due to large quantities of petroleum oils
handled and used.  Sources of the waste are from spillage of crude
oil and refined products and from petroleum refinery operations.
This paper deals with the refinery operation, but the findings are
expected to be applicable for most petroleum sludges.

When large quantities of oil are accidentally spilled on seawater,
inland water, or land, much of the oil can often be recovered, but some
emulsified waste oils are formed.  Removal and disposal of these oily
wastes are separate problems for each accident.  Also, refining of
petroleum crude oil into fuels, solvents, waxes, and asphalts results
in small quantities of oily waste materials.  These oily sludges
obtained on an infrequent basis may be accumulated in a temporary
storage vessel or disposed of on a more or less continuous basis.
Waste oils from refinery processes and from oil spills are similar
in composition.

One method for disposal of oily sludges has been by mixing with the
soil and utilizing soil microorganisms to decompose the oil.  Applica-
tion of this method (referred to also as land spreading operation) at
several refineries has been reported,  and a modification of the method
was used during the Santa Barbara oil spill.2  Wet oil-straw-sand
mixture removed from the beaches was disposed of as solid waste in a
land fill, and presumably the straw and oil decomposed by microbial
action.  Many investigations have shown that some microorganisms
normally present in the soil will attack petroleum hydrocarbons and
utilize them as their sole source of carbon.   The reaction is
dependent upon many interrelated conditions, which have not yet been
fully understood, such as temperature, moisture, soil properties
(physical and chemical, including nutrient content), oil content and
properties, microbial content, acclimation period, and the availability
of oxygen and nutrients.  Poorly aerated soils become anaerobic and
under these conditions the microorganisms decompose organic matter
very slowly.  Aeration of the soil by frequent cultivation is a means
of supplying oxygen essential for the more rapid acting aerobic
microbes.

Although the land spreading or fill method for disposal of oily
sludges has been used at some locations, knowledge of the process is
limited to a few trial-and-error applications.  An exploratory study
of hydrocarbon decomposition rates was made by the Shell Oil Company
for their disposal area at the Houston, Texas, refinery.  Also, spot
samples of their soil were analyzed  for microorganism contents by

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Environmental Protection Agency, National Environmental Research
Center, Cincinnati, Ohio.  Results of the preliminary hydrocarbon
decomposition rate study and microbial counts were reported by
Dotson, Dean, Kenner, and Cooke.*  Sufficient data to permit
evaluation of the process were not obtained and are not available
from other sources.  This paper discusses a further study designed
to demonstrate the effectiveness and cost of the soil cultivation
process for disposal of oily waste from petroleum.

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                           SECTION IV

                           OILY SLUDGE

Almost all operations of the petroleum industry, including exploring,
producing (extracting), storing, transporting, and refining of crude
oil and the storing, distributing, and handling of products are
potential sources of oily sludges.

Accidental spills of crude oil and petroleum products during the hand-
ling, storing, and transporting operations are the principal cause for
the formation of oily sludges in large quantities.  At refineries,
accidents seldom cause oily sludge production.  Common sources are
incoming crude oil, ship ballast water, tank and vessel cleanings,
oil-water separators, and numerous miscellaneous sources such as sewer
boxes and emulsion breaking facilities (demulsifier).  The quantity of
oily sludge for disposal at a refinery does not depend only upon the
nature of the crude and processing units.  Oily sludge formation can
be minimized by prudent operating practices, sensitive attitudes and
suitable control methods.  Generally, most of the oily sludges accumu-
late in oil-water separators and in tank bottoms.

Crude oil shipped to the refinery contains emulsified material which is
commonly referred to as bulk, sediment, and water (BS&W).  The BS&W
concentration in the crude oil ranges from about 0.01 to 0.1 percent
by volume (%v).  This BS&W fraction contains about 307<,v oil, 50%v
water, 10%v carbonaceous matter, and inorganic salts equivalent to
about 10%v.  The carbonaceous material which is the part of BS&W that
could eventually become oily sludge for disposal, amounts to about
0.005 percent of the incoming crude oil to the refinery.  However, in
the refining steps, much of this carbonaceous material is utilized
into fuels.  The quantity of waste sludge from crude oil is small for
the following reasons:  1) Crude oil receipts at a refinery are often
stored in tanks for a few days before being fed to processing units.
During quiescent periods, most of the BS&W material settles out as
tank bottoms and is removed during tank cleanings at three to five-
year intervals.  Fortunately, tank bottoms have been in demand for oil
and wax recoveries by independent operators and for road bed additives.
When crude tank bottoms must be handled at the refinery, a concentra-
tion step is possible with conventional slop oil demulsifying
facilities.  The BS&W emulsion is broken and some of the carbonaceous
material is included with the recovered oil phase and some with the
water phase.  The recovered oil phase is fed to a refining unit and
the water phase flows into an oil-water separator.  2) In the event a
settling time is not possible, the crude oil containing BS&W material
is fed to a crude distilling unit.  The BS&W material is removed in
the pretreatment (heater and desalter) section of the distilling unit.
The oil fraction of the BS&W remains in the crude.  Water fraction in
the desalter effluent flows into the feedwater stream of an oil-water
separator.  Emulsified materials in the oil-water separator will either

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enter the recovered oil stream, settle with the bottom sludge inside
the separator box, or remain in the water phase flowing to secondary
treating facilities.

Ship ballast water-handling facilities vary at different refineries as
does the quantity and quality of both the water and oil phases of the
incoming ballast water.  At Shell Oil Company's Houston Refinery, bal-
last water from docked ships is pumped into an American Petroleum
Institute (API) design ballast tank, and the water drained from this
ballast tank flows by gravity into a holding pond equipped with down-
stream oil retention baffles and skimmers.  Incoming ballast water
sometimes contains oily material which has a density near that of water
and will pass through the ballast tank into the holding pond.  If heavy
oil or emulsion material remains in the holding pond for a few days,
the physical characteristics change from a fluid brownish-black mixture
to a thick congealed mass (sometimes greenish in color) which will
float on the water but will not flow.  This change apparently is caused
by microorganisms.  The congealed mass containing oil may be treated
in a demulsifying unit to recover the oil.  If judged to be low in oil.
content, it may become a part of the oily sludge which is a potential
feed for the soil cultivation process.  The frequency and quality of
oily waste from the ship ballast water source are uncontrolled and
unknown.

Tank bottoms obtained during cleaning of tanks vary in composition and
the residual oil content can usually be recovered or at least reduced
with emulsion breaking facilities.  However, the real bottoms and final
washings from a tank are often oily sludges not suitable for feed to a
demulsifier unit and are potential feed for the soil cultivation pro-
cess.

Oily sludge from oil-water separator and holding pond cleanings is
usually low in oil content and is suitable for disposal by means such
as the cultivation process.  The sludge cleanings from the water box
or pond usually contains about two percent carbonaceous material.  Oil
box cleanings which are mostly oil can be fed to the demulsifying unit,
but sometimes mixtures of oil, straw, grass, and dirt make this mate-
rial suitable only for disposal.

Solids and oily waste at the demulsifying unit usually accumulate in
tanks which must be cleaned occasionally, and these cleanings cannot
be further improved with additional treatment in demulsification
facilities.  Disposal of this oily-sand material can be accomplished
by the soil cultivation process.

Process unit shutdowns include complete cleaning of a piece of equip-
ment and/or vessel before maintenance work begins.  In the shutting
down of a process unit, oil is returned to storage.  Immediately after
shutdown and where possible, the equipment is washed with water which
removes residual traces of oil and carbonaceous material.  This material
is sent to an oil sewer which connects to an oil-water separator.

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                            SECTION V

               PROJECT DESCRIPTION AND OBJECTIVES

The soil cultivation process consists of the treatment of oily waste
material by spreading and cultivating into soil under prevailing
climatic conditions.  The project includes three parallel experiments.
Oily feed materials were selected to represent different combinations
of hydrocarbon types.  These were designated as sludge A, sludge B,
and sludge C.

Sludge A was a crude oil tank bottoms which contained a natural balance
of hydrocarbon types.

Sludge B was a high molecular weight fuel oil (bunker C or No. 6) con-
taining olefinic and aromatic components.

Sludge C was a waxy  raffinate, which is an intermediate waxy oil
product containing highly paraffinic components.

The properties of the three oily materials (simulated sludges) used in
the study are given  in Appendix A.  Sludge A was added to each of the
three separate soil  test plots designated A-l, A-2, and A-3.  These
test plots were treated identically except for soil nutrient additions.
A-l was fertilized heavily, A-2 received a moderate or intermediate
quantity of fertilizer material, and A-3 was the control with zero
addition of fertilizer.  Sludge B was added to three separate plots
also, and these were designated B-l, B-2, and B-3.  Likewise for sludge
C, experimental plots were C-l, C-2, and C-3.  Again, the No. 1 plots
were fertilized heavily, No.  2 plots received intermediate quantity of
fertilizers, and No. 3 plots were not fertilized.  The design and
details of the project are discussed separately.

Objectives of the project were to determine:

1.  The decomposition rate of oily waste sludges in cultivated soil.

2.  The effectiveness of adding nutrient supplements.

3.  Major microbiological species active in soil where oily wastes
    are decomposed.

4.  The cost of the  process for disposal of oily waste.

5.  If the oil infiltrates vertically into the soil at the test site
    and the depth of penetration.

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                   FIGURE 1
DIMENSIONS OF EACH PLOT

  WIDTH, FT        12
  LENGTH, FT      125
  AREA, FT2      1500
  DEPTH, FT       0.5
  VOLUME, FT3     750
HOLDING

 POND
^
/-RAINFALL RUNOFF CONTROL
r
x
• •,


A-l



t





\-2


A- 2
-DITCH

t





J-l


B-2
1


S






B-3


C-l
/



t


C-2
1=

C-3

1
i
A

i-r/
k


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                           SECTION VI

                         SOIL TEST PLOTS

Nine soil test plot areas were needed to conduct three parallel experi-
ments with sludges A, B, and C.  The layout of plots A-l, A-2, A-3, B-l,
B-2, B-3, C-l, C-2, and C-3 is shown in Figure 1.  Each of the nine
plots was 12 feet in width and 125 feet in length (1500 square foot
area).  The plots were located near the Houston Ship Channel within the
Shell Oil Company refinery at Deer Park, Texas.

The plots were separated by levees and were designed so that rainfall
runoff from each plot drained through a pipe to a ditch.  A plug in the
pipe was used to control the runoff.  The levees prevented crossflow of
water from one plot to another and kept oil within the plot area.  Ex-
cess runoff water flowed from the ditch into a holding pond (Figure 1)
and was either discharged to the Ship Channel or transferred by vacuum
truck to the refinery biological treating facilities.

Space between the plots permitted tank truck passageway adjacent to
each plot for convenience in spreading oily sludge evenly over the
surface.  Also, the plots were cultivated separately.  Before moving
from one plot to another, the space between plots was cultivated to
clean the plow and prevent mixing of oil, soil, and fertilizers from
one plot to another.

The experimental plots are located in an area which had been used
previously for oily waste disposal.  This offered the advantage of
having a start of oil-consuming variety of microorganisms in the soil
without an acclimation period.  However, a disturbing disadvantage as
far as the •experimental work was concerned was the presence of residual
oil in the soil at the start of the experiment.  Initial leveling and
grading of the area and preparation of the levees to separate each plot
area are shown in Figure 2.  Identification markers were installed as
shown in Figure 3.

The texture of top soil sample from the plot area was 58 percent sand,
14 percent silt, and 28 percent clay.  The soil textural classification
was sandy clay loam.  At a depth of two feet from the surface, the soil
was classified as loam and at  four-foot depth was sand.

The bulk density of oil-free top soil was 1.6 or equivalent to approxi-
mately 100 pounds per cubic foot.  The cation exchange capacity at the
start was about 10 milliequivalents per 100 grams (me/lOOg) of soil
and at the end of the experiment ranged from 30 to 60 me/lOOg.   (This
increase in exchange capacity  is larger than expected from the effects
of adding fertilizers and oil materials.  The sample taken at the start
of the project may not have been representative as the samples at the
close of the experiment.)  Conductivity was 13 micromhos per centimeter
(Mmhos/cm) at the beginning and ranged from about 4 to 6 Mmhos/cm at
                                11

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                 FIGURE 2
          PREPARATION OF TEST  PLOTS
            LEVELING AND  GRADING
PREPARATION OF LEVEES TO SEPARATE PLOT AREAS
                      12

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                   FIGURE 3
       SOIL PLOT IDENTIFICATION MARKERS
                                         ••

             SIGN AT  PROJECT  SITE
            -
.
,
               -
 1   • •'• 0   ! •  HI   •     • -  •     •

I .
                                 :-  .  — --.p>
      IDENTIFICATION MARKER  FOR  EACH  PLOT
                       13

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the close of the study.  Conductivity measurements were made at room
temperature (20-22°C).  Other properties (including pH, oil, nitrogen,
phosphorus, sulfur and metals contents) of the soil at the beginning
of the project are tabulated and discussed along with results of the
study.
                                14

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                           SECTION VII

                   SOIL NUTRIENTS (FERTILIZER)

Since the disposal of oily waste by the soil cultivation process depends
upon the soil microorganisms to oxidize the oils or convert the oily
waste into cell protoplasm, the ingredients needed for microbial growth
must be present.  The main elements are carbon, hydrogen, oxygen,
nitrogen, potassium, and phosphorus.  Also, key trace elements includ-
ing sulfur, sodium, calcium, magnesium, iron, and others are needed.
Most of the trace elements are abundant in soils, but two of the main
elements, nitrogen and phosphorus, often limit organic cellular growth.
Chemical analyses of protoplasm indicate phosphorus requirement is
about one-fifth the nitrogen requirement.1*

In agricultural crop production, nitrogen and phosphorus are removed
from the soil with the harvested grain or forage.  Replenishing of the
nutrients is required.  For a waste disposal system such as the soil
cultivation process, there should be an equilibrium condition estab-
lished which would minimize the need for supplementing the nutrients.
Endogenous metabolism, autoxidation of cellular protoplasm, results in
the release of nitrogen and phosphorus previously used for synthesis.
The released nutrients are made available for reuse in biological waste
treatment systems.1*  No reference information was found to use as a
guide in selection of appropriate quantities of nitrogen and phosphorus
which should be added to the biological system.  Instead, the selection
of fertilizer quantities was based upon agricultural-oriented expe-
rience.  Grass land is frequently fertilized with 500 pounds of
nitrogen and 100 pounds of phosphorus per acre, and three times these
quantities have been used according to our consultants.  Warnings that
1) excess nitrogen fertilizer elements hinder (poison) bacterial action
and 2) excessive total soluble salts may cause unfavorable osmotic
conditions for bacterial growth, were considered in the selection of
the quantities of nutrient additive.  Excess phosphate was not con-
sidered toxic except for its contribution to the total soluble salt
content.  Initial additions in pounds per acre  (Ibs/ac) of nitrogen
(urea) as N were 1000, 500, and zero to plots 1, 2, and 3, respectively.
The additions of phosphorus (calcium hydrogen phosphate) were 200, 100,
and zero Ibs/ac as P20s to plots 1, 2, and 3, respectively.

Periodic analyses of nitrogen and phosphorus were made, and additional
fertilizer materials were added to plots 1 and 2 during the course of
the experiment.  The types and quantities of fertilizer materials
added during the study are shown in Table 1.
                                15

-------
                                                               TABLE 1

                                                  NUTRIENTS ADDED DURING THE STUDY
                                              (The Number 3 Plots Were Not Fertilized)
                                                           1970
FERTILIZER
Urea, as N
Ammonium
 Nitrate, as N
                    RATIO
                    N-P-K

                    45-0-0
35-0-0
Phosphorus,
 as P00C            0-46-0
Potash, as K-0      0-0-60



Ib/plot
Ib/acre
ppm by wt
Ib/plot
Ib/acre
ppm by wt
Ib/plot
Ib/acre
ppm by wt
Ib/plot
Ib/acre
ppm by wt
MAY 7
PLOTS NO.
1 2
34 17
1000 500
450 225



7 3.5
200 100
95 47



AUGUST 7
PLOTS NO
1
58
1700
775



184
5350
2450




.
2
23
680
305



46
1340
610



                                                                                                          1971
                                                                JAN. 27
                                                                PLOTS NO.
                  APRIL 21
                  PLOTS NO.
                         AUGUST 11
                         PLOTS  NO.
  35
1000
 470
 17
500
225
  9
260
120
  4.5
130
 60
 12
350
160
  6
175
 80
                                                                 30
                                                                875
                                                                400
           15
          438
          200

-------
                          SECTION VIII

                STARTUP, OPERATION, AND APPEARANCE

After the plots had been laid out and the levees prepared, each plot
was cultivated with a "roto-tiller" type plow.  Fertilizer materials
were added by manually spreading the solid granular fertilizer over
the surface of plots No. 1 and 2.  After spreading fertilizer and be-
fore adding oily sludge materials, the plots were cultivated a second
time to mix and distribute the fertilizer salts with the soil.

Simulated oily sludges were transported to the test plot area with a
vacuum tank truck, and the sludge was distributed over the surface of
the soil by manual direction of a discharge hose attached to the tank.
Soil cultivation and addition of oil are shown in Figure 4.  Mixing of
the oil and the soil by cultivation appeared uniform and presented no
problem at the ambient May temperature of about 80°F.

The initial schedule for cultivating the soil plots was once each two
weeks, weather permitting.  However, after the first quarter, the plow-
ing frequency was increased in an attempt to improve oil decomposition
rates.  The plowing frequencies and results are discussed separately.
After about six months of cultivation, the soil was friable, had lost
the oily appearance, and was judged from test results to be suitable
for a second addition of the oily sludge.  Some differences in the
plots were obvious.  For instance, the color of the No. 3 plots was
darker than that for Nos. 1 and 2.  Photographs of these plots just
before and just after the addition of the second application of the
simulated oily sludges are shown for the "A" plots in Figure 5, for
the "B" plots in Figure 6, and for the "C" piots in Fig'ure 7.  The
date for this second" addition of sludge was in early February, and
the temperature was in the low 40's  F.  Congealing and solidification
of the sludges were apparent, and mixing of the viscous oily matter
into the soil was not successful until the ambient temperature had
increased to about 80°F.

At the close of the 18-month experiment, the soil had again returned
to a friable condition with the appearance of a normal agricultural
soil.  A close-up view of soil in plots A-l, B-l, and C-l is shown in
Figure 8.

