See discussions, stats, and author profiles for this publication at: https://www.researchgate.

net/publication/230205528

Biodegradability of hydrocarbons by Cyanobacteria

Article  in  Journal of Phycology · July 2010

DOI: 10.1111/j.1529-8817.2010.00865.x

CITATIONS READS

23 311

1 author:

Ibraheem Borie Mohammad Ibraheem

Faculty of Science, Beni Suef University, Beni-Suef, Egypt
55 PUBLICATIONS   1,185 CITATIONS   

SEE PROFILE

Some of the authors of this publication are also working on these related projects:

algal nano-technology View project

Algae and Treatment of Pollution View project

All content following this page was uploaded by Ibraheem Borie Mohammad Ibraheem on 09 December 2018.

The user has requested enhancement of the downloaded file.

J. Phycol. 46, 818–824 (2010)
 2010 Phycological Society of America
DOI: 10.1111/j.1529-8817.2010.00865.x

BIODEGRADABILITY OF HYDROCARBONS BY CYANOBACTERIA 1

Ibraheem Borie Mohammad Ibraheem2

Botany Department, Faculty of Science, Beni-Suef University, Beni-Suef 62511, Egypt

Five cyanobacterial species (Phormidium sp., Nostoc
sp., Anabaena sp. Aphanothece conferta, and Synecho-
cystis aquatilis) isolated from the Suez Canal coast at
the city of Ismailia (Egypt) were tested for biodegra-
dation of four hydrocarbon (HC) compounds: two
aliphatic compounds (n-octadecane and pristine)
and two aromatic compounds (phenanthrene and
dibenzothiophene). High degradation efficiencies
for the two aliphatic compounds were measured for
A. conferta (64% for n-octadecane and 78% for pris-
tine) and S. aquatilis (85% for n-octadecane and
90% for pristane). However, the other biodegrada-
tion percentages ranged between weak and moder-
ate percentages.
Key index words: Aphanothece conferta; biodegrada-
tion of hydrocarbons; cyanobacteria; marine
aquatic pollution; Suez Canal coast
Abbreviations: BTMA, benzyl-trimethylammonium-
chloride; DCM, dichloromethane; HC, hydrocar-
bon; OCC, organo-clay complexes
Pollution of marine environments with HCs has
become a worldwide problem on the tide of indus-
trialization. The sources of marine HC pollution are
mainly runoff from land and municipal ⁄ industrial
wastes, routine ship maintenance like bilge clean-
ing, air pollution from cars and industry, natural
seeps, tanker accidents, and offshore oil production
(NRC 1985, U.S. Coast Guard 1990). It has long
been known that poly-nuclear aromatic HCs exhibit
serious toxic and carcinogenic effects (Miller and
Miller 1974, McCann et al. 1975). Therefore, the
study of the biotransformation and biodegradability
of these aromatic compounds in the environment is
of basic and particular value.
The main process acting in the cleanup of HC-
contaminated ecosystems is microbial biodegrada-
tion, which has been extensively studied and
reviewed (Atlas 1984, Leahy and Colwell 1990, Chai-
llan et al. 2006). Numerous microorganisms, includ-
ing bacteria, fungi, and yeasts, are known for their
ability to degrade HCs (Oudot et al. 1993, Chaillan
et al. 2004). In tropical crude oil production sites,
1
2
Received 17 November 2009. Accepted 19 April 2010.
Author for correspondence: e-mail ibraheemborie@hotmail.com.
cyanobacterial mats often develop on petroleum-pol-
luted zones including surface soils and water envi-
ronments. After the release of oil during the Gulf
War in Kuwait, a bloom of cyanobacteria intimately
associated with oil was also observed (Sorkhoh et al.
1992).
There is increasing evidence that photosynthetic
microorganisms, particularly cyanobacteria, may
contribute to the oxidation and degradation of
HCs. However, it is important to emphasize that
only in some cases were the tested cyanobacterial
cultures axenic and that many studies have been
carried out on nonaxenic cultures.
Among the earliest studies on the potential of
photosynthetic microorganisms, including cyanobac-
teria, for aromatic HC oxidation is the study of Ellis
(1977). This author investigated the phenol and cat-
echol degradation potential of some microalgae and
cyanobacteria. Most of the work on aliphatic HCs
has focused on the potential of these phototrophs
for the complete utilization of these compounds.
Previous reports have shown the ability of cyano-
bacteria to oxidize oil components. Al-Hasan et al.
(1998) reported that nonaxenic cultures of Microco-
leus chthonoplastes and Phormidium corium, isolated
from oil-rich sediments of the Arabian Gulf, were
able to degrade n-alkanes. Studies on Oscillatoria sp.
and Agmenellum quadruplicatum demonstrated their
ability to oxidize naphthalene to 1-naphthol (Cerni-
glia et al. 1980a). Other studies showed that Oscilla-
toria sp. can oxidize biphenyl to n-hydroxybiphenyl
(Cerniglia et al. 1980b) and that A. quadruplicatum
metabolizes phenanthrene into trans-9,10-dihydroxy-
9,10-dihydrophenanthrene and 1-methoxy-phen-
enthrene (Narro 1985). Several other strains were
reported to degrade crude oil and other complex
organic compounds (Lee et al. 1974, Cerniglia et al.
1984, Yan et al. 1998, Radwan and Al-Hasan 2000,
Raghukumar et al. 2001, Mansy and El-Bestway
2002). However, in most biodegradation studies
with cyanobacteria, it was not clear whether the
strains used were definitively axenic (Abed and
Köster 2005). It is known to be very difficult to culti-
vate cyanobacteria in axenic culture and to clean
them from naturally associated aerobic heterotro-
phic bacteria. Thus, the contribution of aerobic het-
erotrophic bacteria associated with cyanobacteria to
the biodegradation process needs to be carefully
evaluated. In this study, I have addressed this

