Journal of Environmental Management 304 (2022) 114313

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Journal of Environmental Management

journal homepage: www.elsevier.com/locate/jenvman

Bioaugmentation as a green technology for hydrocarbon pollution

remediation. Problems and prospects
Amechi S. Nwankwegu a, b, *, Lei Zhang a, Deti Xie a, Chukwudi O. Onwosi c,
Wada I. Muhammad b, d, Chuks K. Odoh b, e, Kabari Sam f, John N. Idenyi g
a
College of Resources and Environment, Southwest University, 1 Tiansheng Road, Beibei District, Chongqing, 400716, China
b
Organization of African Academic Doctors, Off Kamiti Road, P.O. Box 25305-00100, Nairobi, Kenya
c
Department of Microbiology, Faculty of Biological Sciences, University of Nigeria, Nsukka, Enugu State, Nigeria
d
College of Water Conservancy, Hohai University, No.1 Xikang Road, Gulou District, Nanjing, 210098, China
e
Dalian Institute of Chemical Physics, Chinese Academy of Sciences, Beijing, China
f
Faculty of Environmental Management, Department of Marine Environment and Pollution Control, Nigeria Maritime University, Okerenkoko, Delta State, Nigeria
g
Department of Biotechnology, Ebonyi State University Abakaliki, Nigeria

A R T I C L E I N F O A B S T R A C T

Keywords: Environmental pollution mitigation measure involving bioremediation technology is a sustainable intervention
Environmental pollution for a greener ecosystem biorecovery, especially the obnoxious hydrocarbons, xenobiotics, and other environ­
Bioremediation mental pollutants induced by anthropogenic stressors. Several successful case studies have provided evidence to
Bioaugmentation
this paradigm including the putative adoption that the technology is eco-friendly, cost-effective, and shows a
Hydrocarbon
Microorganism
high tendency for total contaminants mineralization into innocuous bye-products. The present review reports
advances in bioremediation, types, and strategies conventionally adopted in contaminant clean-up. It identified
that natural attenuation and biostimulation are faced with notable limitations including the poor remedial
outcome under the natural attenuation system and the residual contamination occasion following a bio­
stimulation operation. It remarks that the use of genetically engineered microorganisms shows a potentially
promising insight as a prudent remedial approach but is currently challenged by few ethical restrictions and the
rural unavailability of the technology. It underscores that bioaugmentation, particularly the use of high cell
density assemblages referred to as microbial consortia possess promising remedial prospects thus offers a more
sustainable environmental security. The authors, therefore, recommend bioaugmentation for large scale
contaminated sites in regions where environmental degradation is commonplace.

1. Introduction
Environmental contamination (land and water) as a result of the
geometric increase in industrialization and urbanization remains a
global ecosystem challenge threatening environmental security and
sustainability (https://www.sciencedirect.com/science/article/abs/pii
/S0959652621024598Rithuparna et al., 2021; Manral et al., 2021).
Despite the introduction of sustainable and renewable energy forms,
many polluted sites requiring remediation exist. With declines in the
global economic outlook, effective and efficient remediation technolo­
gies are sought (Chatterjee et al., 2017; Nwankwegu and Onwosi, 2017a;
Sivaperumal et al., 2017). Anthropogenic stressors including; oil min­
ing, pipeline vandalism, accidental discharge, and poor municipal waste
management cause adverse effects on public health and the environment
(Nwankwegu et al., 2016a; Varjani et al., 2017). As a result of the acute
and chronic impacts, over time, contaminants generated by anthropo­
genic processes are removed by conventional approaches such as the
physical and chemical methods (Zebra et al., 2018). These techniques
which include; evaporation, photo-oxidation, thermal extraction,
incineration (Agarwal and Liu, 2015), soil washing (Agarwal and Liu,
2015), land farming (Brown et al., 2017a, 2017b), and dissolution are
not wholesomely sustainable. The use of dispersants/inorganic surfac­
tants like detergents in hydrocarbon remediation of water ecosystem to
emulsify the oil into droplets and ease depuration is a notable remedial
step. Its major principle is to increase the solubility of hydrocarbon in
water for consequent rapid disappearance (Zheng and Chai, 2016).

* Corresponding author. College of Resources and Environment, Southwest University, 1 Tiansheng Road, Beibei District, Chongqing, 400716, China.
E-mail address: smetchie@yahoo.com (A.S. Nwankwegu).

