Bioremediation as A Solution for Environmental Cleaning

Abstract

This paper discusses the methodologies adopted to test the ability of members belonging to
Pseudomonas,Nocardioides (especially N. deserti), Dietzia (especially D.  papillomatosis),

Microbacterium, Micrococcus, Arthrobacter, Cellulomonas, Gordonia and related genera to

degrade the complex hydrocarbons from oil spills in desert soils, and the conclusions drawn.
The major objective of the experiment is to check whether bioremediation was best in diluted or
undiluted soils and effective microbial degradation. A sample of pristine desert soil was
contaminated with 17.3% w/w of crude oil and diluted with varying aliquots of pristine garden
and desert soils containing indigenous micro flora. The mixtures were incubated in natural
conditions for a month to acclimatize the microbes to the oil concentration. The soil heaps were
then moistened and aerated to provide enough oxygen for the microbes to degrade the
hydrocarbons, and the residue of oil was measured on a weekly basis. The study concluded that
the efficiency of the bacteria in biodegradation of hydrocarbons is not limited by the
concentration of crude oils in the soil. The sites of action of the hydrocarbonoblasts are
selectively enhanced, and thus provides a means to effectively treat oil spills.

Introduction

Human beings are highly inquisitive by nature. This curiosity about the world around them and
the science behind the working of various is what led to many amazing discoveries and
inventions. One such discovery was the existence of microorganisms, by Anton Van
Leeuwenhoek. This discovery of living beings on every surface surrounding us, till then
unknown to us, was intriguing and exciting. They have been enthusiastically studied ever since.

But some inventions which have contributed to making our lives easier have done so at a great
cost: the environment. Environmental pollution and degradation is one of the greatest concerns
today. There is much discussion and debate on the best ways to preserve and save our
environment. But the actions being taken up are not enough to meet the need of the hour.
Fortunately, some microbes have been doing this for millions of years. We can exploit these
special abilities of our invisible friends, microorganisms for more efficient management of our
environment.

The use of plants, decomposers, microbes or their enzymes to clean up pollutants in the
environment is called bioremediation. Since they are generally present in the environment, this
happens naturally. But if exogenous organisms are deliberately introduced into an environment
for this purpose by humans, it is called bio augmentation. Apart from degrading wastes, this
process also degrades pollutants and raw materials from industries. Usually remediation from air,
water and soil inhibits biological process. Currently bioremediation with inorganic contaminants
has developed, that aiming to improve biological processes.

Though it is a relatively new field of study, it is developing fast and gaining more acclaim, since
conventional remediation has been carried out by digging up the contaminated soil and removing
it to a landfill, or by capping and containing the contaminated areas of a site (Zeyaullah, Md et
al, 2009). The main disadvantages of these techniques are that land-filling leads to contamination
of ground waters. Over the years, it has become increasingly difficult and expensive to find new
landfill sites for the final disposal of the material. The cap and contain method is only an interim
solution since the contamination remains on site, and requires to be monitored and maintained.

This paves way to bioremediation, which is comparatively cost effective and in environment
friendly. This includes two steps. The first is bioaugmentation where new strains of bacteria in
inoculated in polluted areas whereby gene pools are increased in soil. Biostimulation is the
action in soil by management. Self-cleaning is where is bioremediation takes place without any
management.

When these organisms utilize hydrocarbons and other pollutants, over time these compounds are
found within their cells, and are called xenobiotic. A xenobiotic is defined as any unusual
chemical compound found within an organism since it is not expected to be present naturally.

Methodology

The researchers used contaminated with crude oil soil diluted with pristine soil, undertaken in a
outdoor condition. They found that the oil was consumed and a phenomenon called
bioremediation takes place. Even diluted soil sample consumed oil as much as 45-60%. Most of
these bacteria has a tolerance level of about 20% to crude oil of about 20%. For this soils were
added crude oil and diluted with unpolluted soil samples. This mixture was left for incubation for
six months along with undiluted soil.