Although no attempt was made to grow vegetation matter on soils
previously used for oil decomposition during this experiment, native
grass and plants did sprout and grow on top of the levee.  The soil
in the levees was the same as the starting test plots, which contained
about ten percent oily material as discussed later.
                                 17

-------
              FIGURE 4

SOIL CULTIVATION AND ADDITION OF OIL
                  18

-------
                      FIGURE 5

BEFORE AND AFTER SECOND ADDITION  OF OIL TO  "A" PLOTS
                         19

-------
                       FIGURE 6
BEFORE AND AFTER SECOND ADDITION  OF OIL TO "B"  PLOTS
                           20

-------
                      FIGURE 7


BEFORE AND AFTER SECOND ADDITION OF OIL TO  "C"  PLOTS
                          21

-------
                           FIGURE 8
:;; -  /> '-^vl^^V'V*«*ivl\.v^-'^'^'"?-v'>:'','';r..         ^•:M&''^L;'*r« '•
%•-... .\>< •' v -iT***-'' »<*- ' i.  ''•*'.. ,":?•:_: •:'•  \-   *.•*.-•:•''<  -.,•' .    *?,-^
:** •   -fc . \ .    . *W'-ti»  - .vK*--* v 4 . ' ^- .. •. l.i£. •'--•. t*M^S.\*W **»...••    ..  - . .    -*-.*;
                                                 •:V  !,    %*j£.i

                                                 W$*  :^i
                                                 • rr'/'atf'-'t'*., .,v;. " ••• . ;..f
 •-,,-'-'-'>f ^^^CfrSWWX^S&W^ '•"• '•  -^3^". -''-  'i-  -"  i'./**i •'  "   '•  ''•'
 '- v'         ;


                           .,••   	
                           ^ -•-  • •     ••-

          SOIL  AT END OF  THE  DEMONSTRATION  PERIOD
                                 22

-------
                           SECTION IX

                      SAMPLING AND TESTING

Oil and nutrient contents were used as controls for the experimental
work during the 18-month project, and these components were tested
frequently.  (Microbiological analyses, hydrocarbon types of oil added
to and extracted from the soil and the oil content of soil core samples
at depths up to six feet were made to achieve the project objectives.
Also, metals, nitrogen, phosphorus, and sulfur contents of the soil
and infrared absorption and gas chromatography of extracted oils were
examined.)  The samples for analyses, analytical tests, method
references, and the frequency of tests during the experimental period
are summarized in Table 2.  In addition, moisture, pH and temperature
of the soil were measured on a regular basis.  Moisture contents of
the soil were obtained along with the oil content determinations as
described in Appendix E.  Temperature and pH were measured using
mercury thermometers and a glass-calomel electrode meter.  The soluble
nutrients were dissolved from oil-free soil (after CCli^ extraction)
using distilled deionized water  for nitrogen and 1.4 normal (1.4 N)
ammonium acetate (NH^Ac) in hydrochloric acid (HCl), pH 4.2, for
phosphorus (P), sodium (Na), potassium (K), magnesium  (Mg) and calcium
(Ca), according to the procedure recommended by State Soil Testing
Laboratories publication.^  For the  less soluble metals, zinc (Zn),
manganese (Mn), copper (Cu), iron (Fe), and lead (Pb), a 50/50 mixture
of concentrated HCl and concentrated  nitric acid (HN03) was used for
the extraction solvent.  Analyses of  the solutions were by atomic ab-
sorption.  The soil consultant used 1.0 N NHi|Ac at pH 7 for extracting
P, K, Na, Ca, and Mg, and 0.1 N HCl for extracting Zn, Mn, Cu, and Fe.
These latter extraction solvents are  generally considered a good
measure of the quantities of elements available for growth of plant
life.  Rainfall runoff water was tested for oil and nutrient contents.
The method used for oil in water determinations was American Petroleum
Institute (API) method 731.llt  Oxygen determinations of the soil,
although desirable, were not made because a suitable method was not
available.

The obtaining of a representative soil sample was a recognized problem
from the start of the project.  Although cultivation mixed the surface
soil vertically, complete horizontal  and vertical mixing throughout a
test plot was not attained.  In an effort to minimize inconsistencies,
samples were taken immediately after  cultivation and from several loca-
tions, then composited before submitting for tests.  The sampling
procedure included combining portions of soil from three sampling
points to form subsamples a, b, and c, as shown in Figure 9.  Each
subsample was analyzed separately for oil content, and the resulting
data for the 18-month experimental period are given in Appendix A.
For all other tests, including nutrients, microbial and metals con-
tents, and other properties of the soil, subsamples a, b, and c were
                                 23

-------
                                                                        TABLE 2

                                                            SAMPLING AND TESTING FREQUENCY
                         Sample
               Soil, top 6 inches
to
               Soil core, 2, A, & 6 ft.
               Oil extracted from soil
               Oil added to soil
	Analytical Test

Oil content

Nutrients (NH3, NO,,
  P205)

Total nitrogen, phos-
  phorus and sulfur

Microbial content

Metals

Oil content

Nutrients (NH,, NO,,
  P2o5)

Hydrocarbon type

Infrared spectroscopy

Gas chromatography

Physical properties

Metals

Hydrocarbon type

Total nitrogen, phos-
  phorus and sulfur

Infrared spectroscopy

Gas chromatograph
                                                                                            Reference
Appendix E
                                                                                              8
                                                                                                    5,10
Standard methods


Specific methods11'12'13


Appendix B

Atomic absorption
          8
Appendix E

Standard methods '


ASTM D-20079

Appendix D

Shell Oil Co.7

ASTM methods

Emission Spectroscopy

ASTM D-20079
                                                                                    Specific methods
                                                                                    Appendix D

                                                                                    Shell Oil Co.'
                                                                                                    11,12,13
   Approximate Frequency

Bi-weekly

Monthly or bi-weekly
  if needed

Beginning and end
Monthly

Beginning, midway, and end

Beginning, midway, and end

Midway and end


Beginning, midway, and end

Beginning, midway, and end

Beginning and end

Beginning

Beginning

When added

Beginning


Beginning

Beginning

-------
            FIGURE 9
o o  o
000
o o  o
             TEST PLOT 125 FT. X 12 FT.

             PORTIONS FROM THREE SAMPLING
             POINTS WERE COMPOSITED TO
             FORM EACH SUB SAMPLE
             SUB SAMPLE c
             SUB SAMPLE b
             SUB SAMPLE a
     TYPICAL SAMPLING POINTS
             25

-------
composited to represent the whole test plot.  For example, A-l soil
sample was a composite of A-la, A-lb, and A-lc subsamples.

Analytical results are given in Appendices A, B, C, and D.
                                26

-------
                            SECTION X

          EFFECTIVENESS OF THE SOIL CULTIVATION PROCESS

Oil Decomposition Rate and Effect of Fertilizer

Percent weight oil in each soil plot given in Tables 5, 6, and 7 of
Appendix A are shown graphically related with time in Figures 10, 11,
and 12.  Residual oil of about ten percent in the soil was due to
previous use of the'soil area for oily waste disposal as mentioned
earlier.  The increased quantities of oil in all plots for May 1970
and February 1971 were due to the addition of oil to the soil.  Devia-
tion in the oil content data from the linear relationships shown on
each of the figures may indicate more rapid reaction during some
periods than others but is likely due to nonrepresentative samples.
The straight line relationship was calculated by the method of least
squares.  For the "B" plots, Figure 11, inconsistent data during
February to April, 1971, were due to the inability to mix the highly
viscous oil into cool soil with the roto-tiller type cultivator.

Generally and as shown in each of Figures 10, 11, and 12, from the
first addition of oil in May 1970 to about November 1970, the oil con-
tent of the soil decreased markedly.  During the period November 1970
to February 1971, a reduction of oil was not apparent.  From February,
when a second dosage of oil was added, to October 1971, the oil content
again decreased.  The reason (or reasons) for the period of inactivity
is not clearly evident.  During this period, food supply may have been
limiting because the concentration of oil in the soil was in some cases
near the starting content, and this residual oil may be nonreactive or
slowly reactive with soil microorganisms.  Another reason may have
been the lack of available nutrients which ranged from about 10 to 50
ppm ammonia as N during this period.  A third reason may have been
temperatures that were too low for favorable bacterial growth.

The soluble nitrogen (N) and phosphorus (as P205) contents from Table 5
of Appendix A for the "A" plots during the project are shown in Figures
13, 14, and 15.  The trend of these nutrient data for the "B" and "C"
plots is similar and is not shown graphically.  Urea was used as the
nitrogen nutrient at the start of the project as recommended by a soil
consultant.  The reason for selecting urea was to avoid increasing the
salt content of the soil.  Immediately after fertilizing in May 1970
(Figure 13), the ammonia (NHs) content was about 700 ppm by weight as
N in the soil.  This was likely due to inadequate mixing or non-
representative sample because the theoretical amount added was about
460 ppm as N (Table 1).  The concentration rapidly decreased to about
30 ppm as N during June and to zero in July.  A larger dosage was
added in August causing the NHs concentration to reach about 1500 ppm
as N.  During September and October, the NHs content remained about
500 ppm as N.  During this same period, the nitrate (NOs) content was
zero and ammonia was being lost to the atmosphere.  The odor of ammonia
                                 27

-------
       20
       10
o
t/0



5
       20
O
DC:
       20
       10
                          A
                                           FIGURE    10
                                                                                       A-l
         Jan. Feb. Hir. Apr. Niy June July Aug. Sept. Oct. Nov. Dec. Jin. Feb. Kir. Apr. May June July Aug. Sept. Oct. Nov. Dec.





                        TEAR OF 1970                                  »EA8 OF 1971
                                     OIL  CONTENT  OF  SOIL
                                            'A"  PLOTS
                                             28

-------
                                            FIGURE    11
        30
        20
        10
o
oo

z      30

>—i OO
   •—I
_J 00


5 co   20
C5 O
i—i OO
   a:
i— Q
        10
       30
       20
       10 '
         Jan. Feb. Mir. Apr. Kty June July Aug.  Sept. Oct.  Nov. Dec. J«n. Feb. Kjr. Apr. tajr June July Auq. Sept. Oct. Nov. Dec




                        TEAR OF  1970                                  TEAR OF 1971
                                      OIL  CONTENT OF  SOIL
                                             •B"  PLOTS
                                              29

-------
       2C
       10
o
1/1
       20
       10
o:
LU
a.
       20
       10
                                            FIGURE   12
                                                  A A
         Jan.  Fel). Mar. Apr. Hiy June July »U9. Sept. Oct.  Nov. Dec. Jan. Feb. Kjr. Apr. Hsy June July Aug. Sept. Oct. Nov.  Dec.




                        YEAR OF  1970                                  »EAR OF 1971
                                      OIL  CONTENT OF  SOIL
                                            'C"  PLOTS
                                               30

-------
                                            FIGURE    13
    lOOOr
IT)
Q.
Q.
O
O
     200.
                                                                            PLOT A-l
       Dec. Jin. Feb. Mar. Apr. Kay June July Aug. Sept. Oct. Nov. Dec. Jin. Feb. Kir. Apr. Hay June July Aug. Sept. Oct. Nov. Dec



                         TEAR OF 1»70                                 VEAR OF 1971
                                  NITROGEN  CONTENT  OF  SOIL
                                             PLOT  A-l
                                            31

-------
    1000
     800
     600
o.
CL
C5
O
o:
«   400
     200
                                           FIGURE   14
                                                                           PLOT A-2
       Dec. J«n. Frt. Kir. Apr. Miy June July Aug. Sept. Oct. Nov. Dec. Jan. Fib. Kir. Apr. M«y June July Aug. Sept. Oct. Nov. Dec.




                         TEA* OF  1970                                YEAR OF U71
                                 NITROGEN  CONTENT  OF SOIL
                                             PLOT A-2
                                              32

-------
                                             FIGURE   15
    lOOOr
     800
o
 
-------
was strong in the test plot area during cultivation periods.  When this
was observed, the frequency of cultivations was increased from once per
two weeks to once per week or more (Appendix A, Table 5).  The next
addition was in January 1971, and ammonium nitrate (NH* NC>3) was used
instead of urea since the presence of nitrate was desirable according
to our microbiology consultant.  Also, a dosage of potash was recom-
mended and added at this time to eliminate the possibility of a
potassium deficiency.  The use of NH^NC^ and relatively small dosages
applied more frequently (about once per three months) was practiced
for the remainder of the experiment.  The lower concentrations of NHg
for the No. 2 plot shown in Figure 14 did not emit a detectable odor,
but the cultivation frequency was the same for all test plots including
the unfertilized No. 3 plot.  Soluble phosphorus (phosphate expressed
as P20s) contents of the soil, shown in Figure 15, did not appear to
be a limiting factor.  During June and July 1970, the test plots
treated with bunker C (No. 6) fuel oil appeared to be repelling water.
Exploratory tests indicated additional phosphorus fertilizer improved
the wetting characteristic of the oily soil.  Again, with the advice
of a consultant, relatively large quantities (2500 ppm as ^2^5 to tne
No. 1 plots and 600 ppm as T?2®5 to tne No. 2 plots) of phosphorus
fertilizer were added.  Differences in oil decomposition rates for the
No. 1 and No. 2 plots are likely due to the fluctuating nitrogen con-
tents rather than to the relatively constant phosphorus concentrations.
The effects of fertilizer on the oil decomposition rate as related with
temperature are shown in Figures 16, 17, and 18 for plots "A", "B", and
"C", respectively.

The pounds of oil removed per cubic foot of soil (based upon a soil
weight of 100 lbs/ft3) during the project are shown to be about 15
pounds for A-l and A-2, which had been fertilized, and about 10 pounds
for the A-3 plot which was not fertilized (Figure 16).  Oil removal
rates for the "B" plots (Figure 17) were about 25, 20, and 12 lbs/ft3
of soil for B-l, B-2, and B-3, respectively, and for the "C" plots
(Figure 18) were about 25, 20, and 10 lbs/ft3 for C-l, C-2, and C-3,
respectively.  These data indicate a direct relationship between the
quantity of fertilizer added and oil removal rate.  Each of these
plots show the oil removal rate to be minimal during the winter months
when the temperature was below about 70°F (20°C).  However, as dis-
cussed earlier, the concentration of oil in the soil was near the
starting oil content during this same period, and the apparent effect
of temperature may have been coincidental.

The oil decomposition rate expressed in pounds of oil per month per
cubic foot of soil are given in Table 3.  The decomposition rate for
the periods May to November 1970, and February to October 1971,
averaged 0.90, 0.94, and 0.71 lbs/ft3/mo for A-l, A-2, and A-3; 1.79,
1.67, and 0.83 lbs/ft3/mo for B-l, B-2, and B-3; and 1.73, 1.25, and
0.67 lbs/ft3/mo for C-l, C-2, and C-3, respectively.  For the three-
month period of November 1970 to February 1971, the oil decomposition
rate was minimum.  The average decomposition rate per year at these
                                34

-------
                             FIGURE   16
LU
     35
     30
     25
     20
     15
     10
        90
        80
        70
        60
50
LEGEND:

  A-2  x	x
  A-3 A—-A
  AMBIENT   •
Q  o
UJ  10
LU
O
          Apr. H«jr June July Au?. Sept. Oct. Nov. D«. Jan. Feb. Kar. Apr. Hay June July Au9 Seot. Oct. "iov Dec
              TEM OF 1970
                                                   TEAR OF 1971
              TEMPERATURE-DECOMPOSITION  RATE RELATIONSHIP
                                 II AH
                                  A"  PLOTS
                                   35

-------
                             FIGURE   17
     °c


     35



     30



     25



     20



     15


     10
90
80
70
60
50.
Q  o
UJ  00
UJ
O  co
        20
         10
         '"
         0
                                                 LEGEND:
                                                   B-l    .—
                                                   B-2   X—
                                                   B-3   A—
                                                   AMBIENT I
                                                       —x
                                                       --&
          Apr. mj June July Aug. Sept. Oct. No». Dec. Jan. Feb. Kar. «pr. HJy June July Aug. Seot. Oct. Hoi. Dec

              TIM OF 1)70                            TUR OF 1971



              TEMPERATURE-DECOMPOSITION  RATE  RELATIONSHIP



                                 "B"  PLOTS
                                    36

-------
                             FIGURE   18
UJ
Q.
°c


36



30



25



20



15



10
        90
        80
70
        60
        50.
                                              LEGEND:
                                                C-l   •—
                                                C-2   x—
                                                C-3   A—
                                                AMBIENT
   o
   (75
£ °
0£. m
        2°
         10
         0
          •or. my June July Aug. Sect. Oct. Nov. Dec. Jin. Feb  **r. »pr. Kay Jun« July «u9. Sect. Oct. lo»  On
             n« or
                                                 run OF
             TEMPERATURE-DECOMPOSITION  RATE  RELATIONSHIP
                                "C"  PLOTS
                                  37

-------
                  TABLE 3

            OIL DECOMPOSITION RATE
POUNDS OF OIL PER MONTH PER CUBIC FOOT OF SOIL
Plot
A-l
A-2
A-3
B-l
B-2
B-3
C-l
C-2
C-3
1
2
3
May to
Nov. 1970
0.67
0.50
0.17
1.83
1.83
1.16
1.83
1.50
0.83



Feb. to
Oct. 1971
1.12
1.37
1.25
1.75
1.50
0.5
1.62
1.0
0.50



Average
0.90
0.94
0.71
1.79
1.67
0.83
1.73
1.25
0.67



Nov. 1970
to
Feb. 1971
Minimum
Minimum
Minimum
Minimum
Minimum
Minimum
Minimum
Minimum
Minimum



Average
Per Year
0.67
0.70
0.53
1.34
1.25
0.62
1.30
0.94
0.50
1.10
0.96
0.55
                     38

-------
test conditions was 1.10, 0.96, and 0.5 Ibs/ft3/mo for heavily ferti-
lized No. 1, medium fertilized No. 2, and unfertilized No. 3 plots,
respectively.  Therefore, fertilization with the medium quantity added
to the No. 2 plots increased the oil decomposition rate about 75 per-
cent over the unfertilized No. 3 plots, and the larger quantity of
fertilizer added to the No. 1 plots increased the oil decomposition
rate 100 percent.  Variations in the effect of fertilizer are apparent.
For the "A" plots during the period February to October 1971, the oil
decomposition rate for the No. 2 and No. 3 plots exceeded the reduction
for the No. 1 plot, indicating the fertilizer addition to the No. 1
plot was excessive.  Also, for the "B" plots during the period May to
November 1970, the oil removal rate for the No. 2 plot was the same as
for the No. 1 plot, indicating the additional fertilizer applied to
No. 1 plot was not needed.  An optimum fertilization program would
appear to be 1) addition of slightly excess quantities of phosphorus
and potassium, on a one-time basis, and 2) the addition of nitrogen in
the form of ammonium nitrate in small dosages as needed based upon
tests to maintain a positive (10 to 50 ppm) NH3 and/or N03 content.

Metals Contents of Soils
Metals obtained with different soil extracting solvents, total nitrogen,
total sulfur, and total phosphorus are given in Tables 8, 9, and 10 of
Appendix A for the "A", "B", and "C" plots, respectively.  The data
show inconsistencies and real effects of the metal components are not
apparent.  Copper, zinc, and lead are normally considered to exhibit
harmful effects on biological growth.  These elements may have been
insoluble at soil pH conditions and not available to the microorganisms.