https://doi.org/10.1016/j.jenvman.2021.114313
Received 8 August 2021; Received in revised form 26 October 2021; Accepted 13 December 2021
Available online 20 December 2021
0301-4797/© 2021 Elsevier Ltd. All rights reserved.
A.S. Nwankwegu et al.
However, the use of physicochemical remedial approaches for hy­
drocarbon decontamination of the environment has been challenged by
several factors, such as cost, environmental residue generations and
residual risks. The high cost of technologies and applicability spectrum
based on exposure scenarios is also a potential limitation. For example,
strategies like incineration, thermal extraction, photo-oxidation, and
evaporation are not suitable for use if sources of contamination maintain
proximity to residential areas. This is primarily due to the inflamma­
bility of most hydrocarbon fractions which threatens public health and
property security. Soil washing involving the use of a mixture of sol­
ubilizers and surfactants is grossly limited owing to soil characteristics
(e.g., particle distribution), extra cost due to soil pretreatment, as well as
logistics, making the process expensive with a high environmental
burden (Liming et al., 2014; Andreolli et al., 2015). Land farming is
associated with shortcomings such as delay in time required to complete
depuration due to difficulty often encountered in the control of condi­
tions directing biological processes, large space, seasonal variations,
leaves shading and plants dieback (Odoh et al., 2019; Valipour, 2015).
Dispersion can be optimized by the supply of chemical dispersants (e.g.,
detergents) which induce solubility of the oil slicks through a significant
reduction of the oil surface tension (fluid-mediated force), thereby
boosting effective pollutant removal. Some hydrocarbon fractions in
crude oil ranging from the heavier aromatic hydrocarbons like poly­
cyclic aromatic hydrocarbons (PAHs) and polychlorinated biphenyl are
not readily volatile like dual-purpose kerosene-(DPK) (Nwankwegu
et al., 2016a), automobile gas oil-AGO (Nwankwegu et al., 2017), pre­
mium motor spirit (PMS) (Nwankwegu, 2016), and diesel (Nwankwegu
et al., 2016b; Onwosi et al., 2018), thus are rarely removed through the
natural events like; photo-oxidation and evaporation. The disadvantage
of the use of dispersants is that it increases the exposure of the aquatic
community to the oil, which could lead to massive hydrospheric disaster
and a dramatic increase in the nutrient budget beyond the ecosystem
requirement. Specifically, this approach significantly increases the
aquatic ecosystem external phosphorus (P) load while exacerbating
eutrophication and the episodic bloom of harmful algae (Nwankwegu
et al., 2019, 2020a, 2020b). From the foregoing, a potential prudent and
sustainable mitigation measure should offer a smart remedial outcome,
as well as cleaner environmental productivity and conservation of the
biological components at little or no tradeoffs (Harelimana et al., 2020;
Nuamah et al., 2020).
Bioremediation options (Fig. 1), however, are more beneficial than
these conventional approaches. It is suitable for a wide range of envi­
ronmental pollutants and can circumvent the various inadequacies
which characterize the physicochemical treatment methods. Besides,
they are cost-effective, eco-friendly, and sustainable hence often gua­
rantees complete mineralization of the targeted environmental con­
taminants (Rodrigues et al., 2015; Chen et al., 2015a; Brown et al.,
Fig. 1. Illustration of the bioremediation, types, forms and approaches tradi­
tionally adopted in pollution abatement responses.
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Journal of Environmental Management 304 (2022) 114313
2017a, 2017b). As a promising biotechnological approach, it involves
the use of living things (e.g., plants and microorganisms) to decontam­
inate polluted environments. Here, the metabolic potentials, as well as
enzyme streams of microorganisms, are utilized to rapidly degrade
obnoxious wastes in the soil, water, and sludge. Although literature
already exists on bioremediation, there is still a paucity of information
on the clear-cut differences based on technology efficiencies and
post-remedial sustainability among the strategies. Most available opin­
ions are controversial and driven by the clean-up rate rather than the
actual ecosystems ‘post-remedial impacts and functionalities. The pre­
sent review, therefore, evaluates the efficacy and applicability of the
various forms of bioaugmentation options for pollutants e.g., hydro­
carbon removal and soil ecosystem reclamation. Further, it addresses
the numerous bioremediation approaches and identifies the strengths
and shortcomings that characterize each remedial strategy while it
strongly remarks that bioaugmentation offers great insight to potentially
fill these gaps given the successes in large scale applications.
2. Types and approaches of bioremediation
Basically, two types of bioremediation technologies notably; ex-situ
and in situ (Table 1) are popularly employed in clean–up of pollutants e.
g., hydrocarbons (Brown et al., 2017a, 2017b). Ex-situ technology in­
volves the treatment of contaminants away from the point of generation.
It is operationalized when contaminants are treated in a place other than
the point of occurrence owing to some ethical and environmental con­
siderations such as the impact on inhabitants (Sam and Zabbey, 2018;
Zabbey et al., 2017). It involves composting, land farming, bio-piling,
and the process by bioreactors. In situ bioremediation involves the
treatment of pollutants at the point of occurrence. In situ remediation
includes technologies like; bioventing (unsaturated/vadose zone), bio­
sparging (saturated) and phytoremediation (Odoh et al., 2019b; Kempa
et al., 2013; Odoh et al., 2019; Zabbey et al., 2017). Furthermore,
applicable bioremediation approaches are generally (i) natural attenu­
ation, (ii) biostimulation, and (iii) bioaugmentation which, are
conventionally adopted in environmental contaminant depurations
(Ikeda-Ohtsubo et al., 2013; Suja et al., 2014; Hassanshahian et al.,
2014, 2015; Ozbayram et al., 2017; Onwosi et al., 2018). Each approach
shows functional variability in environmental biorestoration efficiency
with few systematic and operational challenges which essentially pre­
empt pollution experts to discern clear-cut effectiveness amongst the
strategies. Natural attenuation pertains to the use of soil’s natural ability
to degrade pollutants (Zabbey et al., 2017). Natural attenuation is a
biological process of contaminants concentration reduction. It is a very
slow but ultimately cheap process. However, since the essence of
bioremediation is to significantly reduce the toxic fraction of the
Table 1
Operational differences between ex situ and in situ bioremediation technology.
Factor Ex sit In situ
Response Remediation is Remediation is
point performed off-site performed on-site
Operational suitable for small suitable for large scale
scale scale contamination
contamination
Cost/area Technology is Technology is cost but requires a
expensive due to effective/no logistics large land area
extra cost -on logistics but
requires small area
Manageability The process can Process is not easily
be effectively controlled
controlled
Effectiveness Process is very Process is less effective consideration
effective often with optional for excavation
involves -excavation with
thorough and
permanent clean-up
A.S. Nwankwegu et al.
contaminants e.g., the total petroleum hydrocarbon (TPH) as soon as
possible following a fresh hydrocarbon spill, this strategy does not
guarantee significant recovery of the polluted site. It relies principally
on natural processes mediated by biological transformation to dissipate
and mineralize pollutants. In most cases, the pollutants often have
enough time to establish their irreversible and chronic ecosystem
toxicity while eliciting permanent sterility which causes most environ­
ments, particularly land to lose their arable potentials. This remediation
strategy is consequently not recommended in polluted land intended for
use for agricultural activities upon recovery (Nwankwegu et al., 2018).
Biostimulation on the other hands, requires the modification of the
environment via nutrient addition or electron acceptor (oxygen) into the
polluted media to activate the autochthonous hydrocarbonoclastic mi­
croorganisms. This approach optimizes the processes and conditions
crucial for remediation of polluted sites (Masy et al., 2016; Nwankwegu
and Onwosi, 2017a). Although, the adoption of biostimulation in the
recovery of polluted environment has attracted considerable attention
(Nikolopoulou et al., 2013a; Nwankwegu et al., 2016a; Onwosi et al.,
2017, 2018), it is still not without challenges. For instance, introduction
of inorganic fertilizer is limited by its high cost. Secondly, there is always
paucity of the chemical formulations and lack of the knowledge on the
application ratio owing to the sites’ complexity and heterogeneity of
pollutants (Odoh et al., 2019). In addition, biostimulation could lead to
increased cases of plant and soil toxicities, as well as certainty of
eutrophication and algal bloom due to abundant supply of nitrogen (N)
and phosphorus (P), particularly in aquatic ecosystem (Nwankwegu
et al., 2016b).
Consequently, given that different microorganisms display differen­
tial preference to hydrocarbon derivatives; the introduction of different
microbial diversities from exogenous sources described as bio­
augmentation represents a more suitable alternative in addressing the
issues related to soil-plant toxicities and eutrophication (Lebeau et al.,
2008; Wang et al., 2014a, 2014b; Herrero and Stuckey, 2015). The
performances and the benefit-impact implication of the potential
bioremediation strategies are summarized in Fig. 2. Bioaugmentation is
the use of autochthonous, allochthonous, or genetically engineered
microbial strains to treat the contaminated environment (Sermany et al.,
2012; Liu et al., 2015; Cycon et al., 2017). It is the introduction or
seeding of the non-indigenous pollutant degraders (pure culture or
mixed) into the polluted media for expedited contaminant removal.
Bioaugmentation technology exists in different forms and each has been
Fig. 2. Total benefit -environmental impact evaluation (percentage) of the
available bioremediation approaches. NA= Natural attenuation, BG = Bio­
augmentation, BS = Biostimulation. ET = Environmental toxicity, RC = Re­
sidual contamination, PRAP = post-remedial arable potential (usability of the
soil/media following clean-up), C = Cost, MPR = Multiple pollutant removal
(tendency to tackle a wide range of pollutants), CS = Clean-up speed.
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Journal of Environmental Management 304 (2022) 114313
extensively studied by several researchers who provided expert and
scientific views. Combinations of more than one strategy have also been
studied in both field and laboratory scales over the years and significant
successes reported.
3. Forms of bioaugmentation
Bioaugmentation exists in various forms. The adoption of each often
depends on both recent and historic knowledge of the polluted sites.
Certain contaminants are recalcitrant and would require the deployment
of two or more bioaugmentation forms for the total removal of such
obnoxious pollutants. The forms of bioaugmentation so far applied
include; (i) use of microorganisms indigenous to the polluted sites, (ii)
use of exogenous microorganisms (either the pure culture of known
microbial species/single strain or collection of individual microorgan­
isms to form a high-density cell mass called microbial consortium (iii)
Use of genetically engineered microorganisms (recombinant
microorganisms).
3.1. Bioaugmentation using autochthonous microorganisms (indigenous
microbes)
The knowledge of the type and contaminants level is necessary but
not enough for successful bioaugmentation operations. Also essential is
the actual microbial species in the contaminated area needing remedi­
ation (Mrozik and Piotrowska–Seget, 2010). Here, organisms are pro­
cedurally isolated from a specific polluted site (e.g., soil) and activated
through culturing under controlled conditions. Following these, the
pre-adapted pure microbial strains are then seeded back to the same soil
(Mrozik and Piotrowska–Seget, 2010; Nikolopoulou et al., 2013b). This
method can also be referred to as soil re-inoculation with autochthonous
species. In a study that involved a single strain-bioaugmentation,
Poorsoleiman et al. (2019) used the indigenous bacterial strains (Aci­
netobacter radioresistens strain KA2) isolated from oil waste sludge
(OWS) and reported the total petroleum hydrocarbons (TPHs) removal
of 67.64–89.56% under eight weeks composting incubation. Again,
previous studies had demonstrated the potentials of bioaugmentation
for efficient removal of hydrocarbon and its congeners. Specifically,
Bento et al. (2005) adopted natural attenuation, biostimulation and,
bioaugmentation, in a comparative study of bioremediation of soil
contaminated with diesel oil and reported the highest hydrocarbon
degradation in both the light and heavy fractions at 72.7% and 75.2%,
respectively of TPH in the re-inoculated single strain-bioaugmentation
system. In the same study, optimal microbial activity (de­
hydrogenases) was also observed in the bioaugmentation system. In a
different study by Suja et al. (2014), the effectiveness of microbial
bioaugmentation and biostimulation was evaluated. Results demon­
strated 97% of TPH removal in the topsoil seeded with inoculum
designated as MC2 in the study. Experimental microcosms were con­
ducted in both tank and field scenarios. It was again discovered that 79%
TPH was degraded in the tank which was composed of an MC3 inoculum
size of 10% and mineral salt medium (MSM). This study, however,
confirms a facilitated contaminant (hydrocarbon) removal due to
polluted media (soil) seeding with indigenous microbial species.
3.2. Allochthonous seeders in bioaugmentation
The selection of efficient microbial culture (pure culture) for
increased hydrocarbon biodegradation should take into consideration
the following microbial characteristics; microbial stability (survivabil­
ity) over a wide range of environmental conditions (stress-tolerant mi­
croorganisms), ability to withstand high concentration of the
contaminants without exhibiting signs of cytotoxicity, fast growth, and
easily culturable. The “heirloom” microorganisms (microbial genera­
tion) which are specifically grown and handed down over the years and
are particularly adapted for bioaugmentation operations often show
A.S. Nwankwegu et al.
remarkable importance (Bento et al., 2005; Singer and Van der Gast.,
2005; Zhang et al., 2015). Abena et al. (2019) recently evaluated the
efficacy of bioaugmentation involving exogenous bacteria in the biore­
mediation of highly polluted sites and reported the removal efficiency of
48.10% under a half-life of 41.76 d. In the same study, the authors
showed that site enrichment with the allochthonous bacteria expedited
the TPH removal in the heavily polluted soil sample with a net TPH
reduction of 59,934 mg kg− 1 (25.34%) of dry soil in 40 d. In a large-scale
treatment of hydrocarbon-contaminated groundwater by Poi et al.
(2018), a significant reduction in the TPH levels of >100 mg kg− 1 was
reported indicating a successful bioremediation outcome as revealed by
the phytotoxicity bioassay. Nwankwegu et al. (2016c) while using the
bacterial isolates from palm oil mill effluent from Southeastern Nigeria
to evaluate the ex-situ biodegradation of crude oil reported 98%
(7653.69–153.07 mg kg− 1), 83% (7653.69–1301.13 mg kg− 1), and 75%
(7653.69–1913.42 mg kg− 1) hydrocarbon removal by Escherichia fer­
gosonii, Klesiella variicola, and Micrococcus luteus, respectively within 15
d incubation relative to the control which showed only 43%
(7653.69–4362.60 mg kg− 1) removal efficiency under similar incuba­
tion period. When bioremediation is targeted to sites polluted with
polynuclear aromatic hydrocarbons and polychlorinated biphenyl
(PAHs and PCBs, respectively), it is often recommended that strains able
to produce biosurfactants are inoculated as this increases the bioavail­
ability of the contaminants for the effective cleanup process. Species of
Pseudomonas and Bacillus are known to produce rhamnolipids and sur­
factin, respectively. Bioaugmentation involving allochthonous micro­
bial sources could be a single strain or collection of different microbial
species known as consortia.
3.2.1. Single strain
In this case, a specific microbial species is introduced into the
polluted media. When contaminants do not include multiple and
recalcitrant hydrocarbon fractions, its removal using single strain hy­
drocarbon degrader proceeds faster with a short half-life and high cor­
relation coefficient (R2) but if contaminants consist of pollutants of
different fractions/congeners, removal using single strain microor­
ganism becomes significantly slow and almost impossible. Varjani et al.
(2020) currently studied the bioremediation of oily sludge polluted soil
and reported 92.97 ± 0.92% hydrocarbon removal efficiency and
80.95% germination index (%IG) of Vigna radiata using a single inoc­
ulum of Pseudomonas aeruginosa in 56 d. The use of single strain
biosurfactant-producing and hydrocarbon-degrading bacteria desig­
nated as P1 in bioremediation of petroleum polluted seawater led to
>70% removal efficiency under 45 d incubation (Shi et al., 2020).
Similarly, Roy et al. (2018) in a different study also reported a 57–75%
reduction of total petroleum hydrocarbon due to bioaugmentation
treatment with biosurfactant producing and hydrocarbon utilizing
indigenous Bacillus strain during bioremediation of oil refinery sludge.
Further, Nwankwegu and Onwosi (2017b) studied bioremediation of
gasoline impacted arable soil using single microbial strains of Micro­
coccus luteus (AJ536198) and Rhizopus arrhizus (JN942919) previously
isolated from a diesel oil impacted soil and reported the removal effi­
ciencies of 75.70% and 71.10%, respectively after 8 weeks incubation.
In the same study, eco-monitoring assessment indexed by dehydroge­
nase activity confirmed optimal soil recovery for immediate cultivation.
Again, several previous studies involving hydrocarbon removal by single
strain microorganisms also have been reported. For instance, Milena
et al. (2011) carried out a feasibility study involving the biorestoration
of polyaromatic hydrocarbon contaminated soil by autochthonous
microflora seeded with Sphingobium chlorophenolicum strain C3R. It was
reported therein that the seeded bacterial strain C3R effectively
removed phenanthrene alone or co-contaminated with molecular
weight PAHs, as well as cadmium and arsenate. A biomonitoring assay
indexed by the denaturing gradient gel electrophoresis (DGGE) revealed
the persistence of the seeded CR3 strain in the inoculated media and
adequately outcompeted the autochthonous bacteria.
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Journal of Environmental Management 304 (2022) 114313
Furthermore, the effect of Pseudomonas sp strain WBC-3 on soil
contaminated with methyl parathion (MP) was evaluated by Liming
et al. (2014). The bacterial strain was seeded into a lab-scale MP
contaminated soil initiating a bio-augmentation treatment. Removal of
MP from the polluted soil impregnated with the bacterial strain was
significantly accelerated compared to treatment without microbial
inoculation. Total removal of 0.36 mg g− 1 dry soil in the bio­
augmentation system with production of secondary metabolites at the
idiophasic microbial growth profile within 15 d was also reported. The
seeded bacterial strain survived without perturbation (undisturbed) in
the microcosm while bioremediation action lasted as confirmed by
DGGE and real-time polymerase chain reaction (PCR) analyses. In the
same vein, monitoring the level of interactions between microbial
community and functional environmental factors through a standard
redundancy analysis demonstrated that the structure of the indigenous
bacterial population was significantly (P = 0.002) influenced by the
inoculation of strain WBC-3.
Again, the effect of bioaugmentation by Paracoccus spp HPD-2 on the
soil microbial diversities, as well as PAHs removal from old PAH
contaminated soil was performed by Teng et al. (2010). Their results
indicated the residual PAHs concentration of 2304 μg kg− 1 dry soil
which is directly equivalent to 23.20% PAH immediate removal from its
initial concentration of 9942 μg kg− 1 to 7638 μg kg− 1 dry soil in four
weeks. This significant reduction in average PAH concentration was
completely attributed to bioaugmentation treatment. Removal effi­
ciencies of 35.10%, 20.70%, and 24.30% PAHs fractions (intermediates)
were also reported for 3–4, 4–5, and 5–6 ringed PAHs, respectively. High
performances of all studied parameters such as enzyme assay (dehy­
drogenase activity), total heterotrophic microbial cell density (cfug− 1)
as well as total culturable PAH degrading bacteria (cfug− 1) were re­
ported in the bio-augmentation system. Average well-colour develop­
ment (AWCD) and specific biodiversity footprint evaluated through the
use of BIOLOG Ecoplate assay and Shannon–Weaver index (H) depicted
a significant increase (p < 0.05) in the microcosm seeded with bacte­
rium relative to the control. The single strain bioaugmentation treat­
ment and control microcosm were clear and elaborate as revealed by the
data generated from the principal component of the BIOLOG. This
further confirmed that the significant biorestoration of the microbio­
logical functions, as well as the total arable recovery of the PAH
contaminated soil, was largely attributed to bioaugmentation treatment.
The study, however, posited that Paracoccus sp. strain HPD-2 is an
effective seeder for aged PAH polluted soil towards effective bioreme­
diation action.
3.2.2. Microbial consortia
Bioremediation is mediated by a biodegradation process involving
the conversion or transformation of pollutants from one intermediate to
another. These intermediates often require different arrays of microbial
enzymes and metabolic mechanisms from different microorganisms for
complete contaminant breakdown through cometabolism. Thus, the
deployment of an assemblage of high-density cell masses called con­
sortia (Table 2) is usually of paramount importance for effective reme­
dial outcomes. During bioremediation, one organism gratuitously
produces a particular intermediate which it does not utilize via micro­
bial metabolism and enzymatic conversion. Such metabolic products
could either be innocuous or deleterious to the functioning of such
primary microbial producers but could constitute a vital source of en­
ergy and carbon for other microorganisms in the complex ecosystem
(Mrozik and Piotrowska–Seget, 2010). For example, during the micro­
bial degradation of cyclohexane by Mycobacterium vaccae (Fig. 3). The
cyclohexane is gratuitously oxidized to cyclohexanone by the M. vaccae
growing on propane as a source of carbon and energy. The bacterium
gains no energy and assimilates no carbon from the metabolic trans­
formation from cyclohexane to cyclohexanone. Instead, the cyclohexa­
none is utilized by other microbial populations within the polluted
community e.g., Pseudomonas sp. as a source of energy and carbon. This
A.S. Nwankwegu et al. Journal of Environmental Management 304 (2022) 114313

