Bioremediation With Horizontal Injection for BTEX and Naphthalene Remediation

In situ remediation is a contact sport, requiring contact between the microbes and contamination.  Clay-rich soils are particularly challenging as the low permeability limits effective distribution of inoculants.  While numerous closely-spaced injection points can improve the distribution of inoculants, this approach is often not possible at active properties and can be costly.  Horizontal drilling was used on the subject site to overcome the combined challenges of low permeability and limited site access to inject Petrox® microbes for bioaugmentation.  The combined technologies reduced the total BTEX concentrations in ground water from 2,771 µg/L to 645 µg/L in less than one year.


The site is a former gas station located in the Florida panhandle.  The site soils are mixed silt and clayey silts that are typical of the coastal setting.  The depth to the water table varied seasonally from 13 to 18 below ground surface.

The underground storage tanks and contaminated soil had been removed prior to the ground water treatment.  Residual ground water contamination exceeded Florida Ground Water Cleanup Target Levels (GCTLs) for benzene, toluene, ethylbenzene, xylene, naphthalene and methylnaphthalene isomers.

The proposed treatment area was approximately 2,500 square feet.  The treatment depth was from 10 to 25 feet below grade.  The treatment depth included the capillary fringe to treat residual contamination above the seasonal low water table.

Horizontal Injection

 Access to the treatment area was limited by the current property use, the proximity of roads on two sides and a building on the third.  Horizontal drilling was selected by the site consultant, Advanced Environmental Technologies ( to deliver Petrox® to the contamination.

The horizontal drill rig was set back from the treatment zone on the opposite side of the building.  The horizontal injection wells were set in four horizontal sets of eight wells. The horizontal layers were at 10, 15, 20 and 25 feet deep.  The wells in each layer were five feet apart.  A total of 32 injection wells were closely spaces for excellent coverage through the treatment zone.

Petrox® was injected into the ground water in two treatment events – November 6, 2008 and June 24, 2009.  Petrox® was delivered in each injection well as the drill stem was withdrawn through the treatment zone.  The injection was monitored for accuracy so that 0.2 gallons of Petrox® was injected per foot of injection zone.  A total of 320 gallons of Petrox® slurry was injected.

Following the Petrox® injection, air was injected periodically through vertical sparging wells to increase the oxygen availability for the microbial metabolism.


Ground water samples were collected from a monitoring well inside the treatment area to track the progress of the bioremediation.  Approximately 60 days after the first Petrox® treatment, analysis of ground water samples showed 84% reduction in the benzene concentration and 35% reduction in the total BTEX compounds concentrations.  There was an apparent increase in the xylene concentration due to ground water mixing and induced increase in solubility due to the bioaugmentation injections.

A second sampling event approximately 30 days after the second injection showed additional reduction in the contaminant concentrations.  After the second injection, the total BTEX concentrations were 23% of the original concentration with xylene decreasing from 1,200 to 95 µg/L.

In addition to the BTEX compounds, naphthalene and methylnaphthalene isomer concentrations decreased through both treatments.

  Sampling Date Benzene Toluene Ethylbenzene Xylene Naphthalene
Pre-treatment 11/11/05 3,000 42 1,100 1,100 230
10/23/08 1,700 18 460 454 280
11/6/08 1,600 41 370 760 310


1/22/09 420 43 140 1,200 43
7/30/09 490 2.6 57 95 55


Horizontal drilling and injection made it possible to remediate ground water at this site of petroleum contamination with limited access and low natural permeability.  Without disturbing the property use, the horizontal injection of Petrox® provided effective distribution of the microbes for bioremediation.  The injection may have also made the contamination more available for bioremediation by increasing the contaminant solubility as shown by temporary increases in concentration.