Oil and Nutrient Infiltration

Oil and nutrient concentrations at depths of 2, 4, and 6 feet (Table 11
of Appendix A) show these constituents did not infiltrate the soil under
the conditions of the project.  As discussed earlier, oil was present
at the two-foot depth at the start of the study, and no change was
noted after 18 months.  The presence of phosphate at the two-foot depth
was expected since the initial grading and leveling operations mixed
the top two feet of soil.
                                 39

-------
                             TABLE 4

      QUANTITY AND COST BASED UPON EXPERIMENTAL CONDITIONS


Calculations For Sludge Containing 100 Percent Oil;
OIL DECOMPOSED/MONTH
  POUNDS/CUB1C FOOT
  POUNDS/SQUARE FOOT
  POUNDS/ACRE
  GALLONS/ACRE
  BARRELS/ACRE
  SPREADING COST/ACRE

FERTILIZER
  NITROGEN, POUNDS/YEAR
            COST/POUND
            COST/MONTH
  PHOSPHORUS, POUNDS/YEAR
              COST/POUND
              COST/MONTH
  SPREADING COST/ACRE

CULTIVATION, HOURS/PLOWING
             COST/HOUR
             COST/2 PLOWINGS
                                  QUANTITY    DOLLARS
     1
     0.5
21,780
 2,904
    69.14
             70.00
 1,000
   500
 0.15
12.50

 0.145
 6.00
60.00
             25.00
                       TOTAL  DOLLARS
                        PER     PER
                       MONTH   BARREL
          70.00
                       12.50
                        6.00
                        5.00
                      400.00
                      493.50
                                                                  7.15
Calculations For Sludge Containing 33 Percent Oil:
DECOMPOSED/MONTH
  BARRELS/ACRE
  SPREADING COST/ACRE

FERTILIZER
  MATERIAL AND SPREADING
  (SAME AS ABOVE)

CULTIVATION
  (SAME AS ABOVE)
             COST/2 PLOWINGS
   210
            210.00
         210.00
                       23.50
                      4.00. QQ
                      633.50    3.00
                                40

-------
                           SECTION XI

             QUANTITY AND COST BASED UPON CONDITIONS
                       DURING THE PROJECT

The pounds of oil decomposed per cubic foot of soil per month from
Table 3 were 1.10, 0.96, and 0.55 for the Nos. 1, 2, and 3 plots,
respectively.  A quantity of 1.0 Ib/ft3/mo was selected as the basis
for determining cost of the process.  The fertilizer requirement was
assumed to be 1,000 pounds of nitrogen and 500 pounds of phosphorus
per acre per year, and costs for spreading oil and fertilizer and
cultivating the soil were based upon the expense incurred during the
project period.  The quantities and costs based on experimental condi-
tions are given in Table 4.  An oil decomposition rate of 1 Ib/ft3/mo
is equal to 21,780 Ibs/acre/mo or about 70 bbls/acre/mo.  The cost for
delivering the oil to the area and distributing the oil over the
surface of the soil was about $l/bbl and for 70 bbls/acre/mo would
cost $70/acre/mo.  The fertilizer material and labor for spreading
costs prorated on a monthly basis was estimated to be about $23.50/mo.
Cultivation would require about 8 hrs/acre at $25.00/hr and two
plowings/mo totals $400.00/mo.  The total cost/acre/mo was estimated
to be $493.50 or about $7.15 per barrel of oil.  On a basis of oily
sludge which may contain 33 percent oil, the total volume to be
delivered and spread would be 210 barrels and would cost $210 instead
of $70 for 100 percent oil.  The cost for disposal of oily sludge
would be about $3.00 per barrel.
                                 41

-------
                         FIGURE    19
                                                     LEGEND:
                                                        A-l   •-
                                                        A-2   x-
                                                        A-3 A
Apr. Hiy June July Aug. Sept. Oct. Nov. Dec. Jin. Fee. Kir. Apr. M«y June July Aug. Sept. Oct. Nov. Dec.


    TEAR OF  1970                               YEAR OF 1971
       MICROBIAL  CONTENT  OF  SOIL  FROM "A"  PLOTS
                          42

-------
                           SECTION XII

                        MICROBIAL ACTION

A natural function of microorganisms found abundantly in soil is to
decompose nitrogenous and carbonaceous materials into microbial cellu-
lar matter.  Side products of gases and partially reacted organics
("humus") are formed.  When oil or hydrocarbon is the only source of
carbon, oil degrading microorganisms survive and become the predominant
species.  An intermediate product is organic acids as discussed later.

Results of monthly determinations of the predominant organisms and a
brief description from literature of individual organisms are given in
Appendix B.

Due to mixing of various depths of soil in the initial grading and
leveling operations, the microbial population was different on all
plots.  However, the microbial population contained oil decomposing
bacteria especially of the Flavobacterium and Pseudomonas species.
During the project period, distribution of the microorganism species
was not greatly different as shown in Tables 1, 2, and 3 of Appendix B
for the three oils used in the experiment.  During the first eight
months, the Flavobacterium, Nocardia, Pseudomonas, and Arthrobacter
appeared to be most prominent in all plots.  During the last nine
months, Corynebacterium increased in prominence and Arthrobacter was
seldom a prominent species.  Also, during the latter.part of the ex-
periment, yeast was found prominent in numerous samples.  The addition
of fertilizer did not appear to materially affect the distribution of
microorganisms but did affect the total aerobic count.  The total
aerobic count for "A", "B", and "C" plots are given in Tables 5, 6,
and 7 of Appendix A and shown graphically in Figures 19, 20, and 21.
The No. 1 plots (heavily fertilized) generally were higher in total
aerobic count than No. 2 (medium fertilized) and No. 3 (not fertilized)
plots.  Also, the addition of oil in May 1970 and February 1971
appeared to upset the soil microbial equilibrium and caused the total
aerobic count to be low.

An exploratory experiment was made to determine the effect of adding
oil (same as used on a specific test plot during the project period)
to the nutrient agar used in the microbial analyses.  The primary
effect appeared to be a lower total cell count  for the oil containing
agar.  Data from this exploratory test, plus summarized data and com-
ments by the consultant concerning the microbial analyses of the total
project period, are given in Appendix C.
                                43

-------
                                  FIGURE  20
   200
    180
   160
   140
x  120

co
UJ
CQ

    100
    80
    60
    40
     20
                                                               LEGEND:
                                                                 B-l   •
                                                                 B-2  x--

                                                                 B-3
	X

— -A
          Apr. Miy June July Aug. Sept. Oct. Nov. Dec. Jan. Feb. Kir. Apr. May June July Aug. Sept. Oct. Nov. Dec.




             YEAR Of 1970                              YEAR OF 1971
                MICROBIAL  CONTENT OF SOIL  FROM "B" PLOTS
                                    44

-------
                                   FIGURE   21
   200



   180



   160



   140

o

x 120
CO

§  100
     80
     60
     40
     20
                                                                LEGEND:
                                                                  01   -
                                                                  C-2  x-
                                                                  C-3  A	A
          Apr. Hay June July Aug. Sept. Oct. Nov. Dec. Jan. Feb. Kir. Apr. Hay June July Aug. Sept, Oct. Nov. Dec.



             YEAR OF  1970         •                      YEAR OF 1971
                MICROBIAL CONTENT  OF  SOIL FROM  "C"  PLOTS
                                    45

-------
                             FIGURE   22
     30
                                A-l
LEGEND:
  SATURATES S
  RESINS    R
  AROMATICS A
UJ
Q_
20


IU

S
R
A

S
R
A
—.^^
~~"~~-^. ^

^~~~~~~~- —
—
-



S


A
~~^^

— — — _
	 . 	

••*.



^^
— -~-^T~I






A-2
30
20

10
n
S
.B
A
S
R
A
^ -«^ ^^
	 	 	
' — •




S
R

A
^
— — __
^^-^^

•—i
_
—

^
__ "^^.
~~"~~
^^
-~
-


    30
                                A-3

20


10

n
Dec. Jan. Feb. Har. Apr.



s
o
K
A
"•>


S

R
A

Ju


~


~- — — — _


-------
                          SECTION XIII

           COMPOSITION OF OIL EXTRACTED FROM THE SOIL

Oils added to and extracted from soil plots were 1) quantitatively
analyzed by a clay gel adsorption method9 for hydrocarbon types (total
saturates, resins, and aromatics); 2) qualitatively analyzed by infra-
red absorption for "fingerprinting" differences in oils from the
various test plots; and 3) qualitatively analyzed by gas chromatography
for retention times relative to normal paraffin carbon numbers.  Also,
the gas chromatography method7 was used to obtain a boiling point pro-
file (up to about 510°C) relative to percent weight of the oil.

HYDROCARBON TYPES

The clay gel method9 utilizes clay to retain the strongly polar con-
stituents (asphaltenes, organic acids, etc.) and designates them as
resins.  Silica gel is then used to separate the saturate and aromatic
fractions.  Hydrocarbon types from Tables 12, 13, and 14 of Appendix A
are related with time and shown in Figures 22, 23, and 24 for Plots
"A", "B", and "C", respectively.  For the "A" plots which were treated
with crude oil tank bottoms, the saturate and aromatic contents de-
creased and the resin content increased, indicating a conversion of
saturates and/or aromatics to resinous material.  The magnitude of the
change appeared greatest for the No. 1 plot and least for the No. 3
plot.  The change for No. 2 plot was intermediate between A-l and A-3,
indicating an effect of added fertilizer material.  For the "B" plots
which were treated with bunker C fuel oil, the saturate and aromatic
contents decreased and the resin content appeared to remain unchanged,
possibly indicating a relatively nonreactive resinous fraction.  The
magnitude of change for B-l and B-2 (Figure 23) appeared to be about
the same and B-3 showed the least reaction.  For the "C" plots which
were treated with waxy raffinate, the saturate content decreased with
time, the resin content appeared to increase slightly, and the aromatic
content was not markedly changed, substantiating the fact that paraf-
fins react more easily than aromatics.  The three figures (22, 23, and
24) indicate more rapid changes in composition for No. 1 and No. 2
plots which were fertilized than for the No. 3 plot.  Further differ-
ences in the oils from fertilized and unfertilized plots were evident
from infrared and gas chromatography examinations.

INFRARED SPECTROSCOPY

The infrared (IR) method for analyses of the oils extracted from the
soil and a discussion of the analysis are given in Appendix D.  The IR
spectra of oil extracted from the soil after eight months and after 17
months of the project are given in Figures 25, 26, and 27 for "A", "B",
and "C" plots, respectively.  These figures are discussed separately
in Appendix D.  General conclusions from the study are as follows:
                                47

-------
                                       FIGURE   23
CJ3
Ul
                                                                         LEGEND:
                                                                                  MES  S
                                                                                         R
B-1
30
i A
:0
o F
R
A
S
R
— *
A

-*•>.
--^.. 	 ^
^^^-


—




B-2
0

0

0 Is
T?
A
n 1 1
-
S
R

A

^*\.
^>>
-»» «>^

~^"~--~ -^^










S
R

A

S
R
—

A


"\^
"^
"""-•


^-
'
^^_









• — ,
—





3MIUKMI LO
RESINS
^ AROMATICS
\
\
^ -v.

">""^-









\^
--^_

"^--^,















B-3
0
0

o [s
I
A
n 	 LJ
S
R
—
A

^-^^^
~~*' — ^^_ ^*~
~~~~~— --~.___^


^™








S
R

A


~-— ^


^^™





•""



^^~^-
^ — — ~
	



—
—
—









        Jan. Fed M«r. Apr. K»y June July Aug. Sept. Oct. Nov Dec. Jan Feb. fir. Apr. fiy June July Aug. Sept. Oct. Nov. Dec


                      YEAR OF 1970                                "EAR OF 1971




                            HYDROCARBON TYPE  FOR "B"  PLOTS
                                         48

-------
     30
     20
     10
     30
     20
o    10
    30
                              FIGURE  24
                                 C-l
LEGEND:
  SATURATES S
  RESINS    R
  AROMATICS A






S
R
IX
A

S
E
A
*\
""^
• —



^^






S

R
A
-^
^\

	

•-.



\
^\
\
— .. „__

J\







                                 C-2







s
R
A
— 1
S

R
A
-^.
•>«.
^-^^

^" 	
^^ ^™



-






S


R
A
^









^•^




.__









                                 C-3
20


10



Dec. Jan. Feb. Mar. Apr.





R
A
Hay


S

R

A
Ju

~~~— — — . 	





ne July Aug. Sept. Oct. Nov. Dec.







Ji







n. Fe

S


R

A
b.
~ ~-






Par. Apr. Hay







Ju
^~~-- •-,






ne July Aug. Sept







0







t. Nov. Oe
                     HYDROCARBON TYPE FOR "C" PLOTS
                               49

-------
4000 3000      3000
                                       9     10    II     12     11     14     \f
                                                                                    HOOfRATf
                                                                                                                   IKFRARED SPECTRA OF  OILS EXTRACTED FROM  "A" PLOTS
                                                                                                                      (SOILS TREATED WITH  CRUDE OIL TANK BOTTOHS)
                                                                                                                   Smple:   8.5  to  15.0 "Icronj -  10J«/v CS? Solution
                                                                                                                            2.0  to   9.0 nUrons -  4lw/v CC14 Solution

                                                                                                                   Reference:  Solvent Covpenuted

                                                                                                                   Cell  TMcknesi:   O.S m. Hid
                                                                                                                                      AFTER 8 MOUTHS

                                                                                                                            1500      CM'      1000    900      BOO
                            WAVELENGTH (MICRONS,
                                                                                                                                            9    10    II     12    13    U     1?

-------
4000 3000      2000      1500
                                                                                                                        INFRARED SPECTRA OF OILS EXTRACTED FROM "B" PLOTS
                                                                                                                             (SOILS TREATED WITH BUNKER C FUEL OIL)
                                                                                                                        Sample:   8.5 to 15.0 microns - lOWv CS? Solution
                                                                                                                                 2.0 to  9.0 microns -  4:w/v CCU Solution
                                                                                                                        Reference:  Solvent Compensjteo

                                                                                                                        Cell Thickness:  O.S ran,

-------
Ln
                                                                                                                          FERTILIZATION
                                                                                                                                                          INFRARED SPECTRA OF  OILS  EXTRACTED FBOH T PLOTS
                                                                                                                                                                 (SOILS TREATED KITH K/UY RAFFIKATE)
                                                                                                                                                          Suvle:   8.5 to 1S.O nicrons -  lOU/v CS2 Solution
                                                                                                                                                                   2.0 to  9.0 ilcrms -  4lw/v CCU Solution
                                                                                                                                                          Reference:   Solvent Covpensited
                                                                                                                                                          Cell Thickness:   01 m. Hid
                                                                                                                                                                           ftnCR 8 MOUTHS
                                                                                                                                                                     1500       CM'     1000   900
                                                                   WAveiENGTH (MICRONS'.
                                                                                                                                                                                          10    II
                                                                                                                                                                         WAVELENGTH (MICRONS!

-------
The IR spectral character of all the oils extracted from the soil plots
are similar to that of the oil found in the soil at the beginning of
the study, despite a substantial variation in the type of oils added to
the soil.  The extracted materials would be grossly characterized from
the IR spectra as highly aromatic oils containing a substantial amount
of condensed multi-ring aromatic structures.  Organic acids were
present in each of the extracted oil samples, but in varying amounts,
based upon the presence of an acid carbonyl (C=0) group (absorbing at
5.85y).  The oils from plots that were fertilized show higher concen-
trations of organic acids, and the concentration appeared to be the
greatest in the heavily fertilized plots.  The increase in organic
acids correlates, in general, with a decrease in the long chain paraf-
fin (absorption at 13.88y) and a decrease in total saturate groups
(absorption at 6.86y and 7.25y).  A substantial reduction of long chain
paraffin groups was observed even in the absence of fertilizer suggest-
ing that such groups are the most readily decomposed under the condi-
tions existing in these studies.

GAS CHROMATOGRAPHY

Gas chromatographs7 were obtained with nonpolar media which presumably
makes no separation according to molecular type and permits passage of
all hydrocarbon components on the basis of their boiling points.  The
retention times and boiling points of normal paraffins were used as a
basis for determining temperatures.  The detector response of a sample
being analyzed indicates the quantity of hydrocarbon in the sample
relative to the carbon number of normal paraffins.  The gas chromato-
graphs and boiling point profiles for the "A", "B", and "C" plots are
shown in Figures 28, 29, and 30, respectively.  Crude oil tank bottoms
(Figure 28) contains components whose carbon numbers, relative to
normal paraffins, range from about 5 to 40.  The normal paraffin con-
tent was substantial, based upon a peak for each normal paraffin carbon
number.  The initial boiling point was less than 50°C, and about 80
percent weight of the sample boiled at a temperature less than 400°C
or equivalent to about 25 carbon numbers  (based upon n-paraffins).
For the highest temperature reached, 511°C  (equivalent to 38 n-paraffin
carbon numbers), about 95 percent weight of the sample had passed
through the gas chromatograph column.  (Residuals were back-flushed
from the column to obtain a material balance.)
                               \
Residual oil at the start of the project contained carbon numbers
ranging from about 20 to >38 relative to normal paraffins.  The boiling
point profile ranged from about 350 to 510°C at about 70 percent weight
of the sample passing through the column.  Detector response peaks did
not coincide with normal paraffin carbon numbers.

Residual oil extracted from soil plots A-l and A-3 at the end of the
project period (Figure 28) appears quite different from the oil added
and the  starting residual oil.  The carbon number range was about 15
to >38 for each of the three oils.  The initial boiling point was
                                53

-------
                               FIGURE  28
                   CRUDE OIL TANK  BOTTOMS
      OIL  ADDED
   20
   10
_
Di.
00

O
CL.
00
CsL
O
LU
O
      RESIDUAL'OIL"AT  START
20
   10
      RESIDUAL OIL AT END
   20
   10
                                A-2
   20


   10
                                A-3
                                          400
                                          200
                                          200
                                      o
                                      o
                                A-l   ^
                                "  I   Q;
                                          400
                                          200
                                          400
                                          200
                                          400
                                       200
          10      20       30

    CARBON NUMBER-OF N-PARAFFIN PEAK
                                                20     40     60,   80
                                                PERCENT WEIGHT'
        GAS CHROMATOGRAPH AND BOILING POINT  PROFILES, "A" PLOTS
                                   54

-------
                              FIGURE   29
   20


   10
      OIL  ADDED
                     BUNKER C FUEL OIL
   20
   10
UJ

o:
UJ
Q-
UJ
00

o
o.
00
O
UJ
a
      RESIDUAL OIL AT START
20
   10
      RESIDUAL OIL AT END
   20



   10
   20



   10
                                B-l
                                B-2
                                B-3
                                      200
                                         200
                                      200
          10       20       30

   CARBON  NUMBER OF N-PARAFFIN  PEAK
                                               20     40    60

                                               PERCENT WEIGHT
       GAS  CHROMATOGRAPH AND BOILING  POINT PROFILES,  "B"  PLOTS
                                  55

-------
   20




   10
      OIL  ADDED
   20
   10
o
DC
o
a.-
a:
o
      RESIDUAL OIL AT START
   20
   10
      RESIDUAL OIL AT END
   20
   10
                               FIGURE  30
  WAXY
RAFFINATE 
-------
about 200°C and the percent weight up to 510°C was about 45 and 65 for
plots A-l and A-3, respectively.  Normal paraffin peaks were not
apparent for plot A-l.  The detector response for oil extracted from
the A-3 plot showed small peaks at carbon numbers 16, 22, 23, 25,  and
28, indicating incomplete microbial reaction.  Also, the residual oil
at the end of the project was different from the residual oil at the
start, indicating the starting residual oil had been changed during
the project.  Therefore, the starting residual oil was not a stable,
nonreactive material.

Chromatographs and boiling point profiles for the "B" plots  (Figure 29)
are similar to the "A" plots discussed above.  Differences appear in
the percent weight boiling less than 510 C due to the higher boiling
bunker C fuel oil which was added to the "B" plots.  Also as expected,
normal paraffin peaks for bunker C fuel oil were not apparent.  Again
the residual oils at  the start and end of the project were different.

