BIOREMEDIATION AND

PHYTOREMEDIATION

Gizel Rivera-Santiago
Graduate Student
Philippine Normal University
Taft Avenue, Manila
1
 OUTLINE

1 Sub Topic 2 Sub Topic

Bioremediati Phytoremediati
on on

2
1 BIOREMEDIATION

What is
Bioremediation?
3
Use of different
biological systems to
destroy or reduce
concentrations of
contaminants from
polluted sites.
https://www.emaze.com/@ALCTRFZ
T/Presentation-Name
(Rittmann, B. E, McCarty, P. L. 2001)

4
 Microbes and plants have a
natural capability to attenuate
or reduce:
Mass
Toxicity
Volume
Concentration of
pollutants
https://explorecuriocity.org/Explore/Articl
without human eId/2833/bioremediation.aspx

interventions (Rittmann, B. E, McCarty, P. L. 2001) 5

Conventional methods of remediation
Dig up and remove it to a
landfill

Cap and contain

Maintain it in the same land but
isolate it
6
Is there a better approach?

7
Better approaches
Destroy them completely, if possible

Transform them into harmless substances

8
But, are they effective?

9
Drawbacks
Technological complexity
The cost for small scale application expensive
Lack of public acceptance especially in
incineration

10
Drawbacks
Fall back on earth and pollute some other
environment.
Dioxin production due to burning of plastics leads
to cancer.
May increase the exposure to contaminants, for both
workers and nearby residents.
11
 BIOREMEDIATION

Use of Microorganisms

Use of plants

12
Bioremediation mediated biodegradation
in general it is bio mediated decomposition of
paper, paint, textiles, hydrocarbons and other
pollutants.
Superior technique over using chemicals
why?
Biodegradation is the initial process that results
to bioremediation. (Marshall, F. M., 2009)
13
 Enzymatic processes in bioremediation
Major types of reactions
Oxidation.
Decarboxylation in which the -CO2H is replaced with
an H atom or OH group.
Hydrolysis which involves the addition of H2O to a
molecule accompanied by cleavage of the molecule
into two species.
Substitution in which one group of atom is replaced by
another (such as OH for Cl- ).
(Marshall, F. M., 2009) 14
 Enzymatic processes in bioremediation
Major types of reactions
Elimination whereby atoms or group of atoms are removed
from adjacent carbon atoms, which remained joined by a
double bond.
Reduction, dehalogenation , demethylation, deamination,
condensation, in which two smaller molecules are joined to
produce a larger one: conversion of one isomer of a
compound to another with a same molecular formula but
different structure ; conjugation; ring cleavage.
(Marshall, F. M., 2009)
15
Biodegradation has at least 3 outcomes:
1. A minor change in an organic molecule leaving the main
structure intact.
2. Fragmentation of a complex organic structure in such a way
that the fragments could be reassembled to yield the
original structure.
3. Complete mineralization, which in the transformation of
organic molecules to mineral forms.
One example to describe all 3 types
2, 6-Dichlorobenzonitrile (Marshall, F. M., 2009)16
Minor change in a molecule (Dehalogenation)
Cl

Cl C N HOH

2, 6-Dichlorobenzonitrile
OH

Cl C N Cl

Cl is replaced with OH (Prasad MNV., 2003)17

Fragmentation
Cl

Cl C N HOH

2, 6-Dichlorobenzonitrile
OH

OH OH Cl

NH2CH2 Cl is replaced with OH

(Prasad MNV., 2003) 18
 Mineralization
Cl

Cl C N

2, 6-Dichlorobenzonitrile

HOH 2Cl NH3

Completely converted into inorganic forms

(Prasad MNV., 2003)
19
 Bioremediation Effectiveness
Depends on:
Microorganisms
Environmental factors
Contaminant type &
state

(Prasad MNV., 2003) 20

 Microorganisms
Aerobic bacteria:
Examples include: Pseudomonas, Alcaligenes,
Sphingomonas, Rhodococcus, and
Mycobacterium.
Methanotrophs:
Aerobic bacteria that utilize methane for
carbon and energy.
Methane monooxygenase has a broad
substrate range. 21
(Bodishbaugh, D.F., 2006)
 Microorganisms
Anaerobic bacteria:
Not used as frequently as aerobic bacteria.
Can often be applied to bioremediation of
polychlorinated biphenyls (PCBs) in river sediments,
trichloroethylene (TCE) and chloroform.
Fungi:
Able to degrade a diverse range of persistent or
toxic environmental pollutants.

