IPTC 11237

Optimising Fluid Choices for Landfarm Applications

John Hall and Simon Seaton, Halliburton and Suzanne
Visser, Dept. of Biological Sciences, University of Methods
Calgary Properties of Test Base Fluids
The first seven fluids listed in Table 1 were tested for both
biodegradability and ecotoxicity. They included a diesel, a
Copyright 2007, International Petroleum Technology Conference petroleum distillate (mineral oil), paraffin-based petroleum oil
This paper was prepared for presentation at the International Petroleum Technology (paraffin), two synthetic olefins (C16/18 IO; C14 LAO) and
Conference held in Dubai, U.A.E., 4–6 December 2007.
two esters (ester; alternate ester). An alkane/alkene blend was
This paper was selected for presentation by an IPTC Programme Committee following review tested only for its biodegradability potential using the
of information contained in an abstract submitted by the author(s). Contents of the paper, as
presented, have not been reviewed by the International Petroleum Technology Conference respiration monitoring approach. With exception of the
and are subject to correction by the author(s). The material, as presented, does not necessarily
reflect any position of the International Petroleum Technology Conference, its officers, or paraffin, all of the test fluids are insoluble in water.
members. Papers presented at IPTC are subject to publication review by Sponsor Society
Committees of IPTC. Electronic reproduction, distribution, or storage of any part of this paper
for commercial purposes without the written consent of the International Petroleum Technology Properties of Receiving Soil
Conference is prohibited. Permission to reproduce in print is restricted to an abstract of not
more than 300 words; illustrations may not be copied. The abstract must contain conspicuous The receiving soil was a loam obtained from a pasture site
acknowledgment of where and by whom the paper was presented. Write Librarian, IPTC, P.O.
Box 833836, Richardson, TX 75083-3836, U.S.A., fax 01-972-952-9435.
near Turner Valley, Alberta. Soil physical/chemical properties
indicate that the receiving soil is a relatively fertile soil with a
neutral pH and low conductivity (Table 2). These properties
Abstract
are conducive to promoting the degradation of hydrocarbons.
A well established method for bioremediation of drilled
cuttings and adhering base fluids is the use of landfarming or
Experimental Methods
composting. All base fluids are amenable to this method.
Biodegradability of Base Fluids
However, degradation of organic materials takes place over
Prior to initiating the research, the soil was passed through a 4-
different timescales, and there is potential for toxic residues to
mm sieve, homogenized and tested for moisture content.
affect the future use of land to which cuttings have been
Moisture content was determined gravimetrically by
applied. This paper describes a project where a degradation of
measuring the weight of three samples of soil before and after
a range of different commonly use base fluids was
drying at 80ºC.
investigated and the use of various commercial accelerants
was also tested to determine their impact on degradation rates.
Respiration Monitoring
It subsequently describes an investigation into plant and
Soil microbial activity is highly dependent on carbon
animal toxicity of soils to which base fluids have been applied.
availability to the microbial biomass and, therefore, is a
valuable tool for monitoring the degradation of bio-available
Introduction
hydrocarbons in the soil.
Recent concern about the environmental risks associated with
Three aliquots of field moist receiving soil, each
diesel-based drilling muds has encouraged consideration of
equivalent to 100 g dwt, were sprinkled with two grams of
alternate base fluids for the production of drilling muds. Of
each of the first seven test fluids listed in Table 1. Three
interest is the development of a drilling mud which performs
replicates were also run with no base fluid additions as
efficiently in the drilling operation and can be treated by land-
controls. Each base fluid replicate was amended with 400 µg
farming with no adverse environmental consequences. The
N/g dwt and 80 µg P/g dwt so that fluid degradation would not
purpose of the present research was to determine the
be constrained by low nutrient conditions. Previous studies
degradability and ecotoxicity associated with several base
have demonstrated that it is essential that available nitrogen be
fluids. The test fluids included synthetic olefins, esters,
sufficient to allow rapid and complete degradation of any
paraffin-based petroleum oil and diesel.
highly bioavailable carbon in the drilling fluids. Soil moisture
was adjusted to approximately 40% (dwt), and each sample
Objectives
was placed in a glass tube and incubated at 22ºC.
The overall objective was to develop a drilling fluid
Respiration (CO2 efflux) was monitored at regular intervals
formulation that degrades readily and rapidly in soil leaving
over 95 days using an Infrared Gas Analyzer (IRGA). This
no ecotoxic residual after the fluid has degraded to a stable
was to determine potential for degradability and to identify the
treatment endpoint. Specific objectives of the present research
biological treatment endpoint. The treatment endpoint was
were to compare the biodegradability of seven candidate base
defined as the point at which respiration stabilizes, and very
fluids in a clay loam soil, and also to evaluate the toxicity of
little difference in CO2-C release remains between successive
the fluid residuals remaining in the soil following
sample times. This endpoint signals the exhaustion of easily
biodegradation to a stable endpoint to a range of ecological
available hydrocarbons and the initiation of a slow decay
receptors (plants, earthworms, springtails).
2 IPTC 11237
phase. In the case of the test base fluids, a treatment endpoint
had been achieved after 95 days incubation.
Respiration Monitoring of Ester, C14 LAO and
Alkane/Alkene Fluids
The respiration profiles obtained from the trial described
above, indicated that the conventional ester degraded very
rapidly, and that peak respiration occurred prior to when
respiration monitoring was initiated (i.e., one day after fluid
application). To determine if this was the case, the respiration
trial with ester was repeated. For comparison, this trial
included re-testing of the C14 LAO fluid and testing of
another fluid, an alkane/alkene blend.
A total of four treatments were tested: control (untreated),
ester, C14 LAO and alkane/alkene blend. The experimental
methods and conditions were identical to those described for
the previous respiration trial, with the exception that
respiration was measured over 18 days, rather than 95 days.
Hydrocarbon analyses were not conducted on the treatments
tested in this trial.
Hydrocarbon (HC) Analyses
In addition to respiration, total extractable hydrocarbons
(TEH) in both control and base fluid treatments were analyzed
at the initiation of the biodegradability study and after 95 days,
when a biological treatment endpoint had been achieved, as
indicated by stable respiration measurements.
Extractable hydrocarbons (C11-C40+) were quantified
using the capillary column gas chromatography/flame
ionization detection (GC/FID) method described in Method
No. G108.0 (Alberta Environment, 1992). This method
involves extraction of soil with methanol and methylene
chloride, centrifugation, and the analysis of the methylene
chloride layer by injection into a gas chromatograph.
Individual carbon groups are separated according to boiling
point and quantified. Carbon scans and chromatograms were
used to quantify changes in the hydrocarbons. All hydrocarbon
data in the fluid treatments were corrected for background
