Informe de vigilancia biorremediación (2)

Biorremediación: aplicación de microorganismos externos

Índice del documento

Prefacio............................................................................................3

1. Introducción................................................................................5

2. Proyectos.....................................................................................6

3. Artículos.......................................................................................8

4. Patentes.....................................................................................45

5. Tesis...........................................................................................66

6. Oferta y demanda tecnológica.................................................70

7. Socios........................................................................................72

8. Otros recursos..........................................................................75

Índice de ilustraciones

1. Figura 1......................................................................................45

2. Figura 2......................................................................................46

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Prefacio
Este informe se elabora con el objetivo de presentar a Bioemprende un informe sobre el estado del arte
en biorremediación: aplicación de microorganismos externos

Las informaciones que se relacionan en este documento tienen carácter referencial. Si desea el
texto completo de cualquiera de ellas, puede solicitarlo al servicio de recuperación de
documentos: se dirija al contacto técnico que figura en la portada de este informe indicando el
título(*s) o referencia(*s) se se indican.

3

Definición del objeto de la búsqueda para la recuperación de
información:

● Búsqueda de artículos relacionados con el tema
● Proyectos que recogen las recientes innovaciones tanto de empresas como de centros
tecnológicos
● Patentes que pueden ser de aplicación en el tema de estudio

Para delimitar el objeto de la búsqueda de información, se definió el ámbito terminológico mediante la
extracción de los identificadores en lenguaje natural:

● [ES] biorremediación, biotratamiento, biorrecuperación, biodegradación, recuperación de suelos.

Los identificadores fueron complementados con la selección de tener controlados multilíngües, sinónimos
y complementarios, utilizando la herramienta IATE. Esta primera acotación resultó en la elección de los
siguientes términos:

● [EN] bioremediation, biotreatment, bioreclamation, biorecovery, biodegradation, soil recovery,
land reclamation

Resultando el análisis anterior en la cadena de interrogación (query) genérica:

bioremediation or bioreclamation

La query genérica fue adaptada a las características de la sintaxis y limitaciones en la busca de cada
fuente de información utilizada, dirigido a obtener los mejores resultados en cada caso.

Definición de las fuentes principales de información especializada:

La búsqueda se hizo en bases de datos especializadas como Web of Science perteneciente a Web of
Knowledge, Cordis entre otras.

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1. Introducción

El término biorremediación es empleado para referirse al conjunto de técnicas utilizadas para la
restauración de ambientes contaminados usando seres vivos.

Las técnicas de biorremediación se pueden clasificar en tres grandes tipos:

-Degradación enzimática
-Fitorremediación
-Remediación microbiana

1. Degradación enzimática

Consiste en el empleo de enzimas en el área contaminada con el objetivo de degradar las sustancias
contaminantes. Dichas enzimas son producidas a escala industrial utilizando a bacterias que las generan
naturalmente o por bacterias genéticamente modificadas

2. Fitorremediación

Se basa en la utilización de plantas en la zona contaminada aprovechando su capacidad para absorber o
acumular las sustancias nocivas.

3. Remediación microbiana

Uso sobre el área contaminada de microorganismos que degradan las sustancias contaminantes. Los
microorganismos utilizados en este proceso pueden estar presentes en el área contaminada o ser
exógenos a la misma.

En este informe nos vamos a centrar en las técnicas de remediación microbianas usando
microorganismos externos.

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2. Proyectos

El siguiente listado recoge tanto proyectos de ámbito nacional como europeo.

Dentro del ámbito europeo, estos proyectos pertenecen al Séptimo Programa Marco de Investigación y
Desarrollo tecnológico (7PM) que agrupa todas las iniciativas comunitarias en materia de investigación.
La búsqueda se hizo en Cordis que es el portal oficial del 7PM.

En el ámbito nacional hay tanto proyectos del Plan Nacional de I+D+i como del Plan Galego de I+D+i
(Incite 2006-2010).

Los proyectos se muestran ordenados alfabéticamente:

Descontaminación microbiológica de fosfoyesos contaminados por
uranio y cadmio
Organismo: Universidad de Granada
Proyectos de Investigación Fundamental no orientada 2010.

Desenvolvemento dun protocolo para a aplicación de técnicas de
descontaminación de solos "in situ" na Comunidade Galega

Solicitante: Novotec consultores, s.a.

INCITE- Programas sectoriais de investigación aplicada, PEME I+D e I+D Suma.Tecnoloxía sectorial:
Medio Natural e Desenvolvemento Sustentable (MDS). Convocatoria 2009

Mining the genome of p. fluorescens f113 to improve agricultural and
bioremediation applications. In silico genomics and functional genomics
approaches to exploit Pseudomonas in biotechnology (MIGENOF113)

Organización: UNIVERSIDAD AUTONOMA DE MADRID
7PM

http://cordis.europa.eu/fetch?
CALLER=FP7_PROJ_ES&ACTION=D&DOC=16&CAT=PROJ&QUERY=012cf3d3d603:7e24:571ae70a&
RCN=95871

6

Soil remediation technique for in situ cleaning of soils contaminated with
heavy hydrocarbons mixtures (SORBENT)

Organización: UAB GROTA
7PM

CALLER=FP7_PROJ_ES&ACTION=D&DOC=12&CAT=PROJ&QUERY=012cf3c40fd5:bcad:5031a325&R
CN=91811

UPSOIL-Sustainable Soil Upgrading by Developing Cost effective,
Biogeochemical Remediation Approaches

Organización: UNIDAD DE CONSTRUCCION DE LABEIN-TECNALIA
7PM

http://www.upsoil.eu/

Using MicroBes for the REgulation of heavy metaL mobiLity at ecosystem
and landscape scAle: An integrative approach for soil remediation by
geobiological processes (UMBRELLA)

Organización:FRIEDRICH-SCHILLER-UNIVERSITAET JENA
7PM

CALLER=FP7_PROJ_ES&ACTION=D&DOC=15&CAT=PROJ&QUERY=012cf3c40fd5:bcad:5031a325&R
CN=92568

UPSOIL-Sustainable Soil Upgrading by Developing Cost effective, Biogeochemical Remediation
Approaches

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3. Artículos
En la presente sección se recoge un listado de artículos de investigación publicados en los últimos
cuatro años. Los artículos están ordenados por año y alfabéticamente.

2011

Biodegradation of commercial gasoline (24% ethanol added) in liquid
medium by microorganisms isolated from a landfarming site
Autor: N.M. Oliveira, F. M. Bento, F. A. O. Camargo, A. J. Knorst, A. Loreiro Dos Santos, T. M.
Pizzolato, M. C. R. Peralba
Fuente: Journal of Enviromental Science and Health, Part A, Volume 46, Issue 1, Pages 86 - 96
Resumen:
Isolation of soil microorganisms from a landfarming site with a 19-year history of
petrochemical residues disposal was carried out. After isolation, the bacteria behavior in
mineral medium with 1% commercial gasoline (24% ethanol) was evaluated. Parameters
employed for microorganism evaluation and selection of those with the greatest
degradation potential were: microbial growth; biosurfactant generation and compound
reduction in commercial gasoline. Starting from bacteria that presented the best
degradation results, consortiums formed by 4 distinct microorganisms were formed. A
microbial growth in the presence of commercial gasoline was observed and, for most of
the bacteria, degradations of compounds such as benzene, toluene and xylenes (BTX)
as well as biosurfactant production was observed. Ethanol was partially degraded by the
bacterial isolates although the data does not allow affirming that it was degraded
preferentially to the aromatic hydrocarbons investigated. The analyzed consortiums
present an efficiency of 95% to 98% for most of the commercial gasoline compounds
and a preferential attack to ethanol under the essay condition was not observed within
72 h.
http://www.informaworld.com/smpp/content~content=a929784912~db=all~jumptype=rss

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2010

A salt tolerant Enterobacter cloacae mutant for bioaugmentation of
petroleum- and salt-contaminated soil
Autor: X. Hua, J. Wang, Z. Wu, H. Zhang, H. Li, X. Xing, Z. Liu
Fuente: Biochemical Engineering Journal, Volume 49, Issue 2, Pages 201-206
Resumen:
A NaCl-tolerant Enterobacter cloacae variant (MU-1) was obtained by mutagenesis
using atmospheric pressure glow discharge (APGD) plasmas. The variant exhibited
regular growth behavior in slurry cultivation and reached a cell density of 5.72 × 108 and
6.44 × 108 colony-forming units (CFU/mL) in the presence and absence of 7.5% NaCl,
respectively, when crude oil was used as the sole carbon source (crude oil/soil = 1.5%).
The total petroleum hydrocarbon (TPH) degradation percentage was 7.94% with mutant
MU-1 in the presence of 7.5% NaCl whereas that of the wild-type strain was 3.17%.
When cultivated in saline medium, MU-1 showed a slight change in membrane
permeability but significant increases in both the K+ concentration inside the cell
membrane (from 234.24 to 1422.88 ppm/g dry cell weight in the first 2 h) and the
exopolysaccharide (EPS) level outside the membrane (from 1350 to 1825 mg/g dry cell
weight). The rapid increase in K+ inside the cell and the simultaneous accumulation of
EPS outside the cell may be responsible for maintaining the osmotic balance during
saline cultivation, and this could facilitate the microbial growth and TPH degradation of
MU-1.
http://www.sciencedirect.com/science?_ob=ArticleURL&_udi=B6V5N-4Y3TXG7-
B&_user=10&_coverDate=04%2F15%2F2010&_rdoc=1&_fmt=high&_orig=search&_origin=searc
h&_sort=d&_docanchor=&view=c&_searchStrId=1585656496&_rerunOrigin=google&_acct=C000
050221&_version=1&_urlVersion=0&_userid=10&md5=103ca0ec5b754b16cc2c94f8c8326099&s
earchtype=a