Results for the "C" plots which received waxy raffinate  (Figure 30)
show predominant normal paraffin peaks in the oil added  to the soil
and some definite peaks in the residual oil at the end of the project.
This was expected since the hydrocarbon type analyses  (Figure 24)
showed unreacted saturates present for all plots  (C-l and C-3).  The
residual oil at the start and end (Figure 30) of  the project appear
different, indicating a reaction of the starting  residual oil has
occurred especially in the carbon number range of 30 to  35.
                                57

-------
                           SECTION XIV

                         RAINFALL RUNOFF

Rainfall accumulated and was trapped in the plot area as discussed
previously.  Drainage of the plots was controlled, and a sample of the
runoff water was obtained for analysis.  On a few occasions during ex-
cess rainfall periods, water overflowed the plots.  Analyses of the
water for oil, ammonia, nitrate, and phosphate contents indicated that
little, if any, of these constituents were present when drained imme-
diately after a rain.  Long standing appeared to cause higher oil
contents.  The oil content ranged from about 30 to 100 mg/1, and the
oil content of water from the fertilized plots as discussed earlier
were found to contain the highest concentration of organic acids.
Each plot was drained about 36 times during the 18-month project, and
many times the plots were only partially full but needed to be dry for
cultivation.  Based upon an average oil content of 60 mg/1, about 0.6
pounds of oil was discharged during the 17-month experimental period.
This quantity equals about 0.03 lbs/ft3 of oil in the soil, and adjust-
ments in earlier decomposition rate determinations for such a small
quantity were not considered justifiable.

Infrared examination of oil recovered  from rainfall runoff water is
discussed in Appendix D and indicated  the oil to be organic acids with
characteristics quite similar with spectra for naphthenic acids.

Ammonia content of the water appeared  to vary with the soil nutrient
content.  During periods when the ammonia was excessive on  the No. 1
plots, discharge water contained up to 150 mg/1 ammonia as N.  Nitrates
and phosphates were generally absent in the discharges.
                                59

-------
                           SECTION XV

                        ACKNOWLEDGMENTS

Facilities, personnel, and partial (30%) financial support were
furnished by Shell Oil Company.  A financial grant (70%) and
guidance were supplied by the Environmental Protection Agency.

Dr. Melvin L. Renquist, now retired,  and Dr. Ben S. Baldwin, Chief
Technologist, served as Grant Directors and provided the necessary
administrative assistance for the project.

Mr. C. Buford Kincannon, Senior Engineer, directed the project
activities and prepared the initial and final reports.

Mr. Wilbert L. Pegues, Laboratory Technician, performed laboratory
tests, supervised the experimental tasks of the plot area, and col-
lected data.

Mr. James M. Martin, Senior Research Chemist, provided infrared
spectra data and interpretation of the infrared results.

Consultants during the project were Dr. Edwin 0. Bennett, Micro-
biologist, University of Houston, and Dr. Warren D. Anderson,
Soil Specialist, Texas A&M University.

From the Environmental Protection Agency, valuable guidance was
provided by Mr. Leon H. Myers, Project Officer, and the final
report was reviewed by Dr. Thomas E. Short, Chemical Engineer.
                                61

-------
                           SECTION XVI

                           REFERENCES

 1.   G.  K.  Dotson,  R.  B.  Dean,  B.  A. Kenner,  and W.  B.  Cooke,  Land
     Spreading,  A Conserving and Non-Polluting Method of Disposing  of
     Oily Wastes, Proceedings of the  Fifth International Water Pollu-
     tion Research Conference,  Vol. 1, Section II,  36/1-36/15,
     Pergamon Press,  1971.

 2.   K.  J.  Stracke,  Cleanup at Santa Barbara,  Paper No.  SPE3197,  1970,
     Society of Petroleum Engineers of AIME.

 3.   J.  B.  Davis, Petroleum Microbiology,  Elsevier  Publishing  Company,
     New York,  N.Y.,  1967.

 4.   Manual on Disposal of Refinery Wastes, Volume  on Liquid Waste,
     Chapter 13, Biological Treatment, First Edition 1969,  American
     Petroleum Institute, Division of  Refining, New York, N.Y.

 5.   Standard Methods  for the Examination of Water  and Wastewater,
     Twelfth Edition,  American Public  Health Association, Washington,
     B.C.

 6.   Robert S.  Breed,  E.  G. D.  Murray, and Nathan R. Smith, Bergey's
     Manual of Determinative Bacteriology, Seventh  Edition, The
     Williams and Wilkins Company, Baltimore,  Maryland.

 7.   L.  E. Philyaw,  A. E. Krc,  and M.  J. O'Neal (Shell Oil Company),
     Anal.  Chem., 43,  #6, 787-89,  May  1971.

 8.   American Society  for Testing and  Materials, ASTM,  Standard Method
     of Test for Sediment in Crude and Fuel Oils by Extraction, ASTM
     D473-65.

 9.   American Society  for Testing and  Materials, ASTM,  Standard Method
     of Test for Characteristic Groups in Rubber Extender and  Process-
     ing Oils by the Clay-Gel Adsorption Chromatographic Method,  ASTM
     D2007-69.                  ,

10.   State Soil Testing Laboratories  in the Southern Region of the
     United States,  Bulletin No. 102,  Southern Cooperative Series,
     June 1965,  Agricultural Experimental Station Publication.

11.   Improved Kjeldal  Method for Nitrate Containing Samples, Official
     Methods of the Association of Official Agricultural Chemist, 1965,
     Pg. 16.

12.   American Society  for Testing and  Materials, ASTM,  Standard Method
     of Test for Sulfur in Petroleum Products, High Temperature Method,
     ASTM D1552-64.
                                 63

-------
13.  M. Gales, Jr.,  E.  Julian,  and R. Kroner, Method for Quantitative
     Determination of Total Phosphorus in Water, Journal AWWA, 58,
     No. 10, 1363 (1966).

14.  American Petroleum Institute, API,  Manual on Disposal of Refinery
     Wastes, Volume IV, Sampling and Analysis of Waste Water, Second
     Edition, 1957,  Volatile and Non-Volatile Oily Material, API,
     Method 731-53.

15.  V. Carlson,  E.  0.  Bennett,  and J. A. Rowe, Microbial Flora and
     Their Relationship to Water Quality, Society of Petroleum
     Engineering  Journal, Volume 1, Pg.  71-80, 1961.

-------
                          SECTION XVII

                           APPENDICES
A.  Analytical Results
    Table  1:
    Table  2:

    Table  3:

    Table  4:

    Table  5:
    Table  6:
    Table  7:
    Table  8:

    Table  9:

    Table 10:

    Table 11:
    Table 12:
    Table 13:
    Table 14:
Oil Properties for Simulated Sludges  A,  B,  and C
Oil Content of Individual Soil Samples  for
"A" Plots
Oil Content of Individual Soil Samples  for
"B" Plots
Oil Content of Individual Soil Samples  for
"C" Plots
Chronological Data for "A" Plots
Chronological Data for "B" Plots
Chronological Data for "C" Plots
Total Nitrogen, Sulfur, Phosphorus,  and
Metals Contents for "A" Plots
Total Nitrogen, Sulfur, Phosphorus,  and
Metals Contents for "B" Plots
Total Nitrogen, Sulfur, Phosphorus,  and
Metals Contents for "C" Plots
Oil and Nutrient Penetration Depths  Into Soil
Hydrocarbon Types for "A" Plots
Hydrocarbon Types for "B" Plots
Hydrocarbon Types for "C" Plots
B.  Microbial Analyses

    Table  1:  Predominant Individual Organisms for
    Table  2:  Predominant Individual Organisms for
    Table  3:  Predominant Individual Organisms for
    Brief Description of Predominant Organisms

C.  Microbiology Consultant's Comments

    Table  1:  Statistics of Microbial Analysis
    Microbiology Consultant Report

D.  Infrared Study of Oils Extracted from Soil
        Infrared Study of Rainfall Runoff Water
    Oil Content in Soil Samples

        Sketch:  Apparatus for Extraction of Oil in Soil
Page
No.

 66

 67

 68

 71

 74
 77
 80
 83

 86

 88

 90
 92
 93
 94
 95

 96
"A" Plots
"B" Plots
"C" Plots

97
99
101
103
                                                   106

                                                   107
                                                   108

                                                   109
                                                   113

                                                   114

                                                   115

-------
                           APPENDIX A

                       ANALYTICAL RESULTS

The three simultaneous experiments using crude oil tank bottoms on the
"A" soil test plots, bunker C (No. 6) fuel oil on the "B" plots, and
waxy raffinate on the "C" plots, were.similar.  Properties of the oils
added to the soils are given in Table 1.  Oil contents of subsamples
from each soil plot described in Section IX are given in Tables 2, 3,
and 4 for the "A", "B", and "C" plots, respectively.  Subsample results
from each plot were averaged, and the averaged values are also given in
Tables 5, 6, and 7.  These tables alsorinclude soil moisture, pH, oil,
soluble nitrogen (ammonia, NHg and nitrate, NO3), soluble phosphorus
(phosphate, PO^, expressed as P20s) and total aerobic microorganism
content and temperature, all listed chronologically.  The number of
plowings between sampling dates and the oil and fertilizer addition
dates have been included with the chronologically recorded data
(Tables 5, 6, and 7).

Metals, nitrogen, phosphorus, and sulfur contents of the soil are given
in Tables 8, 9, and 10 for test plots "A", "B", and "C", respectively.
Oil and nutrient contents of soil taken at depths of 2, 4, and 6 feet
are given in Table 11.

Oil extracted from the soil test plots and oil added to the soil were
analyzed for hydrocarbon types, infrared and gas chromatography charac-
teristics.  Hydrocarbon types for the "A", "B", and "C" plots are given
in tables 12, 13, and 14, respectively.  The infrared and gas chrbmato-
graphic data are presented graphically and discussed in Section XIII.
(Details of the infrared study are given in Appendix D.)
                                66

-------
                           APPENDIX A
                              TABLE 1
                          OIL PROPERTIES
                   SIMULATED  SLUDGES A, B, AND C
SPEC. GRAV., 60/60°F
LBS/GAL
POUR POINT, °F
VISCOSITY, SU, 60°F
          SSF, 122°F
HYDROCARBON TYPE, %w
   SATURATE
   RESINS
   AROMATIC
TOTAL SULFUR, %w
TOTAL NITROGEN, %w
TOTAL PHOSPHORUS, PPM
TOTAL ASH, PPM
CALCIUM
MAGNESIUM
SODIUM
IRON
COPPER
LEAD
TANK
BOTTOMS











31,000
1,880
375
3,135
1,255
94
Nil

A
CRUDE OIL
0.86
7.12
-5
60

36
8
56
0.47
0.09
6







SLUDGE
B
BUNKER C
1.03
8.57
40
19,000
120
18
26
56
1.96
0.41
11
320
10
1
15
20
1
Nil

C
WAXY PRODUCT
0.85
7.08
95
-
59
90
-
10
0.04
O.0005
5
8
Nil
Nil
Nil
Nil
Nil
—
                                67

-------
APPENDIX A
TABLE 2
OIL CONTENT OF INDIVIDUAL SOIL SAMPLES
PERCENT WEIGHT
Plot
Sub -Sample
Date
4-22-70
5-13
5-19
5-27
6-11
6-24
7-10
7-29
8-14
8-27
9-10
9-22
10-16
A-l
a

8.9
8.3
13.1
11.5
13.3
14.0
14.4
13.8
13.1
11.4
11.1
11.9
11.3
b

10.8
10.2
12.1
14.7
16.1
15.7
14.9
15.6
17.7
15.1
15.0
14.2
11.1
11.8
11.3
c avg.

9.6 10
9.4 10
12.5 13
15.4 14
14.3 14
15.4 15
15.3 16
14.2 14
13.9 14
13.9 13
11.0 11
12
11.4 11
a

11.5
12.7
15.0
15.8
16.4
15.5
17.4
15.7
14.3
14,2
13.6
13.6
13.4
FOR "A" PLOTS

A-2
b

10.6
12.4
13.6
16.8
17.6
18.1
16.9
17.7
16.7
15.6
16.2
15.5
13.9
13.4
13.7
c avg.

10.6 11
14.2 13
17.2 16
14.9 16
17.3 17
17.6 17
16.7 17
15.4 16
16.1 16
15.7 15
13.6 14
14
13.4 14
a

11.1
13.6
14.1
15.4
15.3
15.5
—
18.2
17.0
16.3
17.2
17.9
17.4
15.9
16.3


A-3
b

10.6
13.7
15.2
14.9
16.0
17.5
17.5
17.2
17.9
17.3
17.3
17.2
17.2
18.0
18.1
16.0
16.0
c avg.

11.0 11
12.1 13
14.4 15
16.0 16
17.2 17
15.8 17
18.8 18
17.8 17
17.1 17
16.7 17
17.9 18
16
15.5 16
16.3

-------
                                                                       APPENDIX A




                                                                    TABLE 2 (CONTD.)
VO
Plot
Sub-Sample
Date
10-30-70
11-13
12-7
1-6-71
1-27
2-12
2-24
3-10
3-24
4-7
4-21
5-5
5-19
A-l
a
10.1
11.4
11.7
11.2
10.2
18.7
21.9
17.1
17.9
17.1
17.2
16.6
15.8
b
11.3
12.0
11.5
11.2
11.0
20.5
23.5
17.4
19.6
18.8
18.2
17.1
16.0
c
10.3
11.0
10.8
10.8
9.9
30.3
20.9
19.3
17.8
16.9
18.1
16.1
16.1
avg.
11
12
11
11
10
23
22
18
18
18
18
17
16
a
12.7
12.8
12.7
12.5
11.4
23.3
25.2
24.3
22.7
21.9
23.1
20.5
21.6
A-2
b
12.9
13.8
13.7
13.6
13.4
26.8
25.4
22.8
26.3
23.8
24.1
21.4
22.4
c
13.6
14.2
13.8
14.0
12.8
28.6
24.8
27.9
23.9
20.7
22.7
21.1
20.8
avg.
13
14
13
13
13
26
25
25
24
22
23
21
22
a
15.4
15.4
15.1
14.9
14.8
23.4
27.3
33.1
30.1
29.0
25.3
22.6
25.3
A-
b
16.4
16.2
16.7
16.1
16.3
16.0
15.3
15.6
15.6
15.8
26.1
26.1
31.1
32.0
35.2
30.2
36.0
35.7
24.8
24.8
24.4
23.8
23.8
23.1
25.1
26.3
-3
c
15.4
15.7
15.2
15.2
15.1
26.3
26.1
25.6
25.0
21.5
21.9
22.4
21.5

avg
16
16
16
15
15
25
28
30
30
25
24
23
24

-------
                                                       APPENDIX A

                                                    TABLE 2 (CONTD.)
Plot

Sub-Sample

Date
 6-2-71


 6-30


 7-14


 7-28



 8-11


 8-25


 9-23


 9-29
                                A-l
                                                                      A-2
a
15.4
15.4
13.3
14.4
14.4
13.0
13.1
12.5
b
14.8
15.9
15.1
14.4
14.3
12.1
13.7
12.4
c
15.7
16.5
13.9
14.2
13.5
13.0
14.1
12.2
&VR.
15
15
14
14
14
13
14
12
                                                                                                            A-3
18.0    20.6    21.5     20
18.3    19.2    17.9     19
18.1    18.4    18.1     18
19.4    18.7    16.7     18
18.1    17.9    16.5     18
16.7    16.2    15.7     16
15.4    16.2    16.7     16
16.3    15.9    15.8     16
22.8-   24.3    23.9   24
        23.6

21.8    22.0    23.4   23
        21.7

23.9    21.5    19.6   22
        23.4

21.3    22.7
        21.8    19.2   21
22.4    20.7    20.1   21
        20.4

20.7    19.3    17.9   19
        19.9

20.6    20.2    19.9   20
        19.8

22.6    21.2    19.0   21

-------
Plot
Sample Point
Date
4-22-70
5-13
5-19
5-27
6-11
6-24
7-10
7-29
8-14
8-27
9-10
9-22
10-16
APPENDIX A
TABLE 3
OIL CONTENT OF INDIVIDUAL SOIL SAMPLES FOR "B"
PERCENT WEIGHT
B-l B-2
a
10.3
10.2
30.6
25.6
24.8
26.0
25.0
23.9
24.0
22.0
19.7
18.9
17.5
b
10.8
10.6
14.0
29.9
27.2
27.2
24.2
24.4
25.8
24.4
21.9
20.7
19.6
18.9
16.6
11.0
13.2
32.7
24.6
23.8
24.9
24.5
23.4
22.3
21.5
19.5
—
17.9
ave.
11
13
31
26
24
25
25
24
23
21
20
19
17
a
9.4
10.8
29.6
22.2
24.7
23.0
25.4
19.8
19.6
18.7
16.4
16.6
15.8
b
9.5
10.2
10.3
29.4
23.8
18.7
25.0
20.0
19.5
18.8
19.4
18.2
16.8
16.6
15.5
c
11.9
10.9
29.5
23.6
20.3
22.8
20.5
20.3
18.5
19.1
17.1
~
15.4
PLOTS
avg.
10
11
30
23
21
24
22
20
19
19
17
17
16
B-3
a
10.0
12.2
25.6
22.0
24.6
24.1
23.6
23.5
22.1
18.7
19.4
18.8
19.0
19.1
b
9.5
11.8
27.9
27.1
23.4
20.7
24.1
25.3
21.6
19.6
19.7
17.6
17.7
18.8
18.8
19.0
18.5
17.5
c
10.8
13.1
26.6
22.0
18.0
23.4
21.2
19.0
18.9
19.2
18.6
--
18.8
avg.
10
12
28
22
21
24
23
21
19
19
19
19
19

-------
                                                                         APPENDIX A




                                                                      TABLE 3 (CONTD.)
N5
Plot
Sample Point
Date
10-30-70
11-13
12-7
1-6-71
1-27
2-12
2-24
3-10
3-24
4-7
4-21
5-5
5-19
B-l
a
17.3
17.7
15.5
16.2
15.9
35.2
38.0
38.1
48.7
34.2
32.2
31.5
32.5
b
17.4
17.1
15.7
16.2
14.7
33.7
36.4
45.5
41.5
33.3
33.6
31.5
34.4
c
17.3
17.0
17.7
16.3
15.2
29.3
40.0
47.8
43.0
34.6
32.3
36.0
35.3
17
17
16
16
15
33
38
44
44
34
33
33
34
a
14.9
15.7
16.0
14.7
14.3
30.6
34.7
36.5
41.5
33.3
32.6
32.5
34.6
B-2
b
14.6
15.5
15.9
15.0
13.2
38.2
35.5
42.2
38.5
35.0
32.4
33.1
35.4
c
15.3
15.6
15.1
15.4
13.3
30.6
43.4
40.2
37.7
36.4
31.4
32.8
37.7
15
16
16
15
14
33
38
40
39
35
32
33
36
a
16.9
17.8
17.9
17.6
17.2
33.1
33.9
32.9
36.6
31.8
29.0
26.7
37.5
B-
b
17.4
17.3
16.9
17.1
18.4
18.5
17.2
17.4
15.4
15.4
31.9
33.1
34.0
32.3
35.6
33.4
35.5
34.8
34.8
33.9
31.7
30.4
30.7
29.7
35.2
36.6
3
c
16.8
17.0
17.6
17.8
16.5
34.9
34.7
34.0
36.4
36.0
28.5
36.0
34.9