Table 2 90.60% of 4 and 5 ringed PAHs in a total concentration of 9362.1 μg

Potential microbial consortia for bioaugmentation deployment. kg− 1 dry soil of USEPA priority PAHs. Additions of 10% and 20% of
Consortia Pollutant Reference bacterial consortium suspensions resulted in 20.20% and 35.80% TPH
removal, respectively from the soil after 8 weeks. Studies on growth
Micrococcus luteus (AJ536198), Dual purpose Nwankwegu et al.
Rhizopus arrhizus (JN942919) kerosene (DPK) and (2016a) Nwankwegu dynamics depicted an initial increase in the soil microbial population at
gasoline and Onwosi, 2017b the beginning which decreased progressively with the incubation time.
Bacillus srainsB1F, B5A and B3G Mixture of PAHs Gentry et al. (2004) Nucleic acid determination assay via denaturing gradient gel electro­
(anthracene phoresis (DGGE) analysis of polymerase chain reaction (PCR) generated
Bacillus subtilis, Dm – 04, Crude petroleum oil Andreolli et al. (2015)
Pseudomonas hydrocarbon Aeroginosa Mand NM
amplified 16S rRNA gene fragments which remarkably demonstrated
Rhodococcus sp. Acinetobacter PAHs (Fluorene, Gentry et al. (2004) that the soil’s DGGE profile differed clearly from control. Sequencing
sp, Pseudomonas sp pyrene analysis of DGGE bands demonstrated that the preponderance of the
Phenanthrene) dominant bacterial groups in that soil belonged to proteobacteria and
Mycobacterium fortuitum, PAHs (anthracene, Jacques et al. (2008)
sphingobacteria. The study, therefore, reported an increased PAH
Bacillus cereus phenanthrene,
pyrene) degradation due to microbial consortium incorporation into the
Microbacterium sp, Gornodia, Petroleum Mancera-Lopez et al., contaminated soil thus presenting the method as a promising hydro­
Rhizopus sp, Penicillium hydrocarbon 2008 carbon remedial tool. Isis et al. (2009) used native soil microbiota and
funiculosum Aspergillus bioaugmentation with isolated microbial consortia to evaluate the
sydowii, Rhizobiales sp,
Pseudomonassp, Brucella sp,
bioremediation of a PAH-impacted soil. According to the findings, the
Bacillus sp, Rhodococcussp, Crude oil Zhao et al. (2011) removal of PAHs was ascertained in forest soil microcosms performed
Microbacterium sp with or without bioaugmentation involving fungi and bacteria individ­
Roseomonas sp ually and then fungal consortia. The monitored parameters included
Bacillus marisflavi, Petroleum Supaphol et al., 2006
respiratory activity and metabolic intermediates. The PAH degradation
Microbacterium oxydans hydrocarbon
Pseudomonas oleovorans rate and performance indices were evaluated. The result showed that the
Ochrobactrum sp, Crude oil Varjani et al. (2015) rapid rate of removal in the early stage of the study occurred in micro­
Stenotrophomonas maltophila cosms that contained the contaminants PAHs including phenanthrene,
Rhodococcus sp CT451 anthracene, and naphthalene. Bioaugmentation efficiently improved
Methylibium petroleiphilum Gasoline Daghio et al. (2015)
both the rate and extent of PAH degradation in the system amended with
LMG22953
Alcanivorax borkumensis, Crude oil Suja et al. (2014) the consortium. Carbon (IV) oxide (CO2) quotient/evolution which
Thalassolituus Oleivorans varied roughly with the time rate of PAH removal was used to track the
soil’s respiratory activity. The study further confirmed the utilization of
PAHs as the primary source of limiting nutrients (energy and carbon) by
the PAH degraders. The response of the allochthonous microbial di­
versity to the contaminant (PAHs) was rapid following PAH addition
while it consequently displayed an increased affinity to effectively
degrade the PAH intermediates in the contaminated soil. Another study
by Zafra et al. (2017) also demonstrated that the use of microbial con­
sortia by induced selection in the soil during treatment of PAH polluted
soil resulted in high removal efficiency of 92% at the end of the study
period. In addition, the capacity of a specific microbial consortium
involving Mycobacterium fortuitum, Bacillus cereus, Microbacterium
polyisoprenivorans, Microbacteraceae bacterium and Fusarium oxy­
sporum to metabolize phenanthrene and pyrene in soil was evaluated by
Jacques et al. (2008). The study reported the PAH removal efficiency of
96%–99% within 70 d. A study that involved the PAH-degrading mi­
crobial consortium and its pyrene-degrading plasmids from mangrove
sediment samples in Huian, China was carried out by Yi et al. (2008) and
observed that consortium YL showed significant removal efficiencies of
92.1%, 87.6%, 92.3%, and 95.8% for pyrene, fluoranthene, phenan­
Fig. 3. Microbial community interactions through co-metabolism assisting
threne, and, fluorene, respectively at 50 mg L− 1 after 21 d incubation.
complete mineralization of hydrocarbon pollutants into carbon (iv) and water
The Bacillus cereus Py5 and B. megaterium Py6 eventually isolated from
during microbial consortium-bioaugmentation operation.
the consortium were shown to have consumed 65.8% and 33.7% of
pyrene (50 mg L− 1), respectively under 21 d. The enriched bacteria
way, the total removal (mineralization) from such environment of the (Escherichia coli DH5α) bearing the plasmid of consortium YL was re­
cyclohexane which would have been preempted by the generation of the ported to have effectively degraded 85.7% of the original pyrene con­
cyclohexanone intermediate is ensured. The synergistic interactions, centration after 21 d. Xiang et al. (2013) used a consortium consisting of
therefore, promote continuous degradation with little or no toxicity or para-nitrophenol utilizer Pseudomonas sp. strain WBC-3, meta-ni­
delayed clean up even in dealing with recalcitrant contaminants. High trophenol utilizer Cupriavidus necator JMP134 and ortho-nitrophenol
potential for PAHs degradation in contaminated soil by microbial con­ utilizer Alcaligenes sp. strain NyZ215 inoculated into soil contaminated
sortia has been reported (Zafra et al., 2017). Studies that demonstrated with three nitrophenol isomers for bioaugmentation. Accelerated
effective removal of a pollutant from the environment by consortia of removal of all nitrophenols was achieved in the inoculated soils
aromatic hydrocarbonoclastic bacteria very rapidly compared to its compared to un-inoculated soils, with total elimination of nitrophenols
single strain counterpart abound (Goux et al., 2013). Several reports
in the inoculated soils occurring between 2 and 16 d. Real-time poly­
have shown significant success in the depuration of the recalcitrant and merase chain reaction (PCR) targeting nitrophenol-degradation func­
toxic pollutants (e.g., PAHs) from soils using microbial consortia. The
tional genes demonstrated that the three strains survived and were
bioremediation of a PAH-contaminated soil was studied using a bacterial stable over time. Also, Wu et al. (2013) studied the degradation of
consortium enriched from the soil by Jian et al. (2012). Soil composed of
polycyclic aromatic hydrocarbons by microbial consortia enriched from

5
A.S. Nwankwegu et al. Journal of Environmental Management 304 (2022) 114313

three soils using two different culture media and discovered that malt
yeast extract-enriched bacterial consortium demonstrated efficient
PAHs removal rates than the Luria broth (LB) enriched counterpart.
Consortia originally from soil and seeded back to that same soil pre­
sented higher efficiency in degrading PAHs. Thus, the study posited that
microorganisms isolated from the contaminated site, activated by
growth in enrichment media and later transferred back to the polluted
media would demonstrate sufficient removal of such pollutants over
their allochthonous counterparts.
3.3. Bioaugmentation using genetically engineered microorganisms
(GEMs)
Genetic engineering otherwise referred to as recombinant DNA
technology has received global attention as a promising biotechnical
approach towards effective depuration of the environmental pollutants
especially the recalcitrant hydrocarbon fractions (Liu et al., 2019;
Dvorak et al., 2017; Mrozik and Piotrowska–Seget, 2010). This involves
the transfer of genetic materials otherwise referred to as the biomarkers
(Table 3) bearing the desired traits through the use of appropriate
expression vector/plasmids from one organism (wild strain) to another
(host). Several biomarkers are developed to evaluate the potency and
efficiency of particular bacteria that are utilized as suitable inocula in
the biorestoration of polluted sites (Yi et al., 2008). Specifically, the luc
gene, encoding firefly luciferase and the gfp gene, encoding the green
fluorescent protein (GFP) are common biomarkers. In tagging different
microorganisms (bacteria) used in the bioremediation of hydrocarbon
pollutants such as gasoline and chlorophenols, the luc gene is usually
employed. Soil biological activity indexed by luciferase activity on
respiring microbial cells is used as a bioindicator for monitoring bacte­
ria. The chlorophenols monitoring in the soil while applying the prin­
ciple of fluorescence of GPD protein is also performed using the gfp gene.
Different forms of biological indicators or biomarkers can also be
adopted for the same purpose including; dehydrogenase assay, earth­
worm survival, carbon (iv) oxide quotient, and phyto/ecotoxicity by
seed germination assay incorporating germination index and chloro­
phyll content estimations. Different factors influence the decision on the
type of monitoring system to be adopted and these include; required
detection specificity, polluted site history, as well as microbial strain and
specificity to pollutants (Jansson et al., 2000). The autochthonous mi­
croorganisms upon protracted exposure to hydrocarbon pollutants
evolve (metabolic acclimatization) to possess the ability to degrade such
pollutants over time. This metabolic evolution which often involves
mutations and horizontal gene transfer (HGT) regularly occurs but is
relatively slow (Mrozik and Piotrowska–Seget, 2010). As a result,
improving microbial degradative processes using the manipulations of
molecular biology and genetic engineering became crucial globally.
Several studies on hydrocarbon degradation have been carried out using
genetically modified microbial strains (Wei et al., 2008). Such modifi­
cations aimed to enhance the ability of recombinant strains to degrade a
wide range of xenobiotics and increase the effectiveness of removal
relative to the wild strains. Zhao et al. (2017) reported that horizontal
gene transfer of related genes and prophages between Sphingomonas and
Sphingobium strains through transposase and IS coding genes of bph and
xyl gene clusters can result in increased biodegradation of PAHs. Again,
a transcription of either yedQ (diguanylate cyclase) or yhjH (c-di-GMP
phosphodiesterase) from E. coli to P. putida KT2440 was artificially
placed under the tight control of a cyclohexanone-responsive expression
system by Benedetti et al. (2016) and a synthetic operon, tested and
confirmed effective for enhanced 1-chlorobutane biodegradation was a
resulting strain. Similarly, bacterial strains (HLDs) that utilize xenobi­
otic halogenated compounds, and three archetypal HLDs, i.e., DhlA,
DhaA, and LinB, have been extensively studied and improved substrate
specificities, as well as activities during the conversion of halogenated
compounds to their respective alcohols by successful engineering of
HLDs (Nagata et al., 2015). Furthermore, Rodrigues et al. (2006),
engineered Burkholderia xenovorans LB400 using the biphenyl pathway
by inserting in the genome, the operon ohb (oxygonenolytic orth –
dehalogenation), thus providing the organism with the additional ability
to effectively utilize 2– chlorobenzoate and 2 –chlorobiphenyl.
However, the use of genetically engineered microorganisms via re­
combinant DNA technology although a promising and effective remedial
strategy is at present faced with conflicting ethical predispositions.
Considering that a desirable trait of one microorganism is incorporated
into another using expression vectors (e.g., plasmid vector) equipping it
with the desired ability to express the same characteristic as though it
were the original microorganism and most times much more efficient
than the wild strain (Kuppusamy et al., 2017), scientists are perturbed
by its future impacts on the atmosphere and humans (Liu et al., 2019).
The application of recombinant microorganisms (GEMs) in environ­
mental clean-up has gained a controversial world view, a low global
acceptance, and considerable technological drudgery including; the cost
of technology, the paucity of multiple plasmids with potential degra­
dative abilities, rural reachability/availability, its long-term effect on