carbon associated with the soil organic matter.
Chronic Toxicity Assessments of Bioremediated Base Fluid
Residuals
Because of the amount of soil required to conduct the
biological assays, bulk samples of soil with and without the
test fluids were incubated in conjunction with, but separate
from, the respiration samples. Moisture, nutrient and
temperature conditions of the bulk samples were identical to
those of the respiration samples. Longer-term plant (14-21
days), earthworm (70 days) and springtail (28 days) bioassays
were conducted to determine if there was any chronic toxicity
associated with the base fluid treatments following
degradation of the fluids to a stable endpoint.
Plant Bioassays
Three plant species, barley (Hordeum vulgare), alfalfa
(Medicago sativa) and northern wheatgrass (Elymus
lanceolatus) were used in the chronic plant toxicity bioassays.
Control and drilling fluid-treated soil that had been incubated
for 95 days was dispensed into plastic containers at 200 g
wwt/container. For each plant species, there were three
replicates for each of the control and seven fluid treatments,
resulting in a total of 24 containers per plant species.
Barley, alfalfa and northern wheatgrass seeds were planted
at a rate of five seeds/replicate for barley, 10 seeds/replicate
for alfalfa, and six seeds/replicate for northern wheatgrass.
Planting consisted of distributing the seed evenly on the soil
surface, and then embedding the seed to a depth of
approximately twice the diameter of the seed. The soil surface
was tamped lightly to ensure contact between the seed and the
soil, and sprayed with two mls of deionized water to promote
germination. Each container was placed in a plastic bag to
reduce moisture loss, and incubated in a growth chamber at a
temperature of 23-28ºC and a photoperiod of 16 hours light
and eight hours dark. The test units were distributed randomly
in the chamber and soil moisture was adjusted daily with
deionized water if necessary. Seven days after seeding,
emergence was determined and the alfalfa was reduced to five
plants per container.
Shoot and root elongation, and shoot and root dry mass of
barley were determined 14 days after planting, and 21 days
after planting in the case of alfalfa and northern wheatgrass. A
longer growth period was necessary for alfalfa and northern
wheatgrass because they are slower-growing species than
barley. Each plant was separated from the soil and washed
free of soil with deionized water. Shoot and root lengths were
determined on all plants by measuring (cm) from the
shoot/root interface to the tip of the longest shoot or root with
a ruler. Shoots and roots were separated and dried at 80°C for
48 hours to obtain dry shoot and root biomass produced over
14 or 21 days growth.
Earthworm Bioassay
Three replicates aliquots of 300 cc of remoistened soil from
each of the control and base fluid treatments were placed in
900-ml wide-mouthed, glass jars for a total of 24 jars. Ten
large, healthy, mature earthworms were placed on the surface
of the soil in each jar, and the jars were closed with a vented
lid. The test units were incubated at room temperature
(22±2°C), and earthworms were fed cooked oatmeal weekly.
After 42 days exposure, each jar was emptied into a tray
and live earthworms were separated from the soil to determine
percent survival of the adult earthworms. The adult
earthworms from each replicate soil sample were incubated in
deionized water for 24 hours, washed, dried at 80ºC, and
weighed to obtain a measure of earthworm biomass. After
removing the adult earthworms, the test units were incubated
for a further 28 days to allow earthworm cocoons to hatch,
resulting in a total incubation time of 70 days. After 70 days,
soil from each test unit was examined closely for the presence
of juvenile earthworms. After recording the number of
juveniles in each replicate, the combined earthworms from
each replicate were washed with deionized water, dried at
80ºC and weighed to obtain an estimate of juvenile biomass
production.
Springtail Bioassay
Three replicate aliquots of 30 g remoistened soil from each of
the control and base fluid treatments were weighed out and
placed in 125-ml glass jars (total = 24 jars). Using a
stereomicroscope at 10X magnification, 10 healthy, 10 to 12
IPTC 11237
day old springtails (Folsomia candida) were removed from a
stock culture with an aspirator and placed on the surface of the
soil in each jar. The jars were closed with a vented lid and the
test units were incubated at room temperature under a 12-hour
light/12-hour dark photoperiod. The springtails were fed
Baker’s yeast once weekly, and soil moisture was adjusted if
necessary. After 28 days exposure, the adults and juveniles
were heat extracted and counted under 6X magnification.
Statistical Analysis
A one-way analysis of variance (ANOVA) was applied to
each of the measurement endpoints for each of the receptor
species to determine if there were significant differences
amongst the control and base fluid treatments. Bartlett’s test
of equal variances was used to test for homogeneity of
variances. Where the ANOVA revealed a significant
treatment effect, Tukey’s (HSD) comparisons were applied to
determine which treatments were significantly different.
Differences are significant at the p • 0.05 level. Statistix 7
(Analytical Software, Tallahassee, Fl., 2000) was used to
conduct the statistical analyses.
Results and Discussion
Biodegradability: Respiration
Patterns of respiratory response, including the time required
by the microbial biomass to respond to base fluid addition and
the CO2 efflux at the point of peak respiration, can provide
valuable information on the degradability and bioavailability
of the carbon components of base fluids. With regard to
response times, the microbial biomass in the receiving soil
responded to the addition of all the test fluids within one day
of application. The extent of the response, relative to the
untreated soil, was greatest in the ester and alternate ester-
treated soils (Fig. 1; Table 3). Indeed, response to ester
appeared to be immediate since the highest respiration was
measured on the first day following application of this fluid,
and declined thereafter. This suggests, that under the
conditions of this study, ester and alternate ester are more
readily available to the microbial biomass, than are the
hydrocarbons in the other test fluids and, thus would degrade
more rapidly following application to the soil than diesel and
mineral oil, paraffin, C16/18 IO and C14 LAO.
Another indication of substrate availability is time to peak
CO2 efflux following fluid addition to the soil, and the extent
to which respiration is stimulated relative to that in untreated
soil. The amount of time required to attain peak respiration
was shortest in the ester treatment (< 1 day), followed by the
diesel, mineral oil and alternate ester treatments (11 days),
then the paraffin and C14 LAO treatments and lastly the
C16/18 IO olefin treatment (17 days). However, when CO2
efflux at peak respiration is considered also, then the most
carbon evolved as CO2 occurred in the alternate ester, paraffin,
C14 LAO and C16/18 IO fluid treatments (32-36 µg g-1),
followed by the ester treatment (22 µg g-1), and lastly the
mineral oil and diesel treatments (8 -13 µg g-1). Cumulative
CO2-C loss data illustrated in Fig. 2, support the peak
respiration measurements in that the most respiratory carbon
loss over the term of the study occurred in the C16/18 IO, C14
LAO, alternate ester, paraffin and ester-treated soils, while
respiratory carbon loss was much less from the mineral oil and
3
diesel treatments. Based on these respiration patterns, the
paraffin, olefins and esters are most bioavailable and
biodegradable, while hydrocarbons in the diesel and mineral