818
 BIODEGRADABILITY OF HCs BY CYANOBACTERIA
problem by testing the ability of five axenic unicy-
anobacterium strains isolated from Suez Canal,
Egypt, to degrade four petroleum compounds.
MATERIALS AND METHODS
Isolation, identification, and maintenance of cyanobacterial strains.
Five cyanobacterial mat samples were collected from the Suez
Canal coast, city of Ismailia, Egypt, and transferred to the
phycological lab (Faculty of Science, Beni-Suef University, Beni-
Suef, Egypt) within 24 h upon arrival; the mates were incubated
in a glass container filled with aerated artificial seawater. The
mats were incubated at a light regime of 12:12 light:dark (L:D)
with an intensity of 80 lmol photons Æ m)2 Æ s)1. The diversity
and abundance of the cyanobacteria of the mat samples were
microscopically examined as described in Chaillan et al. (2006)
using a Binocular Olympus microscope (serial 8H0222, Medica
Instrument Mfg. Co., Mumbai, India)). Isolation and purifica-
tion of cyanobacteria in a unialgal form were done according to
the methods described by Rippka (1988). Briefly, the cyano-
bacterium was isolated after repeated light migrations on solid
Z-medium prepared in seawater samples, according to Chaillan
et al. (2006). These purified cyanobacterial species were iden-
tified according to Smith (1950) and Desikachary (1959).
Logarithmic growth phase of each cyanobacterium was
detected to obtain the healthy growth mass to be used in this
experiment.
Purification of the cyanobacterial isolates from bacteria. Axenic
cultures of the cyanobacterial isolates were obtained as
described by Pringsheim (1949). These cultures were subjected
to different trials to employ bacteria-free cultures and investi-
gate the growth of bacteria every 20 d throughout the
experimental period, according to Felfoldy and Zsuzsa (1959)
and Hoshaw and Rosewski (1973).
Preparation of organo-clay complexes (OCC). n-Octadecane,
pristane, phenanthrene, and dibenzothiophene KSF (Aldrich,
Steinheim, Germany) were used as model compounds for
petroleum constituents representing straight-chain alkanes,
branched alkanes, aromatic HCs, and organo-sulfur com-
pounds, respectively. Hydrocarboic clay was used as a carrier
substance for the petroleum model compounds to overcome
the low accessibility of the poorly water-soluble compounds
when directly added to the medium.
A 2% aqueous suspension of montmorillonite KSF (Aldrich)
was slowly mixed with a 10 mM solution of benzyl-trimethy-
lammoniumchloride (BTMA, 0.8 mmoln Æ g)1 clay) and stirred
for 24 h (modified after El-Nahhal et al. 2000). Subsequently,
the mixture was washed three times with deionized water to
remove excess BTMA and then dried. To absorb the model
compounds, the hydrophobic clay (BTMA-montmorillonite)
was suspended in n-hexane. The petroleum model compounds
were slowly added under continuous stirring to an n-hexane
suspension in total 20 mg model compounds per 100 mg of
hydrophobic clay. The resulting organo-clay complex slurry
(OCC) was dried in a vacuum rotary evaporator (Roteva
Equitron 63 series Model 87G3.RD0.000, Medica Instrument
Mfg. Co.), which yielded homogenous powder of hydrophobic
clay loaded with 16.67 wt% of petroleum model compounds.
To verify the amount of loaded model compounds, the OCC
was reextracted with dichloromethane (DCM) and analyzed by
gas chromatography according to the method described by
Grötzschel et al. (2002).