Table 3
Biomarkers of environmental monitoring importance from different microbial genome.
Microorganism Biomarker Potential substrate Reference

Sphingobium.
Pseudomonas putida KT2440
Sphingobium chlorophenolicum L-
1
Sphingobium japonicum UT26S
Arthrobacter aurescens (NC008711)
Cupriavidus necatorRW112
Pseudomonas putida PaW 340/
pDH5
Pseudomonas fluorescensRE
Pseudomonas fluorescensMP
Burkholderia xenovoransLB400
(ohb)
Pseudomonas putidaKT2442
enzymes, Tcr
Pseudomonas fluorescens
F113rifpcbrrnBP1::gfpmut3
Intact or partial bph and xyl gene cluster PAHs Zhao et al., 2017
yedQ (diguanylate cyclase),yhjH (c-di-GMP phoshodiesterase) Haloalkanes Benedetti et al. (2016)
2 (1) Pentachlorophenol (PCP) Copley et al. (2012)
2 (3) Hexachlorocyclohexane (HCH) Nagata et al. (2007)
atzA atzB atzC atzD atzE atzF trzN trzD Atrazine Sagarkar et al. (2014)
tcbRCDEF-Genes for chlorocatechol degradation from pBJ4::4 (derived Chlorobenzoates Aroclor 1221 and Wittich and Wolff, 2007
from pP51 of Pseudomonas sp. 51),cbdABC – genes encoding 2-halo­ 1232
benzoate dioxygenase from Pseudomonas cepacia 2CGS located in
pBpc, xylXYZ – genes encoding toluate dioxygenase from TOL plasmid
ofP. putida mt-2
Genes encoding dehalogenase from Arthrobacter sp. FG1 cloned in 4-Chlorobenzoic acid Massa et al. (2009)
pDH5
Chromosomal insertion of dntABDEG genes derived from Burkholderia 2,4-Dinitrotoluene Milena et al. (2011)
sp. DNT
Megaplasmid pJS1with dnt genes from Burkholderia sp. DNT ohb 2, 4-Dinitrotoluene Milena et al. (2011)
Operon cloned into vector pRT1, yielding pRO41,
encoding oxygenalytic ortho-dehalogenation enzymes Naphthalene Romen et al., 2013 pNF142::TnM,od-OTc –
encoding naphthalene
degrading,
F113 with mini-Tn5-Kmr-rrnBP1::gfp [mut3]-T0-T1 Biphenyl, polychlori randomly Chi-Wen et al., 2012
inserted into the chromosome