This case study demonstrates that in situ bioaugmentation may be a feasible solution for sites with limited permeability and access restrictions.  For more information contact CL Solutions at

Improving Long-Term Bioremediation Results with Nutrients

The goal of bioaugmentation is to improve the rate of contaminant removal by adding a high population of beneficial microbes to the contaminated media.  The additional microbes  should provide short-term benefit as the microbes begin metabolizing the contaminants immediately upon injection. But what benefit does bioaugmentation provide in the long term? And how much benefit does bioaugmentation provide over biostimulation by adding nutrients to the native organisms?

A client of CL Solutions completed a bench-scale study to answer these questions.  A bench-scale study was preferred to a field study because it removes the potential distribution and time-lag issues associated with the distances between injection and monitoring locations in the field.

Samples of petroleum-contaminated soils were obtained and separated into split samples for treatment with microbes and nutrients. Some were untreated for comparison.  Samples were tested for petroleum concentrations, including C-fraction concentrations after 30, 40 and 60 days.  Heterotrophic populations were measured at 40 and 60 days.

The tests showed the following results in the early stages:

  • All of the treated samples showed more than 80% total petroleum reduction in the first 30 days.
  • The sample treated with nutrients only had the same level of petroleum removal as the bioaugmented samples in the first 30 days.
  • The heterotrophic population of the biostimulated sample was as high as in the bioaugmented samples at 40 days.

After 30 days the situation changed.

  • The bioaugmented microbial population continued to increase  after 40 days and may have increased by a factor of 100 times.  Meanwhile, the biostimulated population appeared to stall.
  • The petroleum removal continued in the bioaugmented samples and reached as high as 93% removal.  In comparison the biostimulated sample stalled at 82% removal.
  • The difference appears to be that the bioaugmented samples removed the C-21 to C-35 concentrations at a much higher rate than the biostimulated sample.
  • Phenanthrene was target chemical for bioremediation. The biostimulated sample showed 39% removal while the bioaugmented samples showed complete removal to BDL.

Overall, the superior performance of the bioaugmented samples appears to be related to having a greater metabolic range that removed the heavier hydrocarbon fractions.  Microbes with the extended metabolic range could continue to multiply as they grew on the heavy hydrocarbon fraction.  The results are consistent with field results showing the recalcitrance of heavier hydrocarbon fractions and compounds like naphthalene and phenanthrene under natural attenuation.

Contact CL Solutions for more information and insights.






BTEX and Naphthalene Bioremediation to BDL by Petrox Microbes

Case Study:
Orange County Fire and Rescue Station 81, Orlando, Florida

Oculus Number 8520434

Petrox was used to treat the residual petroleum contamination in ground water after the leaking tank was removed.  The petroleum contamination exceeded applicable standards for benzene, toluene, ethylbenzene, and xylene (BTEX) and naphthalene.  Petrox was injected into the ground water through 28 injection points covering a plume area of approximately 6,300 square feet.   Petrox injections were completed on June 18, September 11, and November 1, 2002.

Prior to bioaugmentation 100 pounds of ORC was added to provide supplemental oxygen for the bioremediation.  After verifying the dissolved oxygen concentration was more than 1.0 mg/l, the site was ready for bioaugmentation.

During the first injection 10 drums of Petrox were injected to cover the full plume.  As the area of contamination decreased, less Petrox was injected.  Five drums were injected in a focused application on September 11 and November 1, 2002.

Quarterly monitoring was completed during and after the inoculation to determine the bioremediation results.  The cleanup goals were achieved in the source area after the first inoculation.  The contamination persisted, however, in the down gradient wells.  The cleanup goals were achieved in the down gradient monitoring well after the third injection, but there was subsequent rebound.  After full distribution of Petrox was reached throughout the plume, the BTEX and naphthalene concentration were below detection limits (BDL), and cleanup goals were achieved and maintained. 

Please refer to the table below for the ground water monitoring results.