17
17
18
18
16
33
34
34
36
34
30
31
36

-------
                                                       APPENDIX A

                                                    TABLE 3  (CONTO.)
Plot

Sample Point

Pate
 6-2-71

 6-30


 7-14


 7-28


 8-11


 8-25


 9-23


 9-29
            B-l
                                                  B-2
37.6    37.3    31.3     35

33.2    31.9    30.0     32


34.8    32.2    31.0     33


36.2    34.2    26.9     32


28.9    31.7    32.4     31


29.6    29.2    28.9     29


29.1    25.9    22.6     26


24.2    25.4    25.7     25
a
36.3
36.4
35.7
38.6
26.6
30.0
b
36.2
38.8
37.6
36.5
25.7
20.8
c
38.7
34.0
39.1
31.8
29.5
21.6
avg.
37
36
38
36
27
24
                                                                                         B-3
30.6    19.9    22.2     24
25.4    24.1    22.0     24
35.6    29.8    28.5   31

33,0    31.2    33.3   33
        36.1

33.5    36.1    28.5   33
        35.3

31.1    30.4    32.2   31
        31.1

34.8    30.4    29.6   32
        30.9

34.3    29.7    31.9   32
        26.8

26.4    25.5    26.7   26
        26.2

31.2    26.8    32.0   30
        28.1

-------



Plot
Sample Point
Date
4-22-70
5-13
5-19
APPENDIX A
TABLE 4
OIL CONTENT OF INDIVIDUAL SOIL SAMPLES FOR "C" PLOTS
PERCENT WEIGHT
C-l C-2 C-3
a b c avg. a b c avg. a b c avg.
9.4 10.5 10.6 10 9.4 9.6 9.6 10 10.6 9.9 10.3 10
10.7
10.6 11.9 13.2 11 9.5 12.2 13.8 12 9.0 12.1 12.6 11
12.4
23.4 23.0 24.2 24 16.7 20.8 22.6 20 17.3 18.1 20.5 19
5-27


6-11


6-24


7-10


7-29


8-14


8-27


9-10


9-22
20.3    23.0    23.1     22
        22.8

21.6    21.7    22.7     22
19.2    20.4    16.4     19
19.3

17.3    17.2    25.6     20
16.9    19.1    21.5     19
17.4    18.5    19.1     18
14.8    17.4    18.4     17
15.8    15.5    15.8     16
                                      18.5    22.9    24.6     22
                                      16.2    20.5    22.6     20
                                      12.8    16.0    18.3     16
                                              15.8

                                      24.3    18.1    22.4     22
                                      16.7    18.2    20.1     18
                                              16.7

                                      16.9    15.7    19.3     17
                                      12.8    13.5    18.2     15
                                      14.2    14.3    12.8     14
        18.3

18.3    19.1    19.5   19
17.4    19.8    20.5   19
17.2

15.5    18.7    17.4   17
        18.6

15.4    18.8    17.8   17
15.5

15.6    17.9    19.2   18
15.4


14.2


14.4
17.1
17.2

16.7
16.8

14.4
14.3
14.2    14.2
                         14
                                      12.8    12.9
                                                               13
                                                                            15.3    15.2
17.8   17


16.1   16


15.4   15


       15

-------
                                                                      APPENDIX A




                                                                   TABLE 4 (CONTD.)
Ln
Plot
Sample Point
Date
10-16-70
10-30
11-13
12-7
1-6-71
1-27
2-12
2-24
3-10
3-24
4-7
4-21
5-5
C-l
a
12.8
11.5
12.1
11.2
11.8
11.2
27.8
29.8
27.7
23.9
22.1
21.1
21.7
b
14.6
11.7
13.8
14.1
13.2
12.5
30.9
32.6
25.9
26.7
27. 4
25.9
26.7
c
15.1
13.1
13.8
14.7
13.7
12.1
32.9
37.3
32.2
28.9
25.9
24.6
23.6
14
12
13
13
13
12
31
33
29
27
25
24
24
a
10.9
13.2
10.9
io.s
10.8
10.4
29.5
29.8
31.0
24.7
24.7
26.2
24.6
C-2
b
13.1
12.2
13.5
11.8
12.6
12.4
26.5
29.5
29.9
27.9
26.4
24.6
26.6
c
15.3
14.8
11.7
13.4
13.0
13.7
30.9
33.2
29.8
28.9
27.0
25.4
25.9
13
13
12
12
12
12
29
31
30
27
26
25
26
a
13.8
12.7
13.0
12.8
12.5
12.9
24.0
25.9
26.7
26.4
25.4
24.5
24.9
C-
b
16.3
16.8
14.5
14.9
14.0
14.5
14.4
14.7
14.3
14.7
14.1
14.0
26.3
26.7
28.9
27.9
28.6
27.2
27.5
27.8
26.5
26.6
25.6
25.8
25.7
25.8
•3
c
16.4
14.0
13.2
14.2
14.6
14.5
31.3
30.1
30.5
29.9
28.9
28.9
27.9

16
14
13
14
14
14
27
28
29
28
27
26
26

-------
                                                      APPENDIX

                                                   TABLE 4 (CONTD.)
Plot

Sample Point

Date
 5-19-71


 6-2

 6-30


 7-14


 7-28


 8-11


 8-25


 9-23


 9-29
                                C-l
                                                                      C-2
                                                                                                            C-3
21.1    23.4    24.6     23


19.9    23.7    23.4     22

19.2    22.0    23.3     22


18.3    21.7    22.1     21


17.4    21.1    21.5     20


20.4    23.4    17.9     21


17.6    20.7    18.4     19


16.9    18.2    21.2     19


14.4    18.4    19.6     18
23.9    26.1    25.0     25


24.5    25.4    25.9     25

22.5    23.9    23.5     23


21.5    24.1    24.1     23


21.5    24.2    22.0     23


22.4    25.3    28.2     25


20.6    21.7    21.5     21


22.2    24.5    24.8     24


17.9    15.8    18.5     18
29.4    27.7    30.2   29
        28.6

25.0    26.0    28.9   27

22.0    26.3    27.8   25
        27.2

24.3    27.7    28.4   26
        26.3

27.1    27.4    23.9   26
        28.1

27.1    24.6    28.7   27
        25

21.9    22.0    22.4   22
        22.9

21.8    26.0    26.6   25
        24.8

20.8    23.1    24.2   23
        23.2

-------
          APPENDIX A




            TABLE 5




      CHRONOLOGICAL DATA




CRUDE OIL TANK BOTTOMS, "A" PLOTS
                         1970


pH, Before Extr.


pH, After Extr.


Moisture, 7.w



Oil, %w


NHo as N, ppm



NO 3 as N, ppm


PO as P 0 , ppm
25'

Temperature, °F


Ambient
Total Count
Aerobes x 107
x 10'
x 107
Number Plowings
Between Sample
Da tea 	


A-l
A-2
A-3
A-l
A-2
A-3
A-l
A-2
A-3

A-l
A-2
A-3
A-l
A-2
A-3

A-l
A-2
A-3
A-l
A-2
A-3
A-l
A-2
A-3


A-l
A-2
A-3


APR.
22
6.6
6.9
6.8



11
11
12

10
11
11
Nil
Nil
Nil

Nil
Nil
Nil
100


78


80






MAY
7 13 17
7.3
7.0
7.2



6
6
8 6
o
•g. 10
o 13
•g. 13
1 72°
« 22°
Z Nil %
3 0
TJ N-H *S
flj 1H 1 1 ^
5 Nil 3
< Nil <



77 80


81

3
0.4
2
1

JUNE JULY
19 27 11
6.8
6.8
6.8



17 20 12
20 20 8
19 20 8

13 14 14
16 16 17
15 16 17
30
25
Nil

Nil
Nil
Nil



83 83


80 82 82

170
190
10
2 1 1

24 10
7.0
7.0
7.0



8 18
10 20
10 17

15 16
17 17
17 18
30
20
Nil

3
1
Nil
240
165
95
86


76

38
73
87
1 1

29
7.3





9
6
6

14
16
17
3
Nil
Nil

Nil
Nil
Nil



89


87




1

AUGUST
7 14
6.8
6.8
6.8



7
7
z 7
o
•g. 14
3 16
•a i?
1 1525
. 750
Z Nil
3
•g Nil
3 Nil
< Nil
320
200

89 94


86 86

54
62
6
2


27






6
7
7

13
15
17










94


83




1

          77

-------
          APPENDIX  A




        TABLE 5 (CONTD.)




       CHRONOLOGICAL DATA




CRUDE OIL TANK BOTTOMS,  "A"  PLOTS




SEPT.

pH, Before Extr.


pH, After Extr.



Moisture, %w


Oil, %w


NH, as N, ppm
J


NO as N, ppm
j

PO, as P 0 , ppm
H ft J

Temperature, °F


Ambient
Total
Aerobes x 10.
x 10,
x 10'
Number Plowings
'Between Sample
Dates

A-l
A-2
A-3
A-l
A-2
A-3

A-l
A-2
A-3
A-l
A-2
A-3
A-l
A-2
A-3

A-l
A-2
A-3
A-l
A-2
A-3
A-l
A-2
A-3


A-l
A-2
A-3



10
7.0
7.0
7.0




6
5
5
11
14
18
565
30
Nil

Nil
Nil
Nil



92


82





5

22
6.7
6.7
6.8




5
4
4
12
14
16
500
20
Nil

Nil
Nil
Nil



88


83

48
40
7

0

1970
OCT.
16 31
6.6
6.8
6.9




6 6
8 5
4 5
11 11
14 13
16 16
540
140
Nil

Nil
Nil
Nil
720
370

68 62


62 65

84
104
4

3 3


NOV.
14
6.7
6.9
7.0




15
11
6
12
14
16
Nil
Nil
Nil

Nil
Nil
Nil
630
165
85
63


60

119
32
26

3


DEC.
17



6.2
6.5
6.8

12
9
6
11
13
16
50
40
Nil

Nil
Nil
Nil
680
290

58


61

94
56
7

5



JAN.
6



6.3
6.7
6.7

15
11
4
11
13
15
50
10
Nil

Nil
Nil
Nil
500
290
90
43


38

71
37
122

2

27 2



6.5
6.7
6.8 ^
•H
0
7 T,
5 >g
5 «j.
10
13
15

en
O
sr to
• 3! «_
z o
9)
•O TJ
3 S
570
220
150
64 57
64 58
62 55
60 43





3

1971
FEB.
12 24



6.7
6.9
6.7

23 10
22 15
23 20
23 22
26 25
25 28
390
260
Nil

25
Nil
Nil
540
200
130
61 92
59 62
59 60
58 65

53
11
37

4 2



MARCH
10



6.6
6.7
6.9

20
17
20
18
25
31
255
150
Nil

Nil
Nil
Nil



73
66
66
70

77
52
6

1

24







10
9
12
18
24
30










80
73
70
58





2

           78

-------
           APPENDIX A




         TABLl- 5 (CONTD.)




        CHRONOLOGICAL DATA




CRUDE OIL TANK BOTTOMS, "A" PLOTS







                             1971
APRIL

pH, Before Extr.


pH, After Extr.


Moisture, %w


Oil, 7.w


NH3 aa N, ppm


NO 3 as N, ppm


PO^ as P2°5' PP™


Temperature, °F


Ambient
Total 7
Aerobes x 10_
x 10,
x 10'
Number Plowings
Between Sample
Dates

A-l
A-2
A-3
A-l
A-2
A-3
A-l
A-2
A-3
A-l
A-2
A-3
A-l
A-2
A-3
A-l
A-2
A-3
A-l
A-2
A-3
A-l
A-2
A-3


A-l
A-2
A-3


7
6.2
6.3
6.7



6
6
8
17
22
25
120
85
Nil
0
0
0
470
210
95
84
88
80
67

16
10
0.5
2

21






11
7
7
18
23
24

<*i
*5f
•o
01
•o
3



86
89
86
75




2

MAY
5



6.7
6.8
6.9
b
6
6
17
21
23
140
60
Nil
30
10
0
470
160
95
86
87
82
80




2

19






7
5
7
16
22
24









90
98
94
82

100
30
1
1

JUNE
2 30



6.6
6.7
7.0
5 16
4 13
6 15
15 15
20 19
24 23
165
80
Nil
25
10
0
500
210
95
90 92
90 94
87 90
83

97
6b
8
1 3

JULY AUGUST
14



6.7
6.8
7.0
10
4
6
14
18
22
50
25
Nil
5
Nil
Nil
580
210
95
96
94
92
84

48
14
21
2

28 11
6.4
6.5
6.7
6.7
6.8
6.9
12 21
5 18
6 15
14 14
18 18
21 21

PI
%
£
•a
01
•o
•o
580
210
95
95 88
93 92
91 88
86 83

58
59
35
1 1

25



7.0
7.1
7.3
5
4
4
13
16
20
175
70
Nil
65
40
Nil
580
210
125
99
96
91
84




10

SEPT.
25
6.8
7.0
7.3
6.9
7.1
7.2
16
14
12
14
16
20
120
20
Nil
20
Nil
Nil
410
200
115
80
82
82
78






30









12
16
21



















           79

-------
             APPENDIX A




               TABLE 6




         CHRONOLOGICAL DATA




BUNKER C (NO. 6) FUEL OIL,  "B" PLOTS



pH, Before Extr.


pH, After Extr.


Moisture, 7.w




Oil, 7.w



NH3 as N, ppm



NOj as N, ppm


P04 as P205, ppm


Temperature, °F


Ambient
Total ?
Aerobes x 10
x 10;
x 10'
Number Blowings
Between Sample
Dates



B-l
B-2
B-3
B-l
B-2
B-3
B-l
B-2
B-3


B-l
B-2
B-3

B-l
B-2
B-3

B-l
B-2
B-3
B-l
B-2
B-3
B-l
B-2
B-3


B-l
B-2
B-3



APR.
22
6.9
7.0
7.0



12
13
11


11
10
10

Nil
Nil
Nil

Nil
Nil
Nil
94


78


80








1970



MAY JUNE JULY
7








m
3
p
o
•s.
0)
O
•i.
*o
c
(0
a
«>
ij
3
•a
a)
•o
•o




77




1
1
0.6


13
7.3
7.3
7.3



7
7
6


13
11
12

980
700
Nil

Nil
Nil
Nil



80


81




1

17 19 27 11
6.9
6.9
7.0



677
775
896


31 26 24
30 23 21
28 22 21

220
60
3 Nil
0
U Nil
5 Nil
< Nil



83 83


78 82 82

10
12
5
211

24 10
7.1
7.1
7.2



5 14
6 12
6 13


25 25
24 22
24 23

140
15
Nil

Nil
Nil
Nil
180
150
105
86


76

41
27
11
1 1

29
7.5





4
4
6


24
20
21

10
Nil
Nil

Nil
Nil
Nil



89


87




1

7








CO
3
h
o
•g.
CO
o
i.
•o
§
a]
0)
M
3
•0
4)
•O
"O
<



89


86








AUGUST
14
6.9
6.9
6.9



4
4
5


23
19
19

1100
675
Nil

Nil
Nil
Nil
770
260

94


86

5
6
1
2

27






4
4
5


21
19
19











94


83




1

             80

-------
                                            APPENDIX  A




                                          TABLE 6 (CONTD.)




                                         CHRONOLOGICAL DATA




                                BUNKER C (NO.  6) FUEL OIL,  "B"  PLOTS




                                         1970
                                                                                1971
SEPT.

pH, Before Extr.


pH, After Extr.



Moisture, %w


Oil, Xw


NH, as N, ppm
J



NO. as N, ppm


PO as P 0 , ppm
H £. J

Temperature, 8F



B-l
B-2
B-3
B-l
B-2
B-3

B-l
B-2
B-3
B-l
B-2
B-3
B-l
B-2
B-3


B-l
B-2
B-3
B-l
B-2
B-3
B-l
B-2
B-3
10
7.0
7.1
7.0




5
5
6
20
17
19
655
55
Nil


Nil
Nil
Nil



92


22
6.6
6.8
6.9




5
5
4
19
17
19
420
35
Nil


Nil
Nil
Nil



88


OCT. NOV.
16
6.5
6.7
6.9




6
7
6
17
16
19
220
195
Nil


Nil
Nil
Nil
800
360

68


31 14
6.7
6.9
7.1




5 15
6 14
4 8
17 17
15 16
17 17
Nil
Nil
Nil


Nil
Nil
Nil
920


62 63


DEC.
17



5.8
6.3
6.6

12
8
5
16
16
18
30
20
Nil


Nil
Nil
Nil
1100
285
175
58


JAN.
6



6.1
6.5
6.7

6
8
8
16
15
17
20
10
Nil


Nil
Nil
Nil
900
275
130
43


27 2



6.3
6.5
6.9 3
o
8 1
6 5
4 <
15
14
16

n
O
Z Si
•& to
a: a
z u
0
~O CU
HI
•u -o
•o c
< a
1125
270
140
68 58
64 56
59 54
FEB.
12 24



6.4
6.8
7.0

8 8
9 8
8 7
33 33
33 33
33 33
524
412
Nil


110
120
Nil
1160
300
140
59 60
59 62
59 62
MARCH
10



6.5
6.8
6.9

9
9
9
33
33
33
634
254
Nil


50
35
Nil
825
210
110
68
70
74
24







8
8
9
33
33
33











69
68
68
  Ambient
Total
                         82
83
                                     62
                                           65
                                                  60
                                                         61
                                                                38
                                        60  43
                                                                                  58  65
                                                                                              70
                                                                                                    58
Aerobes x 10' B-l 37 155 80 73 210 3
x 10' B-2 28 27 75 18 21 2
x 10' B-3 3 3| 27 3 53 3 3
Number Plowirigs
Between Sample
Dates
50333523 42 12
                                              81

-------
            APPENDIX A




          TABLE 6 (CONTD.)




         CHRONOLOGICAL DATA




BUNKER C (NO. 6) FUEL OIL, "B" PLOTS




APRIL

pH, Before Extr.


pH, After Extr.


Moisture, %w


Oil, %w


NHo as N, ppm


NOj as N, ppm



P04 as P205> ppm


Temperature, °F


Ambient
Total
Aerobes x 10
xlO7,
x 10'
Number Flowings
Between Sample
Dates

B-l
B-2
B-3
B-l
B-2
B-3
B-l
B-2
B-3
B-l
B-2
B-3
B-l
B-2
B-3
B-l
B-2
B-3

B-l
B-2
B-3
B-l
B-2
B-3


B-l
B-2
B-3


7
6.6
6.8
6.9



9
7
9
34
35
34
630
370
Nil
20
Nil
Nil

930
240
100
90
96
93
67

5
0.1
0.1
2

21






6
8
9
33
32
30


CO
%
g
•o
0)
•o
•0



94
92
87
75




2



MAY
5



6.9
6.9
7.0
6
6
7
33
33
31
440
300
Nil
100
5
Nil

750
220
100
85
84
83
80




2

19






3
3
3
34
36
36










102
103
101
82

4
10
30
1


JUNE
2 30



6.9
7.0
7.1
3 5
3 5
3 4
35 32
37 36
31 33
410
290
Nil
20
1
Nil

840
230
80
92 96
92 94
92 92
83

3
2
3
1 3

1971


JULY AUGUST
14



6.9
7.0
7.1
3
2
2
33
38
33
360
170
Nil
15
1
Nil

900
230
95
93
95
93
84

5
4
1
2

28 11
6.3
6.5
6.7
6.5
6.6
6.7
3 6
3 6
3 6
32 31
36 27
31 32

fl
8,
^
0)
•a
•a

800
230
95
94 90
93 90
93 88
86 83

54
30
0.5
1 1

25



7.1
7.2
7.3
2
2
2
29
24
32
430
230
Nil
145
125
Nil

740
250
130
95
95
94
84




10



SEPT.
25
6.8
7.0
7.2
6.9
7.0
7.2
4
6
4
26
24
26
116
Nil
Nil
Nil
Nil
Nil

680
230
115
82
81
81
78






30









25
22
30




















            82

-------
       APPENDIX  A




          TABLE 7




    CHRONOLOGICAL DATA




WAXY OIL PRODUCT, "C" PLOTS







                       1970


pH, Before Extr.


pH, After Extr.