Biodegradation experiment with cyanobacterial strains. The deg-
radation experiments (three replicates) were carried out in
250 mL sterile Erlenmeyer flasks. Each Erlenmeyer flask
received 99 mL of autoclaved medium, 1 mL of culture
suspension, and 100 mg of OCC. Controls without culture
suspensions were also determined. All flasks were incubated at
28C with constant shaking at 100 rpm, a 12:12 L:D cycle, and
819
80 lmol photons Æ m)2 Æ s)1 light intensity. The experiment
was run for 60 d, and samples for chemical analysis (2 mL
each) were taken every 20 d.
Chemical analysis. Samples of 2 mL from the cultures were
extracted ultrasonically four times with a 1:0.5:0.4 (v ⁄ v ⁄ v)
mixture of methanol (MeOH), DCM, and water (modified
after Bligh and Dyer 1959). DCM and water were added to the
combined extract to yield a solvent mixture of MeOH:DCM:
H2O at a ratio of 1:1:0.9 (v ⁄ v ⁄ v) resulting in phase separation.
The DCM layer was collected, and the methanol-water phase
was washed three times with DCM. The solvent of the
combined DCM phase was removed in the rotary evaporator,
and the extracts were diluted to concentrations appropriate for
gas chromatographic analyses.
Statistical analyses. All the values or readings are the result of
the mean of three replicates in addition to the standard
deviation.
RESULTS AND DISCUSSION
Data recorded in Figures 1 and 2 revealed differ-
ent degradation efficiencies for the studied cyano-
bacteria against the tested model petroleum
compounds. In the case of the control, the degrada-
tion percentages of n-alkanes throughout 60 d were
8% and 7% for n-octadecane and pristane, respec-
tively (Fig. 1A), while they reached 7% in the case
of phenanthrene (on day 20) and 6% in the case of
dibenzothiophene at the end of experiment
(Fig. 2A). This result might be attributed to the nor-
mal reactions between solution contents.
The investigated cyanobacterial isolates exhibited
different degradation efficiencies, which depended
on the cyanobacterial species used and the petro-
leum compound.
Aliphatic oxidation by cyanobacteria. While most of
the work on aromatic HCs was targeted at studying
the potential of cyanobacteria for oxidation of these
compounds, work on aliphatic HCs was focused on
studying the potential of these phototrophs for the
complete utilization of these compounds. Data
recorded in Figure 1 indicated that both filamen-
tous cyanobacteria (Phormidium sp. and Nostoc sp.)
exhibited weak abilities against both n-octadecane
and pristane. Values were 9% (on day 20) and 10%
(on day 40) for n-octadecane and pristane, respec-
tively, by Phormidium sp., and 13% (on day 60) and
11% (on day 40) for n-octadecane and pristane,
respectively, by Nostoc. However, the filamentous cya-
nobacterium Anabaena sp. demonstrated a moderate
biodegradable activity toward the two alkanes: 29%
(on day 60) for n-octadecane and weak activity
(15%) for pristane on day 60 (Fig. 1C).
In contrast, both A. conferta and S. aquatilis had
high efficiencies in degradation of both aliphatic
compounds: 64% (on day 20) and 78% (on day 40)
for n-octadecane and pristane, respectively, by
A. conferta (Fig. 1E), which remained constant until
the end of the experiment, and 85% (on day 20)
and 90% (on day 60) for n-octadecane and pristane,
respectively, by S. aquatilis (Fig. 1F). Thus, the abil-
ity of A. conferta and S. aquatilis to degrade both
820 I B R A H E E M B O R I E M O H A MM A D I B R A H E E M