Bio-Detoxification of Arsenic Laden Ground Water Through a Packed Bed
Column of a Continuous Flow Reactor Using Immobilized Cells
Autor: P. Bag, P. Bhattacharya, R. Chowdhury
Fuente: Soil and Sediment Contamination: An International Journal, Volume 19, Issue 4 , Pages
455 - 466
Resumen:
Under the present study arsenic resistant bacterial strain, Rhodococcus equi
(JUBTAs02), has been used to remove trivalent arsenic from a simulated aqueous
solution of arsenic oxide (As2O3). Batch studies have been conducted to determine the
arsenic-intoxicated growth kinetics of the bacteria. The Monod type kinetic parameters
like saturation constant KS and maximum specific growth rate μmax have been
determined by studying batch mode of growth of microorganisms varying the initial
concentration of limiting substrate, i.e. citrate (carbon source), in absence of arsenic
ions. The kinetic parameter for intoxicated growth, namely the inhibition constant, Ki, has
been determined by varying As3+ concentration for each batch conducted at different

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initial concentrations of citrate. For ground water a first order kinetics with respect to
arsenic concentration has been determined for arsenic uptake rate.

The same microorganism has been used in immobilized form to treat simulated water as
well as naturally occurring arsenic laden ground water in a continuous packed bed
reactor using initial arsenite concentration and inlet flow rate as parameters. A maximum
value of arsenite removal efficiency of 95% has been achieved in this process.
Deterministic mathematical models capable of explaining the trend of removal of arsenic
from simulated and ground water have been developed using the kinetic parameters of
intrinsic growth of the microorganism. The simulated results have been compared with
the experimental ones satisfactorily.

Bioaugmentation efficiency of diesel degradation by Bacillus pumilus
JL(B) and Acinetobacter calcoaceticus LT1 in contaminated soils
Autor: C. Singh, J. Lin
Fuente: African Journal of Biotechnology , Volume 94, Issue 41, Pages 6881-6888
Resumen:
The abilities of diesel-degrading Bacillus pumilus JLB and Acinetobacter calcoaceticus
LT1 were tested in contaminated soils. The effect of nutrient supplementation on
bioaugmented samples was also examined. The results show that bio-augmentation and
biostimulation accelerated significantly (p < 0.05) the diesel degradation in the
contaminated loam soil and sea sand. Supplementing fertilizers to the augmented loam
samples did not result in a significantly higher degradation rate. Furthermore, A.
calcoaceticus LT1 alone failed to stimulate higher degradation rates in sea sand unless
further supplementation of fertilizer. The results from environmental scanning electron
microscopy demonstrate the population increases, then decreases in augmented
samples corresponding to the level of diesel degradation. Fungi-like microorganisms
became dominant in contaminated loam soil at the end of the study but not in sea sand.
The study shows that it is critical not only to understand the physiology of the inoculum
but also how it affects microbial community structure and function before the
microorganism being introduced in the contaminated soil.
http://www.academicjournals.org/AJB/PDF/pdf2010/11Oct/Singh%20and%20Lin.pdf

Biodegradation Potential and Influencing Factors of a Special
Microorganism to Treat Petrochemical Wastewater
Autor: Q. Zhou, B. Shen
Fuente: Petroleum Science and Technology, Volume 28, Issue 2, Pages 135 - 145
Resumen:
The objective of this study is to evaluate the potential ability of an active microorganism
on the biodegradation of petroleum hydrocarbons in the petrochemical wastewater from
Sinopec Shanghai Petroleum and Chemical Company Limited, China. In lab-scale batch
experiments, by using this special functional microorganism to treat the pollutant, the

10

wastewater was purified as one of the applications of the bioaugmentation technique.
Results from this study showed that the biodegradation was quicker and more effective
than the natural and physical degradation. The wastewater was analyzed by gas
chromatography/mass spectrometry (GC-MS), and it indicated that the dominant
pollutants of the wastewater were petroleum-based normal-alkanes (C15-C30). The
concentration of microorganism, aeration time, and temperature of biodegradation all
influence the potential biodegradation ability. Meanwhile, the biodegradation capability to
biodegrade hydrocarbons by this microorganism is the average removal of total
petroleum hydrocarbon and was approximately 85% with chemical oxygen demand
about 65%. This study provides a feasible technology for the treatment of hydrocarbon-
rich wastewater from petrochemical industries and petroleum refineries.

Bioremediation of crude oil-contaminated soil: Comparison of different
biostimulation and bioaugmentation treatments
Autor: Y. Xu, M. Lu
Fuente: Journal of Hazardous Materials, Volume 183, Issues 3-5, Pages 395-401
Resumen:
Biostimulation with inorganic fertilizer and bioaugmentation with hydrocarbon utilizing
indigenous bacteria were employed as remedial options for 12 weeks in a crude oil-
contaminated soil. To promote oil removal, biocarrier for immobilization of indigenous
hydrocarbon-degrading bacteria was developed using peanut hull powder.
Biodegradation was enhanced with free-living bacterial culture and biocarrier with a total
petroleum hydrocarbon removal ranging from 26% to 61% after a 12-week treatment. Oil
removal was also enhanced when peanut hull powder was only used as a bulking agent,
which accelerated the mass transfer rate of water, oxygen, nutrients and hydrocarbons,
and provided nutrition for the microflora. Dehydrogenase activity in soil was remarkably
enhanced by the application of carrier material. Metabolites of polycyclic aromatic
hydrocarbons were identified by Fourier transform ion cyclotron resonance mass
spectrometry.
http://www.sciencedirect.com/science?_ob=ArticleURL&_udi=B6TGF-50J4MM4-
C&_user=10&_coverDate=11%2F15%2F2010&_rdoc=1&_fmt=high&_orig=search&_origin=searc
h&_sort=d&_docanchor=&view=c&_acct=C000050221&_version=1&_urlVersion=0&_userid=10&
md5=146c0f65b353ca6deea4eeb7b9cc1391&searchtype=a

Bioremediation of glyphosate-contaminated soils
Autor: I. T. Ernakova, N. I. Kiseleva, T. Shushkova, M. Zharikov, G. A. Zharikov, A. A.
Leontievsky
Fuente: Applied Microbiology and Biotechnology, Volume 88, Issue 2, Pages 585-594
Resumen:
Based on the results of laboratory and field experiments, we performed a
comprehensive assessment of the bioremediation efficiency of glyphosate-contaminated
soddy-podzol soil. The selected bacterial strains Achromobacter sp. Kg 16 (VKM B-

11

2534D) and Ochrobactrum anthropi GPK 3 (VKM B-2554D) were used for the aerobic
degradation of glyphosate. They demonstrated high viability in soil with the tenfold
higher content of glyphosate than the recommended dose for the single in situ treatment
of weeds. The strains provided a two- to threefold higher rate of glyphosate degradation
as compared to indigenous soil microbial community. Within 1-2 weeks after the strain
introduction, the glyphosate content of the treated soil decreased and integral toxicity
and phytotoxicity diminished to values of non-contaminated soil. The decrease in the
glyphosate content restored soil biological activity, as is evident from a more than
twofold increase in the dehydrogenase activity of indigenous soil microorganisms and
their biomass (1.2-fold and 1.6-fold for saprotrophic bacteria and fungi, respectively).
The glyphosate-degrading strains used in this study are not pathogenic for mammals
and do not exhibit integral toxicity and phytotoxicity. Therefore, these strains are suitable
for the efficient, ecologically safe, and rapid bioremediation of glyphosate-contaminated
soils.
http://www.ncbi.nlm.nih.gov/pubmed/20676632

Bioremediation of heavy metals by growing hyperaccumulaor endophytic
bacterium Bacillus sp. L14
Autor: H. Guo, S. Luo, L. Chen, X. Xiao, Q. Xi, W. Wei, G. Zeng, C. Liu, Y. Wan, J. Chen, Y. He
Fuente: Bioresource Technology, Volume 101, Issue 22, Pages 8599-8605
Resumen:
Heavy metal bioremediation by a multi-metal resistant endophytic bacteria L14 (EB L14)
isolated from the cadmium hyperaccumulator Solanum nigrum L. was characterized for
its potential application in metal treatment. 16S rDNA analysis revealed that this
endophyte belonged to Bacillus sp. The hormesis of EB L14 were observed in presence
of divalent heavy metals (Cu (II), Cd (II) and Pb (II)) at a relatively lower concentration
(10 mg/L). Such hormesis was the side effect of abnormal activities increases of ATPase
which was planned to provide energy to help EB L14 reduce the toxicity of heavy metals
by exporting the cations. Within 24 h incubation, EB L14 could specifically uptake
75.78%, 80.48%, 21.25% of Cd (II), Pb (II) and Cu (II) under the initial concentration of
10 mg/L. However, nearly no chromium uptake was observed. The mechanism study
indicated that its remediation efficiencies may be greatly promoted through inhibiting the
activities of ATPase. The excellent adaptation abilities and promising remediation
efficiencies strongly indicated the superiority of this endophyte in heavy metal
bioremediation at low concentrations, which could be useful for developing efficient
metal removal system.
http://www.sciencedirect.com/science?_ob=ArticleURL&_udi=B6V24-50H224F-
G&_user=10&_coverDate=11%2F30%2F2010&_rdoc=1&_fmt=high&_orig=search&_origin=searc
md5=18a77047bd6b539bb4a9170837991c82&searchtype=a