Moisture, %v


Oil, %w


NH, as N, ppm


NO, as N, ppm
J


PO. as P-0 , ppm
4 2. j

Temperature , °F


Ambient
Total 7
Ae robes x 10-
x 10,
xlO7
Number Plowings
Between Sample
Dates


C-l
C-2
C-3
C-l
C-2
C-3

C-l
C-2
C-3
C-l
C-2
C-3
C-l
C-2
C-3
C-l
C-2
C-3

C-l
C-2
C-3
C-l
C-2
C-3


C-l
C-2
C-3



APR.
22
7.0
7.0
7.0




17
17
13
10
10
10
Nil
Nil
Nil
Nil
Nil
Nil

110


78


80







MAY
7 13 17 19 27
7.4
7.7
7.4


M
o
8 * 17 9
, 7 5 15 8
7 < 16 10
„ 11 24 22
% 12 20 22
°. 11 19 19
a*
| 440
a. 520
1 Nil
3
2 Nil
£ Nil
•o Nil
0)
3


77 80 83


81 80 82

4
5
2

1 21

JUNE
11 24
6.8
6.8
6.9




8 9
8 8
8 9
22 19
20 16
19 17
50
30
Nil
Nil
Nil
Nil




83 86


82 76

16
41
10

1 1

JULY
10
7.2
7.2
7.2




16
13
12
20
22
17
40
35
Nil
Nil
Nil
Nil

140
105
80





39
128
45

1

29
7.5






6
6
8
19
18
18
3
Nil
Nil
Nil
Nil
Nil




89


87





1

AUGUST
7 14
6.9
6.9
6.8




6
5
5
2 18
1 "
1 17
CO
1 1230
770
a Nil
OS
o Nil
3 Nil
•o Nil
0
•o
< 470
160

89 94


86 86

31
25
4

2


27







6
6
5
17
15
16










94


83





1

         83

-------
       APPENDIX  A




     TABLE 7 (CONTD.)




    CHRONOLOGICAL DATA




WAXY OIL PRODUCT, "C" PLOTS
1970
SEPT.

pH, Before Extr.


pH, After Extr.


Mois ture » %v

Oil, Zw


NH, as N, ppm
j


NO. as N, ppm
J


PO as P 0 , ppm
H tf J

Temperature, °F


Ambient
Total
Aerobes x 10,
A 10J
x 10
Number Plowings
Between Sample
Dates

C-l
C-2
C-3
C-l
C-2
C-3
C-2
C-3
C-l
C-2
C-3
C-l
C-2
C-3

C-l
C-2
C-3
C-l
C-2
C-3
C-l
C-2
C-3


C-l
C-2
C-3



10
7.0
7.0
7.0



5
6
16
14
15
380
65
Nil

Nil
Nil
Nil



92


82





5

22
6.6
6.9
7.0



4
4
15
13
15
180
20
Nil

Nil
Nil
Nil



88


83

74
44
67

0

OCT.
16
6.5
6.7
7.0



7
5
14
13
16
115
10
Nil

Nil
Nil
Nil
720
360

68


62





3

31






9
7
12
13
14










62


65

87
19
10

3

NOV.
14
6.7
6.9
7.1

6.6
7.0
8
10
13
12
13
Nil
Nil
Nil

Nil
Nil
Nil
700
320
135
63


60

110
93
38

3

DEC.
17



6.2
6.6
7.0
6
5
13
12
14
20
5
Nil

Nil
Nil
Nil
780
295

58


61

49
34
7

5

JAN.
6



6.2
6.6
7.0
7
6
13
12
14
20
10
Nil

Nil
Nil
Nil
880
270
125
43


38

80
66
15

2

27 2



6.4
6.7
6.9 rt
Q
' 1
5 |
12
12
14

en
O
•z. si
-3- (0
93 O
O
•a a.
•8 *
3 §
780
280
140
64 57
62 55
60 53
60 43





3

1971
FEB.
12 24



6.6
7.0
6.9
•\F\ Q
J.U 7
11 10
9 8
31 33
29 31
27 28
460
310
Nil

40
30
Nil
720
250
150
59 82
58 63
58 63
58 65

17
7
4

4 2



MARCH
10



6.7
6.9
7.1
12
9
29
30
29
460
290
Nil

15
13
Nil
650
210
105
68
68
68
70

34
41
130

1

24





i \
J..3
10
5
27
27
28










70
82
70
58





2

         84

-------
        APPENDIX A




      TABLE 7 (CONTD.)




     CHRONOLOGICAL DATA




WAXY OIL PRODUCT, "C" PLOTS
                          1971
APRIL

pH, Before Extr.


pH, After Extr.


Moisture, %w


Oil, 7.w


NH_ as N, ppm
j


NO 3 as N, ppm


P04 as P205, ppm


Temperature, °F


Ambient
Total
Aerobes x 10,
xlO?
x 10'
Number Flowings
Between Sample
Dates

C-l
C-2
C-3
C-l
C-2
C-3
C-l
C-2
C-3
C-l
C-2
C-3
C-l
C-2
C-3

C-l
C-2
C-3
C-l
C-2
C-3
C-l
C-2
C-3


C-l
C-2
C-3


7



6.2
6.5
6.8
4
4
4
25
26
27
325
120
Nil

Nil
Nil
Nil
750
260
110
88
82
80
67

4
1
0.1
2

21






7
7
5
24
25
26

en
-t
I
•o
o
•D
3



94
86
82
75




2

MAY
5



6.6
6.7
6.9
4
4
4
24
26
26
205
65
Nil

90
20
Nil
620
180
90
86
86
82
80




2

19






9
8
3
23
25
29










97
94
90
82

10
10
9
'l

JUNE
2






4
5
4
22
25
27
120
40
Nil

Nil
Nil
Nil



92
90
86
83




1

30



6.8
7.0
7.2
13
14
9
21
23
25
230
130
Nil

2
2
Nil
620
200
90
96
92
90


4
49
135
3

JULY AUGUST
14



6.8
6.9
7.1
5
6
4
21
23
26
110
35
Nil

Nil
Nil
Nil
780
280
90
98
94
92
84

19
20
10
2

28 11
6.3
6.3
6.6
6.5
6.6
6.9
6 13
5 13
5 8
20 21
23 25
26 27


-------
                                                                       APPENDIX A


                                                                        TABLE 8


                                          TOTAL NITROGEN,  SULFUR,  PHOSPHORUS, AND METALS CONTENTS FOR "A" PLOTS


                 Legend;   Solvent a_ was 1.0 N Ammonium Acetate,  pH 7,  used for soil analysis at Texas A&M University.

                          Solvent b_ was 1.4 N Ammonium Acetate in  1 N  HC1, at pH 4.29 and the analyses were by atomic absorption.
00
CT>
Component START
in ppm April, 1970
Total
Nitrogen 1500
1300
1600



Sulfur 5000


EXTRACTABLE
WITH SOLVENT
a b
Phosphorus 820 36


Sodium AS 700


Potassium 156 930


Magnesium 1490 2050

_^_
Calcium 3300 26900


Plot
No.

A-l

A-2

A-3

A-l
A-2
A-3



A-l
A-2
A-3
A-l
A-2
A-3
A-l
A-2
A-3
A-l
A-2
A-3
A-l
A-2
A-3
DURING
Nov. 13, 1970
Total
2820
2500
2590

1650




EXTRACTABLE
WITH SOLVENT
b
3000


96
146
88
140
140
140
1100
1300
1400
45000
21000
22000

April 21, 1971
Total









EXTRACTABLE
WITH SOLVENT
b



350
1000
510
600
375
210
6850
6700
6700
27500
26500
26250

Sept
Total
3300
3100
3600
3200
2100
1800
2300
2500
3400
END
. 23.











1971










EXTRACTABLE


500
370
430












WITH
a
98
36
21
352
560
550
586
460
236
560
600
560
5900
7300
6550
SOLVENT
b



260
260
360
920
920
740
5750
7250
7120
24800
28000
28000

-------
00
                                                                         APPENDIX A

                                                                      TABLE 8 (CONTO.)

                                            TOTAL NITROGEN, SULFUR, PHOSPHORUS, AND METALS CONTENTS FOR "A" PLOTS

                  Legend:  Solvent c_ was 1 N HC1 used for soil analysis at Texas A&M University.
                           Solvent d was a 50/50 mixture HC1 and HN03 and the analyses were by atomic absorption.
Component
in ppm
Manganese
Iron
Zinc
Copper
Lead
START
April, 1970
EXTRACTABLE
WITH SOLVENT
c d
692 160
3800 10000
1580 1080
21 180
-
Plot
No.
A-l
A-2
A-3
A-l
A-2
A-3
A-l
A-2
A-3
A-l
A-2
A-3
A-l
A-2
A-3
DURING
Nov. 13, 1970
EXTRACTABLE
WITH SOLVENT
d
260
200
270
9750
10000
11500
1000
1140
1150
150
160
160
820
900
920

April 21, 1971
EXTRACTABLE
WITH SOLVENT
d
160
270
130
16500
20000
19800
1200
1560
1640
180
230
200
890
980
1030
END
Sept. 23,
1971
EXTRACTABLE
WITH SOLVENT
c d
146
146
144
1110
1160
1230
685
595
628
37
42
45
-
210
180
240
21500
21500
25300
1200
1500
1220
170
160
170
880
790
920

-------
                                                                       APPENDIX A


                                                                         TABLE 9


                                           TOTAL NITROGEN,  SULFUR,  PHOSPHORUS, AND METALS CONTENTS FOR "B" PLOTS


                  Legend:   Solvent  a was  1.0 N Ammonium Acetate,  pH 7,  used for soil analysis at Texas A&M University.
                           Solvent  b_ was  1.4 N Ammonium Acetate in  1 N  HC1, at pH A.29 and the analyses were by atomic  absorption.
00
00
Component
in ppm

Nitrogen





Sulfur





Phosphorus


Sodium


Potassium


Magnesium


Calcium


START Plot
April, 1970 No.
Total
1500 B-l
1300e
1600f B-2

B-3

5000 B-l
B-2
B-3
EXTRACTABLE
WITH SOLVENT
a b
820 36 B-l
B-2
B-3
45 700 B-l
B-2
B-3
156 930 B-l
B-2
B-3
1490 2050 B-l
B-2
B-3
3300 26900 B-l
B-2
B-3
DURING
Nov. 13, 1970 April 21, 1971
Total Total
3050





5000


EXTRACTABLE EXTRACTABLE
WITH SOLVENT WITH SOLVENT
b b
3810


100 500
330
300
140 550
375
210
1000 , 6850
6250
6850
17000 29000
25000
27250

Sept
Total
3300
3000
3300
3600
2400
2000
4000
3800
5300
END
. 23.











1971










EXTRACTABLE


970
280
360












WITH
a
222
35
16
270
238
278
796
484
258
550
490
550
5200
5550
5900
SOLVENT
b



220
210
220
920
520
520
6250
6750
7120
24440
26440
25200

-------
                                                                       APPENDIX A


                                                                    TABLE 9 (CONTD.)


                                          TOTAL NITROGEN, SULFUR, PHOSPHORUS, AND METALS CONTENTS FOR "B" PLOTS


                 Legend:  Solvent £ was 1 N HCl used for soil analysis at Texas A&M University.

                          Solvent d_ was a 50/50 mixture HCl and HN03 and the analyses were by atomic absorption.
00
vo
Component
in ppm
Manganese
Iron
Zinc
Copper
Lead
START
April, 1970
EXTRACTABLE
WITH SOLVENT
c d
692 160
3800 10000
1580 1080
21 180
-
Plot
No.
B-l
B-2
B-3
B-l
B-2
B-3
B-l
B-2
B-3
B-l
B-2
B-3
B-l
B-2
B-3
DURING
Nov. 13, 1970
EXTRACTABLE
WITH SOLVENT
d
180
6250
850
130
780

April 21, 1971
EXTRACTABLE
WITH SOLVENT
d
130
140
150
16500
20500
21800
1700
1640
1460
200
200
180
1090
1150
1230
END
Sept. 23,
1971
EXTRACTABLE
WITH SOLVENT
c d
138
138
150
1140
960
980
387
376
349
10
22
28
-
180
190
160
22600
22600
37600
1250
1090
860
170
150
150
1000
880
830

-------
                                                                      APPENDIX A


                                                                        TABLE 10


                                          TOTAL NITROGEN,  SULFUR,  PHOSPHORUS,  AND METALS  CONTENTS  FOR "C"  PLOTS


                 Legend:   Solvent  a was  1.0  N Ammonium Acetate,  pH 7, used  for soil  analysis  at  Texas A&M  University.
                          Solvent  b_ was  1.4  N Ammonium Acetate  in  1 N HC1,  at pH 4.29  and the analyses were  by atomic  absorption.
vo
O
Component
in ppm

Nitrogen





Sulfur





Phosphorus


Sodium


Potassium


Magnesium


Calcium


START Plot
April, 1970 No.
Total
1500 C-l
1300
1600 C-2

C-3

5000 C-l
C-2
C-3
EXTRACTABLE
WITH SOLVENT
a b
820 36 C-l
C-2
C-3
45 700 C-l
C-2
C-3
156 930 C-l
C-2
C-3
1490 2050 C-l
C-2
C-3
3300 26900 C-l
C-2
C-3

Nov. 13, 1970
Total


2600

1690

3500


EXTRACTABLE
WITH SOLVENT
b



_
60
59
.
140
140
-
1200
1400
_
20000
22000
DURING
April 21, 1971
Total









EXTRACTABLE
WITH SOLVENT
b



260
280
420
600
440
250
6525
6525
6250
27250
26750
27000

Sept
Total
4200
3500
3000
2500
1900
1900
2700
2200
2200
END
• 23,











1971










EXTRACTABLE


940
280
120












WITH
a
134
38
15
240
246
206
792
554
225
500
630
590
5200
6200
6600
SOLVENT
b



180
220
220
800
720
580
6250
7120
7500
22800
22000
29600

-------
                                                                      APPENDIX A

                                                                    TABLE  10 (CONTD.)

                                          TOTAL NITROGEN,  SULFUR,  PHOSPHORUS, AND METALS  CONTENTS  FOR "C" PLOTS

                Legend:   Solvent  c_ was  1  N  HC1 used  for soil  analysis at  Texas ASM University.
                          Solvent  d_ was  a  50/50 mixture HC1  and  HN03 and the analyses  were  by  atomic  absorption.
vo
Component
in ppm
Manganese
Iron
Zinc
Copper
Lead
START
April. 1970
EXTRACTABLE
WITH SOLVENT
c d
692 160
3800 10000
1580 1080
21 180
-
Plot
No.
C-l
C-2
C-3
C-l
C-2
C-3
C-l
C-2
C-3
C-l
C-2
C-3
C-l
C-2
C-3
DURING
Nov. 13, 1970
EXTRACTABLE
WITH SOLVENT
d
210
200
11250
12500
• 900
900
1AO
150
860
940

April 21, 1971
EXTRACTABLE
WITH SOLVENT
d
100
220
1000
15500
20000
20800
1180
1560
1260
150
170
170
1030
1110
1020
END
Sept. 23,
1971
EXTRACTABLE
WITH SOLVENT
c d
180
162
980
830
842
441
18
48
-
260
250
200
21300
22300
23000
920
1190
1150
140
140
140
900
820
880

-------
                                        APPENDIX A



                                        TABLE  11




                       OIL AND NUTRIENT  PENETRATION  DEPTHS INTO SOIL
OIL


0 - 0.5 ft-


2 ft.


4 ft.


6 ft.


NUTRIENT
NH , 2 ft.
J
4 ft.

NO 2 ft.
J»
4 ft.

P205,2 ft.

4 ft.