oil base fluids are less bioavailable.
Once respiration had peaked, microbial activity in all
treatments subsided quickly. Respiration rates of the base
fluid treatments approached that in the untreated soil
approximately 40 days after the initiation of the study. Thus,
the majority of carbon in all of the base fluids was
metabolized by the microbial biomass within 1.5 months of
addition to the soil. A stable degradation endpoint was
achieved 95 days after fluid application when respiration in all
the fluid treatments (0.8-1.9 µg CO2-C g-1 hr-1) was very
similar to that in the untreated soil (1.2 µg CO2 -C g-1 hr-1).
The low respiration signalled the exhaustion of the majority of
bioavailable carbon in the fluid-treated soils.
In summary, respiration profiles showed that the microbial
biomass responded most rapidly to the esters in the ester and
alternate ester base fluids, and the highest peak CO2 rate and
the greatest cumulative CO2-C loss were measured from soil
treated with C16/18 IO, C14 LAO, alternate ester, paraffin
and ester base fluids. Lower peak respiration rates and
cumulative respiratory carbon losses in the mineral oil and
diesel-treated soils may be a result of lower bioavailability of
the carbon constituents in these fluids, or less carbon available
for microbial utilization as a result of volatile PHCs that
dissipate abiotically.
Respiration Monitoring of Ester, C14 LAO and
Alkane/Alkene Blend
When a second respiration trial was conducted with ester base
fluid, the soil microbial biomass did not respond as rapidly to
the addition of the fluid as observed previously. Slower
response by the soil microbial biomass to ester in the second
trial (Table 4; Fig. 3) is probably due to differences in the
qualitative and quantitative attributes of the microbial biomass
in the receiving soil between the two trials. Soil storage can
reduce the amount of microbial biomass, thereby lengthening
the response time, and this may explain the disparity between
the two trials.
A rapid increase in metabolic activity was initiated
approximately 1.5 days, three days, and four days after
addition of ester, alkane/alkene blend and C14 LAO,
respectively. Within five days, CO2 efflux in all treatments
had peaked with alkane/alkene blend peaking earliest at 3.5
days, and ester and C14 LAO peaking at about the same
time—five days (Table 4, Fig. 3). Peak respiration rate for the
alkane/alkene blend and C14 LAO-treated soils was identical
at 40 µg CO2-C g-1 hr-1. Although the respiration rate of the
ester treatment peaked lower at 27 µg CO2-C g-1 hr-1, the
peak was sustained for a longer period of time than in the case
of the other two base fluids. As a result, the cumulative CO2-
C loss after 18 days of monitoring was greatest for the ester
treatment (7630 µg/g), followed by the C14 LAO and
alkane/alkene blend treatments (7314, 6634 µg C/g,
respectively).
Based on respiration patterns (Fig. 4), it is evident that all
three test fluids (ester, C14 LAO and alkanes/alkene blend) are
readily bioavailable and biodegradable with the majority of
carbon utilization occurring within two weeks after fluid
4 IPTC 11237
application to the soil. The alkane/alkene blend degraded
slightly more rapidly than the esters and olefins in the ester
and C14 LAO, respectively. The respiration pattern of the
ester-treated soil differed somewhat from that observed for the
alkane/alkene blend and C14 LAO treatments, which suggests
that the community of organisms involved in the degradation
of ester may not be the same as that involved in the
degradation of the other two fluids.
Biodegradability: Total Extractable Hydrocarbons (TEH)
The application rate of the test base fluids to the clay loam soil
was 2% by weight. Total extractable hydrocarbons measured
immediately following application of the fluids were similar to
the target 2% amount, ranging from 1.7% in the ester
treatment to 2.2% in the diesel treatment (Table 5). Carbon
scans showed that the hydrocarbons in each fluid were
primarily in the following carbon ranges:
• Diesel: C11 to C25; also some BTEX (benzene, toluene,
ethylbenzene, xylenes) (2.2 mg/kg) and purgeable (C5-
C10) HCs (134 mg/kg).
• Mineral Oil: C11 to C14.
• Paraffin: C12 to C17.
• C16/18 IO: C16 and C18.
• C14 LAO: C14 and C15.
• Ester: C16 to C25.
• Alternate ester: C19 to C25.
Any volatile PHCs in the base fluids would be expected to
dissipate within the first few days after fluid application.
Volatilized PHCs would not be available for microbial
utilization, thus reducing the PHCs available for microbial
uptake in any fluids with a volatile component. In this study,
there was a volatile component associated with the diesel
(BTEX, purgeable PHCs), and possibly with the mineral oil
fluid, although analysis for volatile PHCs was not performed
on this treatment.
Total extractable hydrocarbons measured after a stable
respiratory endpoint had been achieved revealed almost
complete removal of the PHCs after 95 days incubation (Table
5, Fig. 5) of all base fluids. Both carbon scans and
chromatograms showed extensive degradation of the
hydrocarbon constituents in each of the base fluids. The
greatest amount of hydrocarbon degradation occurred in the
ester treatments where almost 100% of the hydrocarbons were
removed. Hydrocarbon loss from the olefin fluids, C16/18 IO
and C14 LAO, was almost identical at 98%, while the mineral
oil and paraffin treatments demonstrated a 95% loss of PHCs.
PHC removal was lowest in the diesel treatment at 90%.
For the majority of base fluids, respiratory patterns and
cumulative CO2-C loss supported the hydrocarbon losses
determined by the analytical method. Fluids in this category
were C16/18 IO, C14 LAO, alternate ester, ester and paraffin,
which all exhibited high CO2-C release and almost complete
elimination of extractable PHCs.
Of interest are the mineral oil and diesel treatments. In
these two treatments, respiration patterns and cumulative
respiratory carbon losses indicated that PHC bioavailability
and degradation was not as great as it was for the other test
fluids. However, TEH analyses revealed that PHC removal
was greater than 90%. The discrepancy between the
respiration and hydrocarbon analysis results for the diesel and
mineral oil fluids may be due to the loss of hydrocarbons
through volatilization resulting in less carbon available for
microbial metabolism as discussed earlier, or possibly due to
irreversible sorption of PHCs to the soil particles, which also
reduces PHC bioavailability.
It can be concluded from both the TEH and respiration
data that under conditions where nutrients, moisture and
temperature are not limiting, the olefins (C16/18 IO, C14
LAO) and the esters (ester, alternate ester) are all highly
bioavailable and readily biodegraded to a negligible residual
(0.2-2% PHC). The paraffin is also readily bioavailable, but
the PHC residual remaining after respiration has stabilized is
slightly greater (approximately 4% PHC) than for the olefins
and esters. Respiration patterns indicate that mineral oil is not
as bioavailable as the olefins and esters, but TEH analyses
indicate that removal is similar to that measured for the
paraffin, paraffin. Diesel is the slowest to degrade as indicated
by the respiration measurements; however, PHC removal from
this fluid was still high at 90%, and this may be partially due
to abiotic losses of volatile PHCs. The largest PHC residual
after degradation has stabililized would be expected for the
diesel, where, based on the results of this study, 10% of the
PHCs were still present after the majority of bioavailable
PHCs had been metabolized.
Chronic Toxicity of Base Fluid Residuals: Plants
That barley productivity in the various base fluid treatments
was not significantly different from that measured in the