Control Phormidium sp.

60

Degradation %
60

Degradation %
50 50
40 40
30 30
20 20
10 10
0 0
0 20 40 60 0 20 40 60
Time (d)
Time (d)

n-Octadecane Pristane n-Octadecane Pristane

Anabaena sp. Nostoc sp.

60

Degradation %
Degradation %

60 50
50
40
40
30
30
20 20
10 10
0 0
0 20 40 60 0 20 40 60
Time (d) Time (d)

n-Octadecane Pristane n-Octadecane Pristane

Aphanothece conferta Synechocystis aquatilis

100
Degradation %

100
Degradation %

80
80
60 60

40 40

20 20
0 0
0 20 40 60 0 20 40 60
Time (d) Time (d)
n-Octadecane Pristane n-Octadecane Pristane

Fig. 1. Biodegradation percentages of n-octadecane and pristane by cyanobacterial cultures. (A) Control (natural seawater), (B) Pho-
rmidium sp., (C) Anabaena sp., (D) Nostoc sp., (E) Aphanothece conferta, and (F) Synechocystis aquatilis.

tested aliphatic substances was demonstrated. In this building, as well as the growth of cyanobacteria
respect, Al-Hasan et al. (1998) reported important (Soto et al. 1975, Schroeder and Rehm 1981). This
evidence for the potential of cyanobacteria to oxi- may imply that crude oil and n-alkanes do not cause
dize alkanes, namely, that fatty acids resulting from any stress to cyanobacteria (Gaur and Singh 1990).
that oxidation were found esterified in lipid classes In this respect, Gamila et al. (2003) evaluated the
specified of the thylakoids, monogalactosyldiacylgly- biodegradation efficiency of Oscillatoria agardhii
cerols, digalactosyldiacylglycerols, and sulfoquinovo- (nonheterocystous) and Anabaena spharica (hetero-
syldiacylglycerols. In a supporting study, Kuritz and cystous) for petroleum HCs. They reported that
Wolk (1995) showed that two filamentous cyanobac- both cyanobacterial strains revealed a high algal bio-
teria, Anabaena sp. and Nostoc ellipsosporum, have the mass (expressed as chl a lg Æ L)1) in comparison
potential to degrade the highly chlorinated pesti- with that obtained by the control culture and that
cide lindane. The two cyanobacteria could also be the n-alkanes (C10–C24) were more readily degraded
genetically engineered to degrade 4-chlorobenzoate. than polycyclic aromatic HCs (PAHs) by both
A number of studies reported that HCs might strains. On the other hand, they determined that
stimulate the process of photosynthesis and chl n-alkanes were reduced to 99.5% and PAHs to
 BIODEGRADABILITY OF HCs BY CYANOBACTERIA 821

Control Phormidium sp.

60 60

Degradation %
Degradation %
50 50
40
40
30
30
20
20
10
10 0
0 0 20 40 60
0 20 40 60 Time (d)
Time (d)

Phenanthrene Dibenzothiophene Phenanthrene Dibenzothiophene

Anabaena sp. Nostoc sp.

60 60
Degradation %

Degradation %
50
50
40
40
30
20 30
10 20
0 10
0 20 40 60 0
Time (d) 0 20 40 60
Time (d)
Phenanthrene Dibenzothiophene Phenanthrene Dibenzothiophene

Aphanothece conferta Synechocystis aquatilis

60
Degradation %

60
Degradation %

50 50

40 40

30 30

20 20
10 10
0 0
0 20 40 60 0 20 40 60
Time (d) Time (d)

Phenanthrene Dibenzothiophene Phenanthrene Dibenzothiophene

Fig. 2. Biodegradation percentages of phenanthrene and dibenzothiophene by cyanobacterial cultures. (A) Control (natural seawater),
(B) Phormidium sp., (C) Anabaena sp., (D) Nostoc sp., (E) Aphanothece conferta, and (F) Synechocystis aquatilis.