(Bodishbaugh, D.F., 2006)

22
 How Microbes Use the Contaminant
Contaminants may serve as:
Primary substrate
enough available to be the sole energy source.
Secondary substrate
provides energy, not available in high enough concentration.
Co metabolic substrate
Utilization of a compound by a microbe relying on some other
primary substrate.
(Bodishbaugh, D.F., 2006)23
(Bodishbaugh, D.F., 2006) 24
(Bodishbaugh, D.F., 2006)
25
 Environmental Factors

Environmental Optimum conditions Condition required

Factor for microbial Activity

Available soil 25-85% water holding capacity 25-28% of water holding

moisture capacity
Oxygen >0.2 mg/L DO, >10% air-filled pore Aerobic, minimum air-filled
space for aerobic degradation pore space of 10%
Redox potential Eh > 50 milli volts
Nutrients C:N:P= 120:10:1 molar ratio N and P for microbial
growth
pH 6.5-8.0 5.5 to 8.5
Temperature 20-30 C 15-45C
Contaminants Hydrocarbon 5-10% of dry weight Not too toxic
of soil
Heavy metals 700ppm Total content 2000ppm
26
(Vidali , 2007)
 Type of contaminants
Bio-degradable
petroleum products (gas, diesel, fuel oil)
crude oil compounds (benzene, toluene,
xylene, naphthalene)
some pesticides (malathion) some industrial
solvents
coal compounds (phenols, cyanide in coal
tars and coke waste)
27
 Type of contaminants
Partially degradable / Persistent
TCE (trichlorethane) threat to ground water
PCE (perchloroethane) dry cleaning solvent
Arsenic, Chromium, Selenium
Not degradable / Recalcitrant
Uranium
Mercury
DDT

28
Some m.o. involved in the biodegradation of organic pollutants

Organic Pollutants Organisms

Phenolic - Achromobacter, Alcaligenes,

compound Acinetobacter, Arthrobacter,

Azotobacter, Flavobacterium,
Pseudomonas putida
- Candida tropicalis
Trichosporon cutaneoum
- Aspergillus, Penicillium

Benzoate & related Arthrobacter, Bacillus spp.,

compound Micrococcus, P. putida

29
Organic Pollutants Organisms
Hydrocarbon E. coli, P. putida, P. Aeruginosa
Surfactants Alcaligenes, Achromobacter,
Bacillus, Flavobacterium,
Pseudomonas, Candida

Pesticides P. Aeruginosa
DDT Arthrobacter, P. cepacia

BHC P. cepacia

Parathion Pseudomonas spp., E. coli,

P. aeruginosa

30
(Vidali, 2007)
Criteria for Bioremediation Strategies
1. Organisms must have necessary catabolic
activity required for degradation of
contaminant at fast rate to bring down the
concentration of contaminant.
2. The target contaminant must have
bioavailability.
31
Criteria for Bioremediation Strategies

3. Soil conditions must be favourable for

microbial/plant growth and enzymatic
activity.
4. Cost of bioremediation must be less than
other technologies of removal of
contaminants.
32
 Bioremediation Strategies

(Barathi S and Vasudevan N, 2001) 33

 Bioremediation Strategies

In situ Bioremediation
(at the site)

Ex situ Bioremediation
(away from the site)
(Barathi S and Vasudevan N, 2001) 34
 In Situ Bioremediation
In situ biodegradation involves supplying oxygen
and nutrients by circulating aqueous solutions
through contaminated soils to stimulate naturally
occurring bacteria to degrade organic contaminants.
It can be used for soil and groundwater.
It is the most commonly used type of bioremediation
because it is the cheapest and most efficient, so its
generally better to use.
(Wood TK , 2008) 35
 Types of In situ Bioremediation
Doing nothing