6
A.S. Nwankwegu et al.
humans, the problem of plasmid compatibility, as well as delayed mi­
crobial reproduction due to excessive plasmid insertions (Liu et al.,
2019). These inadequacies work in tandem to substantially sabotage the
technology sustainability and security. A more prudent recombinant
DNA biotechnological design is, therefore, imperative to circumvent the
controversies associated with GEMs-mediated environmental remedial
convention.
4. Microbial stimulation by electric field exposures
The novel approach for the manipulation of microorganisms towards
optimizing rapid biomass productions leading to effective pollutants
depuration involves the use of extremely low-frequency electromagnetic
fields (ELF). Although this strategy has been rarely adopted among re­
searchers, especially in field-scale applications, it shows potential
prospects as a useful biotechnological tool. ELF is a bioenergetic stim­
ulation technology adopted in bioremediation, particularly to stimulate
the uptake rates of a wide range of environmental pollutants available in
the waste streams (Hunt et al., 2009). It primarily involves the direct
activation of the allochthonous/autochthonous microbial diversity via
stimulating their growth and metabolic behaviour in a system under­
going bioaugmentation through electric/electromagnetic exposures.
During the field exposure regimes, several conditions often regulate the
biological outcomes including length of exposure, pulse shape, field
strength, frequency, magnetic intensity, and type of modulation. The
potential positive effects of ELF exposures include biomass increase,
increased resistance to environmental pollutants, stimulation of the
degradation of contaminants, substrate uptake kinetics accelerations,
increased denitrification/nitrification activities, increase in metabolic
activity/specific enzymes actions e.g., catalase, dehydrogenases, urease
etc. (Beretta et al., 2019).
However, as useful as ELF applications in environmental recovery
may be, organisms excited by ELF regimes could be affected either
negatively or positively. The processes of metabolism and microbial
growth often show high vulnerability to ELF exposures. The negative
effects usually include inhibition of bacterial growth, reduction of the
degradation of pollutants and/or substratum consumption, reduction in
metabolic activity, and cellular membrane damage. Further, Beretta
et al. (2019) identified that some of these adverse effects could be in­
direct rather than direct and occur due to the modifications in the
environmental conditions e.g., the electrochemical release of toxic
species/radicals and extremes in pH conditions. Under the elec­
tric/magnetic field-induced effects, the growth and performance of
bacteria are inhibited or stimulated. Specifically, Justo et al. (2006)
previously reported that E. coli exposed to extremely low-frequency
electromagnetic fields at 0.1 mT under 6.5 h showed some changes
relative to the unexposed cells. Significant decreases in biofilm forma­
tions under exposure to extremely low-frequency magnetic fields
(ELF-MF) have been demonstrated to cause a substantial reduction in
the bacterial adherence to substratum with altered permeability of the
ionic channels of the cell membrane. Generally, exposure to the
extremely low-frequency magnetic field (ELF-MF) increases bacterial
growth, while it decreases both biofilm formation and viability (Justo
et al., 2006). In the same study, inhibition of cell viability of E. coli at the
electromagnetic field of 100 mT was reported. The adverse effects are
associated with the numerous latent parameters which are often difficult
to control notably sunspot events, cosmic, the orientation of Earth’s
geomagnetic field, local intensity radiations, and solar winds (Hunt
et al., 2009). In effect, the use of ELF in pollution abatement could have
some environmental and public health consequences as exposure to
these radiations, especially high voltage powerlines could trigger
adverse environmental modifications, thymine-dimer related mutations,
and the proliferation of tumour and cancer cells.
7
Journal of Environmental Management 304 (2022) 114313
5. Principle of bioremediation
Environmental pollutants e.g., crude oil is a composite mixture of
varying chemical compounds while the TPHs include the hydrocarbon
derivatives that bioaccumulate and bioconcentrate in the food chain
from the environment (Nwankwegu et al., 2018). Each composite is
unique and being acted upon differently by specific microbes or flora.
Bioremediation proceed naturally or can be expedited by microbes or
nutrients augmentation or both. The microbes present in the polluted
media recognize the pollutants (e.g., oil) and their congeners. The mi­
crobes then secrete their solubilizing agent like; biosurfactants other­
wise referred to as bioemulsifiers (Table 4). They attach themselves and
utilize the pollutants (hydrocarbon) as a source of energy and carbon
(Chen et al., 2015b; Liu et al., 2016). The biosurfactants, biologically
surface-active agents (bioemulsifers), lower the surface tension of liq­
uids, the intermolecular (interfacial) forces acting between liquid and
soil. These can act as dispersants, solubilizing agents, or emulsifiers. For
instance; Pseudomonas species produces a glycolipid based biosurfactant
known as rhamnolipid (Rekadwad et al., 2019; Sood et al., 2019; Zheng
et al., 2019) while a lipoprotein based biosurfactant known as surfactin
is produced by Bacillus subtilis (Thérien et al., 2020; Paraszkiewicz et al.,
2017; Alonso and Martin, 2016; Rahman et al., 2003). Under the natural
attenuation approach, the bioavailability to microorganisms of the high
molecular weight hydrocarbon is often limited due to the low adsorption
and solubility. The bioemulsification offered by biosurfactants signifi­
cantly increases solubility and removal efficiencies of the pollutants,
thus, improving biodegradation actions (Anaukwu et al., 2016). The
incidence of starvation stress and consequent push of contaminant
degrading microorganisms into the idiophasic or stationary phase of
growth dynamics thus a remarkable delay in biodegradation action is a
strong limitation synonymous to natural attenuation. To address the
concern, pollutants utilizing microorganisms are delivered into the
polluted environment in immobilized bioaugmentation systems
(Nwankwegu and Onwosi, 2017a). The immediate resumption of
degradation activity by microbial systems in suspension during bio­
augmentation directly affects positively the removal efficiency, enzyme
assay (e.g., dehydrogenase activity), degradation half-life, as well as
overall rate and extent of contaminant removal. This is because the
seeded inocula already activated in their respective primary micro­
systems quickly slide into the exponential growth profile when trans­
ferred into a polluted environment resulting in effective contaminant
utilization. In this scenario, the loss of cellular respiration often dem­
onstrates linear overlap with the disappearance rate of the targeted
pollutants. It is established that no known environmental contaminant
removal option shows 100% removal efficiency. This is due to the
metabolite production by the competing microbial diversities that
attempt to modify the microenvironment to suit its singular metabolic
style slowing down the actions of other microbial populations. Apart
from this, another reason for the non-total contaminant degradation is
that some environmental contaminants (e.g., hydrocarbon) at the level
of ALARP point (As Low As Reasonably Practicable Point) become
recalcitrant and technologically inaccessible though usually nontoxic at
this point. As Low As Reasonably Practicable (ALARP) point has been
Table 4
Different biosurfactants/bioemulsofers produced by microorganisms.
Microorganism Biosurfactant Reference
Pseudomonas aeruginosa rhamnolipid Nwankwegu and Onwosi (2017a)
Bacillus subtilis surfactin Nitschke et al., 2004
Mycobacterium spp Trehalose lipids Xia et al. (2011)
Yeast sophorolipds Souza et al. (2017)
Thiobacillus Thioxidans phospholipids Souza et al. (2017)
Brevibacterium spp polymixins Varjani et al., 2017
Serratia marcescens serrewettin Lai et al. (2009)
Leuconostoc messenteroids viscosin Banat et al. (2010)
Aspergillus ustus glycolipoprotein Alejandro et al. (2011)
A.S. Nwankwegu et al.
described as the point at which no further significant breakdown or
reduction in the residual concentration of a specific contaminant can be
achieved both economically and substantially (Nwankwegu et al., 2018;
Nwankwegu, 2016). In other words, the residual fraction of such
contaminant automatically loses its bioavailability and toxicity to
microbiota. Therefore, to ensure significant contaminant reduction, as
well as obtaining a sufficiently low ALARP point, bioremediation by
bioaugmentation becomes ultimately imperative. The hydrocarbon
fractions show strong variability in both physical and chemical char­
acteristics including solubility, volatility and susceptibility to biodeg­
radation. Some compounds have a shorter half-life and are degraded
easily (lower molecular weight hydrocarbon fractions), most resist
degradation under normal conditions (higher molecular weight hydro­
carbon e.g., PAHs), while others are non-biodegradable (Sublette, 2001;
Agarwal and Liu, 2015). The removal of hydrocarbon pollutants does
not occur evenly, because different species of microbes often show
preference to different fractions such that the first to be degraded is a
function of which congeners show a wide uptake preference among the
degraders (Nwankwegu et al., 2016b). In the presence of carbon sources,
the process of enzymes production is triggered in degraders and these
enzymes quickly catalyze the efficient degradation of the hydrocarbon
molecules (Gao et al., 2014; Thaoti et al., Bilal et al., 2017) into carbon
(iv) oxide and water. The hydrocarbon degradation mechanism is,
however, regulated by the streams of enzymes and metabolic pathways.
The efficient pollutant removal is preempted by the lack of appropriate
enzymes involved in the metabolic breakdown of pollutants into their
chemical intermediates during environmental depuration.
6. Successful application of bioaugmentation across regions
Bioaugmentation though can be influenced by abiotic and biotic
components (temperature, pH, humidity, microbial predation, and
bioavailability), success in large scale application could vary across
Table 5
Case studies of successful bioaugmentation, potential pollutants, and remedial achievements across regions.
Country Case study/compound
United Waste water treatment
Kingdom
Estonia Phenolic compounds
Italy Diesel oil, phenanthrene, isoprenoids and total hydrocarbons
France Phenanthrene, benzene, toluene, ethylbenzene, xylene, and
naphthalene
India polycyclic aromatic hydrocarbons (PAHs)
India refinery oily sludge
Total petroleum hydrocarbon
China Clay-loam soil contaminated with 16,300 mg/kg of total
petroleum hydrocarbons (TPH); comprising of 51% saturated
hydrocarbons and 31% aromatic
USA pentachlorophenol (PCP) and creosote contaminated soil
USA contaminated aquifers
USA Trichloroethene (TCE)-contaminated aquifer
USA Chlorinated Solvents in trichloroethene [TCE], dichloroethene
Groundwater ([DCE], vinyl chloride [VC])
Journal of Environmental Management 304 (2022) 114313
regions. The regional variabilities in the bioaugmentation functionality
could also be linked to other potential background factors including;
ecosystem heterogeneity, environmental-specific microbial diversity,
pollutant characteristics, and the environmental acceptability of the
foreign microorganisms which, are regrettably not considered and
identified in most cases. Inherently, the existence of a variety of stra­
tegies by microbes such as cellular membrane modification, surface-
active compounds (surfactant), use of efflux pumps, and the mainte­
nance of biological functions, could favour compound detoxification
inside the microbial cells (de Carvalho et al., 2009). It has also been
highlighted that Burkholderia pickettii, P. putida PaW340/pDH5 (Massa
et al., 2009), and P. fluorescens CS are promising agents for successful
bioaugmentation. Studies have also shown the collaborative influence of
soil components such as insoluble organic, water content and organic
matter for BTEX (benzene, toluene, ethylbenzene and xylenes) degra­
dation. Table 5 presents the cases of the successful bioaugmentation
application across regions.
7. Prospects of bioaugmentation
The application of specialized microorganisms at the contaminated
site to sufficiently biodegrade contaminants to an environmentally safe
level shows potential prospects vis-à-vis environmental sustainability
and flexibility. This technology over the years has been proven effective,
fast, and affordable greener cleanup option (Guo et al., 2010; Wang
et al., 2013; Chen et al., 2015c; Zabbey et al., 2017). In a situation where
biodiversity lacks the expression of appropriate enzymes and metabolic
pathways to quickly tackle the hydrocarbon contaminant following
fresh pollution, allochthonous populations with tested and effective
metabolic capacity are introduced. This method has been described by
researchers as the most effective intervention for fresh hydrocarbon
spills. The seeded microbial suspensions quickly metabolize the pollut­
ants converting them into carbon (iv) oxide and water with little or no
Achievement Reference
Waste water biaugmentation using reactors and selection of valuable strains Herrero and
or mixed cultures to improve the catabolism of specific compounds such as Stuckey (2015)
refractory organics, or overall carbon oxygen demand (COD).