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Bioremediation of marine sediments contaminated by hydrocarbons:
Experimental analysis and kinetic modeling
Autor: F. Beolchini, L. Rocchetti, F. Regoli, A. Dell’Anno
Resumen:
This work deals with bioremediation experiments on harbor sediments contaminated by
aliphatic and polycyclic aromatic hydrocarbons (PAHs), investigating the effects of a
continuous supply of inorganic nutrients and sand amendments on the kinetics of
microbial growth and hydrocarbon degradation. Inorganic nutrients stimulated microbial
growth and enhanced the biodegradation of low and high molecular weight
hydrocarbons, whereas sand amendment increased only the removal of high molecular
weight compounds. The simultaneous addition of inorganic nutrients and sand provided
the highest biodegradation (>70% for aliphatic hydrocarbons and 40% for PAHs). A
semi-empirical kinetic model was successfully fitted to experimental temporal changes of
hydrocarbon residual concentrations and microbial abundances. The estimated values
for parameters allowed to calculate a doubling time of 2.9 d and a yield coefficient
biomass/hydrocarbons 0.39 g C biomass g-1C hydrocarbons, for the treatment with the
highest hydrocarbon biodegradation yield. A comparison between the organic carbon
demand and temporal profiles of hydrocarbons residual concentration allowed also to
calculate the relative contribution of contaminants to carbon supply, in the range 5–32%.
This suggests that C availability in the sediments, influencing prokaryotic metabolism,
may have cascade effects on biodegradation rates of hydrocarbons. Even if these
findings do not represent a general rule and site-specific studies are needed, the
approach used here can be a relevant support tool when designing bioremediation
strategies on site.
http://www.sciencedirect.com/science?_ob=ArticleURL&_udi=B6TGF-50BJNV7-
1&_user=10&_coverDate=10%2F15%2F2010&_rdoc=1&_fmt=high&_orig=search&_origin=searc
md5=73239833b47a07920be394cf93e4e227&searchtype=a

Bioremediation of polyaromatic hydrocarbon contaminated soils by native
microflora and bioaugmentation with Sphingobium chlorophenolicum
strain C3R: A feasibility study in solid- and slurry-phase microcosms
Autor: M. Colombo, L. Cavalca, S. Bernasconi, V. Andreoni
Fuente: International Biodeterioration & Biodegradation, Article in Press
Resumen:
The aim of the research was to verify if a Sphingobium chlorophenolicum strain C3R
was effective in the degradation of phenanthrene (Ph) in agricultural soil co-
contaminated by metals and mixtures of PAHs. The presence of PAHs in mixtures
produced interactive effects which could either increase or decrease the utilization rate
of Ph by C3R and by the native bacterial microflora. Bioaugmentation significantly
improved the biodegradation rate of Ph in the presence of both cadmium and arsenic
and PAH mixtures. The augmented C3R strain persisted in inoculated microcosms as
monitored by the DGGE analysis and outcompeted some indigenous bacteria. The
potential role of the soil bacteria in PAH degradation could be envisaged. The results
indicate the applicability of S. chlorophenolicum C3R toward in situ bioremediation of

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sites contaminated with phenanthrene alone or co-contaminated with low molecular
weight PAHs and with cadmium and arsenate.
http://www.sciencedirect.com/science?_ob=ArticleURL&_udi=B6VG6-51N7C7C-
md5=bb97041431bb3d13217984c403a3c2f8&searchtype=a

Bioremediation of wastewaters with recalcitrant organic compounds and
metals by aerobic granules
Autor: A.M. Maszenan, Y. Liu, W. J. Ng
Fuente: Biotechnology Advances, Volume 29, Issue 1, Pages 111-123
Resumen:
Compared to activated sludge flocs, aerobic granules have a regular shape, and a
compact and dense structure which enhances settleability, higher biomass retention,
multi-microbial functions, higher tolerance to toxicity, greater tolerance to shock loading,
and relatively low excess sludge production. The potential for improved process
efficiency and cost-effectiveness can be attractive when it is applied to both municipal
and industrial wastewaters. This review discusses potential applications of aerobic
granulation technology in wastewater treatment while drawing attention to relevant
findings such as diffusion gradients existing in aerobic granules which help the biomass
cope with inhibitory compounds and the ability of granules to continue degradation of
inhibitory compounds at extreme acid and alkaline pHs.
http://www.sciencedirect.com/science?_ob=ArticleURL&_udi=B6T4X-516MRBY-
md5=b7923f55b3a59275219a677d86acbf72&searchtype=a

BTEX biodegradation by bacteria from effluents of petroleum refinery
Autor: D. E. Mazzeo, C. E. Levy, D. de F. de Angelis, M. A. Marin-Morales
Fuente: The Science of the Total Environment, Volume 408, Issue 20, Pages 4334-4340
Resumen:
Groundwater contamination with benzene, toluene, ethylbenzene and xylene (BTEX)
has been increasing, thus requiring an urgent development of methodologies that are
able to remove or minimize the damages these compounds can cause to the
environment. The biodegradation process using microorganisms has been regarded as
an efficient technology to treat places contaminated with hydrocarbons, since they are
able to biotransform and/or biodegrade target pollutants. To prove the efficiency of this
process, besides chemical analysis, the use of biological assessments has been
indicated. This work identified and selected BTEX-biodegrading microorganisms present
in effluents from petroleum refinery, and evaluated the efficiency of microorganism
biodegradation process for reducing genotoxic and mutagenic BTEX damage through
two test-systems: Allium cepa and hepatoma tissue culture (HTC) cells. Five different
non-biodegraded BTEX concentrations were evaluated in relation to biodegraded
concentrations. The biodegradation process was performed in a BOD Trak Apparatus

14

(HACH) for 20 days, using microorganisms pre-selected through enrichment. Although
the biodegradation usually occurs by a consortium of different microorganisms, the
consortium in this study was composed exclusively of five bacteria species and the
bacteria Pseudomonas putida was held responsible for the BTEX biodegradation. The
chemical analyses showed that BTEX was reduced in the biodegraded concentrations.
The results obtained with genotoxicity assays, carried out with both A. cepa and HTC
cells, showed that the biodegradation process was able to decrease the genotoxic
damages of BTEX. By mutagenic tests, we observed a decrease in damage only to the
A. cepa organism. Although no decrease in mutagenicity was observed for HTC cells, no
increase of this effect after the biodegradation process was observed either. The
application of pre-selected bacteria in biodegradation processes can represent a reliable
and effective tool in the treatment of water contaminated with BTEX mixture. Therefore,
the raw petroleum refinery effluent might be a source of hydrocarbon-biodegrading
microorganisms.

Characterization of a bacterial strain capable of degrading DDT congeners
and its use in bioremediation of contaminated soil
Autor: H. Fang, B. Dong, H. Yan, F. Tang, Y. Yu
Resumen:
A bacterial strain DDT-6 (D6) capable of utilizing dichlorodiphenyltrichloroethane (DDT),
dichlorodiphenyldichloroethane (DDD), and dichlorodiphenyldichloroethylene (DDE)
(DDTs) as its sole carbon and energy source was isolated and identified as
Sphingobacterium sp. The degradation of DDTs by strain D6 in mineral salt medium and
in field soil was investigated. The half-lives of the degradation of DDTs increased with
increasing concentration ranging from 1 to 50 mg L−1. Favorable degradation conditions
for DDTs by strain D6 were found to be pH 7.0 and 30 °C. The degradation of DDTs by
strain D6 was found to be statistically significantly enhanced (p ≤ 0.05) by the addition of
glucose. Based on the metabolites detected, a pathway was proposed for DDT
degradation in which it undergoes dechlorination, hydrogenation, dioxygenation,
decarboxylation, hydroxylation, and phenyl ring-cleavage reactions to complete the
mineralization process. The addition of strain D6 into the contaminated soils was found
to statistically significantly enhance (p ≤ 0.05) the degradation of DDTs. The results
indicate that the isolate D6 can be used successfully for the removal or detoxification of
residues of DDTs in contaminated soil.
http://www.sciencedirect.com/science?_ob=ArticleURL&_udi=B6TGF-50T41PS-
md5=4e0ae670c69f4a8e2b85c43782ad48d1&searchtype=a

15

Characterization of a fenpropathrin-degrading strain and construction of a
genetically engineered microorganism for simultaneous degradation of
methyl parathion and fenpropathrin
Autor: Y. Hong, J. Zhou, Q. Hong, Q. Wang, J. Jiang, S. Li
Fuente: Journal of Environmental Management, Volume 91, Issue 11, Pages 2295-2300
Resumen:
A gram-negative fenpropathrin-degrading bacterial strain Sphingobium sp. JQL4-5 was
isolated from the wastewater treatment sludge of an insecticide factory. Strain JQL4-5
showed the ability to degrade other pyrethroid insecticides, but it was not able to
degrade methyl parathion. To enhance its degrading range of substrate, a methyl
parathion hydrolase gene (mpd) was successfully introduced into the chromosome of
strain JQL4-5 with a mini-Tn-transposon system. A genetically engineered
microorganism (GEM) named JQL4-5-mpd resulted, which was capable of
simultaneously degrading methyl parathion and fenpropathrin. Soil treatment results
indicated that JQL4-5-mpd is a promising multifunctional bacterium in the bioremediation
of multiple pesticide-contaminated environments.