untreated soil (Table 6; Fig. 6), demonstrates that the HC
residuals remaining in the soil after more than 90% of the
hydrocarbons had been removed by volatilization, microbial
metabolism or irreversible sorption were not toxic to barley
growth. Slightly better growth in the diesel and paraffin
treatments than in the C14 LAO treatment may be due to
slight differences in fertility amongst these treatments. More
microbial activity and biomass production in the C14 LAO
treatment may have led to greater immobilization of nitrogen
and phosphorus in this treatment than in the diesel and paraffin
treatments where microbial activity was not stimulated to the
same extent.
No significant effects of HC residuals on emergence and
shoot and root production by alfalfa were detected in any of
the base fluid treatments, relative to that measured in the
untreated control (Table 7; Fig. 7). Rather, growth tended to
be better in the base fluid treatments than in the control,
particularly in soils containing mineral oil, C16/18 IO and
alternate ester hydrocarbon residuals.
As was observed for barley and alfalfa, no significant
inhibitory effects of base fluid residuals were detected on
emergence and shoot and root growth of northern wheatgrass
(Table 8).
In this study, test base fluids were applied to a clay loam
soil at 2% by weight and bioremediated under optimal
moisture, temperature and nutrient conditions to negligible HC
residuals (>90% removal). Results from the plant bioassays,
show that, under these circumstances, HC residuals ranging
from 45-79 mg/kg for ester and alternate ester, 297-423 mg/kg
for C14 LAO and C16/18 IO olefins, 777 mg/kg for the
paraffin, 923 mg/kg for mineral oil HCs and 2217 mg/kg for
IPTC 11237
diesel PHCs were not observed to significantly inhibit plant
growth relative to that measured in an untreated control soil.
Chronic Toxicity of Base Fluid Residuals: Earthworms
Of the bioassays generally conducted on PHC-contaminated
soils, the earthworm reproduction bioassay is often the most
sensitive to the presence of both fresh and weathered HCs, and
this was evident in the present study. Whereas there were no
significant inhibitory effects of base fluid HC residuals
detected in the plant bioassays, the earthworm bioassay
revealed a significant reduction in the number of juveniles
produced in the soil with diesel residuals (Table 9). However,
no significant effects of HC residuals on adult earthworm
survival and dry mass were evident in any of the base fluid
treatments. Mineral oil also tended to reduce juvenile
production relative to that in the control, but not statistically
so, probably because of high variability.
The earthworm bioassay revealed that HC residuals from
the majority of base fluids that were tested did not
significantly affect earthworm reproduction. Exceptions were
the diesel PHC residual, which significantly depressed the
number of juveniles produced, and the mineral oil PHC
residual, which also tended to reduce earthworm reproduction.
Chronic Toxicity of Base Fluid Residuals: Springtails
Adult springtail survival was high and not significantly
different amongst the treatments after 28 days exposure (Table
10). Springtail reproduction was not significantly affected by
the HC residuals in the various base fluid treatments, relative
to that measured in the untreated control soil (Fig. 9). Instead,
reproduction in soil with diesel, paraffin, C16/18 IO, C14
LAO, ester and alternate ester residuals was significantly
greater than that measured in the control.
Many springtails are fungal feeders, and it is suspected that
higher fungal biomass generated during the bioremediation of
the base fluids served as a food source for the springtails and
stimulated reproduction. An exception was the mineral oil
treatment, where springtail reproduction was not stimulated
and was, instead, slightly less than reproduction in the control
soil. Since cumulative CO2-C loss was lowest in the mineral
oil treatment, fungal biomass production was probably not as
great in this treatment as in the other treatments, which may
have reduced springtail reproductive capacity. Also, it is
possible that some of the petroleum distillates in the mineral
oil fluid inhibit not only springtail reproduction, but also
earthworm reproduction as discussed previously.
Conclusions
1. Respiration profiles over the 95-day period of the
study showed that highest peak CO2 rate and the
greatest cumulative CO2 -C loss occurred in the
C16/18 IO, C14 LAO, alternate ester, paraffin and
ester treatments. This indicates that bioavailability of
base fluid carbon was highest in these treatments.
2. Lower peak respiration rates and cumulative
respiratory carbon losses in the mineral oil and diesel
treatments suggests lower bioavailability of
hydrocarbons in these fluids. However, any volatile
PHCs potentially associated with these fluids would
5
dissipate abiotically and would not be accounted for
in the cumulative respiratory carbon measurements.
3. Based on cumulative CO2-C losses over the term of
the study, the order of the fluids in terms of % carbon
lost through respiration (most to least) was: C16/18
IO > C14 LAO > alternate ester > paraffin > ester >
diesel > Mineral Oil.
4. In a second respiration trial with ester and C14 LAO
and alkanes/alkene Mixture, it was evident that all
three base fluids were readily bioavailable and
biodegradable with the majority of carbon being
metabolized within the first two weeks after fluid
application. The alkane/alkene blend degraded
slightly more rapidly than the ester and C14 LAO
respectively. The respiration pattern of the ester-
treated soil varied from that observed for the
alkane/alkene blend and C14 LAO treatments, which
suggests that the microbial community involved in
the degradation of ester differed from that involved in
the degradation of the other two fluids.
5. TEH analyses after 95 days incubation showed that
there was over 90% loss of hydrocarbons in all base
fluid treatments with the order of degradability
(highest to lowest) as follows: Alternate ester
(99.8%) > ester (99.5%) > C14 LAO (98.4%) >
C16/18 IO (98%) > paraffin (95.7%)> mineral oil
(94.9%) > diesel (90.1%). Thus the smallest HC
residual remaining in the soil after incubation
occurred in the alternate ester base fluid treatment
(0.2%), while the highest PHC residual was present
in the diesel treatment (10%).
6. In general, biodegradability potential based on
respiration measurements supported the HC removal
data calculated from TEH analyses, with alternate
ester, ester, C14 LAO, C16/18 IO and paraffin base
fluids demonstrating the greatest respiratory carbon
loss and the greatest reduction in TEH. Exceptions
were mineral oil and diesel base fluid treatments,
which exhibited much lower respiratory carbon
losses, but relatively high TEH removal (90-95%).
The discrepancy between the respiratory carbon and
extractable hydrocarbon loss data, suggest that there
was abiotic removal of hydrocarbons, possibly
through volatilization, from the mineral oil and diesel
treatments. Volatilized PHCs would not be
accounted for in the cumulative respiratory carbon
losses, but would be included in the TEH
measurements.
7. Laboratory bioassays showed that hydrocarbon
residuals ranging from 45-79 mg/kg for ester and
alternate ester esters, 297-423 mg/kg for C14 LAO
and C16/18 IO olefins, 777 mg/kg for the paraffin,
923 mg/kg for mineral oil HCs and 2217 mg/kg for
diesel PHCs did not significantly affect survival and
growth by barley, alfalfa and northern wheatgrass.
8. There was no significant toxicity of HC residuals to
adult earthworm and springtail survival in any the
base fluid treatments. Also, the majority of the base
fluids did not significantly influence earthworm and
springtail reproduction. Exceptions were the diesel
6 IPTC 11237