Engineered Bioremediation Intrinsic Bioremediation

(Wood TK , 2008) 36
Intrinsic Bioremediation

(Barathi S and Vasudevan N., 2001)37

 Engineered Bioremediation
The second approach involves the introduction
of certain microorganisms to the site of
contamination.
When site conditions are not suitable,
engineered systems have to be introduced to
that particular site.
38
(Barathi S, Vasudevan N., 2001)
 Insitu Engineered bioremediation types
Bioventing

39
(Vidali,M., 2001)
Biosparging

40
(Vidali,M.2001)
Bioaugmentation

41
(Rittmann B.E and McCarty, P.L. 2001)
Ex situ engineered bioremediation Strategies

(Source: http://ndpublisher.in/ndpjournal.php?j=IJAEB) 42
Solid phase system Ex Situ Bioremediation

43
(Source: https://www.google.co.in/search?q=bioremediation+images)
Land farming Operation

(Rittmann, B.E and McCarty, P.L, 2001) 44

Biopile System

45
(Rittmann,B.E and McCarty,P.L.2001)
Bioremediation using bioreactor System

46
(Rittmann,B.E and McCarty,P.L.2001)
 Case study: Oil degradation
Prof. Chakrabarty discovered a method for genetic
cross-linking that fixed all four plasmid genes in place
and produced a new, stable, bacteria species (now
called pseudomonas putida) capable of consuming oil
one or two orders of magnitude faster than the previous
four strains of oil-eating microbes.
The new microbe, which Chakrabarty called "multi-
plasmid hydrocarbon-degrading Pseudomonas," could
digest about two-thirds of the hydrocarbons that would
47
 By use of genetic engineering:
a). Plasmid transfer:
CAM OCT XYL NAH

Recombination Non-recombination

CAM + OCT XYL + NAH

SUPERBUG (Dowling, DN and Doty, SL. 2009)

48
Biodegradation of hydrocarbons and petroleum

Source: https://www.google.co.in/search?q=bioremediation+images 49
Use of bioremediation strategies over different years by developed
countries ( in percent)
80

70

60

50

40

30

20
2001 2002 2003 2004 2005 2006 2007 2008 2009 2010 2011 2012
50
Source: http://ndpublisher.in/ndpjournal.php?j=IJAEB
Review of bioremediation strategies

51
(Rittmann B E and McCarty P L, 2001)
2 PHYTOREMEDIATION

What is
Phytoremediati
on?
52
Phyto= Plant (in
Greek)
Remediare= To
remedy (in Latin)

53
NEED FOR THE NEW REMEDIATION TECHNIQUE

Microbial/ Biological Measures

Chemical Measures

54
 VARIOUS PHYTOREMEDIATION PROCESSES

Phytoextraction
Rhizofiltration
Phytostabilization
Phytotransformation
Rhizosphere
Bioremediation

55
PHYTOEXTRACTION

56
PHYTOSTABILIZATION

57
PHYTOTRANSFORMATION

58
RHIZOFILTRATION

59
RHIZOSPHERE BIOREMEDIATION

60
 PHYTOREMEDIATION BY AQUATIC PLANTS

Water Hyacinth (Eichhornia crassipes ) Fool's Watercress (Apium nodiflorum)

Eurasian Watermilfoil (Myriophyllum spicatum) Duckweed (Lemna trisulca L.) 61

 Water Hyacinth
Floating plant with broad
,thick, and glossy leaves
that the plant body can
grow as much as 1m
high.
Able to phytoaccumulate
metal pollutants contain
Ag, Pb, Cd and Zn in
municipal and
agricultural wastewater. 62
Case Study: Removal of Cadmium and Zinc by Water Hyacinth