Start
Midway
End
Start
Midway
End
Start
Midway
End
Start
Midway
End

Midway
End
Midway
End
Midway
End
Midway .
End
Midway
End
Midway
End

1
10
10
12
8
6
7
<0.5
<0.5
<0.5
<0.5
<0.5
	

NIL
	
NIL
_
NIL
	
NIL
	
240
	
NIL
	
A
2
13
13
16
18
11
21
<0.5
<0.5
<0.5
<0.5
<0.5
	














3
13
15
21
21
13
25
<0.5
<0.5
<0.5
<0.5
<0.5
	


NIL

NIL

NIL

NIL

150

NIL

1
13
15
25
11
11
15
<0.5
<0.5
<0.5
<0.5
<0.5
	

96
NIL
NIL
NIL
NIL
NIL
NIL
NIL
160
150
NIL
NIL
B
2
11
14
24
11
11
13
<0.5
<0.5
<0.5
<0.5
<0.5
	














3
12
16
30
11
16
12
<0.5
<0.5
<0.5
<0.5
<0.5
	














1
11
12
18
11
12
13
<0.5
<0.5
<0.5
<0.5
<0.5
	

NIL
NIL
NIL
NIL
NIL
NIL
NIL

190
200
NIL

C
2
12
12
18
11
11
12
<0.5
<0.5
<0.5
<0.5
<0.5
	




NIL



50


NIL


_3_
ll
14
23
11
16
18
<0.5
<0.5
<0.5
<0.5
<0.5
	













                                           92

-------
                                     APPENDIX A

                                       TABLE  12

                           HYDROCARBON TYPE FOR "A"  PLOTS
AT START

 Residual oil in
 soil

  Oil added
AFTER 8 MONTHS

 Residual oil in
 soil

  Oil added
A-l
A-2
A-3
  After addition   A-l
   (calculated)    A-2
                   A-3
A-l
A-2
A-3
  After addition   A-l
   (calculated)    A-2
                   A-3
AFTER 14 MONTHS

 Residual  oil  in
 soil

AT END (17 MONTHS)
 Residual  oil  in
 soil
A-l
A-2
A-3
A-l
A-2
A-3

Sats.
40
40
40
36

16
27
33
44

24
30
35
14
21
35
Basis
Resin
21
21
21
8

50
29
34
5

32
22
18
44
42
26
Oil
Arom.
39
39
39
56

32
44
33
51

44
48
47
A2
37
39

Total
100
100
100
100

100
100
100
100

100
100
100
100
100
100

Sats.
4.0
5.2
5.2

6.2
7.0
7.0
1.8
3.5
5.3

6.3
9.2
10.6
3.8
6.3
8.4
1.7
3.1
6.3
Basis
Resin
2.1
2.7
2.7

2.6
3.1
3.1
5.5
3.8
5.4

6.1
4.5
6.0
5.1
4.6
4.3
5.3
6.3
4.7
Soil
Arom.
3.9
5.1
5.1

7.3
7.9
7.9
3.5
5.8
5.3

8.6
12.4
11.4
7.1
10.2
11.3
5.0
5.6
7.0

Total
10
13
13

17
18
18
11
13
16

21
26
28
16
21
24
12
15
18
                                         93

-------
                                APPENDIX A

                                 TABLE 13

                     HYDROCARBON TYPE FOR "B" PLOTS
                                               Percent Weight
                                  Basis Oil
                                            Basis Soil
                          Sats.  Resin  Arom.  Total   Sats.   Resin   Arom.   Total
AT START

  Residual oil in
  soil


  Oil added

  After addition
   (calculated)


AFTER 8 MONTHS

  Residual oil in
  soil

  Oil added

  After addition
   (calculated)


AFTER 14 MONTHS
B-l
B-2
B-3
B-l
B-2
B-3
B-l
B-2
B-3
B-l
B-2
B-3
40
40
40

18
10
15
26
21
21
21
21

26
39
36
25

27
39
39
39
56
51
49
49
52
100
100
100

100
100
100
100

100
Residual oil in
soil

B-l
B-2
B-3
14
13
21
29
23
23
57
65
56
100
100
100
4.3
4.2
6.7
9.0
7.4
7.4
17.7
20.8
17.9
31
32
32
AT END (17 MONTHS)

  Residual oil in
  soil
B-l
B-2
B-3
13
17
23
40
33
26
47
50
51
100
100
100
5.2
4.4
4.8
7.9
6.9
7.0
1.7
2.1
4.4
6.3
7.4
8.2
4.3
4.2
6.7
3.3
4.6
6.9
2.7
2.3
2.5
6.6
5.9
5.6
6.6
.5.0
4.3
12.6
11.7
9.2
9.0
7.4
7.4
10.0
8.9
7.8
5.1
4.3
4.7
13.5
12.0
11.4
8.7
6.9
8.3
20.1
19.9
17.7
17.7
20.8
17.9
11.7
13.5
15.3
13
11
12
                                                    28
                                                    25
                                                    25
17
14
17
                                                    39
                                                    39
                                                    35
25
27
30
                                     94

-------
            APPENDIX A

             TABLE 14

  HYDROCARBON TYPE FOR "C"  PLOTS
                            Percent Weight
               Basis  Oil
                                    Basis Soil
       Sats.   Resin  Arom.   Total    Sats.   Resin  Arom.  Total
C-l
C-2
C-3
C-l
C-2
C-3
C-l
C-2
AT START

 Residual oil in
 soil

  Oil added

  After addition
   (calculated)


AFTER 8 MONTHS

 Residual oil in
 soil

  Oil added

  After addition   C-l
   (calculated)    C-2
                   C-3

AFTER 14 MONTHS

 Residual oil in   C-l
 soil              C-2
                   C-3

AT END (17 MONTHS)

 Residual oil in   C-l
 soil              C-2
                   C-3
40
40
40

90
27
38
42

90
        51
        60
        62
        34
        54
        61
 21
 21
 21

Nil
 35
 26
 23

Nil
       19
       14
       10
       33
       18
       13
39
39
39

10
38
36
35

10
        30
        26
        28
        33
        28
        26
100
100
100
100

100
100
100
100

100
100
100
100
100
100
4.4
4.8
4.4

15.2
12.9
11.6
3.5
4.6
5.9

18.8
19.9
18.5
11.7
14.4
16.1
5.8
11.3
14.6
2.3
2.5
2.3

2.3
2.5
2.3
4.6
3.1
3.2

4.6
3.1
3.2
4.4
3.4
2.6
5.6
3.8
3.1
4.3
4.7
4.3

5.5
5.6
5.1
4.9
4.3
4.9

6.6
6.0
6.3
6.9
6.2
7.3
5.6
5.9
6.3
11
12
11
                                                   23
                                                   21
                                                   19
13
12
14
                                                           30
                                                           29
                                                           28
                                     23
                                     24
                                     26
                                     17
                                     21
                                     24
                  95

-------
                           APPENDIX B

                       MICROBIAL ANALYSES


                            PROCEDURE

Soil sample was mixed thoroughly and one gram was weighed aseptically
and placed in a dilution bottle containing 99 ml saline.  The sample
was shaken vigorously for one minute then placed on a rotary shaker
for 25 minutes.  At the end of this period the bottle was removed and
one ml was pipetted into another 90 ml saline blank and mixed 30
seconds.  One-tenth ml of this sample was pipetted onto the surface
of a nutrient agar plate and spread with a sterile glass spreader.
Further serial dilutions and platings were made of the sample in order
to obtain statistically accurate plates (those with 30-300 colonies).
Duplicate platings were made of each sample.

After four days' incubation at 30 C the plates were counted, and the
four most numerous organisms were isolated for identification.  Gram
strains and normal biochemical tests were performed on the isolates
and identification was made through Bergey's Manual of Determinative
Bacteriology.6

Predominant individual organisms determined monthly throughout the
project for "A", "B", and "C" plots are given in following Tables 1,
2, and 3, respectively.  A.brief description from literature3*6 of
each predominant organism follows these tables.
                                96

-------
                                                       APPENDIX B

                                                         TABLE 1

                                     PREDOMINANT INDIVIDUAL ORGANISMS FOR "A" PLOTS

                                                         LEGEND
                                                         a = A-l
                                                         b = A-2
                                                         c = A-3

                                             1970	
                                                                           1971
MICROORGANISM

Achromobacter, Sp
   cycloclastes
   delmarvae
   pestifer
   xerosIs

Arthrobacter, Sp
   tumescens

Bacillus, Sp
   cereus var.mycoides

Corynebacterium, Sp

Flavobacteriura, Sp
   aquatile
   arborescens
   balustinura
   breve
   diffusum
   ferruglneura
   lutescens
   peregrinum
   rhenanum
   rigense
   solare

Micrococcus,  Sp
   roseus
MAY   JUNE  JULY  AUG.  SEPT. OCT.  NOV.  DEC.  JAN.  FEB.  MAR.  APR.  MAY   JUNE  JULY  AUG.  SEPT.

                              a                             t                             b
abc   abc   ac
                        abc
                                    ac    be
                        ab          be    a

                  abc         abcc
c

be
                                                                              ab
be    ace         bee   abc   aabc

      ab    abc   be          a
be
ab
c
             be
                                     abc

-------
vo
                                                                       APPENDIX B




                                                                     TABLE  1  (CONTD.)




                                                      PREDOMINANT INDIVIDUAL  ORGANISMS  FOR "A" PLOTS




                                                             1970
1971
MICROORGANISM
Nocardia, Sp
actinomorpha
alba
corallina
flavescens
maculata
minima
opaca
paraffinae
po lychromogenes
rubroportincta
salmonicolor
Pseudomonas, Sp
aeruginosa
arvilla
boreopolis
crucivlae
dacunhae
denitrificans
effusa
oleovorans
oval is
putida
rathonls
striata
stutzeri
Rhodotorula, Sp
Saccharomyces , Sp
analatus
Sarcina, Sp
barker i
flava
Yeast
MAY JUNE JULY AUG. SEPT. OCT. NOV. DEC. JAN. FEB. MAR. APR. MAY JUNE JULY AUG. SEPT.
b a
c t c
ac ab t c be
be
abc
be ab be ac be ac
be a
ac a
ab ab
ac
c a be a ab abc abc t abc c c
a abcc
c
ab abc ab
c
ab ac a
ab ab
b b
ab a
b a- a-
b abc t bbc b
b
a
be bbc abc abbe

-------
VO
VO
                                                                       APPENDIX B

                                                                         TABLE 2

                                                     PREDOMINANT INDIVIDUAL ORGANISMS FOR "B" PLOTS

                                                                          LEGEND
                                                                          e = B-l
                                                                          h = B-2
                                                                          c = B-3

                                                            1970
MICROORGANISM

Achromobacter, Sp
   eyeloclastea
   delmarvae
   pestifer
   xerosis

Arthrobacter, Sp
   turoescens

Bacillus, Sp
   cereus var.mycoides

CorynebacCerium, Sp

Flavobacterium, Sp
   aquatile
   arborescens
   balustinum
   breve
   diffusum
   ferrugineum
   lutescens
   peregrinum
   rhenanum
   rigense
   solare

Micrococcus, Sp
   roseus
                                                                                                                  1971
                                       MAY   JUNE  JULY  AUG.  SEPT. OCT.  NOV.  DEC.  JAN.  FEB.  MAR.  APR.  MAY   JUNE  JULY  AUG.  SEPT.
be
be
b
c
                                                         bee
b
c
be
a

abc
                                                               ab
                  abc   ab
                                                               be
                                                                     ac
                                                                     be

                                                                     ab
                                    be

                                    ab
                                                                                 bbc
                                    abc

                                    be
                                                                           bee
                                                                                       abc
                                                                                                               abc
                                                                              abc   abc

                                                                              b     b
                                                                                                                     be
                                                                                                                                 aac

                                                                                                                                 b
                                                                                                                                       aabb
                                                                                                be

-------
o
o
MICROORGANISM

Nocardia, Sp
   actinomorpha
   alba
   corallina
   flavescens
   maculata
   minima
   opaca
   paraffinae
   polychromogenes
   rubroportincta
   salmonicolor

Pseudomonas, Sp
   aeruginosa
   arvilla
   boreopolis
   crucivlae
   dacunhae
   denitrificans
   effusa
   oleovorans
   ovalis
   putida
   rathonis
   scriata
   stutzeri

Rhodotorula, Sp

Saccharomyces, Sp
   analatus

Sarcina, Sp
   barker!
— flava
                                                                      APPENDIX B

                                                                   TABLE 2  (CONTD.)

                                                    PREDOMINANT  INDIVIDUAL ORGANISMS FOR "B" PLOTS

                                                    	1970	  	
                                                                                                  1971
                                      MAY   JUNE  JULY  AUG.  SEPT. OCT.  NOV.  DEC.  JAN.  FEB.  MAR.  APR.  MAY   JUNE  JULY  AUG.  SEPT.
                                            be
                                      abc
                                                         ab
                                                         b
                                                              abc
                                                                                            abc
                                                                                abc
                                                                                                  ab
                                                                                            ab
                                                                                abc
                                                                          ab
                                                                                       c      abc
abc
abbe
b     a
                                                                                                              be

                                                                                                              c
            ab

            ac    aac
               Yeast
                                                                                                                          bbc   b     ab

-------
                                                            APPENDIX B

                                                              TABLE 3

                                          PREDOMINANT INDIVIDUAL ORGANISMS FOR "C" PLOTS

                                                              LEGEND
                                                                  C-l
                                                                  C-2
                                                                  C-3
                                           1970
                                                                                                1971
MICROORGANISM

Achromobacter, Sp.
  cycloclastes
  delmarvae
  pestifer
  xerosis

Arthrobacter, Sp
  tumescens

Bacillus, Sp
  cereus var.mycoides

Corynebacterium, Sp

Flavobacterium, Sp
  aquatile
  arborescens
  balustinum
  breve
  diffusum
  ferrugineum
  lutescens
  peregrinum
  rhenanum
  rigense
  solare

Mlcrococcus, Sp
  roseus
MAY   JUNE  JULY  AUG.  SEPT. OCT.  NOV.  DEC.  JAN.  FEB.  MAR.  APR^  MAY

                        a     c           c
      a                                   ac
                                                                                                    JUNE  JULY  AUG.   SEPT.
            ab
                  ab
                        be
            be
                  bcc
abc
c
                                                be    abc

                                                abcc
                              be

                              t
                                                                        bb
                                                                              ab
                                                                                    be
                                                                                               be
abc
      ab
                                                                              abc

-------
o
NJ
MICROORGANISM

Nocardia, Sp
  actinomorpha
  alba
  corallina
  flavescens
  maculata
  minima
  opaca
  paraffinae
  polychromogenes
  rubroportincta
  salmonicolor

Pseudomonas, Sp
  aeruginosa
  arvilla
  boreopolis
  crucivlae
  dacunhae
  denitrificans
  effusa
  oleovorans
  ovalis
  putida
  rachonis
  striata
  stutzeri

Rhodotorula, Sp

Saccharomyces, Sp
  analatus

Sarcina, Sp
  barkeri
  f lava
                                                                             APPENDIX B

                                                                          TABLE 3 (CONTD.)

                                                           PREDOMINANT INDIVIDUAL ORGANISMS FOR "C"  PLOTS

                                                            1970
                                                                                                                 1971
                                       MAY   JUNE  JULY  AUG.  SEPT. OCT.  NOV.  DEC.  JAN.  FEB.  MAR.  APR.  MAY   JUNE  JULY  AUG. SEPT.
                                       ab
                                             ab
                                                   be
                                                                     abc
                                                                                       ab
                                                                                       ab
                                                               abc   ab    aab   ac
                                                                                             ac
                                                         ab    be          be          b           c
                                                         b     c     ab          b           b
                                                         a
                                                                                                   abc
aabbc a     abcc  ab    be
abbcc
c
                                                                                                                           abbe  ab    abbbcc
                 Yeast

-------
                             APPENDIX B
                         MICROBIAL ANALYSES
             BRIEF DESCRIPTION OF PREDOMINANT ORGANISMS
Achromobacterium, Sp

   cycloclastes

   delmarvae

   pestifer
   xerosis
Arthrobacter, Sp
   tumescens

Azotobacter, Sp

   agilis
Bacillus , Sp

   cereus var.mycoides

Corynebacterium, Sp

Flavobacterium, Sp

   aquatile
   arborescens
   balustinum
   breve
   dif f us urn
   ferrugineum

   lutescens
   peregrinum
Attacks polysulfide polymer and found  in used
oil3
Attack phenol and naphthalene,  organic acids
not formed6
Produces acid from glucose, nitrites from
nitrates6
Organic acids from hydrocarbons not formed6
Produces nitrites from nitrates6
Citrates not utilized, nitrates and ammonia not
sole source of N6
Fixes atmospheric nitrogen, gives off C02,  opt
temp 25-28C6
Attacks polysulfide polymers, found in jet air
fuel, utilize paraffins but not cycloparaf fins3
Optimum pH 5.2, temp 40 °C, starch, sugar hydro-
lizes and produces acetylmethylcarbonol6
Seldom produces acid, glucose reacts to C02 +
     , utilizes paraffin tetradecane3
Acids not developed when nitrogen containing
compounds are in the medium6
pH 6.5 - 7.8, 10-30°C, produce acid from sugar6
Opt temp 30°C6 survive in jet fuel3
20-25°C6,
35° found in sewage6
25-30°C6, attacks polysulfide polymer3
pH 7-7.5 min. 6.5, 22-39°C, decomposes carbo-
hydrates6
30-35 °C
Opens benzene rings, destroys 2,4 dichloro-
phenoxyacetic acid
                                103

-------
   rhenanuro

   rigense

   solare
Hyphomicrobium, Sp
   vulgare
Micrococcus, Sp

   roseus
Nocardia, Sp

   actinomorpha

   alba
   corallina

   florescens
   maculata
   minima
   opaca

   paraffinae
   polychromogenos
   rubropertineta

   salmonicolor
Pseudomonas, Sp

   aeruginesa

   arvilla
   boreopolis
   crucivlae
   dacunhae
   denitrificans
   effusa
30°C produces acid from sugar, no reaction with
starch6
30°C produces acid from glucose and starch
hydrolized6
30° C
pH 7-7.5, 20-37°C, utilizes organic acids6
Utilizes paraffins, oxidizes phenols and found
in used oil3
25°C
Highly efficient paraffin to cellular matter
conversion3
pH 7.8 - 8.5, 25-30°C, utilizes phenols and
naphthalene6, oxidizes alkyl cyclic hydrocarbon6
Hydrolizes sugar, starch and organic acids6
pH 6.8 - 8, 22-25°C, utilizes phenol, cresol,
and naphthol6
28-38°C, hydrolizes starch6
Hydrolizes starch6
22-25°C, utilizes paraffin6
pH 6.8 - 7.3, 30°C, utilizes phenol and
naphthalene6
Min. pH 4.4, utilizes paraffin wax6
22-25°C6
pH 6.8 - 7.2, 20-37°C, utilizes benzene,
paraffin and mix petroleum6
20-22°C, utilizes paraffins6
Oxidizes paraffins, cycloparaffins, hydrocarbons,
used oil3, oxidizes hydrocarbons6
37-42°C, oxidizes toluene, asphalt, produces
odor trimethylamine (TMA)6
37°C, attacks naphthalene6
35-37°C, attacks naphthalene6
30-35°C, attacks phenols and m-cresol6
37°C, attacks phenols6
25 °C6
37°C6
                                104

-------
   olevorans
   ovalis
   putida
   rathonis
   striata
   stutzeri
Rhodotorula, Sp
Saccharomyces, Sp
   analatus
Sarcina, Sp
   barker!
   f lava
Streptomyces, Sp
   anulutus
   bikinierisis
Yeast
25-37°C,  attacks cutting oil  and  starch6
25-36°C6
25-27°C,  putrify material and form  TMA6
35°C, attacks phenol,  cresol  and  naphthalene5
25-36°C6
pH 7-9, 35°C, anaerobic N03 to N6
Found in used oil3
pH 7, 30°C, produces methane from carbonic acid6

Utilize paraffins and found in used oil3
Antagonistic mycobacteria
Strongly antagonistic, produces streptomycin
                               105

-------
                           APPENDIX C

               MICROBIOLOGY CONSULTANT'S COMMENTS

Many studies have been carried out on the identity of organisms
capable of oxidizing hydrocarbons.  The results obtained in this
project are very similar to the results obtained in other research
projects.15  The major species present are members of the genus
Pseudomonas, Flavobacterium, Nocardia, Corynebacterium, and Arthro-
bacter.

The nature of the hydrocarbon substrate does not appear to influence
the types of organisms present; however, it does have an effect upon
the number of bacteria in the soil samples.  Generally, the same types
of organisms predominated in the three plots.   Crude oil tank bottom
sludge produced the highest counts, the waxy oil product produced
intermediate counts, while bunker C fuel exhibited the lowest
microbial population.

Some evidence of ecological progression was noted during the 18-month
test period.  Members of the genera Achromobacter and Arthrobacter
were present in the early stages of the project but gradually dis-
appeared from the plots.  These genera were generally absent from all
plots during the second half of the study.  Members of genus Coryne-
bac terium began to appear at about the same time as Achromobac ter and
Arthrobacter species disappeared from the soil samples.  Members of
the genera Flavobacterium, Nocardia, and Pseudomonas were generally
present throughout the test period.  Unidentifiable yeasts began to
invade the test plots during the last three months of the project.

The presence of Bacillus, Sarcina, and Streptomyces species is prob-
ably not significant and has no relationship to the disposal of hydro-
carbons.  These organisms may have invaded the plots at variable
times due to dust contamination of the plots.   The dust may have come
from other areas.