residual, which significantly depressed juvenile
production by earthworms and the mineral oil
residual which, albeit not significant, tended to
reduce earthworm and springtail reproduction.
Enhanced springtail reproduction, relative to the
control, in many of the base fluid treatments is
attributed to extensive fungal development resulting
from degradation of the base fluids. An exception is
the mineral oil treatment, where springtail
reproduction was not stimulated and was, instead,
slightly less than reproduction in the control soil.
Since cumulative CO2-C loss was lowest in the
mineral oil treatment, fungal biomass production was
probably not as great in this treatment as in the other
treatments, and this may have reduced springtail
reproductive capacity. Also, it is possible that some
of the petroleum distillates in the mineral oil fluid
inhibit not only springtail reproduction, but also
earthworm reproduction.
Acknowledgements
The authors thank their respective organisations for giving
them permission to publish this paper.
References
Alberta Environment. 1992. Methods manual for chemical analysis
of trace organics and pesticides in environmental samples. AECV
92-M2.

TABLE 1-PROPERTIES OF TEST BASE FLUIDS AS SUMMARIZED FROM MATERI AL SAFETY DATA SHEETS
Base Fluid Properties
Diesel Clear, colourless liquid with diesel odour; 60-100% diesel; potentially carcinogenic.
Clear water-white liquid with mild mineral spirits odour; comprised of 100% petroleum distillates that are hydrotreated;
Mineral Oil
approximately 99% C11-C15 saturated hydrocarbons and 1% C11+ aromatics; no PCBs.
Paraffin Clear liquid with mild hydrocarbon odour; paraffin-based petroleum oil (60-100%).
Clear, colourless to pale yellow liquid; 100% C16/C18 Alpha Olefins, Isomerized; contains hexadecene (<80%) and
C16/18 IO
octadecene (<80%); soluble in hydrocarbon solvents.
Colorless liquid with slight hydrocarbon odour; >80% 1-tetradecene, <20% 1-hexadecene, 1-5% 1-dodecene, 1-5%
C14 LAO
octadecene (by weight).
Ester Yellowish liquid with mild odour; 60-100% fatty acid ester.
Alternate Ester Yellow liquid; readily biodegradable according to OECD classification.
Slightly yellow liquid with a mild hydrocarbon odour; light paraffin belonging to the acyclic hydrocarbon family; readily
Alkanes/Alkene Mixture
biodegradable (98% in 28d); relatively non-toxic.

TABLE 2-PHYSICAL/CHEMICAL CHARACTERISTICS OF RECEIVING SOIL PRIOR TO FLUID APPLICATION

Soil Texture Loam to clay loam (27-34% sand; 39-43% silt, 26-33% clay)
pH 6.6-7.0
Conductivity (dS/m) 0.6-1.1
Sodium Adsorption Ratio SAR (meq/100g) 0.1
Cation Exchange Capacity (CEC) 37-45
NH4-N, NO3-N (ug/g) 14.4
Organic matter (%) 12-15

TABLE 3. RESPIRATION RATES OF CLAY LOAM SOIL LEFT UNTREATED (CONTROL) OR TREATED WITH VARIOUS DRILLING BASE
FLUIDS AT A RATE OF 2% (WT:WT). RESPIRATION WAS MEASURED OVER PERIOD OF 95 DAYS. DAT A ARE MEANS (N=3) ± SD.
Treatment Respiration Rate (µg CO2 -C/g dm/hr)
D1 D3 D5 D7 D9 D11 D13 D15 D17 D19 D21 D23 D25 D33 D42 D64 D95
Control 3.6 3.2 2.9 2.8 2.8 2.7 2.5 2.6 2.4 2.9 2.2 2.3 2.5 2.7 2.8 1.8 1.2
± 0.2 ±0.2 ±0.5 ±0.2 ±0.2 ±0.5 ±0.1 ±0.1 ±0.1 ±0.1 ±0.1 ±0.03 ±0.5 ±0.2 ±0.5 ±0.1 ±0.04
Diesel 3.7 7.5 7.3 7.9 8.1 8.3 8.0 7.7 7.2 7.1 6.8 6.3 5.8 5.1 4.8 3.8 1.6
±0.04 ±0.3 ±0.2 ±0.2 ±0.2 ±0.3 ±0.2 ±0.1 ±0.2 ±0.2 ±0.2 ±0.1 ±0.1 ±0.3 ±0.2 ±0.4 ±0.1
Mineral Oil 3.6 6.9 7.8 10.4 12.6 12.8 10.0 7.6 6.4 5.6 4.9 4.3 3.8 2.9 2.3 1.6 0.8
±0.1 ±0.3 ±0.7 ±0.6 ±0.7 ±0.6 ±0.8 ±0.7 ±0.5 ±0.4 ±0.3 ±0.5 ±0.4 ±0.2 ±0.1 ±0.1 ±0.1
Paraffin 3.8 6.9 8.2 11.2 14.8 20.1 26.4 33.7 31.7 19.7 11.8 9.0 7.6 5.4 3.4 2.0 1.0
±0.2 ±0.2 ±0.1 ±0.2 0.4 ±0.8 ±1.6 ±0.5 ±6.5 ±4.9 ±1.5 ±0.9 ±0.6 ±0.6 ±0.3 ±0.1 ±0.1
C16/18 IO 3.6 6.7 9.0 11.8 15.5 18.2 25.0 29.5 31.9 28.1 21.0 15.4 13.2 9.4 5.9 3.4 1.7
±0.1 ±0.3 ±0.4 ±0.6 0.7 ±0.8 ±1.7 ±2.5 ±3.8 ±1.4 ±1.9 ±1.7 ±1.7 ±0.7 ±0.4 ±0.3 ±0.4
C14 LAO 3.8 7.8 11.5 16.2 21.4 22.6 29.9 32.7 30.4 26.0 18.5 12.0 9.0 5.9 3.9 2.1 1.0
±0.04 ±0.1 ±0.2 ±0.4 0.6 ±0.6 ±0.8 ±1.0 ±0.4 ±0.9 ±1.5 ±1.2 ±0.6 ±0.5 ±0.2 ±0.02 ±0.1
Ester 22.1 20.6 20.9 18.4 15.6 9.1 12.2 11.0 10.9 10.8 10.1 8.6 8.9 8.2 5.3 5.3 1.3
±0.7 ±0.4 ±0.4 ±0.1 0.3 ±0.3 ±0.5 ±1.1 ±1.3 ±1.9 ±1.9 ±1.4 ±1.0 ±0.9 ±0.7 ±1.9 ±0.3
Alternate 13.1 13.7 18.2 24.4 31.4 35.6 24.6 15.4 12.0 10.7 8.9 7.9 7.9 7.7 5.3 3.3 1.9
ester
±0.2 ±0.3 ±0.3 ±0.3 0.8 ±0.4 ±1.6 ±0.2 ±0.5 ±0.3 ±0.3 ±0.2 ±0.3 ±0.8 ±0.3 ±0.1 ±0.4
IPTC 11237 7