Comparative bioremediation potential of four rhizospheric microbial
species against lindanestar
Autor: P.C. Abhilash, S. Srivastava, N. Sing
Fuente: Chemosphere, Volume 82, Issue 1, Pages 56-63
Resumen:
Four microbial species (Kocuria rhizophila, Microbacterium resistens, Staphylococcus
equorum and Staphylococcus cohnii subspecies urealyticus) were isolated from the
rhizospheric zone of selected plants growing in a lindane contaminated environment and
acclimatized in lindane spiked media (5–100 μg mL−1). The isolated species were
inoculated with soil containing 5, 50 and 100 mg kg−1 of lindane and incubated at room
temperature. Soil samples were collected periodically to evaluate the microbial
dissipation kinetics, dissipation rate, residual lindane concentration and microbial
biomass carbon (MBC). There was a marked difference (p < 0.05) in the MBC content
and lindane dissipation rate of microbial isolates cultured in three different lindane
concentrations. Further, the dissipation rate tended to decrease with increasing lindane
concentrations. After 45 d, the residual lindane concentrations in three different spiked
soils were reduced to 0%, 41% and 33%, respectively. Among the four species, S.
cohnii subspecies urealyticus exhibited maximum dissipation (41.65 mg kg−1) and can
be exploited for the in situ remediation of low to medium level lindane contaminated
soils.
http://www.sciencedirect.com/science?_ob=ArticleURL&_udi=B6V74-51C9PG1-
md5=9ff326e65e53ac16b2f169e08f167b04&searchtype=a

16

Construction of a genetically engineered microorganism with high
tolerance to arsenite and strong arsenite oxidative ability
Autor: C. Yang, l. Xu, L. Yan, Y. Xu
Fuente: Journal of Environmental Science and Health, part A: Toxic/hazardous substances and
environmental engineering, Voluem 45, Issue 6, Pages 732-737
Resumen:
Genetically engineered microorganisms (GEMs) have shown great potential for use in
environmental bioremediation. In this study, the TTHB128 and TTHB127 genes, which
encode the small and large subunits of arsentie oxidase in Thermus thermophilus HB8,
respectively, were cloned into the broad-host-range vector pBBR1MCS-5 to produce the
recombinant plasmid, TTHB127-pBBR1MCS-5-TTHB128. This resulted in successful
construction of a GEM with high tolerance to arsenite and strong arsenite oxidative
ability. Culture of the GEM in media containing arsenite for 28 h resulted in 87.6% of the
arsenite being oxidized. Overall, the oxidative ability of the GEM was much stronger than
that of the wild type host strain. Gentamicin was necessary to maintain the stability of the
recombinant plasmid, TTHB127-pBBR1MCS-5-TTHB128, in the GEM. The oxidative
ability of the GEM remained unchanged when it was grown in medium containing
gentamicin (60 mg/L) for 30 growth cycles, after which its activity gradually decreased.

Evolution of bacterial community during bioremediation of PAHs in a coal
tar contaminated soil
Autor: C. Lors, A. Ryngaert, F. Périé, L. Diels, D. Damidot
Fuente: Chemosphere, Voluem 81, Issue 10, Pages 1263-1271
Resumen:
The monitoring of a windrow treatment applied to soil contaminated by mostly 2-, 3- and
4-ring PAHs produced by coal tar distillation was performed by following the evolution of
both PAH concentration and the bacterial community. Total and PAH-degrading bacterial
community structures were followed by 16S rRNA PCR–DGGE in parallel with
quantification by bacterial counts and 16 PAH measurements. Six months of biological
treatment led to a strong decrease in 2-, 3- and 4-ring PAH concentrations (98, 97 and
82% respectively). This result was associated with the activity of bacterial PAH-
degraders belonging mainly to the Gamma-proteobacteria, in particular, the
Enterobacteria and Pseudomonas genera, which were detected over the course of the
treatment. This group was considered to be a good bioindicator to determine the
potential PAH biodegradation of contaminated soil. Conversely, other species, like the
Beta-proteobacteria, were detected after 3 months, when 2-, 3- and 4-ring PAHs were
almost completely degraded. Thus, presence of the Beta-proteobacteria group could be
considered a good candidate indicator to estimate the endpoint of biotreatment of this
type of PAH-contaminated soil.
http://www.sciencedirect.com/science?_ob=ArticleURL&_udi=B6V74-5172K72-
md5=28791308a39b846e7d8cc2038399f468&searchtype=a

17

Genetically engineered oil-eating microbes for bioremediation: Prospects
and regulatory challenges
Autor: O. C. Ezezika, P. A. Singer
Fuente: Technology in Society, Volume 32, Issue 4, Pages 331-335
Resumen:
The use of genetic engineering to enhance the natural capacity of microorganisms for
remediation has become very promising with new scientific discoveries occurring every
year. Unfortunately, the application and commercialization of this technology has not
kept pace with these research discoveries. This article uses two examples of genetically
engineered microorganisms that were designed but never deployed in the clean-up of
wastes to show how the application of genetically engineered microbes for
bioremediation has not progressed in line with other biotechnological innovations. We
argue that a more risk-based regulatory environment that fosters commercialization is
important. In addition, we show how scientists could foster the commercialization of
genetically engineered microbes for bioremediation through the use of technical
safeguards and the consideration of regulatory challenges at the onset of their research.
The lessons provided by these challenges could be applicable to current
biotechnological innovations that face similar regulatory challenges.
http://www.sciencedirect.com/science?_ob=ArticleURL&_udi=B6V80-51H5HDD-
md5=3c38a518c6e17cf9ac055b439212b564&searchtype=a

Isolation of pesticide-degrading actinomycetes from a contaminated site:
Bacterial growth, removal and dechlorination of organochlorine pesticides
Autor: M.S. Fuentes, C.S. Benimeli, S.A. Cuozzo, M.J. Amoroso
Fuente. International Biodeterioration & Biodegradation, Volume 64, Issue 6, Pages 434-441
Resumen:
Organochlorine pesticides are notorious, due to their high toxicity, persistence in the
environment and their tendency to bioaccumulate. Their extensive use in the northwest
of Argentina has left residues in the environment.
Microbial degradation is an important process for pesticide bioremediation and
actinomycetes have a great potential for that.
The current study examined organochlorine pesticides in contaminated soil. Indigenous
actinomycetes were isolated from contaminated samples to evaluate bacterial growth as
well as pesticide removal and release of chloride ions as a result of degradation.
Most of the isolated microorganisms belonged to the Streptomyces genus, except one,
which belonged to Micromonospora. Bacterial growth depended on the microorganism
and the pesticide present (chlordane, lindane or methoxychlor). Highest growth and
pesticide removal were observed with chlordane. Twelve out of 18 studied strains
released chloride into culture supernatants, and percentages were higher with chlordane
as carbon source than with lindane or methoxychlor. These results are supported by
principal component analysis.

18

This is the first report about actinomycetes isolated from an illegal storage of
organochlorine pesticide in Argentina with capacity to growth, remove and use different
organochlorine pesticide.
http://www.sciencedirect.com/science?_ob=ArticleURL&_udi=B6VG6-50819MG-
050221&_version=1&_urlVersion=0&_userid=10&md5=14654fe09c06cb45f679cadbde7245a0&s
earchtype=aç

Mercury-resistant bacterial strains Pseudomonas and Psychrobacter spp.
isolated from sediments of Orbetello Lagoon (Italy) and their possible use
in bioremediation processes
Autor: M. Pepi , C. Gaggi, E. Bernardini, S. Focardi, A. Lobianco, M. Ruta, V. Nicolardi, M.
Volterrani, S. Gasperini, G. Trinchera, P. Renzi, M. Gabellini, S. E. Focardi
Fuente: International Biodeterioration & Biodegradation, Article in Press
Resumen:
This study was aimed to isolate Hg-resistant bacteria from contaminated sediments of
the Orbetello Lagoon in Italy and to assess their possible use as biofilms in
bioremediation processes. Enrichment cultures prepared from contaminated sediments
in the presence of 0.05 mM of mercury and under aerobic conditions allowed the
isolation of five heterotrophic bacterial strains. 16S rDNA gene sequencing assigned the
isolated strains to the genera Pseudomonas and Psychrobacter. For the first time
mercury-resistant bacterial strains belonging to the genus Psychrobacter were
evidenced. Minimum inhibitory concentrations in the presence of HgCl2 and of CH3HgCl
showed high levels of resistance. EC50 values for the isolated bacterial strains in the
presence of HgCl2 and of CH3HgCl confirmed the adaptation to the metal. Hg-resistant
strains ORHg1, ORHg4 and ORHg5 showed the capacity to volatilize inorganic and
organic mercury to elemental mercury, and formed biofilms on pumice particles, and
were shown to play a role in the removal of mercury from sediment leachates. This study
reports isolation and characterization of new Hg-resistant bacterial strains and provides
novel insight into their possible use in bioremediation processes of mercury polluted
sediments.
http://www.sciencedirect.com/science?_ob=ArticleURL&_udi=B6VG6-51B8G56-
md5=d0b5dfe665186b825d77b1e95979c23a&searchtype=a