The stock solution was prepared in distilled water with

analytical grade CdCl2. 2 H2O and ZnSO4.7H2O which
was later diluted as required. The plants were maintained in
tap water with concentrations of 0.5, 1, 2, 4 mg/L of Cd and
5, 10, 20, 40 mg/L of Zn.
The test durations were 0 (two hours), 4, 8 and 12 days.
Relative growth, metal accumulation, and bioconcentration
factor (BCF) are evaluated.
63
Case Study: Removal of Cadmium and Zinc by Water Hyacinth

Relative growth
(above) and BCF
(below)

64
 Relative plant Metal BCF
growth Accumulation

C C
Cd d d
Z Z Z
n n n

(Lu et al., 2004)

65
Duckweed
Has a very simple
structure that lacks
obvious stems or
leaves, with small
plate-shaped
structure floating on
water surface.
(Kara and Kara, 66
2004)
Case Study: Removal of Cadmium by Duckweed
The duckweed obtained from natural lake
was acclimatized to laboratory conditions for
one week before starting research.
Solution of Cadmium was prepared using
Cd(NO3)2 and contact with plant sample for
different length.
After absorption, water samples were
(Kara and Kara,

analyzed by AAS at 228.8nm. 2004) 67

Case Study: Removal of Cadmium by Duckweed

68
Cd removal efficiencies (Kara and Kara,
 ADVANTAGES

Cost effective when compared to other more

conventional methods.
natural method, more aesthetically pleasing.
minimal land disturbance.
reduces potential for transport of contaminants by
wind/soil erosion.
multiple contaminants can be removed with the same
plant.
It is one of the least harmful method.
The possibility of recovery and reuse of valuable
metals.(Phyto mining) 69
 DISADVANTAGES
Disadvantages
Slow rate and difficult to achieve acceptable levels of
decontamination.
Possibility of contaminated plants entering the food chain.

Possible spread of contaminant through falling leaves.

Trees and plants require care.

Contaminant might kill the tree.

Degradation product could be worse than original contaminant.

Only surface soil (root zone) can be treated.

Cleanup takes several years. 70

Although much remains to be studied,
Phytoremediation will clearly play some role
in the stabilization and remediation of many
contaminated sites.
The main factor driving the implementation of
Phytoremediation projects are low costs with
significant improvements in site aesthetics
and the potential for ecosystem restoration.
71
REFERENCES:
Majeti Narasimha Vara Prasad,et al, Metal hyperaccumulation in plants - Biodiversity
prospecting for phytoremediation technology, Electronic Journal of Biotechnology
6(3),2003,450 -458.

Yeim Kara,et al, Removal of Cadmium from Water Using Duckweed (Lemna
trisulca L.), International Journal Of Agriculture & Biology 4,2005,660-662.

Angela Pierre Vitria,et al, Structural and Ecophysiological Alterations of the Water
Hyacinth [Eichhornia crassipes (Mart.) Solms] Due to Anthropogenic Stress in
Brazilian Rivers, Brazilian Archives Of Biology And Technology 54(5),2011, 1059-
1068.

Mesania Rizwana,et al, Phytoremediation of Textile Waste Water Using Potential

Wetland Plant: Eco Sustainable Approach, International Journal of Interdisciplinary
and Multidisciplinary Studies (IJIMS) 1(4),2014,130-138.

72
 REFERENCES

Divya Singh,et al, Phytoremediation of lead from wastewater using aquatic plants,
Journal of Agricultural Technology 8(1),2012,1-11.

Honey Aggarwal,et al, Phytoremediation of some heavy metals by agronomic crops,

Developments in Environmental Science 5,2007,79-98.

Hiromichi Morikawa,et al, Basic processes in phytoremediation and some applications

to air pollution control, Chemosphere 52,2003, 1553-1558.

Hongyu Wang,et al, An Application of Phytoremediation to River Pollution

Remediation, Procedia Environmental Sciences 10 ,2011 ,1904 1907.

73
The Earth is the LORDS and all that is in it, ~Psalm 24:1
LET US BE GOOD ENVIRONMENTAL STEWARDS.

THANK YOU AND GOD

BLESS

Gizel Rivera-Santiago
Graduate Student
Philippine Normal University
Taft Avenue, Manila
74