TABLE 4-MEAN RESPIRATION RATES (µG CO2 -C/g DM/Hr) OF CLAY LOAM SOIL LEFT UNTREATED (CONTROL) OR TREATED WITH
ESTER, C14 LAO AND BIOBASE BASE FLUIDS AT A RATE OF 2% (WT:WT). RESPIRATION MEASURED OVER A PERIOD OF 18 DAYS.
DATA ARE MEANS (N=3) ± SD.
Control Ester C14 LAO Alkanes/Alkene Mixture
Day Mean SD Mean SD Mean SD Mean SD
0.25 2.05 0.05 3.85 0.26 2.55 0.15 5.41 0.28
0.5 2.08 0.11 4.31 0.19 2.33 0.13 4.54 0.10
0.75 1.92 0.20 4.53 0.23 2.12 0.11 4.26 0.26
1 1.99 0.07 5.27 0.10 2.21 0.07 4.11 0.06
1.25 1.88 0.23 5.19 0.83 2.07 0.41 4.32 0.02
1.5 1.72 0.15 4.83 1.10 1.84 0.58 3.99 0.21
1.75 1.79 0.10 7.00 0.10 2.15 0.06 4.25 0.42
2 2.05 0.03 8.57 0.04 2.70 0.07 5.31 0.56
2.25 1.72 0.17 10.19 0.38 2.68 0.09 6.45 0.93
2.5 1.68 0.08 12.24 0.44 3.07 0.05 9.80 2.31
2.75 1.71 0.12 14.70 0.42 3.63 0.05 17.38 5.54
3 1.91 0.07 17.55 0.22 4.94 0.14 29.93 6.95
3.25 1.71 0.03 20.06 0.11 6.45 0.24 38.88 3.23
3.5 1.81 0.08 21.23 0.34 8.89 0.38 40.14 2.38
3.75 1.80 0.13 22.63 0.40 13.12 0.47 39.26 2.40
4 1.53 0.02 23.80 0.37 19.34 0.45 38.42 2.84
4.25 1.80 0.21 25.33 0.17 26.78 0.11 35.75 3.46
4.5 2.42 0.13 26.64 0.32 34.54 1.23 33.28 4.42
4.75 1.55 0.17 26.00 0.59 37.52 1.71 30.44 5.24
5 1.51 0.07 25.74 0.17 39.61 1.05 29.17 3.94
5.25 1.79 0.11 27.57 1.28 38.15 2.31 27.42 3.74
5.5 1.43 0.08 26.75 1.32 36.51 2.01 26.27 3.41
5.75 0.93 0.08 25.67 1.17 34.81 2.03 25.15 4.37
6 2.05 0.07 26.67 1.13 34.70 2.55 24.78 3.07
6.25 1.46 0.12 26.77 0.89 33.15 2.26 23.60 2.77
6.5 2.41 0.03 26.49 0.81 31.67 2.61 23.96 3.63
6.75 1.98 0.21 25.19 0.72 30.47 2.43 22.58 3.49
7 1.57 0.23 24.04 0.50 29.27 2.26 21.32 3.49
7.25 1.43 0.52 23.33 0.55 29.45 3.17 20.70 3.59
7.5 1.81 0.21 23.48 0.30 28.43 2.12 19.15 2.50
7.75 1.62 0.25 22.08 0.15 27.18 1.95 17.72 2.48
8 0.82 0.42 21.26 1.31 27.39 1.96 17.02 2.45
8.25 1.53 0.21 21.04 0.12 27.52 3.20 16.32 3.05
8.5 1.75 0.56 20.75 0.09 25.22 4.62 15.27 2.91
8.75 1.51 0.21 20.39 0.12 25.38 3.03 14.45 2.89
9 1.55 0.21 20.11 0.22 23.97 2.96 13.66 2.73
9.25 1.35 0.17 19.24 0.39 22.16 2.75 12.68 2.66
11 1.71 0.16 17.79 0.21 17.62 1.52 11.17 1.70
14 0.99 0.02 13.96 0.81 10.33 1.25 9.72 2.25
18 1.50 0.07 13.57 0.87 8.25 0.93 8.16 1.75
8 IPTC 11237

TABLE 5- REMOVAL OF EXTRACTABLE PHCS FROM CLAY LOAM SOIL TREATED WITH VARIOUS DRILLING BASE FLUIDS AND
BIOREMEDIATED FOR 90 DAYS. DATA HAVE BEEN CORRECTED FOR BACKGROUND INDIGENOUS SOIL CARBON AS DETERMINED
FROM UNTREATED SOIL
Drilling Base Fluid Initial PHC mg kg-1 Final PHC mg kg-1 PHC Loss after 90 d mg kg-1 % PHC Removal
Diesel 22295 2217 20078 90.1
Mineral Oil 17951 923 17028 94.9
Paraffin 17989 777 17212 95.7
C16/18 IO 20737 423 20314 98
C14 LAO 18370 297 18073 98.4
Ester 17179 79 17100 99.5
Alternate Ester 19192 45 19147 99.8

TABLE 6- EMERGENCE, SHOOT AND ROOT LENGTHS, AND SHOOT AND ROOT DRY MASSES OF BARLEY PLANTED IN CLAY LOAM
SOIL CONTAINING VARIOUS DRILLING BASE FLUIDS INITIALLY ADDED AT 2% (WT:WT) AND BIOREMEDIATED FOR 95 DAYS. PLANTS
WERE HARVESTED AFTER 14 DAYS. DATA ARE MEANS (N = 3) ± SD.

Test Fluid Emergence % Shoot Length Root Length Shoot Mass Root Mass Total plant mass
cm/plant cm/plant mg plant mg/plant mg dm/plant
Untreated 100 30.2 ± 1.1 23.4 ± 0.7 56.7 ± 6.5 16.7 ± 1.3 73.5 ± 7.1
a ab ab ab ab
Diesel 100 26.8 ± 1.1 20.2 ± 2.6 61.9 ± 6.7 20.5 ± 3.2 82.4 ± 9.9
b bc ab a a
Mineral Oil 100 28.0 ± 1.1 18.4 ± 1.5 60.2 ± 4.9 16.4 ± 0.3 76.5 ± 5.2
ab c ab ab ab
Paraffin 100 29.1± 0.7 26.2 ± 1.6 67.0 ± 4.4 20.7 ± 3.3 87.7 ± 7.7
ab a a a a
C16/18 IO 100 29.3 ± 1.2 21.7 ± 1.7 60.2 ± 3.3 19.5 ± 0.5 79.7 ± 3.5
ab abc ab ab ab
C14 LAO 100 27.3 ± 1.1 20.6 ± 1.7 49.3 ± 2.7 14.1 ± 1.0 63.4 ± 3.3
b bc b b b
Ester 100 28.9 ± 1.0 21.9± 1.1 61.9 ± 7.1 14.3 ± 2.3 76.2 ± 9.4
ab abc ab b ab
Alternate Ester 100 28.0 ± 0.7 23.6 ± 1.9 55.5 ± 4.4 17.7 ± 2.0 73.2 ± 2.6
ab ab ab ab ab

Data analyzed by one-way ANOVA and differences detected by Tukey (HSD) comparisons. Means for each response variable,
followed by the same letter(s) are not significantly different (p • 0.05). Where means are not followed by a letter(s), no significant
differences were detected.

