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Bioremediation of arsenic using algae,
bacteria and fungi
Mohamed Wahby Hussein
M.Sc. Candidate
Faculty of science - Alexandria University
16 May 2013
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Introduction
The global water budget contains about 97.2% saltywater, mainly in oceans, and only 2.8% is available as
fresh water at any time on the planet. Out of this
2.8% of fresh water, about 2.2% is available as
surface water and 0.61% as ground water. Water that
collects below the land surface in soil, sediment, and
permeable rock strata is called groundwater(Ehrlich,
H.L., Newman, D.K., 2009).
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Introduction (contd.)
Out of those 0.61% of stored ground water, only
about 0.25% can be economically extracted with the
present drilling technology (the remaining being at
greater depths). Thus, it can be said that the groundwater is the most possible accessible water source
on earth (Raghunath, 2006)
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Arsenic definition
Arsenic is a toxic metalloid found in rocks, soil, water,
sediments, and air. Despite its low crustal abundance
(0.0001%), it is widely distributed in nature and is
commonly associated with the ores of metals likecopper, lead, and gold (Johri, 2012).
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Mobilization and accumulation
Mobilization of arsenic due to the oxidation of
arsenic bearing pyrite minerals. Insoluble arsenic
bearing minerals such as arsenopyrite (FeAsS) are
rapidly oxidized when exposed to atmosphere,releasing soluble arsenite As(III), sulphate (SO4-2),
and ferrous iron Fe(II).
FeAsS +13Fe+3 + 8H2O = 14 Fe+2 + SO4
-2 +13H+ +H3AsO4 (Aq.)
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Mobilization and accumulation
Dissolution of arsenic rich iron oxyhydroxides
(FeOOH) due to onset of reducing conditions in the
subsurface.
Under oxidizing conditions, and in the presence ofFe, inorganic species of arsenic are predominantly
retained in the solid phase through interaction with
FeOOH coatings on soil particles.
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Mobilization and accumulation
Release of arsenic sorbed to aquifer minerals by
competitive exchange with phosphate (H2PO-4) ions
that migrate into aquifers from the application of
fertilizers to surface soil. The second mechanism involving dissolution of
FeOOH under reducing conditions is considered to
be the most probable reason for excessive arsenic
accumulation in groundwater.
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Mobilization and accumulation
Volcanoes are also considered as a geological source
of arsenic to the environment with the total
atmospheric annual emissions from volcanoes being
estimated at 31,000 mg/year.
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Sources of arsenic
From anthropogenic activities, arsenic discharged
onto land originate from commercial wastes (~40%),
coal ash (~22%), mining industry (~16%), and the
atmospheric fallout from the steel industry (~13%).
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Sources of arsenic
Biological sources
contribute only small
amounts of arsenic
into soil and waterecosystems. Arsenic
accumulates readily in
living tissues because
of its strong affinity forproteins, lipids, and
other cellular
components .Mohamed Wahby - M.Sc. candidate -
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Accumulation and transportation in
food chain
Aquatic organisms canaccumulate arsenic easily,thus accumulatingconsiderably higher
concentrations than theirsurroundings (i.e.,biomagnification). Theseorganisms contribute to
environmentalcontamination uponconsumption ordisposal/degradation.
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Effect of arsenic on human health
Arsenic problem was first identified in West Bengal in
the 1990s when people started showing typical signs
of arsenicosis, beginning with skin rashes and leading
to cancers of major organs such as the lungs,kidneys, and bladder.
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Effect of arsenic on human health
(Contd)
There are millions of people are at risk of developing
health effects associated with the ingestion or
arsenic. A number of large aquifers in various parts
of the world have been identified with arsenic
occurring at concentrations above 10 g/L, the
maximum concentration limit (MCL) recommended
for drinking water by the World Health Organization.
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Algae in bioremediation
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Algae in bioremediation
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Bioremediation (Algae)
The alga releases metallothioneins which chemically
binds to the metal as a defense mechanism to
remove the metal from its regular cellular activity.
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Bioremediation (Algae)
Chlorella sp. and Scenedesmus sp. are the two most
common algae species used for metal uptake.
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Bioremediation (Algae)
Chlorella sp. was exposed to concentrations of arsenite
ranging from 0 to 100 g/mL. The cell growth ofChlorella sp.
was not affected by the arsenite until it was exposed to
concentrations higher than 50 g/mL.
At concentrations greater than 50 g As/mL, the cell growth
of the species was suppressed. It was concluded that
Chlorella sp. retained approximately 50% of arsenite from a
solution. Also, it was observed that most of the biosorption
was rapid and occurred in the first 15 min by the Chlorellasp.
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Scenedesmusabundans as a possible cost effective
method of bioremediation of arsenic from water
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Bioremediation (Algae)
Scenedesmusabundans biomass (40 mg/L) wasexposed to varying concentrations of As(III) i.e. 1, 5,10, 20, 50 and 100 mg/L and samples were obtainedat certain time intervals to analyze for residual
arsenic concentrations.
The removal of As(III) was found to be around 70%.Algal morphology changed in presence of arsenic.Sorption of arsenic with algae could be modelled by
the conventional Langmuir isotherm. The isothermconstants indicate a high adsorptive capacity of theselect alga for arsenic.
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Bioremediation (Algae)
It was studied that As (V) biosorption using dried
algae (Lessonia nigrescens) collected in Valparaiso
bay, Chile. The experiments were performedusing
laboratory solutions (200 mg/L, pH 2.5, 4.5 and 6.5). Lessonia nigrescens showed very good adsorption
capacities and its use may be interesting for small
scaledrinking water treatment, deserving further
investigation.