19

Microbial communities involved in the bioremediation of an aged
recalcitrant hydrocarbon polluted soil by using organic amendments
Autor: M. Ros, I. Rodríguez, C. García, T. Hernández
Resumen:
An 8-month field bioremediation experiment using fresh (FS) and composted (CS)
sewage sludge and unamended soil (US) was carried out on an aged hydrocarbon
contaminated semi-arid soil. FS treatments led to the highest percentage of hydrocarbon
degradation (46%) and the highest bacterial and fungal population. Denaturing gradient
gel electrophoresis analysis demonstrated differences in bacterial and fungal community
structure of treated compared to uncontaminated soil (control). Time of sampling
accounted for most of the differences than type of treatment. The principal phyla
observed in bioremediation treatments were Actinobacteria and Ascomycota. Results
pointed to the addition of organic amendments, particularly sewage sludge, as an useful
strategy for improving the effectiveness of landfarming biodegradation processes in
hydrocarbon polluted soils.
http://www.sciencedirect.com/science?_ob=ArticleURL&_udi=B6V24-4YX0BKB-
md5=68e98f4defbfc757b6dbf23bcf3dfc02&searchtype=a

Microbial degradation of tetrachloromethane: mechanisms and
perspectives for bioremediation
Autor: C. Penny, S. Vuilleumier, F. Bringel
Fuente: FEMS Microbiology Ecology, Volume 74, Issue 2, Pages 257-275
Resumen:
Toxic man-made compounds released into the environment represent potential nutrients
for bacteria, and microorganisms growing with such compounds as carbon and energy
sources can be used to clean up polluted sites. However, in some instances,
microorganisms contribute to contaminant degradation without any apparent benefit for
themselves. Such cometabolism plays an important part in bioremediation, but is often
difficult to control. Microbial degradation of tetrachloromethane (carbon tetrachloride,
CCl4), a toxic ozone-depleting organic solvent mainly of anthropogenic origin, is only
known to occur by cometabolic reduction under anoxic conditions. Yet no microbial
system capable of using CCl4 as the sole carbon source has been described. Microbial
growth based on CCl4 as a terminal electron acceptor has not been reported, although
corresponding degradation pathways would yield sufficient energy. Known modes for the
biodegradation of CCl4 involve several microbial metabolites, mainly metal-bound
coenzymes and siderophores, which are produced by facultative or strictly anaerobic
bacteria and methanogenic Archaea. Recent reports have demonstrated that CCl4
dechlorination rates are enhanced by redox-active organic compounds such as humic
acids and quinones, which act as shuttles between electron-providing microorganisms
and CCl4 as a strong electron acceptor. The key factors underlying dechlorination of
CCl4, the practical aspects and specific requirements for microorganism-associated

20

degradation of CCl4 at contaminated sites and perspectives for future developments are
discussed.
http://onlinelibrary.wiley.com/doi/10.1111/j.1574-6941.2010.00935.x/abstract

Mineralization of PCBs by the genetically modified strain Cupriavidus
necator JMS34 and its application for bioremediation of PCBs in soil
Autor: J. M. Saavedra, F. Acevedo, M. González, M. Seeger
Resumen:
Polychlorobiphenyls (PCBs) are classified as "high-priority pollutants." Diverse
microorganisms are able to degrade PCBs. However, bacterial degradation of PCBs is
generally incomplete, leading to the accumulation of chlorobenzoates (CBAs) as dead-
end metabolites. To obtain a microorganism able to mineralize PCB congeners, the bph
locus of Burkholderia xenovorans LB400, which encodes one of the most effective PCB
degradation pathways, was incorporated into the genome of the CBA-degrading
bacterium Cupriavidus necator JMP134-X3. The bph genes were transferred into strain
JMP134-X3, using the mini-Tn5 transposon system and biparental mating. The
genetically modified derivative, C. necator strain JMS34, had only one chromosomal
insertion of bph locus, which was stable under nonselective conditions. This modified
bacterium was able to grow on biphenyl, 3-CBA and 4-CBA, and degraded 3,5-CBA in
the presence of m-toluate. The strain JMS34 mineralized 3-CB, 4-CB, 2,4'-CB, and 3,5-
CB, without accumulation of CBAs. Bioaugmentation of PCB-polluted soils with C.
necator strain JMS34 and with the native B. xenovorans LB400 was monitored. It is
noteworthy that strain JMS34 degraded, in 1 week, 99% of 3-CB and 4-CB and
approximately 80% of 2,4'-CB in nonsterile soil, as well as in sterile soil. Additionally, the
bacterial count of strain JMS34 increased by almost two orders of magnitude in PCB-
polluted nonsterile soil. In contrast, the presence of native microflora reduced the
degradation of these PCBs by strain LB400 from 73% (sterile soil) to approximately 50%
(nonsterile soil). This study contributes to the development of improved biocatalysts for
remediation of PCB-contaminated environments.

Rapid Biodegradation of Benzo[a]pyrene by Bacillus subtilis BUM Under
Thermophilic Condition
Autor: Z. Zhao, J. W.-C. Wong
Fuente: Environmental Engineering Science. Voluem 27, Issue 11, Pages 939-945
Resumen:
Two thermophilic bacterial strains, Bacillus subtilis BUM (BUM) and Mycobacterium
vanbaalenii BU42 (BU42), were tested for their potential in biodegradation of
benzo[a]pyrene. Neither BUM nor BU42 utilized benzo[a]pyrene as the sole substrate
under thermophilic condition. In the presence of 50, 250, and 500mg L−1 phenanthrene,
the biodegradation of benzo[a]pyrene by BUM occurred and removals in 30 days were
14.8%, 38.8%, and 63.6%, respectively. This is the first report on the biodegradation of

21

benzo[a]pyrene by isolated thermophilic microorganism. The BUM strain was further
tested for its ability in a soil composting system. Within a composting period of 42 days,
removal of benzo[a]pyrene in the absence of BUM was 30.3%. Treatment with the
inoculation of BUM significantly increased the removal of benzo[a]pyrene to 52.2%.
Maximum zero-order degradation rates of benzo[a]pyrene by BUM in aqueous
biodegradation experiment and soil composting system were 12.3mg L−1 day−1 and
9.7mg kg−1 day−1, respectively, which were significantly greater than most of the
reported degradation rates by mesophiles. Experimental results affirmed that the strain
BUM can effectively degrade benzo[a]pyrene under thermophilic condition.
http://www.liebertonline.com/doi/abs/10.1089/ees.2010.0101?journalCode=ees

Screening of PAH-degrading bacteria in a mangrove swamp using PCR-
RFLP
Autor: H. Liu, C. Yang, Y. Tian, G. Lin, T. Zheng
Fuente: Marine Pollution Bulletin, Volume 60, Issue 11, Pages 2056-2061
Resumen:
There are abundant PAH-degrading bacteria in mangrove sediments, and it is very
important to screen the high efficiency degraders in order to perform bioremediation of
PAH polluted environments. In order to obtain the more highly efficient PAH-degrading
bacteria from a mangrove swamp, we first obtained 62 strains of PAH-degrading
bacteria using traditional culture methods and based on their morphological
characteristics. We then used the modern molecular biological technology of PCR-
RFLP, in which the 16S rDNA of these strains were digested by different enzymes.
Based on differences in the PCR-RFLP profiles, we obtained five strains of
phenanthrene-degrading bacteria, five strains of pyrene-degrading bacteria, four strains
of fluoranthene-degrading bacteria, five strains of benzo[a]pyrene-degrading bacteria
and two strains of mixed PAH-degrading bacteria (including phenanthrene, pyrene,
fluoranthene and benzo[a]pyrene). Finally, a total of 14 different PAH-degrading bacteria
were obtained. The 16S rDNA sequences of these strains were aligned with the BLAST
program on the NCBI website and it was found that they belonged to the α-
proteobacteria and γ-proteobacteria, including four strains, where the similarities were
no more than 97% and which were suspected therefore to be new species. This study
indicated that PCR-RFLP was a very important method to screen degrading-bacteria,
and also a significant molecular biological tool for the rapid classification and accurate
identification of many different strains. On the other hand, it also showed that rich
bacterial resources existed in mangrove areas, and that exploring and developing the
functional microorganism from these mangrove areas would have wide use in the study
of bioremediation of contaminated environments in the future.