Using cultivation-based Box-PCR genomic fingerprints, fingerprinting of Juhanson et al.
the 16S rRNA and lmPH genes, Pseudomonas strains was successfully apply (2009)
in the degradation of Phenolic compounds
Complete degradation of diesel oil and phenanthrene. Alisi et al. (2009)
60% and 75% reduction of isoprenoids, and total hydrocarbons
respectively.
Successful degradation of pollutants using Mycobacterium, Rhodococcus, Farhadian et al.
Haemophilus, Mycobacterium, Paenibacillus and Ralstonia as microbial (2008)
agents
Degradation of polycyclic aromatic hydrocarbons (PAHs) Haritash and
Kaushik (2009)
Bioaugmentation presented a positive tide in the oil degradation at 90% Jasmine and
confidence level in a microcosm study using AcinetobacterSZ-1 strain K Mukherji (2014)
F453955 (TPH-degrader). Wu et al. (2016)
AcinetobacterSZ-1 strain K F453955 shows 34% at degradation plateau of
sixth and seventh weeks of incubation
degraded by yeast Candida tropicalis SK21 isolated from petroleum Fan et al. (2014)
contaminated soil
treated with the fungus Phanerochaete sordida Lamar et al.
(1994)
Aquifers contaminated with chlorinated ethenes can be contained by filling Lendvay et al.
the site with competent microbial inoculants. (2003)
de Wildeman
et al., 2004
Through anaerobic bio-augmentation techniques, trichloroethene (TCE)- Ellis et al. (2000)
contaminated aquifer and cis-1,2-dichloroethene (cDCE) in groundwater
were successfully converted to ethane through microbial enrichment
in situ bio-augmentation by treatment with Burkholderia cepacia ENV435 Steffan et al.
(1999).
8
A.S. Nwankwegu et al.
lag in growth dynamics (Wang et al., 2013; Diaz et al., 2016). The
rationale behind this strategy is that the autochthonous microbial di­
versities although present may not possess the metabolic potentials to
degrade broad varieties of the potential substrates present in complex
mixtures such as petroleum over a wide range of temperature and pH
(Liu et al., 2017). Specifically, the commercial bacterial inoculum was
estimated to achieve 99% depuration efficiency of the targeted recalci­
trant chlorinated volatile organic compounds (CVOCs) which, are
resistant to degradation by the native bacterial diversity (U.S. EPA,
2010). Again, the condition under which bioaugmentation may be
considered is when there is a paucity of indigenous hydrocarbonoclastic
microorganisms at the site of hydrocarbon contamination to effectively
attack the contaminants following a fresh spill. This is common to a fresh
spill in a pristine ecosystem where the allochthonous diversity often
requires a long period of adaptation through a slow genetic recognition
before utilizing the contaminants as the sole source of energy and car­
bon. The rate and extent of decontamination are an important footprint
in evaluating and measuring the magnitude of technology effectiveness.
These microorganisms once seeded into polluted media automatically
drive the microbial growth dynamics into its active troph­
ophasic/logarithmic profile with little or no lag period while dealing
with the contaminants. The operational conditions for enhanced
degradation during bioaugmentation centrally include; ease of micro­
bial adaptability, genetic stability, microbial motility, as well as mass
transfer mechanisms through the polluted system. These operational
conditions are optimized by microbial cell immobilization of the bio­
augmentation system. The process of cell entrapment, adsorption, and
encapsulation are vital immobilization techniques used to deliver the
microbial system into the polluted environment while reducing in­
adequacies (environmental perturbations) associated with initial
cellular adaptation and the ambient toxicity of the polluted environment
to the allochthonous microbial species. Polymeric materials such as;
agarose, polyacrylamide gel, K-carrageenan alginate, clay, anthracite,
porous glass, and ceramics are suitable immobilizing agents referred to
as hydrogel materials for cell immobilization in a bioaugmentation
system (Nwankwegu and Onwosi, 2017a). These operational advances
are currently been used to significantly increase the remedial outcome of
bioaugmentation options during the environmental pollutant cleanups
(Dai et al., 2020; Ma et al., 2020; Tao et al., 2019; Shi et al., 2018; Wang
et al., 2014a, 2014b). The immobilizing media are also suitable
attachment surfaces for optimized microbial actions while improving
contact between the microorganisms and the pollutants.
The science of microorganisms in bioremediation systems directly
simplifies the selection of microbial population(s) with potential for the
degradation and production of the product(s) with biotechnological
applications in the oil hydrocarbon and petrochemical industries (Her­
rero and Stuckey, 2015; Diaz et al., 2016). Microorganisms are ubiqui­
tous such that environmental pollution experts engaged in
environmental decontamination processes involving bioaugmentation
technology would, therefore, require no cost in mobilizing these mi­
croorganisms. The use of biostimulation (chemical formulations) as a
source of additional nutrients apart from its residual negative ecological
impact also threatens land reclamation for potential arable functions
(Nwankwegu et al., 2018) due to altered eco-stoichiometry by effective
but toxic biostimulants. The natural attenuation approach is the
cheapest and easy ton setup pollution remedial strategy; although the
use of soil’s natural ability to degrade contaminants is usually slow such
that the total sterility of polluted systems is possible before half the
contaminant is removed (Bento et al., 2005; Nwankwegu et al., 2016b;
Nwankwegu, 2016). Indeed, the essence and goal of bioremediation is to
remove fresh pollutants from the environments soon after contamina­
tion events occur so that it does not establish the process of permanent
sterility. Therefore, to decide which bioremediation strategy to be
adopted for rapid contaminants removal while eliminating the chances
of eutrophication and the deleterious toxicity to microbiota, bio­
augmentation shows a promising possibility in preserving
9
Journal of Environmental Management 304 (2022) 114313
environmental footprint.
However, it must be noted that authors have not exclusively claimed
that bioaugmentation operation is without any limitation rather we
essentially posit that bioaugmentation inadequacies are better
controlled under practical scenarios relative to other bioremediation
strategies. For instance, previous failed attempts/or poor remedial out­
comes during bioaugmentation have been linked to a few factors such as
biotic e.g., the initial adaption of the exogenous microbial species in the
presence of their native counterparts including predation and competi­
tion between autochthonous and allochthonous communities, abiotic
influences e.g., pH and temperature. These are crucial factors that could
preempt bioaugmentation performances if not properly controlled.
Interestingly, recent bioaugmentation steps have adequately resolved
these perturbations through the process of microbial cell immobilization
(Nwankwegu and Onwosi, 2017a).
8. Conclusion and future research perspectives
Anthropogenic activities are a driving force for the generation of
enormous environmental pollutants, particularly hydrocarbons and
their derivatives severely threatening the global environment. The
present review extensively evaluated the potential bioremediation types
ranging from ex-situ to in situ technologies, as well as the approaches
including; natural attenuation, biostimulation, and, bioaugmentation. It
remarks that the traditional physicochemical remedial steps are costly
and create chances for secondary and residual pollution possibilities.
The natural attenuation though essentially cost-effective is often very
slow that the polluted environment e.g., soil loses its arable function
before a remedial attempt is completed. The biostimulation strategy is
exceptionally effective and fast leading to successful remedial in­
terventions within a short time. However, aside from its high cost
associated with material procurements, most chemical formulations
used as “amendment” are composed of inorganic stimulants which are
themselves more toxic than the primary pollutants. In other words, it
posits that the contaminants eventual removal often does not guarantee
ecosystem eventual recovery for potential agronomic functions, partic­
ularly when artificial additives capable of severely altering soil’s phys­
icochemical characteristics (e.g., pH, cation exchange capacity,
conductivity etc.) are used to expedite remedial actions. This thus makes
most biostimulation involving inorganic stimulants parochially appli­
cable in environmental reclamation (e.g., land) intended for non-
agricultural/non-residential uses e.g., industrial establishment.
Further, some nitrogenous and phosphorus-based “amendment” agents
are flushed into the aquatic system via runoff with a dramatic exacer­
bation of eutrophication and the episodic outbreak of harmful algal
bloom (HAB) consequently threatening water quality and public health
while debunking the strategy effectiveness. The use of the novel ELF
approach for cellular stimulation during bioaugmentation was also
advised but the problems associated with ELF exposures to both envi­
ronment and public health raise a threatening concern. It, therefore,
posits that the bioaugmentation approach involving the use of micro­
organisms, which are ultimately ubiquitous and innocuous be priori­
tized. Consequently, the use of high cell density microbial assemblages
referred to as the microbial consortia owing to its wide range of enzy­
matic and degradative pathways suited for different pollutant in­
termediates is the most promising bioaugmentation option which
demonstrates high efficiency in ecosystem biorecovery and the eventual
land use for arable purposes. The use of genetically engineered micro­
organisms would also suggest a prudent and sustainable remedial design
but at present challenged by low public acceptance and associated
controversies due to constrained ethical considerations thus, future
research perspectives should be focused on developing ethical free re­
combinant DNA technology for more efficient microbial engineering
delivery that could beat the present progress and by-pass the ethical
issues while addressing its associated high-cost implication.
A.S. Nwankwegu et al.
Credit author statement
Amechi S. Nwankwegu: Conceptualization, Data curation, Formal
analysis, Investigation, Methodology, Writing – original draft. Lei Zhang
and Deti Xie: Funding acquisition, Supervision. Chukwudi O. Onwosi:
Visualization, Writing – review & editing. Wada, I. Muhammad, Chuks
K. Odoh, Kabari Sam, John N. Idenyi: Formal analysis and Visualization.
Declaration of competing interest
The authors declare that they have no known competing financial
interests or personal relationships that could have appeared to influence
the work reported in this paper.
Acknowledgements
This work was financially supported by China Postdoctoral Fellow­
ship, Southwest University, Chongqing, China (station; No.289550) and
the Ontario–China Research and Innovation Funding project “Innova­
tive monitoring and prediction of non-point source pollution and water
quality in the Three Gorges Reservoir catchment” (2013DFG92520).
References
Abena, M.T.B., Li, T., Shah, M.N., Zhong, W., 2019. Biodegradation of total petroleum
hydrocarbons (TPH) in highly contaminated soils by natural attenuation and
bioaugmentation. Chemosphere 234, 864–874.
Agarwal, A., Liu, Y., 2015. Remediation technologies for oil-contaminated sediments.
Mar. Pollut. Bull. 101, 483–490.
Alejandro, C.S., Humberto, H.S., Maria, J.F., 2011. Production of glycolipids with
antimicrobial activity by Ustilago maydis FBD12 in submerged culture. Afr. J.
Microbiol. Res. 5, 2512–2523.
Alisi, C., Musella, R., Tasso, F., Ubaldi, C., Manzo, S., Cremisini, C., Sprocati, A.R., 2009.
Bioremediation of diesel oil in a co-contaminated soil by bioaugmentation with a
microbial formula tailored with native strains selected for heavy metals resistance.
Sci. Total Environ. 407, 3024–3032.
Alonso, S., Martin, P.J., 2016. Impact of foaming on surfactin production by Bacillus
subtilis: implications on the development of integrated in situ foam fractionation
removal systems. Biochem. Eng. J. 110, 125–133.
Anaukwu, C.G., Ezemba, C.C., Anakwenze, V.N., Agu, K.C., Nwankwegu, A.S., Okeke, B.
C., Awah, N.S., 2016. Influence of anionic, cationic and non-ionic surfactants on
growth of hydrocarbon utilizing bacteria. Amer. J. Cur. Microbiol. 4 (1), 1–7.
Andreolli, M., Lampis, S., Brignoli, P., Vallini, G., 2015. Bioaugmentation and
biostimulation as strategies for the bioremediation of a burned woodland soil
contaminated by toxic hydrocarbons: a comparative study. J. Environ. Manag. 153,
121–131.
Banat, I.M., Franzetti, A., Gandolfi, I., Bestetti, G., Martinotti, M.G., 2010. Microbial
biosurfactants production, applications and future potential. Appl. Microbiol.
Biotechnol. 87, 427–444.
Benedetti, I., de Lorenzo, V., Nickel, P.I., 2016. Genetic programming of catalytic
Pseudomonas putida biofilms for boosting biodegradation of haloalkanes. Metab.
Eng. 33, 109–118.
Bento, F.M., Flavio, A.O.C., Benedict, C.O., William, T.F., 2005. Comparative
bioremediation of soil contaminated with diesel oil by natural attenuation,
biostimulation and bioaugmentation. Bioresour. Technol. 96, 1049–1055.
Beretta, G., Mastorgio, A.F., Pedrali, L., Saponaro, S., Sezenna, E., 2019. The effects of
electric, magnetic and electromagnetic fields on microorganisms in the perspective
of bioremediation. Rev. Environ. Sci. Biotechnol. 18, 29–75.