TABLE 7- EMERGENCE, SHOOT AND ROOT LENGTHS, AND SHOOT AND ROOT DRY MASSES OF ALFALFA PLANTED IN
CLAY LOAM SOIL CONTAINING VARIOUS DRILLING BASE FLUIDS INITIALLY ADDED AT 2% (WT:WT) AND
BIOREMEDIATED FOR 95 DAYS. PLANTS WERE HARVESTED AFTER 21 DAYS. DATA ARE MEANS (N = 3) ± SD.
Test Fluid Emergence Shoot Length Root Length Shoot Mass Root Mass Total plant mass
% cm/plant cm/plant mg plant mg/plant mg dm/plant
Untreated 73.3 ± 30.6 10.5 ± 2.0 13.1 ± 2.7 17.7 ± 6.8 bc 3.2 ± 1.7 20.9 ± 8.6
c bc
Diesel 70.0 ± 17.3 13.7 ± 0.4 16.9 ± 1.3 26.1 ± 2.8 4.0 ± 0.2 30.2 ± 3.0
abc abc abc
Mineral Oil 70.0 ± 17.3 14.3 ± 2.2 11.2 ± 2.6 28.0 ± 2.3 ab 4.4 ± 1.4 32.4 ± 3.7
abc ab
Paraffin 73.3 ± 11.5 11.9 ± 3.0 11.6 ± 2.3 16.9 ± 5.2 c 2.1 ± 0.8 19.0 ± 5.6
bc c
C16/18 IO 56.7 ± 30.6 15.5 ± 1.0 13.7 ± 0.4 30.6 ± 2.5 a 4.0 ± 0.1 34.6 ± 2.6
ab a
C14 LAO 66.7 ± 11.5 16.6 ± 1.1 14.2 ± 1.6 26.9 ± 0.9 abc 2.8 ± 0.5 29.7 ± 0.4
ab abc
Ester 83.3 ± 5.8 16.9 ± 1.4 13.1 ± 1.8 27.3 ± 1.4 abc 2.7 ± 0.6 29.9 ± 1.9
a abc
Alternate Ester 76.7 ± 15.3 16.4 ± 1.4 15.1 ± 3.1 31.6 ± 3.8 4.3 ± 0.6 35.9 ± 4.2
ab a a

Data analyzed by one-way ANOVA and differences detected by Tukey (HSD) comparisons. Means for each response variable,
followed by the same letter(s) are not significantly different (p • 0.05). Where means are not followed by a letter(s), no significant
differences were detected.
IPTC 11237 9

TABLE 8-EMERGENCE, SHOOT AND ROOT LENGTHS, AND SHOOT AND ROOT DRY MASSES OF NORTHERN
WHEATGRASS PLANTED IN CLAY LOAM SOIL CONTAINING VARIOUS DRILLING BASE FLUIDS INITIALLY ADDED AT
2% (WT:WT) AND BIOREMEDIATED FOR 90 DAYS. PLANTS WERE HARVESTED AFTER 21 DAYS. DATA ARE MEANS (N
= 3) ± SD.
Emergence Shoot Length Root Length Shoot Mass Root Mass Total plant mass
Test Fluid
% cm/plant cm/plant mg plant mg/plant mg dm/plant
Untreated 88.9 ± 9.6 25.6 ± 1.8 19.3 ± 0.1 23.4 ± 0.5 5.7 ± 0.5 29.1 ± 0.9
Diesel 88.9 ± 19.3 23.9 ± 2.5 22.0 ± 3.5 28.2 ± 6.3 8.5 ± 1.7 36.7 ± 7.6
Mineral Oil 88.9 ± 9.6 23.7 ± 1.0 19.8 ± 3.0 25.4 ± 2.2 6.5 ± 0.9 31.9 ± 3.0
Paraffin 83.3 ± 16.7 24.0 ± 1.6 22.0 ± 2.3 26.3 ± 5.3 6.8 ± 1.3 33.1± 6.4
C16/18 IO 88.9 ± 19.3 24.0 ± 1.5 21.4 ± 1.9 24.8 ± 1.6 6.8 ± 1.6 31.6 ± 2.9
C14 LAO 100 25.7 ± 0.4 24.1 ± 4.4 26.4 ± 0.6 7.5 ± 0.5 33.8 ± 0.4
Ester 83.3 ± 16.7 25.4 ± 1.8 21.3 ± 3.2 27.3 ± 2.5 7.0 ± 1.6 34.3 ± 1.7
Alternate Ester 94.4 ± 9.6 24.2 ± 1.5 20.8 ± 4.4 27.1 ± 1.7 6.3 ± 1.4 33.3 ± 2.2

Data analyzed by one-way ANOVA and differences detected by Tukey (HSD) comparisons. Means for each response variable,
followed by the same letter(s) are not significantly different (p • 0.05). Where means are not followed by a letter(s), no significant
differences were detected.

TABLE 9-ADULT SURVIVAL, ADULT MASS, JUVENILE PRODUCTION AND JUVENILE MASS OF THE EARTHWORM,
EISENIA ANDREI, FOLLOWING EXPOSURE TO UNTREATED SOIL AND SOIL TREATED WITH VARIOUS DRILLING BASE
FLUIDS AT 2% BY WT AND BIOREMEDIATED FOR 90 DAYS. DATA ARE MEANS (N = 3) ± SD. MEANS FOR EACH
RESPONSE VARIABLE FOLLOWED BY THE SAME LETTER(S) ARE NOT SIGNIFICANTLY DIFFERENT (P•0.05).
Treatment Adult survival Adult dry mass Juveniles produced Juvenile dry mass
(%) (mg /individual) (no./adult) (mg/total individual)
Control (no fluid) 100 ± 0 68.1 ± 8.2 9.7 ± 2.7 193 ± 45
ab ab ab
Diesel 100 ± 0 54.7 ± 7.2 0.3 ± 0.1 11 ± 9
ab c c
Mineral Oil 100 ± 0 51.9 ± 1.6 5.8 ± 0.8 62 ± 10
b b bc
Paraffin 93 ± 12 53.4 ± 6.6 13.9 ± 1.7 240 ± 86
ab a a
C16/18 IO 97 ± 6 53.3 ± 13.0 9.8 ± 1.7 211 ± 62
ab ab a
C14 LAO 100 ± 0 55.4 ± 4.1 10.3 ± 1.2 155 ± 24
ab ab ab
Ester 100 ± 0 74.7 ± 4.7 10.8 ± 3.7 196 ± 77
a ab ab
Alternate Ester 100 ± 0 55.9 ± 12.0 7.9 ± 1.0 131 ± 30
ab b abc

Data analyzed by one-way ANOVA and differences detected by Tukey’s HSD test.