22

Synthesis and Utilization of E. coli-Encapsulated PEG-Based Microdroplet
Using a Microfluidic Chip for Biological Application
Autor: K.G. Lee,T. Jung Park, S. Y. Soo, K. W. Wang, B. I.I. Kim, J. H. Park, C.-S. Lee, D. H.
Kim, S. J. Lee
Fuente: Biotechnology and Bioengineering, Volume 107, Pages 747–751
Resumen:
We report herein an effective strategy for encapsulating Escherichia coli in polyethylene
glycol diacrylate (PEGDA) microdroplets using a microfluidic device and chemical
polymerization. PEGDA was employed as a reactant due to the biocompatibility, high
porosity, and hydrophilic property. The uniform size and shape of microdroplets are
obtained in a single-step process using microfluidic device. The size of microdroplets
can be controlled through the changing continuous flow rate. The combination of
microdroplet generation and chemical polymerization techniques provide unique
environment to produce non-toxic ways of fabricating microorganism-encapsulated
hydrogel microbeads. Due to these unique properties of micro-sized hydrogel
microbeads, the encapsulated E. coli can maintain viability inside of microbeads and
green fluorescent protein (GFP) and red fluorescent protein (RFP) genes are efficiently
expressed inside of microbeads after isopropyl-β-D-thiogalactopyranoside induction,
suggesting that there is no low-molecular weight substrate transfer limitation inside of
microbeads. Furthermore, non-toxic, gentle, and outstanding biocompatibility of
microbeads, the encapsulated E. coli can be used in various applications including
biotransformation, biosensing, bioremediation, and engineering of artificial cells.
http://onlinelibrary.wiley.com/doi/10.1002/bit.22861/abstract

The role of salicylate and biosurfactant in inducing phenanthrene
degradation in batch soil slurries
Autor: Gottfried, A., N. Singhal, R. Elliot, S. Swift
Resumen:
The majority of polycyclic aromatic hydrocarbons (PAHs) sorb strongly to soil organic
matter posing a complex barrier to biodegradation. Biosurfactants can increase soil-
sorbed PAHs desorption, solubilisation, and dissolution into the aqueous phase, which
increases the bioavailability of PAHs for microbial metabolism. In this study,
biosurfactants, carbon sources, and metabolic pathway inducers were tested as
stimulators of microorganism degradation. Phenanthrene served as a model PAH and
Pseudomonas putida ATCC 17484 was used as the phenanthrene degrading
microorganism for the liquid solutions and soil used in this investigation. Bench-scale
trials demonstrated that the addition of rhamnolipid biosurfactant increases the apparent
aqueous solubility of phenanthrene, and overall degradation by at least 20% when
combined with salicylate or glucose in liquid solution, when compared to solutions that
contained salicylate or glucose with no biosurfactant. However, salicylate addition, with

23

no biosurfactant addition, increased the total degradation of phenanthrene 30% more
than liquid systems with only biosurfactant addition. In soil slurries, small amounts of
biosurfactant (0.25 g/L) showed a significant increase in total removal when only
biosurfactant was added. In soil slurries containing salicylate, the effects of biosurfactant
additions were negligible as there was greater than 90% removal, regardless of the
biosurfactant concentration. The results of experiments performed in this study provide
further evidence that an in situ enhancement strategy for phenanthrene degradation
could focus on providing additional carbon substrates to induce metabolic pathway
catabolic enzyme production, if degradation pathway intermediates are known.

24

2009

Biodegradation of Stored jet Fuel by a Nocardia sp. Isolated from
Contaminated Soil
Autor: E. de Barros Gomes, A. Ururahy Soriano, R. de Cássia Mendonça de Miranda, M. de F.
Vieira de Queiroz Sousa, N. Pereira Jr.
Fuente: Brazilian Archives of Biology and Technology, Volume 52, Issue 5, Pages 1279-1284
Resumen:
The aim of this study was to investigate the potential of degradation of an autochthonous
bacterial strain, isolated from petroleum derivatives contaminated soil samples against
jet fuel hydrocarbons. The autochthonous bacterial strain was characterized as Nocardia
sp. Evaluation of their degrading abilities was carried out by presumptive assays as
redox indicator test and by observations of surface tension decreases in aqueous
medium. Degradation of jet fuel hydrocarbons was evaluated by chromatographic
methods. Experiments were performed in flasks at two biostimulation rates. A bacterial
strain of Pseudomonas aeruginosa UFPEDA 39 was utilized as a reference
microorganism. The bacterial strain, identified as Nocardia sp, demonstrate high ability
to degrade jet fuel compounds as well as to produce surface active compounds when
compared to the reference microrganism.
http://www.scielo.br/pdf/babt/v52n5/v52n5a27.pdf

Biological treatment of saline wastewater using a salt-tolerant
microorganism
Autor: S.I. Abou-Elela , M. M. Kamel, M. E. Fawzy
Fuente: Desalination, Volume 250, Issue 1, Pages 1-5
Resumen:
Biological aerobic treatment of saline wastewater provides the material of this study. A
salt-tolerant microorganism (Staphylococcus xylosus) was isolated from a vegetable
pickled plant containing about 7.2% salt. Selection, identification and characterization of
the microorganism were carried out. The isolated microorganism was used as inoculum
for biodegradation. An activated sludge reactor operated in a fed-batch mode was used
for the treatment of synthetic saline wastewater using three different microbial cultures
namely: activated sludge (100%), a mixture of Staphylococcus supplement by activated
sludge (1:1) and pure S. xylosus (100%) at different salt concentrations ranging from 0.5
to 3% NaCl. The results obtained showed that at low NaCl concentration (1%), the
removal efficiency of chemical oxygen demand (COD) using different microbial cultures
were almost the same (80–90%). However, increasing the NaCl concentration to 2% and
using Staphylococcus-supplemented mixture by activated sludge and S. xylosus alone
improved the treatment performance as indicated by COD removal rates which reached
91% and 93.4%, respectively, while the system performance started to deteriorate when
activated bacterial culture was used alone (74%). Furthermore, the increase in NaCl
concentration up to 3% and with the inclusion of Staphylococcus-supplemented mixture
by activated sludge increased the COD removal to 93%, while the use of S. xylosus
alone further improved the COD removal rate up to 94%. Also, the use of S. xylosus

25

alone proved to be capable for biological treatment of a real case study of a vegetable
pickled wastewater containing 7.2% salinity; the removal efficiency of COD reached 88%
at this very high concentration of NaCl.
http://www.sciencedirect.com/science?_ob=ArticleURL&_udi=B6TFX-4XG3D6G-
050221&_version=1&_urlVersion=0&_userid=10&md5=598bbaf10f741cb6c5a6b3f8ee56b57b&se
archtype=a

Bioremediation of Fenvalerate by Pseudomonas aeruginosa in a Scale up
Bioreactor
Autor: M. H. Fulekar
Fuente: Romanian Biotechnological Letters, Volume 14, Issue 6, Pages 4900-4905
Resumen:
The potential of microorganisms has been assessed for bioremediation of fenvalerate
using the scale up process followed by bioremediation in bioreactor under controlled
environmental conditions.
Pseudomonas aeruginosa isolate NCIM 2074 was adapted to varying concentration of
fenvalerate Viz. 10,25,50,75,100 mg/l in an incubator shaker at 37º C and 150 rpm. An
initial 10 mg/l concentration of fenvalerate was added in a minimal salt medium (MSM)
and subjected to incubation for 14 days with Pseudomonas aeruginosa and thereafter
the culture was scaled up to higher concentration of fenvalerate by transferring one
milliliter of MSM to 25,50,75,100 over a period of 70 days at a frequency of 14 days. The
fenvalerate concentration after every 14 days was assessed by GC-MS for the
adaptability of microorganism for biodegradation. The research findings show that 10
mg/l fenvalerate was completely degraded within a period of 14 days in MSM; whereas
in the concentration ranging from 25 to 100 mg/l fenvalerate in MSM, the bioremediation
rate was found decreasing with increasing concentration. Fenvalerate at 50 and 100
ppm concentration was found inhibiting to the microorganism. The adapted
microorganism, after scale up process was bioremediated in a flask shaker method at
10, 25 50 mg/l in MSM separately under controlled environmental conditions. The parent
compound was found biodegraded into the primary metabolite 4-chloro-α (1-methylethyl)
benzene acetic acid and α -cyano-3-phenoxybenzyl alcohol and 3-Phenoxy benzoic
acid. These intermediates in long run on acclimatization with Pseudomonas aeruginosa
might converts into environment friendly compounds. The research findings show that
Pseudomonas aeruginosa has the potential to degrade the toxic compound such as
fenvalerate ranging from 10 to 25 ppm. This technology would be beneficial to pesticides
industry for bioremediation of pesticide fenvalerate.
http://www.rombio.eu/rbl6vol14/Lucr%2018%20Fulekar.pdf

26

Bioremediation of soil heavily contaminated with crude oil and its
products: Composition of the microbial consortium
Autor: J. S. Milic, V. P. Beskoski, M. V. Ilic, S. A. M. Ali, G. D. Gojic-Cvijovic, M. Vrivic
Fuente: Journal of the Serbian Chemical Society, Volume 74, Issue 4, Pages 455-460
Resumen:
Bioremediation, a process that utilizes the capability of microorganism to degrade toxic
waste, is emerging as a promising technology for the treatment of soil and groundwater
contamination. The technology is very effective in dealing with petroleum hydrocarbon
contamination. The aim of this study was to examine the composition of the microbial
consortium during the ex situ experiment of bioremediation of soil heavily contaminated
with crude oil and its products from the Oil Refinery Pančevo, Serbia. After a 5.5-month
experiment with biostimulation and bioventilation, the concentration of the total
petroleum hydrocarbons (TPH) had been reduced from 29.80 to 3.29 g/kg (89 %). In
soil, the dominant microorganism population comprised Gram-positive bacteria from
actinomycete-Nocardia group. The microorganisms which decompose hydrocarbons
were the dominant microbial population at the end of the process, with a share of more
than 80 % (range 107 CFU/g). On the basis of the results, it was concluded that a stable
microbial community had been formed after initial fluctuations.
http://www.doiserbia.nb.rs/img/doi/0352-5139/2009/0352-51390904455M.pdf