Techniques such as monitoring oil consumption could be done were soil heaps are regularly
monitored by performing gas liquid chromatography. The amount of microorganisms were then
calculated by isolation and culturing and then analyzed for different gene pools. 17 microbial
isolates were obtained in this experiment out of which 13 of the isolates were tolerant to crude
oil. And finally a statistical data was prepared to analyze which strain of bacteria show higher
rate of consumption of oil and show effective bioremediation.

Result

The experimenters concluded that highly soil polluted areas need to diluted so that bacteria can
break down hydrocarbons and show tolerance. Around 73-79% of hydrocarbons was found to be
removed by bioremediation which was initially at a rate of 18%. A peak of 63% was reached,
after which the rate of degradation slowed down. Different dilution gave different dynamics of
microorganisms. The oil saturated soil showed Pseudomonas songnenensis at initial and later
Nocardioides solisilvae stabilized.

In lowest dilution Blastococcus aggregatus predominated

In moderate dilution Kocuria dechangensis predominated

And in highest dilution predominated.

In course of six months of experiment each month an organism predominates causing

bioremediation

At lowest level of dilution the following bacteria were high the sample for each month:

March-Acinetobacter radioresistens

April-Acinetobacter radioresistens

May-Microbacterium lacusdiani

June-Dietzia papillomatosis
July and September-Dietzia papillomatosis

Likewise each dilution level gave each predominant organism in each month of the experiment.

Discussion

17 isolates were prepared containing different bacterial species and some in common too. Group
A in this show higher level of tolerance to 20% of oil as the increase in oil concentration showed
growth. When broad beans are cultivated in this soil they attained maturity. Whereas group B on
the other side had less tolerance level to the soil and showed less growth and no germination of
seedlings. Initially, only 1-6% of oil for these experiments. But increasing the oil concentration
gave new results and raised some questions too. It was discovered that bacteria are able to live in
such soils that lacks aeration and water flow only due to the fact that they form biofilms. These
biofilms acts as channels that allows passage of water and nutrients and air that retains the soil
quality. The polluted soil with oil automatically invite microorganisms with a range that
corresponds to toxicity in soil, so that they in turn could be tolerant. The initial 16-18% of oil
consumption was indeed due to volatilization as bacteria show evasion and adaptation in that
period.

The bioremediation by bacteria gives a peak and active with heat due to increase in thermophilic
bacteria. These hydrocarbanoclastic bacteria possess mono oxygenases and dioxygenases which
catalyze in breaking of oxygen into their corresponding atoms that are fitted aromatic or
aliphatic amino acids via beta oxidation. The alkanes are converted to alkanols which further
broken down to corresponding amino acids for cell sources. The Krebs cycle which contain some
these fatty acid as intermediates provide the ATP required.

Pseudomonas aeruginosa degrades the complex hydrocarbons in crude oils by releasing a bio
surfactant that reduces their molecular weight to a suitably lower form, where it can easily utilize
it. In a study conducted in desert oil spills (Ali, N., Dashti, N., Khanafer,. M et al, 2020), it was
found that the organisms belonging to or related to the genera
Nocardioides (especially N.  deserti), Dietzia (especially D. papillomatosis), Microbacterium, 

Micrococcus, Arthrobacter, Cellulomonas, Gordonia  also contributed a great deal in the

degradation of hydrocarbons in the soil.
Conclusion

The amount of oil lost due to volatility during the exposure to natural environment during the
course of the experiment has been admitted to be significant. This may have had an effect on the
final conclusions drawn. In addition, the effects of the oil spill in the soil which would have
resulted in groundwater contamination has not been studied. If studied, bioremediation could
provide a much awaited solution to the bigger picture of oceanic oil spills.

Regardless of the multifarious uses of microorganisms, we have a responsibility towards the

environment which sustains us. Each one of us has to do our bit, however small, towards
sustaining the environment and preventing its further degradation. Although technology and
research open up new and innovative methods for environmental management, real progress can
only be made only if each individual makes a conscious effort.

References

Ali, N., Dashti, N., Khanafer, M. et al. (2020) Bioremediation of soils saturated with spilled
crude oil

Orji Frank Anayo, et al (2019) The Beneficial Roles of Pseudomonas in Medicine, Industries,
and Environment: A Review, pp 7-9

Mohammed Zeyaullah et al (2009), Bioremediation: A Tool for Environmental Cleaning