97.5% in Anabaena culture, and a significant (on day 40) and 30% (on day 60) for phenanthrene
(P < 0.05) negative correlation was observed and dibenzothiophene, respectively, by Anabaena
between the microbial growth of Anabaena and the (Fig. 2C), and 43% and 44% at the end of the
total concentration of petroleum HCs. They con- experiment for phenanthrene and dibenzothioph-
cluded that the biodegradation rate of saturated ene, respectively by Nostoc (Fig. 2D).
and aromatic fractions was time dependent, depend- Although high degradation activity was exhibited
ing on the cyanobacterial strains. by Aphanothece and Synechocystis against the two alk-
Aromatic oxidation by cyanobacteria. Data recorded anes, they exhibited different sensitivities against
in Figure 2 demonstrate different biodegradable the aromatic compounds. They exhibited moderate
behaviors in between moderate and weak ability degradation percentages against phenanthrene,
according to cyanobacterium species and the which reached 40% and 46% (on day 40) by
aromatic compound. Phormidium exhibited weak Aphanothece and Synechocystis, respectively (Fig. 2, E
degradation for phenanthrene (12%) and dibenzo- and F). However, they demonstrated weak degrada-
thiophene (17%) on day 60 (see Fig. 2B). Both tion percentages against dibenzothiophene, stopped
Anabaena and Nostoc had moderate degradation at 14% and 21% by Aphanothece and Synechocystis,
percentages for both aromatic compounds: 51% respectively (Fig. 2, E and F). The high efficiency of
822 I B R A H E E M B O R I E M O H A MM A D I B R A H E E M
some investigated cyanobacteria to degrade some of
the tested HCs might contribute to the dominance
of cyanobacteria in many polluted sites including
the contaminated coasts of the Arabian Gulf after
the Gulf War in 1992 in Kuwait, which has given the
impression that cyanobacteria possess the potential
to break down oil components. In this respect,
Sorkhoh et al. (1992), Radwan et al. (1999),
Raghukumar et al. (2001), and De Oteyza et al.
(2004) reported that the cyanobacterial mats were
often reported in oil-contaminated environments,
suggesting that cyanobacteria are able to tolerate oil
contamination and have strong potential to degrade
HCs. This finding is in agreement with previous
research that reported that some aromatic HCs can
be oxidized by the cyanobacterial strains (Ellis
1977). Ellis investigated the phenol and catechol
degradation potential by Anabaena cylindrical and
Phormidium foveolarum. He determined that both
cyanobacteria degrade phenol and catechol. In this
respect, Cerniglia et al. (1979, 1980a) studied the
oxidation of naphthalene by Oscillatoria sp. and
Agmenellum quadruplicatum, which oxidized naphtha-
lene under photautotrophic conditions to 1-naph-
thol, cis-1,2-dihydroxy-12-dihydronaphthalene, and
4-hydroxy-1-tetralonen. They concluded that cyano-
bacteria probably possess both the monooxygenase
and dioxygenase systems that catalyze the initial
step of naphthalene oxidizing. However, some
researchers have reported that these results were
ambiguous and, consequently, that no real ability of
cyanobacteria to degrade crude oil was demon-
strated (Radwan and Al-Hasan 2000). Nevertheless,
some cyanobacteria have been reported to metabo-
lize simple bicyclic aromatic HCs like naphthalene
(Cerniglia et al. 1980a, Narro et al. 1992, De Oteyza
et al. 2004).
Studies have also been conducted on the oxida-
tion of two other aromatic HCs, methylnaphthalene
and biphenyl phenanthrene, by photosynthetic
microorganisms (Smith and Rosazza 1974, Meyer
and Scheline 1976, Wiebkin et al. 1976, Dodge et al.
1979, Cerniglia et al. 1980b). They demonstrated
that the oxidation of methylnaphthalenes and
biphenyl by cyanobacteria is probably similar to that
mediated by fungal and mammalian enzyme sys-
tems and differs from that carried out by bacteria
(Catelani et al. 1971, Gibson et al. 1973). In a sup-
porting study, Cerniglia et al. (1981) reported that
two autotrophically grown cyanobacteria, Agmenellum
quadruplicatum and Oscillatoria sp., metabolize
aniline (aromatic HCs) producing formanilide, acet-
anilide, and p-aminophenol. It is known that bacte-
ria and mammals also metabolize aromatic amines
by N-formylating them (Gothoskar et al. 1979,
Alexander 1981).
Using hydrophobic montmorillonite clay as a car-
rier system in the present work is suitable to accom-
plish close contact of the petroleum compounds
with cyanobacteria to enable biodegradation (Abed