Temperature appeared to have no effect upon the organisms present.
Some of the highest counts were obtained during the winter months.
                               106

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            APPENDIX C




              TABLE 1




 STATISTICS OF MICROBIAL ANALYSIS






PERCENT TIMES ISOLATED FROM SAMPLES
Genus
Ps eudomonas s p .
Flavobacterium sp.
Nocardia sp.
Corynebacterium sp.
Arthrobacter sp.
Unidentified yeasts
Sac char omyces sp.
Micrococcus sp.
Achromobacter sp.
Rhodotorula sp.
Sarcina sp.
Bacillus sp.
Streptomyces sp.
A
88.8
72.2
77.7
38.8
31.4
22.2
16.6
9.2
9.2
5.5
3.7
3.7
1.8
Plots
B
72.2
92.5
62.9
53.7
25.9
14.8
9.2
16.6
16.6
9.2
3.7
1.8
3.7
C
94.4
61.1
77.7
33.3
14.8
27.7
18.5
16.6
12.9
3.7
9.2
3.7
3.7
Mean
85
75
72
41
24
21
14
14
12
6
5
3
3
                 107

-------
                           APPENDIX C

                 MICROBIOLOGY CONSULTANT REPORT


Report of sampling of plots B-2 and C-2 with comparison counts on N.A.
and 17<> H.C. in nutrient agar media.

          B-2   -   1%   #6 Fuel oil
          C-2       1%   Paraffinate oil

     Nutrient Agar                    Fuel


     31~X 106 cells/gm                17~X 106 cells/gm

       Co                              c* *y
      ™ S                              \J ~ £,
     ~T~X 106 cells/gm                Ti~X 105 cells/gm

Organisms - N.A.

B-2  Flavobacterium lutescens
     Corynebacterium
     Yeast IV
     Co ryneb ac t e r ium

C-2  Yeast I
     Yeast II
     Yeast IV
     Corynebacterium


Hydrocarbons

B-2  Only three isolated, checked for characteristics by streaking
     on N.A. plate

          Corynebacterium sp. I
          Corynebacterium sp. II
          Yeast sp. IV

C-2  Only three org. isolated

          Corynebacterium sp. IV
          Pseudomonas sp. (unlike others)
          Yeast sp. Ill
                               108

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                           APPENDIX D

           INFRARED STUDY OF OILS EXTRACTED FROM SOIL


                         Method Summary

The infrared (IR) spectra of diluted oil samples were obtained on a
Perkin-Elmer, Model 137, double-beamed spectrophotometer.   Wavelength
accuracy is ±0.03 microns.  The oils were examined in matched NaCl
absorption cells of 0.5 mm thickness.

The oil samples were prepared for IR scans by first dissolving a small
portion of the oil in carbon tetrachloride (CC14).  The sample was then
centrifuged in a high-speed unit (5000 rpm) to remove very fine parti-
cles of soil entrained during the original extraction of the soil.  The
solvent was evaporated from the decanted solution under a nitrogen
atmosphere at 250°F.  Two solutions were prepared gravimetrically for
scanning.  The 2.0 to 9.0 micron (y) region was scanned on a 4%w/v
CCl^ solution, and the 8.5y to 15.Op region was scanned on a 10%w/v
carbon disulfide (€82) solution.  Solvent absorption was cancelled by
scanning the solvent simultaneously in the reference beam of the
spectrophotometer.

                       IR Band Assignments

The various structures from which the IR absorption bands arise are
noted at the bottom of each of Figures 25-27.  These assignments are
well known but require some explanatory comments.  The absorption near
2.9y is assigned to OH groups other than those in organic acids.  This
absorption could also arise from NH groups; however, such materials
are not present in waxy raffinate, yet the 2.9y band is the most
prominent in this material.  The absorption intensity of this band in
all of the samples is weak and appears to vary randomly.  Since no
special steps were taken to dry the oil samples or exclude small
amounts of moisture during handling, the 2.9y band may arise from
small amounts of water.  No further consideration will be given to
this band.

There are four absorption bands that have been assigned to organic
acids.  The OH portion of the carboxyl group (COOH), when present as
the dimer, exhibits a broad absorption on the long-wavelength side of
the strong CH absorption band.  This absorption is weak and difficult
to see at low concentrations of organic acids.

The sharp absorption band at 5.85y has been attributed to the C=0 of
the acid group.  This assignment has been verified by examination of a
single oil sample in p-dioxane solution resulting in a shift of the
C=0 band to 5.75y.  The effect of polar solvent alone should produce a
shift of the band to longer wavelength.  The shift of the C=0 band in
                               109

-------
question to a shorter wavelength is indicative of dissociation of the
acid dimer.  Moreover, the actual shift to a shorter wavelength is less
than would have been predicted by about the amount of reverse shift
expected from the polar solvent.  This prediction is based on Silver-
stein, R.M., and Bassler, G.C., Spectrometric Identification of Organic
Compounds, Second Edition, John Wiley and Sons, Inc., New York, 1967.
The assignment has been further verified by treating the oil sample
with triethylamine in chloroform solution.  This treatment resulted in
a shift of the C=0 band to about 6.3y by conversion to the carboxylate
ion, and the occurrence of a new band at 4.05y indicative of the
"ammonium band", also described by Silverstein and Bassler.

The absorption bands at 7.75y and 10.6y are assigned to the acid C=0
stretching band and the OH bending of the dimer, respectively.  At first
glance, absorption will be noted in the 7.65   7.75y range in all of
the samples including the waxy raffinate.  Absorption at 7.65y is
attributed to saturate groups; however, careful examination will reveal
that, in those samples where the acid C=0 absorption is strongest, this
weak band is shifted to 7.75y and becomes more pronounced.

The absorption band near 6.2y is assigned to C=C groups in aromatic
structures.  This band can be used as a gross measure of aromatic con-
tent, but widely varying intensities among aromatic types limits its
usefulness for quantitative measurements.

The absorption bands at 6.85y and 7.25y are assigned to CH2 and CH3
groups, respectively.  It should be pointed out that these bands
include such groups in both saturate molecules, per se, and in alkyl
appendages on aromatic nuclei.  The band at 13.88y is the well known
absorption arising from long paraffin chains; i.e., where four or more
CH2 groups are linked together in an uninterrupted sequence.  Again, a
five carbon sequence attached to an aromatic ring would also qualify.

The three strong absorption bands near 11.4y, 12.2y, and 13.2y all
arise from aromatic structures.  The structures shown on the figures
are intended only to illustrate the different configurations of the
hydrogen atoms around the aromatic nucleus, as these absorption bands
actually arise from the in-phase, out-of-plane vibrations of the
hydrogen atoms.  The position of the bands are actually dependent upon
the number of hydrogen atoms that are adjacent to each other, and the
aromatic nucleus can be a single or multi-ring system.  Thus, in a
polyaromatic ring system such as         H                  , where all
hydrogen vacancies are occupied by R groups, all three configurations
of hydrogen atoms are present.
                               110

-------
                             Results

The three different types of oils added to the soil test plots  in the
demonstration of oily waste disposal by the soil cultivation process,
oil extracted from the soil at the start of the project, and oils ex-
tracted from the soil at the end of eight months (midway) and again at
the end of the experiment (after 17 months), were examined by infrared
spectroscopy.  In between the 8-month and 17-month sampling periods,
additional oils were added to the plots.  The spectra of the oil samples
where crude oil tank bottoms was added to the soil plots are designated
as "A" plots and are shown in Figure 25.  The "B" plots received bunker
C oil (No. 6 fuel oil) and are shown in Figure 26.  The "C" plots
which were treated with waxy raffinate are shown in Figure 27.
Different degrees of fertilization, which was a principal variable in
the experiment, are noted on the figures.

Oil Residue in Soil

The IR spectrum of the oil extracted from the soil at the beginning of
this study is repeated in each figure  for convenience,  and has the
gross character of a polyaromatic oil.  Considerable evidence of
saturate groups is observed at 6.85y and 7.25y  together with a small
amount of long chain paraffin groups at 13,88y.  The weak C=0 band at
5.85y indicates a small amount of organic acids.  The other  organic
acid bands are not apparent, but treatment of this oil  with  tri-
ethylamine confirmed the presence of acid C=0 structures.

"A" Plots, Figure 25

The IR spectrum of the crude oil tank  bottoms  is  typical of  a saturate-
aromatic oil mixture.  The aromatic content  is  probably less than  that
in the oil extracted from the soil  originally.  This oil appears to
contain simpler aromatic structures as  evidenced by  the differences  in
the ll-14y region.  As expected, there  is no evidence  of CO structures.

The gross aromatic character of  the oils extracted  from all  of  the "A"
plots are similar to that of the original oil  extracted from the soil,
indicating that the remaining oil  in each case  was more polyaromatic.
There does not appear to be any  significant  effect of  fertilizer upon
the aromatic structures.

There is evidence of a small amount of long  chain paraffin  groups  at
13.88y, but the amount diminishes as the degree of  fertilization is
increased.  After 17 months there  is still some evidence of  paraffin
chains.

The most significant differences among the oils from the "A" plots are
found in the organic acid content.  After  17 months  there  is only  a
small amount of C=0 structure in A-3,  and  the  absorption for saturate
groups at 6.85y and 7.25y are still quite strong.  A-2  shows a  large
                                111

-------
increase in C=0 band intensity and the other acid absorption bands are
evident.  There is a further small increase in organic acids in A-l,
and the saturate groups at 6.85y and 7.25y decrease in both A-l and A-2
as the organic acid groups increase.  There is little difference in the
results obtained on the samples from the "A" plots after eight months,
except that the increase in organic acids was not as great with
moderate fertilization as was observed for the sample after 17 months.

"B" Plots, Figure 26

The IR spectrum of the bunker C oil is typical of a polyaromatic oil
and is quite similar to the oil extracted originally from the soil,
except that there is less long chain paraffin content and an absence
of organic acids.

After eight months' residence time, the aromatic characteristics of the
oils from the "B" plots are quite similar and also similar to the
original oil extracted from the soil.  The results are similar to
those in the "A" plots in that the long chain paraffin band is small .
and decreases with increased degree of fertilization.  The organic acid
content is evident but low in the spectrum of B-3; the other spectra
show a progressive increase in plots B-2 and B-l as the amount of
fertilizer was increased.  There is a concomitant decrease in the
saturate groups at 6.85y and 7.25y with the increase in organic acids.

After 17 months, the IR spectra of the oils from the "B" plots indi-
cate that something unique has occurred, since the organic acid content
does not appear to increase in those plots where fertilizer was applied.
There is also little change in the long chain paraffin groups in going
from plots B-3 to B-l.  An explanation for these results is offered in
the body of this report.

The absorption at 9.65y, attributed originally to aromatic materials,
shows a broadening and an increase in intensity in the spectra of  the
original bunker C oil and also of plot B-2 after 17 months.  This
behavior is probably due to contamination by traces of dirt particles
as silicas display absorption in this region of the IR spectrum.

"C" Plots, Figure 27

The IR spectrum of the waxy raffinate is typical of a nonaromatic  oil.
There is evidence of a very small amount of aromatics near 6.2y.   How-
ever, the principal absorption arises from the saturate groups at
6.85y and 7.25v and a large concentration of long paraffin chains  at
13.88y.

Oil extracted from each of the "C" plots is similar to that of the
original oil in the soil.  The spectra are typical of polyaromatic oils
even though the added oil was principally paraffinic.  This is not to
suggest that the paraffin oils are converted to aromatics, but that
                               112

-------
the paraffinic portion of the added oils was largely decomposed,  even
in the absence of fertilizer.

The amount of long chain paraffins remaining after 17 months in the
unfertilized plot (C-3) was greater than that after eight months  as
evidenced by the 13.88y band intensity.  There .was a higher concentra-
tion of long chain paraffins in both C-3 plots than in the oil present
originally in the soil, indicating that some paraffin material survived
the 17 months' residence in the soil.  The amount of paraffinic
material appeared to diminish in those plots where fertilizers were
used.

The use of fertilizer enhanced the production of organic acids in pro-
portion to the amount of fertilizer used as evidenced by the increased
CO band intensity at 5.85y.  The increase with moderate amounts  of
fertilizer was greater in the eight months sample.  A significant
decrease in saturate groups at 6.85p and 7.25y parallels the increase
in organic acid C=0 structures.  The organic acid content was quite
low in C-3 after 17 months but greater in C-3 after eight months.
This difference inversely paralleled the change in long chain paraffin
content as would be expected.

             INFRARED STUDY OF RAINFALL RUNOFF WATER

Infrared spectra of the oils recovered from samples of runoff water by
CClij extraction were obtained as conventional scans on the neat oils
following the evaporation of the solvent.

Three nonvolatile oils (NVO) and two volatile oils  (VO) recovered from
samples of runoff water were selected  for IR examination.  These oils
included NVO from plots A-l, B-l, and  C-l, and VO from plots A-2 and
C-l.  The IR spectra of all of the oils were very similar and were
identified as naphthenic acid concentrates by comparison with an IR
spectrum of a naphthenic acids mixture obtained from Eastman Kodak
Company.  This identification has been confirmed  from examination of
the NVO from plots A-l and C-l by high-resolution mass spectiometry,
which shows that both samples are very similar and  that they consist
of mixtures of naphthenic acids in the 200-300 molecular weight range.
Small amounts of other saturated organic acids may be present.

The long chain paraffin based at 13.88 per  is absent from  the IR
spectra in all but one of the oils (VO from plot  C-l), and no sig-
nificant amounts of aromatics are seen in any of  the oil.  Ultraviolet
(UV) spectra of NVO from plots A-l and C-l  (other oils were not
examined by UV) are completely lacking in characteristic UV absorp-
tion, and thus tend to confirm the absence of significant amounts of
aromatic structures.
                               113

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                           APPENDIX E

                   OIL CONTENT IN SOIL SAMPLES

                   Modified ASTM Method D-473,
            "Sedimentation in Fuel Oil by Extraction"

Modifications of ASTM method D-4738 were made to adapt the method for
determination of oily matter in soil.  A change in the apparatus con-
sisted of a stainless steel wire cone in place of the extraction
thimble of D-473.  A sketch of the modified apparatus is attached. For
the test, a folded filter paper placed inside the cone held a weighed
quantity of soil in a position beneath the condenser coil such that
solvent condensate (condensed vapors) trickled through the soil and
accomplished the extraction of oil from the soil.  The solvent was
carbon tetrachloride, C.P. grade, and the source of heat was an
electrical heater (Ful-Kontrol, 750 watts).

The procedure included 1) placing a weighed soil sample inside the
filter-paper lined, conical-wire basket, 2) drying the soil by placing
the basket in an oven at 110°C for one hour, and 3) weighing the dry
soil before extraction of the oil.  The difference in weight before
and after drying was considered to be water.  The dried weighed soil
was then placed in the extraction flask and extracted with carbon
tetrachloride according to D-473 method procedure.  After a period of
two to three hours, and/or when the drops from the bottom of the wire
basket appeared colorless, the heater was turned off and the sample
allowed to cool.  Then the condenser was removed and the carbon tetra-
chloride was evaporated, leaving the oily residue in the flask.  Final
evaporation was accomplished by introducing a stream of nitrogen gas
into the heated oil-containing flask.  Heat lamps were used for this
final step.  The flask containing the oil was weighed, then the oily
residue was washed from the flask with carbon tetrachloride and a tare
weight of the dry flask without the oil was obtained.

An alternate method was to oven-dry the soil sample after the extrac-
tion step and obtain the weight of the dry, oil-free soil.

For oily soil samples containing volatile components at the initial
drying temperature of 110°C, the undried soil sample was weighed and
extracted with carbon tetrachloride.  The extracted soil was then oven
dried and the difference in the initial weight and final weight was
the total oil plus water.  Oil was determined from the differences in
the flask weights by the above described procedure and the water con-
tent was calculated by difference.
                               114

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                                APPENDIX E (CONTD.)
      COOLANT
                                                           STAINLESS STEEL
                                                           WIRE BASKET
                                                                      SOLVENT
                      APPARATUS FOR EXTRACTION  OF OIL  IN SOIL
                                        115
OU.S. GOVERNMENT PRINTING OFFICE:1973  514-151/137 1-j

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1
Accession Number
w
5
2

Subject Field & Group
10A
SELECTED WATER RESOURCES ABSTRACTS
INPUT TRANSACTION FORM
Organization
OIJT7T T OTT rnvCDAVTV UnTTCTY^M CTTPTNTi'PV
         DEER PARK,  TEXAS
    Title
         OILY WASTE DISPOSAL BY  SOIL CULTIVATION PROCESS
1 Q Authors)
Kincannon, C. Buford
16

21
Project Designation
EPA, Project No.
12050 EZG
Note
 22
    Citation
                       Environmental Protection Agency report
                       number EPA-R2-72-110, December 1972.
 23
     Descriptors (Starred First)
      *0il Waste, *Sludge Treatment, *Soil  Disposal  Fields,  *Soil Microbiology, *Sludge Di:
posal, Disposal, Soils Oil Industry, Oil, Water  Pollution Sources, Industrial Waste, Waste;
Bacteria, Decomposing Organic Matter, Biodegradation,  Waste  Disposal, Microorganisms,
Aerobic Bacteria, Soil Bacteria Pseudomonas Fertilization, Drainage Effects, Fertilizers,
Soil Treatment Nutrient Requirements, Ammonium Compounds, Nitrates, Phosphates, Potash,
Ureas, Chromatography, Separation  Techniques,  Solvent  Extraction Fertilizer.
 25
     Identifiers (Starred First)
      Arthrobacter, Corynebacterium, Flavobacterium,  Nocardia,  Infrared Adsorption
      Oil Analyses
 27
     Abstract
      Three oily materials were used  in parallel experiments to demonstrate oily waste dis
posal by a soil cultivation  process at  prevailing climatic conditions.  The 18-month exper:
ments conducted with nine soil test plots  at Deer Park,  Texas, showed average oil decompos:
tion rates of 0.5  Ibs/ft^ of soil  per month without fertilizers and about 1.0 Ib/ft^/month
when fertilized.   Results of semi-monthly  oil determinations for each plot are given.  Maj<
microbial species  active in  the soil  were  members of the genus Arthrobacter, Corynebacterii
Flavobacterium, Nocardia, and Pseudomonas.  Predominant  species in each soil test plot are
reported on a monthly basis.
      Differences  in decomposition rate and microbial species due to hydrocarbon type as
present in the three feedstocks,  i.e.,  crude oil, bunker C fuel oil, and waxy raffinate oi
were minimal.  Infrared and  gas chromatography examinations of oil extracted from fertiliz<
and unfertilized soils showed differences  in organic acid contents and boiling ranges.
      Oil and fertilizer chemicals did  not infiltrate vertically into the soil at the test
location under prevailing conditions.
      Rainfall runoff water  contained 1) up to 100 ppm extractable oils found to be naph-
thenic acids and 2) up to 150 mg/1 ammonia as N when the nitrogen nutrients were excessive
in the soil.
      Photographs  show preparation of soil test plots, spreading of oil on  the soil, and
cultivation.  Data are tabulated  and  shown graphically.    (Kincannon-Shell)
Abstractor
        C. Buford Kincannon
                              Institution
                                         SHELL OIL COMPANY
 WR:I02 (REV. JULY 1969)
 WRSIC
                             SEND, WITH COPY OF DOCUMENT, TO: WATER RESOURCES SCIENTIFIC INFORMATION CENTER
                                                       U.S. DEPARTMENT OF THE INTERIOR
                                                       WASHINGTON, D. C. 20240

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