Enrichment and isolation of endosulfan-degrading microorganism from
tropical acid soil
Autor: S. S. Kalyani, J. Sharma, S. Singh, P. Dureja, P. Lata
Fuente: Journal of Environmental Science and Health, Part B, Pesticides, food contaminants, and
agricultural wastes, Volume 44, Issue 7, Pages 663-672
Resumen:
Endosulfan (6,7,8,9,10,10-hexachloro-1,5,5a,6,9,9a-hexahydro-6,9-methano-2,3,4-
benzo-dioxathiepin-3-oxide) is a cyclodiene organochlorine currently used as an
insecticide all over the world and its residues are posing a serious environmental threat.
This study reports the enrichment and isolation of a microbial culture capable of
degrading endosulfan with minimal production of endosulfan sulfate, the toxic metabolite
of endosulfan, from tropical acid soil. Enrichment was achieved by using the insecticide
as sole sulfur source. The enriched microbial culture, SKL-1, later identified as
Pseudomonas aeruginosa, degraded up to 50.25 and 69.77 % of alpha and beta
endosulfan, respectively in 20 days. Percentage of bioformation of endosulfan sulfate to
total formation was 2.12% by the 20th day of incubation. Degradation of the insecticide
was concomitant with bacterial growth reaching up to an optical density of 600 nm
(OD600) 2.34 and aryl sulfatase activity of the broth reaching up to 23.93 microg
pNP/mL/hr. The results of this study suggest that this novel strain is a valuable source of
potent endosulfan-degrading enzymes for use in enzymatic bioremediation. Further, the
increase in aryl sulfatase activity of the broth with the increase in degradation of
endosulfan suggests the probable involvement of the enzyme in the transformation of
endosulfan to its metabolites.

27

Isolation and characterization of an SDS-degrading Klebsiella oxytoca
Autor: M.Y. Shukor, W.S.W. Husin, M.F.A. Rahman, N.A. Shamaan, M.A. Syed
Fuente: Journal of Environmental Biology, Volume 30, Issue 1, Pages 129-134
Resumen:
Sodium dodecyl sulfate (SDS) is one of the main components in the detergent and
cosmetic industries. Its bioremediation by suitable microorganism has begun to receive
greater attention as the amount of SDS usage increases to a point where treatment
plants would not be able to cope with the increasing amount of SDS in wastewater. The
purpose of this work was to isolate local SDS-degrading bacteria. Screening was carried
out by the conventional enrichment-culture technique. Six SDS-degrading bacteria were
isolated. Of these isolates, isolate S14 showed the highest degradation of SDS with 90%
degradation after three days of incubation. Isolate S14 was tentatively identified as
Klebsiella oxytoca strain DRY14 based on carbon utilization profiles using Biolog GN
plates and partial 16S rDNA molecular phylogeny. SDS degradation by the bacterium
was optimum at 37oC. Ammonium sulphate; at 2.0 g l-1, was found to be the best
nitrogen source for the growth of strain DRY14. Maximum growth on SDS was observed
at pH 7.25. The strain exhibited optimum growth at SDS concentration of 2.0 g l-1 and
was completely inhibited at 10 g l-1 SDS. At the tolerable initial concentration of 2.0 g l-
1, almost 80% of 2.0 g l-1 SDS was degraded after 4 days of incubation concomitant
with increase in cellular growth. The Km (app) and Vmax (app) values calculated for the
alkylsulfatase from this bacterium were 0.1 mM SDS and 1.07 >mol min-1 mg-1 protein,
respectively.
http://www.jeb.co.in/journal_issues/200901_jan09_spl/paper_20.pdf

Mitigation of Ca, Fe, and Mg loads in surface waters around mining areas
using indigenous microorganism strains
Autor: E. Fosso-Kankeu, A.F. Mulaba-Bafubiandi , B.B. Mamba, T.G. Barnard
Fuente: Physics and Chemisty of the Earth, Parts A/B/C, Voluem 34, Issues13-16, Pages 825-
829
Resumen:
In attempting to achieve acceptable minimum concentration levels of excess calcium,
iron and magnesium in surface waters around mining areas, experiments conducted at
laboratory scale to remove these metals from synthetic solutions (30 ppm and 50 ppm)
using indigenous strains of Shewanella sp., Bacillus subtilis sp. and Brevundimonas sp.
revealed varying abilities of these microorganisms. B. subtilis and Shewanella sp.
absorbed the highest amount (14% Ca, 8% Mg and 8% Fe) of each of the three metals,
recorded from solutions containing the metals at 30 ppm concentration, with calcium
being most easily removed metal species. The purpose of this study was to investigate a
cost-effective solution based on indigenous microorganisms for the bioremediation of
toxic metallic species in the mine dumps where small scale mining operations occur.
Metal removal from solution decreased when their concentration in solution was at 50
ppm. It was also found that combining the metals in one solution affected the
microorganisms’ affinity for the metals thus reducing their removal efficiency. There was
also a tendency for microorganisms to release the absorbed metal into solution after a

28

certain period of time, most probably due to an efflux transport mechanism. It was further
concluded that the metal removal efficiency is dependent on the biomass, and the
percentage removals obtained in this study suggest that we could achieve better
removal rates of targeted metals and reduce their concentrations to below
recommended values through the optimization of the biomass. The success of this study
has prompted a broader research project into the removal of metal species in mine
dumps before contamination of water resources occurs so that the water in the disused
mine pits is suitable for irrigation, farming and washing.
http://www.sciencedirect.com/science?_ob=ArticleURL&_udi=B6X1W-4WRD3KF-
050221&_version=1&_urlVersion=0&_userid=10&md5=68fd7b658910206fe55287ab1cbcc473&s
earchtype=a

Multifunctional properties of phosphate-solubilizing microorganisms
grown on agro-industrial wastes in fermentation and soil conditions
Autor: M. Vassileva, M. Serrano, V. Bravo, E. Jurado, I. Nikoleva, V. Martos, N. Vassilev
Fuente: Applies Microbiology and Biotechnology, Volume 85, issue 5, Pages 1287-1299
Resumen:
One of the most studied approaches in solubilization of insoluble phosphates is the
biological treatment of rock phosphates. In recent years, various techniques for rock
phosphate solubilization have been proposed, with increasing emphasis on application
of P-solubilizing microorganisms. The P-solubilizing activity is determined by the
microbial biochemical ability to produce and release metabolites with metal-chelating
functions. In a number of studies, we have shown that agro-industrial wastes can be
efficiently used as substrates in solubilization of phosphate rocks. These processes were
carried out employing various technologies including solid-state and submerged
fermentations including immobilized cells. The review paper deals critically with several
novel trends in exploring various properties of the above microbial/agro-wastes/rock
phosphate systems. The major idea is to describe how a single P-solubilizing
microorganism manifests wide range of metabolic abilities in different environments. In
fermentation conditions, P-solubilizing microorganisms were found to produce various
enzymes, siderophores, and plant hormones. Further introduction of the resulting
biotechnological products into soil-plant systems resulted in significantly higher plant
growth, enhanced soil properties, and biological (including biocontrol) activity.
Application of these bio-products in bioremediation of disturbed (heavy metal
contaminated and desertified) soils is based on another important part of their
multifunctional properties.
http://www.springerlink.com/content/25271645484r5748/

Specific dechlorinase activity in lindane degradation by Streptomyces sp
M7
Autor: S. A. Cuozzo, G. G. Rollán, C. M. Abate, M. J. Amoroso

29

Fuente: World Journal of Microbiology and Biotechnology, Volume 25, Issue 9, Pages 1539-1546

Resumen:
Synthesis of dechlorinase in Streptomyces sp. M7 was induced when the microorganism
was grown in the presence of lindane (γ-hexachlorocyclohexane) as the only carbon
source. Activity of cells grown with lindane was about four and half times higher
compared to cells grown with glucose. Maximum dechlorinase activity was observed at
30°C in alkaline conditions pH (7.9) and the enzyme did not show cation dependency.
Sodium dodecyl sulfate polyacrylamide gel electrophoresis revealed one differential
band with a molecular weight similar to serum albumin (M r 66,200), which corresponded
to polynucleotide phosphorylase, an enzyme that plays an important role in the
regulation system and could be involved in the regulation of the dechlorinase gene.
Detected in cell-free extracts were γ-pentachlorocyclohexene and 1,3,4,6-tetrachloro-
1,4-cyclohexadiene, both being products of the dechlorinase activity. This is the first time
that the presence of an enzyme with dechlorinase activity has been demonstrated in an
actinomycete strain isolated in Tucumán, Argentina. Characteristics of this enzyme
revealed that Streptomyces sp. M7 could be useful in the future in bioremediation of soil
or as a biosensor.
http://www.springerlink.com/content/l8q231lh42378714/