Bilal, M., Igbal, H.M.N., Hu, H., Wang, W., Zhang, X., 2017. Development of horseradish
peroxidase-based cross- linked enzyme aggregates and their environmental
exploitation for bioremediation purposes. J. Environ. Manag. 188, 137–143.
Brown, D.M., Samson, O.S., Gils, J.V., Spanning, R.V., Bonte, M., Hutchings, T.,
Linden, O., Egbuche, U., Bruun, K., Smith, J.W.N., 2017a. Comparison of
landfazarming amendments to improve bioremediation of petroleum hydrocarbons
in Niger Delta soils. Sci. Total Environ. 596–597, 284–292.
Brown, L.D., Cologgi, D.L., Gee, K.F., Ulrich, A.A., 2017b. Bioremediation of oil spills on
land. Oil Spill Sci. Technol. 699, 729.
Chatterjee, S., Deb, U., Datta, S., Walther, C., Gupta, D.K., 2017. Common explosives
(TNT, RDX, and HMX) and their fate in the environment: emphasizing
bioremediation. Chemosphere 184, 438–451.
Chen, M., Xu, P., Zeng, G., Yang, C., Huang, D., Zhang, J., 2015a. Bioremediation of soils
contaminated with polycyclic aromatic hydrocarbons, petroleum, pesticides,
chlorophenols and heavy metals by composting: applications, microbes and future
research needs. Biotechnol. Adv. 33, 745–755.
Chen, Q., Ni, J., Ma, T., Liu, T., Zheng, M., 2015b. Bioaugmentation treatment of
municipal wastewater with Heterotrophic-aerobic nitrogen removal bacteria in a
pilot-scale SBR. Bioresour. Technol. 183, 25–32.
Chen, W.C., Juang, R., Wei, Y., 2015c. Applications of a lipopeptide biosurfactant,
surfactin, produced by Microorganisms. Biochem. Eng. J. 103, 158–169.
10
Journal of Environmental Management 304 (2022) 114313
Chi-Wen, L., Chi-Hung, W., Chen-Ting, T., Shih-Hsien, C., 2012. Novel oxygen-releasing
immobilized cell beads for bioremediation of BTEX-contaminated water. Bioresour.
Technol. 124, 45–451.
Copley, S.D., Rokicki, J., Turner, P., Daligault, H., Nolan, M., Land, M., 2012. The whole
genome sequence of Sphingobium chlorophenolicum L-1: insights into the evolution
of the pentachlorophenol degradation pathway. Genome Biol. Evol. 4, 184–198.
Cycon, M., Mrozik, A., Piotrowska-seget, Z., 2017. Bioaugmentation as a strategy for the
remediation of pesticide- polluted soil: a review. Chemosphere 172, 52–71.
Daghio, M., Tatangelo, V., Franzetti, A., Gandolfi, I., Papacchini, M., Careghini, A.,
Sezenna, E., Saponaro, S., Bestetti, G., 2015. Hydrocarbon degrading microbial
communities in bench scale aerobic biocarriers for gasoline contaminated
groundwater treatment. Chemosphere 130, 34–39.
Dai, X., Lv, J., Yan, G., Chen, C., Guo, S., Fu, P., 2020. Bioremediation of intertidal zones
polluted by heavy oil spilling using immobilized laccase-bacteria consortium.
Bioresour. Technol. 123305 https://doi.org/10.1016/j.biortech.2020.123305.
de Wildeman, S., Linthout, G., Van Langenhove, H., Verstraete, W., 2004. Complete lab-
scale detoxification of groundwater containing 1, 2-dichloroethane. Appl. Microbiol.
Biotechnol. 63, 609–612.
Diaz, J.M.C., Delgado-Moreno, L., Nunez, R., Nogalesv, R., Romero, E., 2016. Enhancing
pesticide degradation using indigenous microorganisms isolated under high
pesticide load in bioremediation systems with vermicomposts. Bioresour. Technol.
214, 234–241.
Dvorak, P., Nikel, P.I., Damborsky, J., de Lorenzo, V., 2017. Bioremediation 3.0:
engineering pollutant-removing bacteria in the times of systemic biology.
Biotechnol. Adv. 35, 845–866.
Ellis, D.E., Lutz, E.J., Odom, J.M., Buchanan, J., Ronald, J., Bartlett, C.L., Lee, M.D.,
Harkness, M.R., Deweerd, K.A., 2000. Bioaugmentation for accelerated in situ
anaerobic biorremediation. Environ. Sci. Technol. 34, 2254–2260.
Fan, M.Y., Xie, R.J., Qin, G., 2014. Bioremediation of petroleumcontaminated soil by a
combined system of biostimulationbioaugmentation with yeast. Environ. Technol.
35, 391–399.
Farhadian, M., Vachelard, C., Duchez, D., Larroche, C., 2008. In situ bioremediation of
monoaromatic pollutants in groundwater: a review. Bioresour. Technol. 99,
5296–5308.
Gao, Y., Truong, Y.B., Cacioli, P., Butlen, P., Kyratzis, I.L., 2014. Bioremediation of
pesticide contaminated water using an organophosphate degrading enzyme
immobilized on nonwoven polyester textiles. Enzym. Microb. Technol. 54, 38–44.
Gentry, T.J., Rensing, C., Pepper, I.L., 2004. New approaches for bioaugmentation as a
remediation technology. Crit. Rev. Environ. Sci. Technol. 34, 447–494.
Goux, S., Shapir, N., E l, Fantroussi, S., Lelong, S., Agathos, S.N., Pussemier, L., 2013.
Long term maintenance of rapid atrazine degradation in soil inoculated with atrazine
degraders. Water Air Soil Pollut. Focus 3, 131–142.
Guo, J., Wang, J., Cui, D., Wangv, L., Ma, F., Chang, C., Yang, J., 2010. Application of
bioaugmentation in the rapid start-up and stable operation of biological processes
for municipal wastewater treatment at low temperatures. Bioresour. Technol. 101,
6622–6629.
Harelimana, V., Gao, Z.J., Nyiranteziryayo, E., Nwankwegu, A.S., 2020. Identification of
weaknesses in the implementation of environmental impact assessment regulations
in industrial sector: a case study of some industries in Rwanda, Africa. J. Clean. Prod.
258, 120677.
Haritash, A.K., Kaushik, C.P., 2009. Biodegradation aspects of polycyclic aromatic
hydrocarbons (PAHs): a review. J. Hazard Mater. 169, 1–15.
Hassanshahian, M., Emtiazi, G., Caruso, G., Cappello, S., 2014. Bioremediation
(bioaugmentation/biostimulation) trials of oil pollutedseawater: a mesocosm
simulation study. Mar. Environ. Res. 95, 28–38.
Hassanshahian, M., Bayat, Z., Cappello, S., Smedile, F., Yakimov, M., 2015. Comparison
the effects of bioaugmentation versusbiostimulation on marine microbial community
by PCR–DGGE: a mesocosm scale. J. Environ. Sci. 43, 136–146.
Herrero, M., Stuckey, D.C., 2015. Bioaugmentation and its application in wastewater
treatment: a review. Chemosphere 140, 119–128.
Hunt, R.W., Zavalin, A., Bhatnagar, A., Chinnasamy, S., Das, K.C., 2009. Electromagnetic
biostimulation of living cultures for biotechnology, biofuel and bioenergy
applications. Int. J. Mol. Sci. 10, 4515–4558.
Ikeda-Ohtsubo, W., Miyahara, M., Kim, S., Yamanda, T., Matsuoka, M., Watanabe, A.,
Fushinobu, S., Wakagi, T., Shoin, H., Miyauchi, K., Endo, G., 2013. Bioaugmentation
of a wastewater bioreactor system with the nitrous oxide-reducing denitrifier
Pseudomonas stutzeri strain TR2. J. Biosci. Bioeng. 115 (1), 37–42.
Isis, S.S., Elder, D.D., Andreia, F.D., Elisangela, F., Matthew, G., Lucia, R.D., 2009.
Bioremediation of a polyaromatic hydrocarbon contaminated soil by native soil
microbiota and bioaugmentation with isolated microbial consortia. Bioresour.
Technol. 100, 4669–4675.
Jacques, R.J.S., Okeke, B.C., Bento, F.M., Teixeira, A.S., Peralba, M.C.R., Comargo, F.A.
O., 2008. Microbial consortium bioaugmentation of a polycyclic aromatic
hydrocarbon contaminated soil. Bioresour. Technol. 99, 2637–2643.
Jansson, J.K., Elvang, A.M., Jorgensen, K.S., 2000. Biomarkers for monitoring efficacy of
bioremediation by microbial inoculants. Environ. Pollut. 107, 217–223.
Jasmine, J., Mukherji, S., 2014. Evaluation of bioaugmentation and biostimulation
effects on the treatment of refinery oily sludge using 2n full factorial design. Environ.
Sci. Process Impact. 16, 1889–1896.
Jian, M., Yongming, L., Ying, T., Zhengao, L., 2012. Bioremediation of polycyclic
aromatic hydrocarbon- contaminated soil by a bacterial consortium and associated
microbial community changes. Int. Biodeterior. Biodegrad. 70, 141–147.
Juhanson, J., Truu, J., Heinaru, E., Heinaru, A., 2009. Survival and catabolic
performance of introduced Pseudomonas strains during phytoremediation and
bioaugmentation field experiment. FEMS Microb. Ecol. 70, 446–455.
A.S. Nwankwegu et al.
Justo, O.R., Perez, V.H., Alvarez, V.C., Alegre, R.M., 2006. Growth of Escherichia coli
under extremely low- frequency electromagnetic fields. Appl. Biochem. Biotechnol.
134, 155–163.
Kempa, T., Marschalko, M., Yilmaz, I., Lacková, E., Kubečka, K., Stalmachová, B.,
Bouchal, T., Bednárik, M., Drusa, M., Bendová, M., 2013. In-situ remediation of the
contaminated soils in Ostrava city (Czech Republic) by steam curing/vapor. Eng.
Geol. 154, 42–55.
Kuppusamy, S., Thavamni, P., Venkateswarlu, K., Lee, Y.B., Naidu, R., Megharaj, M.,
2017. Remediation approaches for polycyclic aromatic hydrocarbons (PAHs)
contaminated soils: technological constraints, emerging trends and future directions.
Chemosphere 168, 944–968.
Lai, C.C., Huang, Y.C., Wei, Y.H., Chang, J.S., 2009. Biosurfactant-enhanced removal of
total petroleum hydrocarbons from contaminated soil. J. Hazard Mater. 167,
609–614.
Lamar, R.T., Davis, M.W., Dietrich, D.M., Glaser, J.A., 1994. Treatment of a
pentachlorophenol- and creosote-contaminated soil using the lignindegradating
fungus Phanerochaete sordida: a field demonstration. Soil Biol. Biochem. 26,
1603–1611.
Lebeau, T., Braud, A., Jezequel, K., 2008. Performance of bioaugmentation-assisted
phytoextraction applied to metal contaminated soils: a review. Environ. Pollut. 153,
497–522.
Lendvay, J.M., Loffler, F.E., Dollhopf, M., 2003. Bioreactive barriers: a comparison of
bioaugmentation and biostimulation for chlorinated solvent remediation. Environ.
Sci. Technol. 37, 1422–1431.
Liming, W., Xian, Q., Jun, J.Z., Dong, L.S., Ning, Y.Z., 2014. Bioaugmentation of a methyl
parathion contaminated soil with Pseudomonas sp. strain WBC-3. Int. Biodeterior.
Biodegrad. 87, 116–121.
Liu, X., Chen, Y., Zhang, X., Jiang, X., Wu, S., Shen, J., Sun, X., Li, J., Lu, L., Wang, L.,
2015. Aerobic granulation strategy for bioaugmentation of a sequencing batch
reactor (SBR) treating high strength pyridine wastewater. J. Hazard Mater. 295,
153–160.
Liu, B., Liu, J., Ju, M., Yu, Q., 2016. Purification and characterization of biosurfactant
produced by Bacillus licheniformis Y-1 and its application in remediation of
petroleum contaminated soil. Mar. Pollut. Bull. 107, 46–51.
Liu, S., Zeng, G., Niu, Q., Liu, Y., Zhou, L., Jiang, L., Tan, X., Xu, P., Zhang, C., Cheng, M.,
2017. Bioremediation mechanisms of combined pollution of PAHs and heavy metals
by bacteria and fungi: a mini review. Bioresour. Technol. 224, 25–33.
Liu, L., Bilal, M., Duan, X., Iqbal, H.M.N., 2019. Mitigation of environmental pollution by
genetically engineered bacteria — current challenges and future perspectives. Sci.
Total Environ. 667, 444–454.
Ma, H., Wei, M., Wang, Z., Hou, S., Li, X., Heng, X., 2020. Bioremediation of cadmium
polluted soil using a novel cadmium immobilizing plant growth promotion strain
Bacillus sp. TZ5 loaded on biochar. J. Hazard Mater. 122065 https://doi.org/
10.1016/j.jhazmat.2020.122065.
Mancera – Lopez, M.E., Esparza – Garcia, F., Chavez – Gomez, B., Rodriguez –
Vazquez, R., Saucedo – Castaneda, B., Cortes, J., 2008. Bioremediation of an aged
hydrocarbon soil by a combined system of biostimulation – bioaugmentation with
filamentous fungi. Int. Biodeterior. Biodegrad. 61, 151–160.
Manral, A., Bajpai, P.K., Ahmad, F., Joshi, R., 2021. Processing of sustainable
thermoplastic based biocomposites: a comprehensive review on performance
enhancement. J. Clean. Prod. 316, 128068.
Massa, V., Infantino, A., Radice, F., Orlandi, V., Tavecchio, F., Giudiuci, R., 2009.
Efficiency of natural and engineered bacterial strains in the degradation of 4-chlor­
obenzoic acid in soil slurry. Int. Biodeterior. Biodegrad. 63, 112–125.
Masy, T., Eche, S.D., Tromme, O., Thonart, P., Jacquess, P., Hiligsmann, S., Vogel, T.M.,
2016. Hydrocarbon biostimulation and bioaugmentation in organic carbon and clay-
rich soils. Soil Biol. Biochem. 99, 66–74.
Milena, C., Lucia, C., Silvana, B., Vincenza, A., 2011. Bioremediation of polyaromatic
hydrocarbon contaminated soils by native microflora and bioaugmentation with
Sphingobium chlorophenolicum strain C3R: a feasibility study in soil and slurry- -phase
microcosms. Int. Biodeterior. Biodegrad. 65, 191–197.
Mrozik, A.M., Piotrowska–Seget, Z., 2010. Bioaugmentation as a strategy for cleaning up
of soils contaminated with aromatic compounds. Microbiol. Res. 165, 363–375.
Nagata, Y., Endo, R., Ito, M., Ohtsubo, Y., Tsuda, M., 2007. Aerobic degradation of
lindane (γ- hexachlorocyclohexane) in bacteria and its biochemical and molecular
basis. Appl. Microbiol. Biotechnol. 76, 741.
Nagata, Y., Ohtsubo, Y., Tsuda, M., 2015. Properties and biotechnological applications of
natural and engineered haloalkane dehalogenases. Appl. Microbiol. Biotechnol. 99,
9865–9881.
Nikolopoulou, M., Eickenbusch, P., Pasadakis, N., Venieri, D., Kalogerakis, N., 2013a.
Microbial evaluation of autochthonous bioaugmentation to combat marine oil spills.
N. Biotech. 30, 734–742.
Nikolopoulou, M., Passadakis, N., Kalogerakis, N., 2013b. Evaluation of autochthonous
bioaugmentation and biostimulation during microcosm-simulated oil spills. Mar.
Pollut. Bull. 72, 165–173 (A&B).
Nitschke, M., Ferraz, C., Pastore, G., 2004. Selection of microorganisms for biosurfactant
production using agroindustrial wastes. Braz. J. Microbiol. 35, 81–85.
Nuamah, L.A., Li, Y., Pu, Y., Nwankwegu, A.S., Haikuo, Z., Norgbey, E., Banahene, P.,
Bofah-Buoh, R., 2020. Constructed wetlands, status, progress, and challenges. The
need for critical operational reassessment for a cleaner productive ecosystem.
J. Clean. Prod. 269, 122340.
Nwankwegu, A.S., 2016. Sawdust assisted bioremediation of PMS hydrocarbon impacted
agricultural soil in Niger Delta Nigeria. Int. J. Pharma Bio Sci. 7, 47–57.
Nwankwegu, A.S., Onwosi, C.O., 2017a. Microbial cell immobilization: a renaissance to
bioaugmentation inadequacies. A review. Environ. Technol. Rev. 6, 186–198.
11
Journal of Environmental Management 304 (2022) 114313
Nwankwegu, A.S., Onwosi, C.O., 2017b. Bioremediation of gasoline contaminated
agricultural soil by bioaugmentation. Environ. Technol. Innovat. 7, 1–11.