et al. 2002, Grötzschel et al. 2002). It may contrib-
ute to the ability of A. conferta and S. aquatilis to
degrade alkanes and aromatic HCs. On the other
hand, the pigmentation loss and subsequent death
of Phormidium and Nostoc species confirm that petro-
leum compounds can be toxic to certain cyanobac-
terial strains. Many studies have demonstrated that
crude oils are inhibitory to cyanobacteria even at
low concentrations (Vandermeulem and Ahern
1976, Winters et al. 1976, Batterton et al. 1978).
These compounds, particularly the aromatic ones,
which are considered more toxic than the alkanes,
might inhibit photosynthesis and growth and reduce
enzyme activity and microbial biomass (Megharaj
et al. 2000).
In contrast, some researchers have demonstrated
that aerobic heterotrophic bacteria associated with
the mucilaginous coating of cyanobacteria and
macroalgae are actually the principal degraders
(Sorkhoh et al. 1995, Radwan et al. 2002, Abed and
Köster 2005, Chaillan et al. 2006). They reported
that it is difficult to cultivate cyanobacteria in axenic
culture and to clear the culture from naturally asso-
ciated aerobic heterotrophs, which reflects their
dependence on each other. Therefore, they sug-
gested that the ability of axenic cyanobacteria to
degrade petroleum compounds needs to be care-
fully examined.
Other researchers have suggested that cyanobac-
teria play an indirect role in mixed microbial popu-
lations like mats and biofilms (Cerniglia et al. 1984,
Singer and Finnerty 1984). In this respect, Abed
and Köster (2005) suggested that cyanobacteria and
associated bacteria seem to form a consortium favor-
able for biodegradation and cleanup of polluted
sites. They reported that cyanobacteria fuel the
aerobic heterotrophic bacteria with oxygen and
nitrogen as well as organic matter essential for their
activity. In return, aerobic heterotrophs degrade
petroleum compounds, thus reducing their toxic
effect on cyanobacteria. Furthermore, the complete
degradation of petroleum compounds leads to the
regeneration of CO2, which is used by cyanobacteria
for photosynthesis. Abed (2010) reported that addi-
tion of the cyanobacterial exudates to aerobic het-
erotrophic bacteria plays a role in stimulating their
degradative activities. He concluded that the aerobic
heterotrophic bacteria–cyanobacteria consortia can
be very useful for bioremediating oil-polluted sites,
circumventing the costly use of organic and inor-
ganic fertilizers.
In addition, Safonova et al. (1999) have shown
that the presence of alkane-utilizing bacteria in
association with algae and cyanobacteria restores
the growth of sensitive phototrophic strains exposed
to black oil and stimulates the growth of the
tolerant species. They reported that this associa-
tion degraded black oil more efficiently than a
pure culture of bacteria alone. Therefore, they
suggested that phototrophic and heterotrophic
 BIODEGRADABILITY OF HCs BY CYANOBACTERIA
microorganisms form a consortium that increases
their potential to degrade petroleum compounds.
Moreover, several studies have reported that cyano-
bacteria protect the associated bacteria and fungi
from adverse ecological conditions like excessive
light or dryness (Garcia-Pichel and Pringault 2001,
Des Marais 2003).
I would like to thank all of the general practitioners who par-
ticipated in this study. I also greatly appreciate the efforts of
investigators regarding data collection.
Abed, R. M. M. 2010. Interaction between cyanobacteria and aer-
obic heterotrophic bacteria in the degradation of hydrocar-
bons. Int. Biodeterior. Biodegrad. 64:58–64.
Abed, R. M. M. & Köster, J. 2005. The direct role of aerobic
heterotrophic bacteria associated with cyanobacteria in the
degradation of oil compounds. Int. Biodeterior. Biodegrad.
55:29–37.
Abed, R. M. M., Safi, N. M. D., Köster, J., de Beer, D., El-Nahhal, Y.,
Rullkötter, J. & Garcia-Pichel, F. 2002. Microbial diversity of a
heavily polluted microbial mat and its community changes
following degradation of petroleum compounds. Appl. Envi-
ron. Microbiol. 68:1674–83.
Alexander, M. 1981. Biodegradation of chemicals of environmental
concern. Science 211:132–8.
Al-Hasan, R. H., Al-Bader, D. A., Sorkhoh, N. A. & Radwan, S. S.
1998. Evidence for n-alkane consumption and oxidation by
filamentous cyanobacteria for oil-contaminated coasts for the
Arabian Gulf. Mar. Biol. 130:521–7.
Atlas, R. M. 1984. Petroleum Microbiology. Macmillan Publishing
Company, New York, 692 pp.