Monitoring Well Date Benzene Toluene Ethylbenzene Total Xylenes Naphthalene
MW-7 Source Area 6/12/2000 5.0 46.0 18.0 90.0 24.0
6/13/2001 39.0 272.0 167.0 526.0 26.0
3/14/2002 21.0 19.0 102.0 266.0 39.0
6/5/2002 ORC Added
6/18/02 10 Drums of Petrox Added
7/14/2002 <1.0 <1.0 <1.0 <2.0 <1.0
9/11/2002 <1.0 <1.0 <1.0 <2.0 <1.0
9/11/02 5 Drums of Petrox Added
10/7/2002 <1.0 <1.0 <1.0 <2.0 <1.0
11/1/02 5 Drums of Petrox Added
12/2/2002 <1.0 <1.0 <1.0 <2.0 <1.0
1/9/2003 <1.0 <1.0 <1.0 <3.0 <5.0
4/17/2003 <1.0 <1.0 <1.0 <2.0 <1.0

Down Gradient

6/12/2000 2.0 72.0 21.0 109.0 39.0
6/13/2001 14.8 677.0 207.0 1292.0 113.0
3/14/2002 71.0 1198.0 357.0 2408.0 193.0
6/5/2002 ORC Added
6/18/02 10 Drums of Petrox Added
7/15/2002 19.0 515.0 170.0 690.0 39.0
9/11/2002 <1.0 3.7 3.8 16.4 <1.0
9/11/02 5 Drums of Petrox Added
10/7/2002 6.4 2.3 4.7 23.8 15.0
11/1/02 5 Drums of Petrox Added
12/2/2002 38.0 515.0 95.0 460.0 10.0
1/9/2003 <1.0 <1.0 <1.0 <3.0 <5.0
4/17/2003 <110.0 1060.0 285.0 1120.0 65.0
4/29/2004 <1.0 <1.0 <1.0 <1.0 <1.0
9/1/2004 <1.0 <1.0 <1.0 <1.0 <1.0
12/1/2004 <1.0 <1.0 <1.0 <1.0 <1.0
3/21/2004 <1.0 <1.0 <1.0 <1.0 <1.0

In Situ TCE Bioremediation Rates

The following chart is a compilation of CL-Out bioremediation results from eleven different sites where ground water was contaminated by trichloroethylene (TCE).  The sites represent all kinds of geological conditions with the full range of TCE concentrations in ground water.

Overall, the results show an average removal rate of 97% .  However, the final TCE concentration reached below detection limits on two sites, and an additional three had greater than 99% removal.  

Keyhole Source Treatment to Reduce PCE Ground Water Plume

CL-Out bioremediation was used in a keyhole treatment to reduce the mass of contamination near the source and down gradient concentrations in the plume.  At a former manufacturing facility in Ohio the concentration of PCE near the source was over 100,000 ug/L.  Down gradient of the source the PCE concentrations were less than 10% of the source concentration.  Aggressive treatment in the source area reduced the source concentration and in the down gradient plume.

After one treatment with CL-Out bioremediation, the concentrations decreased as follows:

PCE was reduced from 120,000 to 12 ug/L.

TCE was reduced from 2,000 to 12 ug/L

Cis 1,2-DCE was reduced from 9,500 to 8,100 ug/L.

Vinyl chloride, however, increased from 1,200 to 22,000 ug/L.

The vinyl chloride increased as the aggressive cometabolic treatment stimulated some incomplete reductive dechlorination.

Down gradient from the source, the concentrations decreased with slight to no increase in daughter products.  The following results were measured in the down gradient plume:

PCE reduced from 5,000 to 1,600 ug/L.

TCE reduced from 43 ug/L to BDL.

Cis 1,2-DCE was reduced from 140 to 23 ug/L.

Vinyl chloride  was not detected before or after treatment.

Keyhole treatment was a cost effective approach to reducing the mass of contamination in a ground water plume by focusing aggressive treatment on the source area.  Concentrations in the rest of the plume decreased as the microbes and treated water dispersed through the plume.