Nwankwegu, A.S., Onwosi, C.O., Orji, M.U., Anaukwu, C.G., Okafor, U.C., Azi, F.,
Martins, P.E., 2016a. Reclamation of DPK hydrocarbon polluted agricultural soil
using a selected bulking agent. J. Environ. Manag. 172, 136–142.
Nwankwegu, A.S., Orji, M.U., Onwosi, C.O., 2016b. Studies on organic and in-organic
biostimulants in bioremediation of diesel-contaminated arable soil. Chemosphere
162, 148–156.
Nwankwegu, A.S., Ikpe, E.M., Chukwura, E.I., Irondi, C.R., Obika, E.I., 2016c. Ex Situ
biodegradation of crude oil using bacterial isolates from palm oil mill effluent. Am.
J. Life Sci. 4, 71–75.
Nwankwegu, A.S., Onwosi, C.O., Azi, F., Azumini, P., Anaukwu, C.G., 2017. Use of rice
husk as bulking agent in bioremediation of automobile gas oil impinged agricultural
soil. Soil Sediment Contam. 26, 96–114.
Nwankwegu, A.S., Li, Y., Jiang, L., Lai, Q., Weng Jin, W., Acharya, K., 2018. Kinetic
modeling of total petroleum hydrocarbon in spent lubricating petroleum oil
impacted soil under different treatments. Environ. Technol. https://doi.org/
10.1080/09593330.2018.1498543.
Nwankwegu, A.S., Li, Y., Huang, Y., Wei, J., Norgbey, E., Sarpong, L., Lai, Q., Wang, K.,
2019. Harmful algal blooms under changing climate and constantly increasing
anthropogenic actions: the review of management implications. 3 Biotech 9, 1–19.
Nwankwegu, A.S., Li, Y., Huang, Y., Wei, J., Norgbey, E., Lai, Q., Sarpong, L., Wang, K.,
Ji, D., Yang, Z., Paerl, H.W., 2020a. Nutrient addition bioassay and phytoplankton
community structure monitored during autumn in Xiangxi Bay of Three Gorges
Reservoir, China, Chemosphere. https://doi.org/10.1016/j.chemosphere.2020
.125960.
Nwankwegu, A.S., Li, Y., Huang, Y., Wei, J., Norgbey, E., Ji, D., Pu, Y., Nuamah, L.A.,
Yang, Z., Jiang, Y., Paerl, H.W., 2020b. Nitrate repletion during spring bloom
intensifiesphytoplankton iron demand in Yangtze River tributary, China. Environ.
Pollut. https://doi.org/10.1016/j.envpol.2020.114626.
Odoh, C.K., Zabbey, N., Sam, K., 2019. Status , progress and challenges of
phytoremediation-An African scenario Status , progress and challenges of
phytoremediation - an African scenario. J. Environ. Manag. 237, 365–378.
Onwosi, C.O., Odibo, F.J.C., Enebechi, C.K., Nwankwegu, A.S., Ikele, A.I., Okeh, O.C.,
2017. Bioremediation of diesel-contaminated soil by composting with locally
generated bulking agents. Soil Sediment Contam. 26, 438–456.
Onwosi, C.O., Nwankwegu, A.S., Enebechi, C.K., Odimba, J.N., Nwuche, C.O.,
Igbokwe, V.C., 2018. Bioremediation of Soil Contaminated with Diesel Using
Inorganic Nitrogen Sources: Incorporating Nth- Order Algorithm in the Evaluation of
Process Kinetics. https://doi.org/10.1080/15320383.2018.1423023. Soil and
sediment contamination. 1 – 9.
Ozbayram, E.G., Kleinsteuber, S., Nikolausz, M., Ince, B., Ince, O., 2017. Effect of
bioaugmentation by cellulolytic bacteria enriched from sheep rumen on methane
production from wheat straw. Anaerobe 46, 122–130.
Paraszkiewicz, K., Bernat, P., Siewiera, P., Moryl, M., Paszt, L.S., Trzciński, P., Płaza, G.,
2017. Agricultural potential of rhizospheric Bacillus subtilis strains exhibiting varied
efficiency of surfactin production. Sci. Hortic. 225, 802–809.
Poi, G., Shahsavari, E., Aburto-Medina, A., Mok, P.C., Ball, A.S., 2018. Large scale
treatment of total petroleum- hydrocarbon contaminated groundwater using
bioaugmentation. J. Environ. Manag. 214, 157–163.
Poorsoleiman, M.S., Hosseini, S.A., Etminan, A., Abtahi, H., Koolivand, A., 2019. Effect
of two-step bioaugmentation of an indigenous bacterial strain isolated from oily
waste sludge on petroleum hydrocarbons biodegradation: scaling-up from a liquid
mineral medium to a two-stage composting process. Environ. Technol. Innovat.
100558 https://doi.org/10.1016/j.eti.2019.100558.
Rahman, K.S.M., Thahira-Rahman, J., Lakshmanaperumalsamy, P., Banat, I.M., 2003.
Towards efficient crude oil degradation by a mixed bacterium consortium.
Bioresour. Technol. 85, 257–261.
Rekadwad, B., Maske, V., Khobragade, C.N., Kasbe, P.S., 2019. Production and
evaluation of mono- and di- rhamnolipids produced by Pseudomonas aeruginosa
VM011. Data Brief 24, 103890. https://doi.org/10.1016/j.dib.2019.103890.
Rithuparna, R., Jittin, V., Bahurudeen, A., 2021. Influence of different processing
methods on the recycling potential of agro-waste ashes for sustainable cement
production: a review. J. Clean. Prod. 316, 128242.
Rodrigues, J.L.M., Kachael, A., Aiello, M.R., Quesen, J.F., Maltseva, O.V., Tsoi, T.V.,
2006. Degradation of aloclor 1242 dechlorinatuion products in sediments by
Burkholderiazenovorans LB400 (ohb) and Rhodococcus sp strain RHA1 (fcb). Appl.
Environ. Microbiol. 72, 2476–2482.
Rodrigues, E.M., Kalks, K.H.M., Totolav, M.R., 2015. Prospect, isolation, and
characterization of microorganisms for potential use in cases of oil bioremediation
along the coast of Trindade Island, Brazil. J. Environ. Manag. 156, 15–22.
Roy, A., Dutta, A., Pala, S., Guptab, A., Sarkar, J., Chatterjee, A., Saha, A., Sarkar, P.,
Sarb, P., Kazya, S., 2018. Biostimulation and bioaugmentation of native microbial
community accelerated bioremediation of oil refinery sludge. Bioresour. Technol.
2533, 22–32.
Sagarkar, S., Nousiainen, A., Shaligram, S., Bjorklof, K., Lindstrom, K., Jorgensen, K.S.,
Kapley, A., 2014. Soil mesocosm studies on atrazine bioremediation. J. Environ.
Manag. 139, 208–216.
Sam, K., Zabbey, N., 2018. Contaminated land and wetland remediation in Nigeria:
opportunities for sustainable livelihood creation. Sci. Total Environ. 639,
1560–1573.
Sermany, S., Favier, L., Djelal, H., Taha, S., Amrane, A., 2012. Bioaugmentation: possible
solution in the treatment of bio-refractory organic compounds (Bio-ROCs). Biochem.
Eng. J. 69, 75–86.
A.S. Nwankwegu et al.
Shi, J., Zhao, S., Yu, X., Zhou, T., Khan, A., Yu, Z., Li, X., 2018. Enhanced performance of
sediment microbial fuel cell by immobilization of Shewanella oneidensis MR-1 on an
anode surface. Int. J. Hydrogen Energy 44, 10091–10101.
Shi, K., Zhang, Q., Xue, J., Chen, X., Chen, Y., Qiao, Y., Sun, J., 2020. Study on the
degradation performance and bacterial community of bioaugmentation in
petroleum-pollution seawater. J. Environ. Chem. Eng., 103900 https://doi.org/
10.1016/j.jece.2020.103900.
Singer, A.C., Van der Gast, C.J., 2005. Thompson IP. Perspectives and vision for strain
selection in bioaugmentation. Trends Biotechnol. 23, 74–77.
Sivaperumal, P., Kamala, K., Rajara, R., 2017. Bioremediation of industrial waste
through enzyme producing marine microorganisms. Adv. Food Nutr. Res. 80,
165–179.
Sood, U., Singh, D.N., Hira, P., Lee, J.-K., Kalia, V.C., Lal, R., Shakarad, M., 2019. Rapid
and solitary production of mono-rhamnolipid biosurfactant and biofilm inhibiting
pyocyanin by a taxonomic outlier Pseudomonas aeruginosa strain CR1. J. Biotechnol.
https://doi.org/10.1016/j.jbiotec.2019.11.004.
Souza, K.S.T., Gudina, E.J., Azevedo, Z., de Freitas, V., Schwan, R.F., Rodrigues, L.R.,
Dias, D.R., Teixera, J.A., 2017. New glycolipid biosurfactants produced by the yeast
strain Wickerhamomyces anomalus CCMA 0358. Colloids Surf. B Biointerfaces 154,
373–382.
Steffan, R.J., Sperry, K.L., Walsh, T., Vainberg, S., Condee, C.W., 1999. Field-scale
evaluation of in situ bioaugmentation for remediation of chlorinated solvents in
groundwater. Environ. Sci. Technol. 33, 2771–2781.
Sublette, K.L., 2001. Fundamentals of Bioremediation of Hydrocarbon Contaminated
Soils. The University of Tulsa, Continuing Engineering and Science Education,
Houston,TX.
Suja, F., Fazil, R., Mohd, R.T., Nuraini, H., Rizal, M.R., Alia, K., Ainon, H., 2014. Effects
of local microbial bioaugmentation and biostimulation on the bioremediation of
total petroleum hydrocarbons (TPH) in crude oil contaminated soil based on
laboratory and field observations. Int. Biodeterior. Biodegrad. 90, 115–122.
Supaphol, S., Panichsakpatana, S., Trakulnaleamsai, S., Tungkananuruk, N.,
Roughjanajirapa, P., O’Donnell, A.G., 2006. The selection of mixed microbial
inocula in environmental biotechnology: example using petroleum contaminated
tropical soils. J. Microbiol. Methods 65, 432–441.
Tao, K., Zhang, X., Chen, X., Liu, X., Hu, X., Yuan, X., 2019. Response of soil bacterial
community to bioaugmentation with a plant residue-immobilized bacterial
consortium for crude oil removal. Chemosphere 222, 831–838.
Teng, P.Y., Luo, Y., Sun, M., Liu, Z., Christie, P., 2010. Effect of bioaugmentation by
Paracoccus sp. strain HPD-2 on the soil microbial community and removal of
polycyclic aromatic hydrocarbons from an aged contaminated soil. Bioresour.
Technol. 101, 3437–3443.
Thérien, M., Kiesewalter, H.T., Auria, E., Charron-Lamoureux, V., Wibowo, M.,
Maróti, G., Beauregard, P.B., 2020. Surfactin Production Is Not Essential for Pellicle
and Root-Associated Biofilm Development of Bacillus Subtilis. Biofilm, p. 100021.
https://doi.org/10.1016/j.bioflm.2020.100021.
U.S. EPA, March 2010. Green Technology. Green Remediation Best Management
Practices: Bioremediation. EPA 542-F-10-006.
Valipour, M., 2015. Variations of irrigated agriculture indicators in different continents.
Adv. Water Sci. Technol. 1, 1–14.
Varjani, S.J., Rana, D.P., Jain, A.K., Bateja, S., Upasani, V.N., 2015. Synergistic ex-situ
biodegradation of crude oil by halotolerant bacterial consortium of indigenous
strains isolated from on shore sites of Gujarat, India. Int. Biodeterior. Biodegrad.
103, 116–124.
Varjani, S.J., Gnansounu, E., Pandey, A., 2017. Comprehensive review on toxicity of
persistent organic pollutants from petroleum refinery waste and their degradation by
Microorganisms. Chemosphere 188, 280–291.
Varjani, S., Upasani, V.N., Pandey, A., 2020. Bioremediation of oily sludge polluted soil
employing a novel strain of Pseudomonas aeruginosa and phytotoxicity of petroleum
12
Journal of Environmental Management 304 (2022) 114313
hydrocarbons for seed germination. Sci. Total Environ. 139766. https://doi.org/
10.1016/j.scitotenv.2020.139766.
Wang, J., He, H., Wang, M., Wang, S., Zhang, J., Wei, W., Xu, H., Lv, Z., Shen, D., 2013.
Bioaugmentation of activated sludge with Acinetobacter sp. TW enhances nicotine
degradation in a synthetic tobacco wastewater treatment system. Bioresour. Technol.
142, 445–453.
Wang, L., Chi, X., Zhang, J., Sun, D., Zhou, N., 2014a. Bioaugmentation of a methyl
parathion contaminated soil with Pseudomonas sp. strain WBC-3. Int. J. Biodeter.
Biodegrad. 87, 116–121.
Wang, T., Sun, H., Jiang, C., Mao, H., Zhang, Y., 2014b. Immobilization of Cd in soil and
changes of soil microbial community by bioaugmentation of UV-mutated Bacillus
subtilis 38 assisted by biostimulation. Eur. J. Soil Biol. 65, 62–69.
Wei, M.J., Wang, H., Liu, C., Ning, D.L., 2008. Bioaugmentation with immobilized
genetically engineered microorganisms (GEM)/CAS process for treatment of atrazine
wastewater. Huanjing Kexue 6, 1555–1560.
Wittich, R.M., Wolff, P., 2007. Growth of the genetically engineered strain Cupriavidus
necator RW112 with chlorobenzoates and technical chlorobiphenyls. Microbiol. 153,
186–195.
Wu, M., Chen, L., Tian, Y., Ding, Y., Dick, W.A., 2013. Degradation of polycyclic aromatic
hydrocarbons by microbial consortia enriched from three soils using two different
culture media. Environ. Pollut. 178, 152–158.
Wu, M., Dick, W.A., Li, W., Wang, X., Yang, Q., Wang, T., 2016. Bioaugmentation and
biostimulation of hydrocarbon degradation and the microbial community in a
petroleum-contaminated soil. Int. Biodeterior. Biodegrad. 107, 158–164.
Xia, W., Dong, H.P., Yu, L., Yu, D., 2011. Comparative study of biosurfactant produced by
microorganisms isolated from formation water of petroleum reservoir. Colloids Surf.
A Physicochem. Eng. Asp. 392, 124–130.
Xiang, Q.C., Jun, J.Z., Shuo, Z., Ning, Y.Z., 2013. Bioaugmentation with a consortium of
bacterial nitrophenol- degraders for remediation of soil contaminated with three
nitrophenol isomers. Environ. Pollut. 172, 33–41.
Yi, L., Li, X.C., 2008. PAH-degrading microbial consortium and its pyrene-degrading
plasmids from mangrove sediment samples in Huian, China. Mar. Pollut. Bull. 57,
703–706.
Zabbey, N., Sam, K., Onyebuchi, A.T., 2017. Remediation of contaminated lands in the
Niger Delta, Nigeria: prospects and challenges. Sci. Total Environ. 586, 952–965.
Zafra, G., Absalon, A.E., Anducho-Reyes, M.A., Fernandez, F.J., Cortes-Espinosa, D.V.,
2017. Construction of PAH-degrading mixed microbial consortia by induced
selection in soil. Chemosphere 172, 120–126.
Zebra, A., Dubey, M.K., Meena, M., Aamir, M., Patel, C.B., Upadhyay, R.S., 2018. Role of
penicillium species in bioremediation processes. New Future Dev. Microb.
Biotechnol. Bioeng. 247–260.
Zhang, Z., Lo, I.M.C., Yan, D.Y.S., 2015. An integrated bioremediation process for
petroleum hydrocarbons removal and odor mitigation from contaminated marine
sediment. Water Res. 83, 21–30.
Zhao, D., Liu, C., Liu, L., Zhang, Y., Liu, Q., Wu, W.M., 2011. Selection of functional
consortium for crude oil contaminated soil remediation. Int. Biodeterior. Biodegrad.
65, 1244–1248.
Zhao, Q., Yue, S., Bilal, M., Hu, H., Wang, W., Zhang, X., 2017. Comparative genomic
analysis of 26 Sphingomonas and Sphingobium strains: dissemination of
bioremediation capabilities, biodegradation potential and horizontal gene transfer.
Sci.Total Environ. 609, 1238–1247.
Zheng, Y., Chai, L., 2016. Combined alkali precipitation and bioaugmentation process for
the treatment of black liquor under alkaline condition. Procedia Environ. Sci. 31,
715–724.
Zheng, H., Fan, S., Liu, W., Zhang, M., 2019. Production and separation of Pseudomonas
aeruginosa rhamnolipids using coupling technology of cyclic fermentation with foam
fractionation. Chem. Eng. Process 107776. https://doi.org/10.1016/j.
cep.2019.107776.