Batterton, J., Winters, K. & Van Baalen, C. 1978. Anilines: selective
toxicity to blue-green algae. Science 199:1068–70.
Bligh, E. G. & Dyer, W. J. 1959. A rapid method of total
lipid extraction and purification. Can. J. Biochem. Physiol.
37:911–7.
Catelani, D., Sorlini, C. & Treccani, V. 1971. The metabolism of
biphenyl by Pseudomonas putida. Experientia 27:1173–4.
Cerniglia, C. E., Freeman, J. P. & Van Baalen, C. 1981. Biotrans-
formation and toxicity of aniline and aniline derivatives in
cyanobacteria. Arch. Microbiol. 130:272–5.
Cerniglia, C. E., Gibson, D. T. & Van Baalen, C. 1979. Algal oxi-
dation of aromatic hydrocarbons: formation of 1-naphthol
from naphthalene by Agmenellum quadruplicatum, strain PR-6.
Biochem. Biophys. Res. Commun. 88:50–8.
Cerniglia, C. E., Gibson, D. T. & Van Baalen, C. 1980a. Oxidation of
naphthalene by cyanobacteria and microalgae. J. Gen. Micro-
biol. 116:495–500.
Cerniglia, C. E., Lambert, K. J., Miller, D. W. & Freeman, J. P. 1984.
Transformation of 1- and 2-methylnaphthalene by Cuning-
hamella elegans. Appl. Environ. Microbiol. 47:111–8.
Cerniglia, C. E., Van Baalen, C. & Gibson, D. T. 1980b. Oxidation
of biphenyl by the cyanobacterium Oscillatoria sp. strain JCM.
Arch. Microbiol. 125:203–7.
Chaillan, F., Gugger, M., Saliot, A., Coutè, A. & Oudot, J. 2006. Role
of cyanobacteria in the biodegradation of crude oil by a
tropical cyanobacterial mat. Chemosphere 62:1574–82.
Chaillan, F., Le Flèche, A., Bury, E., Phantavong, Y., Grimont, P.,
Saliot, A. & Oudot, J. 2004. Identification and biodegradation
potential of tropical aerobic hydrocarbon-degrading microor-
ganisms. Res. Microbiol. 155:587–95.
De Oteyza, T. G., Grimalt, J. O., Diestra, E., Sole, A. & Esteve, I.
2004. Changes in the composition of polar and a polar crude
oil fractions under the action of Microcoleus consortia. Appl.
Microbiol. Biotechnol. 66:226–32.
Des Marais, D. J. 2003. Biogeochemistry of hypersaline microbial
mats illustrates the dynamics of modern microbial ecosystems
and the early evolution of the biosphere. Biol. Bull. 204:160–
7.
823
Desikachary, T. V. 1959. Cyanophyta. PhD thesis, Botany Depart-
ment, Faculty of Science, Madras University, India, Part II,
pp. 71–669.
Dodge, R. H., Cerniglia, C. E. & Gibson, D. T. 1979. Fungal
metabolism of biphenyl. Biochem. J. 178:223–30.
Ellis, B. E. 1977. Degradation of phenolic compounds by fresh-
water algae. Plant Sci. Lett. 8:213–6.
El-Nahhal, Y., Nir, S., Serban, C., Rabinovitch, O. & Rubin, B. 2000.
Montmorillonite-phenyltrimethyl ammonium yields environ-
mentally improved formulations of hydrophobic herbicides.
J. Agric. Food. Chem. 48:4791–801.
Felfoldy, L. J. M. & Zsuzsa, F. K. 1959. Some methodical observa-
tions on the use of antibiotics for preparing bacteria-free algal
cultures. Acta Biol. Acad. Sci. Hung. 10:95.
Gamila, H. A., Ibrahim, M. B. M. & Abd El-Ghafar, H. H. 2003. The
role of cyanobacterial isolated strains in the biodegradation of
crude oil. Int. J. Environ. Stud. 60:435–44.
Garcia-Pichel, F. & Pringault, O. 2001. Microbiology: cyanobacteria
track water in desert soils. Nature 413:380–1.
Gaur, J. P. & Singh, A. K. 1990. Growth, photosynthesis
and nitrogen fixation of Anabaena doliolum exposed to
Assam-crude extract. Bull. Environ. Contam. Toxicol. 144:
494–500.
Gibson, D. T., Roberts, R. L., Wells, M. G. & Kobal, V. M. 1973.
Oxidation of biphenyl by a Beijerinckia species. Biochem.
Biophys. Res. Commun. 50:211–9.
Gothoskar, S. V., Benjamin, T., Roller, P. P. & Weisburger, E. K.
1979. N-Formylation of an aromatic amine as a metabolic
pathway. Xenobiotica 9:533–7.
Grötzschel, S., Köster, J., Abed, R. M. M. & de Beer, D. 2002.
Degradation of petroleum compounds immobilized on clay by
a hypersaline mat. Biodegradation 13:273–83.
Hoshaw, R. W. & Rosowski, J. R. 1973. Methods for microscopic
algae. In Stein, J. R. [Ed.] Handbook of Phycological Methods.
Cambridge University Press, London, pp. 56–64.
Kuritz, T. & Wolk, P. 1995. Use of filamentous cyanobacteria for
biodegradation of organic pollutants. Appl. Environ. Microbiol.
61:234–8.
Leahy, J. G. & Colwell, R. R. 1990. Microbial degradation of
hydrocarbons in the environment. Microbiol. Mol. Biol. Rev.
54:305–15.
Lee, C. C., Craig, W. K. & Smith, P. J. 1974. Water-soluble hydro-
carbons from crude oil. Bull. Environ. Contam. Toxicol. 12:
212–6.
Mansy, A. E. & El-Bestway, E. 2002. Toxicity and biodegradation
of fluometuron by selected cyanobacterial species. World J.
Microbiol. Biotechnol. 18:125–31.
McCann, J., Choi, E., Yamasaki, E. & Ames, B. N. 1975. Detection of