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Lessonia nigrescens
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Bioremediation (Algae)
It was investigated the effectiveness and suitability of
driedmacro-algae (Spyrogira spp.) in removing
arsenic from acid mine drainage(AMD) and other
waters from the Poopo lake basin (Bolivia, Andean
highlands)finding higher efficiency i.e. 8090% of As
removal was attained within 4 days.
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Bioremediation (Bacteria)
Corynebacterium glutamicum, which is used for the
industrial production of amino acids and nucleotides,
is one of the most arsenic-resistant microorganisms
described to date (up to 12 mM arsenite and >400mM arsenate).
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Analysis of the C. glutamicum genome revealed the
presence of two complete ars operons ( ars1 and
ars2 ) comprising the typical three-gene structure
arsRBC, with an extra arsC1located downstream
from arsC1 ( ars1 operon), and two orphan genes (
arsB3 and arsC4).
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It was confirmed that the involvement of both ars
operons in arsenic resistance in C. glutamicum by
disruption and amplification of those genes.
The strains obtained by them were resistant to up to60 mM arsenite, one of the highest levels of
bacterial resistance to arsenite so far described.
They are attempting to obtain C. glutamicum mutant
strains able to remove arsenic from contaminatedwater.
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Bioremediation (Bacteria)
Also, It was reported that arsenic removal by three
bacterial strains namely, Ralstonia eutropha MTCC
2487, Pseudomonas putida MTCC 1194 and Bacillus
indicus MTCC 4374, from wastewater (pH 7.1, 29o
C)containing 15 mg/L arsenic were 67%, 60% and 61%,
respectively. It was also observed that arsenic
concentration of 15 mg/L prolonged the stationary
phase of these strains.
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Genetically engineered organisms
Genetically engineering methods could be used to
enhance intracellular accumulation of both As(III)
and As(V).
It was found that Escherichia coliover expressing
ArsR accumulated 5- and 60-fold-higher levels of
As(III) and As(V) than cells withoutArsR over
expression. The level of arsenic accumulation was
1.5 2.2 nmol/mg dry weight (110173 m gAs/g dw).The engineered cells removed 98% of 0.05 mg/L
As(III) from contaminated water after 1 h.
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Bioremediation (fungi)
It was investigated the bioaccumulation of arsenic inthree filamentous fungi,Aspergillus niger, Serpulahimantioides and Trametes versicolorand theirpossible application in remediation of arsenic.
They were exposed to arsenopyrite (FeAsS) inconcentrations 0.2%, 0.4%, 0.6% and 0.8% (W/V).
T. versicolorwas the most efficient in accumulationwith all amounts, accumulating up to 15 times the
amounts accumulated byA . nigerwhich was the leasteffective in accumulation.
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Bioremediation (fungi)
It was reported that the maximum adsorption
capacities of As(III) onto Penicillium purpurogenum
fungal biomass reached 35.6 mg/g under non-
competitive conditions and 3.4 mg/g undercompetitive conditions by other ions [e.g., Cd(II),
Pb(II), Hg(II)] after 4 h (pH 5, 20C). The fungus could
be used for ten cycles for biosorption.
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Penicillium purpurogenum
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Bioremediation (fungi)
It was reported that Penicillium chrysogenum (a
waste byproduct from antibiotic production) pre-
treated with surfactants (hexadecyl-
trimethylammonium bromide and dodecylamine)and a cationic polyelectrolyte was able to remove
significant amounts of As(V) from waters. At pH 3,
the removal capacities of the modified biomass
ranged from 33.3 to 56.1 mg arsenic/g biomass.
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Methodology
1. Site description and sample collection
2. Chemical analysis for determination of arsenicconcentration
3. Enrichment and isolation of arsenic-resistantbacteria
4. Determination of isolates ability of arsenictransformation and bioaccumulation:
Screening of the arsenate reducing and oxidizingbacteria
Ability of bacteria on bioaccumulation arsenic
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Methodology
5. Identification of most promising organism
5.1. Phenotypic identification
5.1.1. Morphological characterization
5.1.2. Biochemical characterization
5.1.3. Physiological characterization
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Methodology
5.2. Genetic identification of isolated
microorganisms
5.2.1. Isolation of bacterial DNA
5.2.2. PCR amplification of 16S ribosomal
(r)RNA
5.2.3. Analysis of sequence data
6. Determination of maximum tolerance
concentration
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References
Ehrlich, H.L., Newman, D.K. (2009). Geomicrobiology(Fifth edition ed.). NewYork, USA: Taylor & Francis Group
Raghunath, H. (2006). Hydrology, Principles, analysis, design. New Delhi: NewAge International
Johri, B. P. (2012). Microorganisms in environmental management, microbesand environment. New York: Springer
Bolan, N. S. (2008). Manipulating bioavailability to manage remediation ofmetal-contaminated soils. Developments in Soil Science, 32 , 657-678.
Ferguson, J. F., & Gavis, J. (1972). A review of the arsenic cycle in naturalwaters. Water research, 6(11) , 1259-1274.
WHO. (2001).Arsenic in drinking-water, background document fordevelopment of WHO guidelines for drinking-water. Geneva: WHO.
WHO. (1996). Guidelines for drinking water; Health criteria and othersupporting information. Geneva, Switzerland: World Health Organization.
Wang, S., & Zhao, X. . (2009). On the potential of biological treatment forarsenic contaminated soils and groundwater.Journal of environmentalManagement, 2367-2376.