Synergic degradation of phenanthrene by consortia of newly isolated
bacterial strains
Autor: Y. M. Kim, C. K. Ahn, S. H. Woo, G. Y.l Jung, J. M. Park
Fuente: Journal of Biotechnology, Volume 144, Issue 4, Pages 293-298
Resumen:
Three different bacteria capable of degrading phenanthrene were isolated from sludge of
a pulp wastewater treatment plant and identified as Acinetobacter baumannii, Klebsiella
oxytoca, and Stenotrophomonas maltophilia. Phenanthrene degradation efficiencies by
different combinations (consortia) of these bacteria were investigated and their
population dynamics during phenanthrene degradation were monitored using capillary
electrophoresis-based single-strand conformation polymorphism (CE-SSCP). When a
single microorganism was used, phenanthrene degradation efficiency was very low
(48.0, 11.0, and 9.0% for A. baumannii, K. oxytoca, and S. maltophilia respectively, after
360 h cultivation). All consortia that included S. maltophilia degraded 80.0% of
phenanthrene and reduced lag time to 48 h compared to the 168 h of pure A. baumannii
culture. CE-SSCP analysis showed that S. maltophilia was the predominant species
during phenanthrene degradation in the mixed culture. The results indicate that mixed
cultures of microorganisms may effectively degrade target chemicals, even if the
microorganisms show low degradation activity in pure culture.
http://www.sciencedirect.com/science?_ob=ArticleURL&_udi=B6T3C-4XDD013-
050221&_version=1&_urlVersion=0&_userid=10&md5=429f34626e16fa0e9461e75f67837302&s
earchtype=a

30

2008

Adaptive and cross resistance to cadmium(II) and zinc(II) by
Pseudomonas aeruginosa BC15
Autor: C. E. Raja, S. Sasikumar, G. S. Selvam
Fuente: Biologia, Volume 63, Issue 4, Pages 461-465
Resumen:
Cadmium and zinc appear in the combined forms and they are co-pollutants. Cd is the
most hazardous metal ion for human beings and causes renal dysfunction, liver and
lungs damage, bone degeneration and blood damage. Though Zn is an essential
nutrient, excess of Zn is toxic. Biological process was more important because
conventional methods fail to remediate these pollutants due to high costs and less
affordability. The screening and understanding of the functioning of microorganism plays
an important role in removal and recovery of metals from heavy-metal-polluted water
and soil. In our study, the strain Pseudomonas aeruginosa BC15 was isolated from oil-
mill-treated waste water and it showed to be highly resistant to 6 mM Cd and 20 mM Zn
in the solid and liquid media. The growth studies of BC15 strain in the medium without
induction exhibited high tolerable capacity when compared to other microbes.
Pretreatment of P. aeruginosa BC15 with sub-lethal concentrations of Cd induced
adaptive resistance to lethal doses of Cd. Cadmium-induced cells also showed cross
resistance to lethal concentration of zinc. The organism had high resistance against Cd
and Zn. This has been clearly proven through biosorption studies: Cd was absorbed up
to 62% and Zn about 60% in single solution, whereas in binary solution Cd was
biosorbed up to 82% and Zn 85%. In conclusion, this study reveals the significance of
using the strain P. aeruginosa BC15 in the bioremediation of Cd and Zn from industrial
waste water and contaminated soil.
http://www.springerlink.com/content/j3k83662k323uq34/

Bioremediation of trichloroethylene contaminated groundwater using
anaerobic process
Autor: C. Chomsurin, J. Kajorntraidej, K. Kuangmuang
Fuente: Water Science and Technology, Volume 58, Issue 11, Pages 2127-2132
Resumen:
Anaerobic remediation of trichloroethylene (TCE) contaminated soil and groundwater
was studied in laboratory setups. In this process fermentation of polymeric organic
materials (POMS) produced volatile fatty acids (VFAs) that were electron donors in
reductive dechlorination of TCE. Shredded peanut shell was selected as low cost POM
and the experiments were set up in 500 ml Erlenmeyer flasks. In the setups,
approximately 25 mg of leachate contaminated soil was used as the main source of
microorganisms and about 5 g of shredded peanut shell (0.5-2.36 mm) was added to
produce VFAs for dechlorination of TCE. In the first set of experiments, fermentation of
soil and shredded peanut shell was studied and it was found that VFAs were produced
continuously with increasing concentration (5.63 mM as CH3COOH from the first day to
17.17 in the 10th day of the experiment). During the fermentation, concentration of

31

ammonia-nitrogen was 22-50 mg/L, the ratio of VFA to NH3 was 15.29-23.44 and pH
was 5.24-6.00. These results show that the system was appropriate for microorganism
activities. In the second set of experiments, TCE (approximately 48 mg/L) was added to
the fermentation system and remediation of TCE by reductive dechlorination was
studied. It was found that 0.04(+/-0.01) mg TCE adsorbed to a gram of soil and peanut
shells at the beginning of the experiment and based on mass balance of the system,
TCE concentration in water was linearly reduced at the rate of 0.0098 mg/hr.

Degradation of mixtures of phenolic compounds by Arthrobacter
chlorophenolicus A6
Autor: M. Unell, K. Nordin, C. Jernberg, J. Stenström, J. K. Jansson
Fuente: Biodegradation, Volume 19, Issue 4, Pages 495-505
Resumen:
In this study the chlorophenol-degrading actinobacterium, Arthrobacter chlorophenolicus
A6, was tested for its ability to grow on mixtures of phenolic compounds. During the
experiments depletion of the compounds was monitored, as were cell growth and
activity. Activity assays were based on bioluminescence output from a luciferase-tagged
strain. When the cells were grown on a mixture of 4-chlorophenol, 4-nitrophenol and
phenol, 4-chlorophenol degradation apparently was delayed until 4-nitrophenol was
almost completely depleted. Phenol was degraded more slowly than the other
compounds and not until 4-nitrophenol and 4-chlorophenol were depleted, despite this
being the least toxic compound of the three. A similar order of degradation was observed
in non-sterile soil slurries inoculated with A. chlorophenolicus. The kinetics of
degradation of the substituted phenols suggest that the preferential order of their
depletion could be due to their respective pKa values and that the dissociated phenolate
ions are the substrates. A mutant strain (T99), with a disrupted hydroxyquinol
dioxygenase gene in the previously described 4-chlorophenol degradation gene cluster,
was also studied for its ability to grow on the different phenols. The mutant strain was
able to grow on phenol, but not on either of the substituted phenols, suggesting a
different catabolic pathway for the degradation of phenol by this microorganism.
http://www.springerlink.com/content/t668777902668v86/

Development of a Bacterial Preparation Based on Immobilized Cells
Autor: K. A. Ausheva, D. A. Goncharuk, E. S. babusenko, S. A. Nekhaev, Z. S. Sultygova, n. S.
Markvichev
Fuente: Theoretical Foundations of Chemical Engineering, Volume 42, Issue 5, Pages 767-773
Resumen:
A new method for removing thin oil films from a water surface with the use of the oil-
degrading microorganism Acinetobacter valentis immobilized in calcium alginate gel has
been developed. It has been demonstrated that n-alkanes emulsified into calcium
alginate granules impart positive buoyancy to the granules. The parameters of obtaining
calcium alginate granules with immobilized oil-degrading cells affect the characteristics

32

of biological preparation. It has been demonstrated that the use of the biological
preparation containing emulsified n-alkanes makes it possible to increase the
bioremediation rate due to localization of immobilized and free cells in the upper water
level. The new form of biological preparations makes it possible to decrease the amount
of oil hydrocarbons by as much as 97% for 21 days at a temperature of 10–22°C versus
63% in the control variant.
http://www.springerlink.com/content/y31721331m6u8xj7/

Enhancement of bioremediation by Ralstonia sp HM-1 in sediment
polluted by Cd and Zn
Autor: Y.-J. Park, J.-J. Ko ,S.-L. Yun, E. Y. Lee, S.-J. Kim, S.-W. Kang, B.-C. Lee, S.-K. Kim
Resumen:
In this study, the potential for the application of the bioaugmentation to Cd and Zn
contaminated sediment was investigated. A batch experiment was performed in the lake
sediments augmented with Ralstonia sp. HM-1. The degradation capacity of 18.7 mg-
DOC/l/day in the treatment group was bigger than that of the blank group (4.4 mg-
DOC/l/day). It can be regarded as the result of the reduction of the metal concentration
in the liquid phase due to adsorption into the sediments, with the increased alkalinity
resulting from the reduction of sulfate by sulfate reducing bacteria (SRB). The removal
efficiency of cadmium and zinc in the treatment group was both 99.7% after 35 days.
Restrain of elution to water phase from sediment in the Ralstonia sp. HM-1 added
treatment group was also shown. In particular, the observed reduction of the
exchangeable fraction and an increase in the bound to organics or sulfide fraction in the
treatment group indicate its role in the prevention of metal elution from the sediment.
Therefore, for bioremediation and restrain of elution from the sediment polluted by metal,
Ralstonia sp. augmentation with indigenous microorganism including SRB, sediment
stabilization and restrain of elution to surface water is recommended.
http://www.sciencedirect.com/science?_ob=ArticleURL&_udi=B6V24-4S92XKV-
050221&_version=1&_urlVersion=0&_userid=10&md5=f11bcc6aa5329751cbe816da2225754b&s
earchtype=a

Enhancement of PCB degradation by Burkholderia xenovorans LB400 in
biphasic systems by manipulating culture conditions
Autor: L. Rehmann, A. J. Daugulis
Fuente: Biotechnology and Bioengineering, Volume 99, Issue 3, Pages 521-528
Resumen:
Two-phase partitioning bioreactors (TPPBs) can be used to biodegrade environmental
contaminants after their extraction from soil. TPPBs are typically stirred tank bioreactors
containing an aqueous phase hosting the degrading microorganism and an immiscible,
non-toxic and non-bioavailable organic phase functioning as a reservoir for hydrophobic
compounds. Biodegradation of these compounds in the aqueous phase results in
thermodynamic disequilibrium and partitioning of additional compounds from the organic
phase into the aqueous phase. This self-regulated process can allow the delivery of

33

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