carcinogens as mutagens in the Salmonella microsome test: assay
of 300 chemicals. Proc. Natl. Acad. Sci. U. S. A. 72:5135–9.
Megharaj, M., Singleton, L., McClure, N. C. & Naidu, R. 2000.
Influence of petroleum hydrocarbon contamination on mic-
roalgae and microbial activities in a long-term contaminated
soil. Arch. Environ. Contam. Toxicol. 38:439–45.
Meyer, T. & Scheline, R. R. 1976. The metabolism of biphenyl. II.
Phenolic metabolites in the rat. Acta Pharmacol. Toxicol.
39:419–41.
Miller, E. C. & Miller, J. A. 1974. Biochemical mechanisms of
chemical carcinogenesis. In Bush, H. [Ed.] Molecular Biology of
Cancer. Academic Press, New York, pp. 377–402.
Narro, M. L. 1985. Oxidation of aromatic hydrocarbons by
marine cyanobacteria. PhD thesis, University of Texas at
Austin, 73 pp.
Narro, L. N., Cerniglia, C. E., Van Baalen, C. & Gibson, D. T. 1992.
Evidence for an NIH shift in oxidation of naphthalene by
the marine cyanobacterium Oscillatoria sp. strain JCM. Appl.
Environ. Microbiol. 58:1360–3.
NRC. 1985. Oil in the Sea. National Academy Press, National
Research Council, Washington, D.C., 601 pp.
Oudot, J., Dupont, J., Haloui, S. & Roquebert, M. F. 1993. Bio-
degradation potential of hydrocarbon-assimilation tropical
fungi. Soil Biol. Biochem. 25:1167–73.
824 I B R A H E E M B O R I E M O H A MM A D I B R A H E E M
Pringsheim, E. G. 1949. Pure Culture of Algae, Their Preparation and
Maintenance. Cambridge University, Cambridge, UK, xii + 119
pp.
Radwan, S. S. & Al-Hasan, R. H. 2000. Oil pollution and cyano-
bacteria. In Whitton, B. A. & Potts, M. [Eds.] The Ecology of
Cyanobacteria. Kluwer, the Netherlands, pp. 307–19.
Radwan, S. S., Al-Hasan, R. H., Al-Awadhi, H., Salamah, S. &
Abdullah, H. M. 1999. Higher oil biodegradation potential at
the Arabian Gulf coast than in the water body. Mar. Biol.
135:741–5.
Radwan, S. S., Al-Hasan, R. H., Salamah, S. & Al-Dabbous, S. 2002.
Bioremediation of oily sea water by bacteria immobilized
in biofilms coating macroalgae. Int. Biodeterior. Biodegrad.
50:55–9.
Raghukumar, C., Vipparty, V., David, J. J. & Chandramohan, D.
2001. Degradation of crude oil by marine cyanobacteria. Appl.
Microbiol. Biotechnol. 57:433–6.
Rippka, R. 1988. Isolation and purification of cyanobacteria. Meth-
ods Enzymol. 167:3–27.
Safonova, E. T., Dmitrieva, I. A. & Kvitko, K. V. 1999. The interac-
tion of algae with alcanotrophic bacteria in black oil decom-
position Resources. Conserv. Recycling 27:193–201.
Schroeder, E. & Rehm, H. J. 1981. Degradation of long chain
n-alkanes by Chlorococcales. Eur. J. Appl. Microbiol. Biotechnol.
12:36–8.
Singer, M. E. & Finnerty, W. R. 1984. Microbial metabolism of
straight-chain and branched alkanes. In Atlas, R. M. [Ed.]
Petroleum Microbiology. Macmillan, New York, pp. 1–60.
Smith, G. M. 1950. The Freshwater Algae of the United States. McGraw-
Hill, New York, 719 pp.
View publication stats
Smith, R. & Rosazza, J. 1974. Microbial models of mammalian
metabolism. Aromatic hydroxylation. Arch. Biochem. Biophys.
161:551–8.
Sorkhoh, N. A., Al-Hasan, R. H., Khanafer, M. & Radwan, S. S. 1995.
Establishment of oil-degrading bacteria associated with
cyanobacteria in oil-polluted soil. J. Appl. Bacteriol. 78:194–9.
Sorkhoh, N., Al-Hasan, R., Radwan, S. & Hopner, T. 1992. Self-
cleaning of the Gulf. Nature (Lond.) 359:109.
Soto, C., Hellebust, J. A., Hutchinson, T. C. & Sawa, T. 1975. Effect
of naphtha and aqueous crude oil extracts on the green flag-
ellated Chlamydomonas angulosa. I. Growth. Can. J. Bot. 53:109–
17.
U.S. Coast Guard. 1990. Update of Inputs of Petroleum Hydrocarbons
into the Oceans Due to Marine Transportation Activities. National
Academy Press, Washington, D.C., 13 pp.
Vandermeulem, J. H. & Ahern, T. P. 1976. Effect of petroleum
hydrocarbons on the algal physiology: review and progress
reports. In Lockwood, A. P. M. [Ed.] Effects of Pollution on
Aquatic Organisms. Cambridge University Press, London,
pp. 107–25.
Wiebkin, P., Fry, J. R., Jones, C. A., Lowing, R. & Bridges, J. W. 1976.
The metabolism of biphenyl by isolated viable rat hepatocytes.
Xenobiotica 6:725–43.
Winters, K., O’Donnell, R., Batterton, J. C. & Van Baalen, C. 1976.
Water-soluble compounds of four fuel oil: chemical charac-
terization and effects on growth of microalgae. Mar. Biol.
36:269–76.
Yan, G. A., Jiang, J. W., Wu, G. & Yan, X. 1998. Disappearance of
linear alkylbenzene sulfonate from different cultures with An-
abaena sp. HB 1017. Bull. Environ. Contam. Toxicol. 60:329–34.