TABLE 10-ADULT SURVIVAL AND JUVENILE PRODUCTION BY THE SPRINGTAIL, FOLSOMIA CANDIDA, FOLLOWING
28 DAYS EXPOSURE TO UNTREATED SOIL AND SOIL TREATED WITH VARIOUS DRILLING BASE FLUIDS AT 2%
(WT:WT) AND BIOREMEDIATED FOR 90 DAYS.
Treatment Adult Survival (%) Juvenile Production (No./10 adults)
Control 90 ± 10 633 ± 33
a d
Diesel 80 ± 26 1210 ± 50
a ab
Mineral Oil 97 ± 6 497 ± 117
a d
Paraffin 93 ± 6 1464 ± 105
a a
C16/18 IO 90 ± 0 918 ± 55
a c
C14 LAO 90 ± 10 991 ± 56
a bc
Ester 90 ± 10 1161 ± 203
a bc
Alternate Ester 97 ± 6 1023 ± 56
a bc

Data are means ± SD. Means for each response variable followed by the same letter(s) are significantly different (p • 0.05). Data for
each variable analyzed by one-way ANOVA and differences detected by Tukey’s HSD test.
10 IPTC 11237

40
Control
Diesel
35
Mineral Oil
Paraffin
30 C16\18 IO
ug CO2-C/g dm/hr

C14 LAO
25 Ester
Alt. Ester
20

15

10

0
0 10 20 30 40 50 60 70 80 90 100
Day

Fig. 1 Respiration rates from clay loam soil treated with various base fluids at a rate of 2% (wt: wt).

20000 18882

18000 17653
Control
16651
Diesel
16000 15093 Mineral Oil
15106 Paraffin
14000 C16\18 IO
C14 LAO
ug CO2-C/g

12000
Ester
Alt. Ester
10000
9729
8000
7899
6000
4979
4000

2000

0
0 10 20 30 40 50 60 70 80 90 100
Day

Fig. 2 Cumulative CO2-C losses from clay loam soil treated with various base fluids at a rate of 2% (wt: wt). Values in the cumulative CO2-C
loss graph are the total amount of carbon lost as CO2 from the various base fluid treatments over a period of 95 days.
IPTC 11237 11

45

40 Control
Ester
35
C16/18 IO
30
Alk Mixture
ug CO2C/gdm/hr

25

20

15

10

0
0 5 10 15 20
Day

Fig. 3 Respiration rates for clay loam left untreated (control) or treated with ester, C14 LAO and alkane/alkene blend base fluids at 2% (wt:wt).
Measurements were taken over a period of 18 days, and are means of 3 replicate readings.

9000

8000 Control
7630
Ester 7314
Cumulative CO2-C loss (ug/g)

7000
C16/18 IO 6634
6000
Alk Mixture
5000

4000

3000

2000

1000
699
0
0 5 10 15 20
Day

Fig. 4 Cumulative CO2 -C loss from clay loam soil left untreated (control) or treated with ester, C14 LAO and alkane/alkene blend base fluids at
2% by weight. Data are means (n=3).
12 IPTC 11237

24000 20000
A Initial E Initial
22000 18000
Final Final
20000
16000
18000
14000
16000
mg PHC/kg

mg PHC/kg
14000 12000

12000 10000
10000
8000
8000
6000
6000
4000 4000

2000 2000
0
0

l
21

25

29
11

3
15

7
19

23

7
7

9
ta
C1

C1

C3

C3
C2

C3

C3

C3

l
11

13

15

19

21

23

27

31

33

35

39
To

7
C

ta
C

C1

C2

C2

C3

To
C

C
Carbon No. Carbon No.

20000 20000
B Initial F Initial
18000 18000
Final Final
16000 16000

14000 14000
mg PHC/kg

mg PHC/kg
12000 12000
10000 10000
8000 8000
6000 6000
4000
4000
2000
2000
0
0
11

13

15

21

27

31

33

35

37

39

l
7

ta
C1

C1

C2

C2

C2

11

15

21

23

25

27

35

37

l
3

9
To
C

ta
C1

C1

C1

C2

C3

C3

C3
To
C

C
Carbon No. Carbon No.

20000 20000
C G
18000 Initial 18000 Initial
Final Final
16000 16000

14000 14000
mg PHC/kg

mg PHC/kg

12000 12000

10000 10000

8000 8000
6000 6000
4000 4000

2000 2000

0 0
13

17

25

33
1
23

27

35

7
39

l
1

ta

13

21

29

37

l
11

15

7
19

23

5
27

31

39
5
C2

C2

C3

ta
C1

C1

C1

C3

C1

C2

C3

C3
C

To
C

To
C

C
C

Carbon No. Carbon No.

22000
D
20000 Initial
Final
18000

16000

14000
mg PHC/kg

12000

10000

8000

6000

4000

2000

0
l
13

19

25

27

31
1

9
ta
C1

C1

C1

C2

C2

C2

C3

C3

C3

C3
To
C

Carbon No.

Fig. 5 Extractable hydrocarbons in clay loam soil immediately following application of A) diesel, B) Mineral Oil, C) paraffin, D) C16/18 IO, E) 14
LAO F) ester and G) alternate ester base fluids at 2% by weight (initial) and after 95 days bioremediation (final).
IPTC 11237 13

120

100

80
mg dm/plant

60

40

20

0
Control Diesel Mineral Oil Paraffin C16\18 IO C14 LAO Ester Alt. Ester

Fig. 6 Barley production (shoot and root) in clay loam soil containing base fluid HC residuals remaining in the soil after 95 days
bioremediation. Data are means (n=3) ± SD. Data analyzed by one-way ANOVA and differences detected by Tukey (HSD) comparisons.
Means for each response variable, followed by the same letter(s) are not significantly different (p • 0.05). Where means are not followed by a
letter(s), no significant differences were detected.

45

40

35

30
mg dm/plant

25

20

15

10

0
Control Diesel Mineral Oil Paraffin C16\18 IO C14 LAO Ester Alt. Ester

Fig. 7 Alfalfa production (shoot and root) in clay loam soil containing base fluid HC residuals remaining in the soil after 95 days
bioremediation. Data are means (n=3) ± SD.
14 IPTC 11237

18

16

14
No./adult earthworm

12

10

0
Control Diesel Mineral oil Paraffin C16\18 IO C14 LAO Ester Alt. Ester

Fig. 8 Earthworm juvenile production in clay loam soil containing base fluid HC residuals remaining in the soil after 95 days bioremediation.
Data are means (n=3) ± SD.

1800

1600

1400
Juvenile no./10 adults

1200

1000

800

600

400

200

0
Control Diesel Mineral Oil Paraffin C16\18 IO C14 LAO Ester Alt. Ester

Fig. 9 Springtail juvenile production in clay loam soil containing base fluid HC residuals remaining in the soil after 95 days bioremediation.
Data are means (n=3) ± SD.