Research Methods on

Bioremediation of
contaminated
agricultural soil

Reginawanti Hindersah
Jurusan Ilmu Tanah
Faperta-Unpad
 Contaminants:
Organic: (bio) degradable
Inorganic: non degradable

Contaminants in Agricultural soil

Pesticide
Heavy Metal
Might be Polyaromatic hydrocarbon
 Source of contaminants
Irrigation water, presipitation
Solid substances (ex. Flooding material, animal excretion,
domestic/industrial waste)
Liquid contaminants (slurry and sludge, from manufacture)
Agricultural Inputs including fertilizer and organic matter
Impact of contaminants on agricultural land
Threaten food web
Accumulate in edible parts of crops
Non tariff barrier for food export
How do contaminants affect soil health ?

Xenobiotic substances are not naturally degradable;

become soil pollution
Exogenous organic matters are not easily
decomposed; distrubance on soil nutrient availability
Increase nonessential elements (heavy metal); plant
toxicity
Change in soil microbology (microbial composition
and activity)
Soil physic alteration
How to remediate contaminated soil ?
Heat method
Physical method Expensive
Chemical method Shift the pollution:
air/water pollution

versus

Cost-efficient
Safe
Biological method Nature-based
Convenient
Easy
 BIOREMEDIATION
The use of microorganisms to degrade/control
contaminants that pose environmental and especially
human risks.

The use of biological mechanisms to destroy, transform, or

immobilize environmental contaminants in order to protect
potential sensitive receptors (Bioremediation Discussion
Group, 2006)
Bioremediation Treatment Techniques
in situ remediation:
leaving the soil in its original place and bringing the
biological mechanisms to the soil
more cost effective
less effective
Less controlable
ex situ remediation:
removing the soil from the subsurface to treat it.
less effective
more thorough (accurate)
 Bioremediation Principle

In situ-ex situ

Microbial innoculation (bioaugmentasi)

Adjusment microbial nutrition: Adding Nutrients, controlling
pH, etc.
Controlling physical environment: Increasing humidity and
aeration

Increasing organic matter degradation rate

Enhancing microbial activities to degrade organic
pollutants or to change the availability of (heavy)
metal.
 Agricultural Land: Shallow soil
In situ bioremediation: PHYTOREMEDIATION
the use of plant to degrade or accumulate organic/
anorganic contaminants and stabilize/sequestrate it in
their biomass

Hyperaccumulator plants
In situ bioremediation PHYTOREMEDIATION:
Contaminant mobilization: organic acid, pH change,
redox potention alternation
Contaminant adsorption and translocation
Nurtrients supply and humadity
Induce contaminant degradation In rhizosphere and
around the roots: Rhizodegradation
Contaminat metabolisms in the plant tissues:
Phytodegradation
Rhizosphere effects

Plant: Phytoextraction, Rhizofiltration, Dissipitation (phytovolatilisation)

Microbes: Mobilization, Immobilization, Biodegradation
In situ bioremediation strategy
Microbe (Bioremediator) Plant (phytoaccumulator)
interaction
Organic contaminat degradation
Inorganic contaminant
mobilization/imobilization
Increase plant growth
Bioprotectant

Contaminant-resistance Plant
Growth Promoting Microbe ?
Ex situ bioremediation
Composting, landfarming, biopiling, and processing by
bioreactors along with thermal, chemical, and physical
processes.
Difficult part of bioremediation
1. No standard criteria for evaluating among methods
2. The applicability of bioremediation techniques requires
close evaluation of each site
3. Treatability studies take a time and money
Determine soil conditions such as porosity, pH,
moisture content, organic matter, terminal electron
acceptors, nutrent
Contouring
The presence of microbes, and pollutants
Water availability
Animal trafficking
Soil contaminants Mapping
= Geochemical Mapping
 Geochemical Mapping (cont.)
Formulate the aim of geochemical mapping clearly and
precisely
Define/ describe the working area
Use Grid Sampling method to collect soil samples,
determined by
o Extent and distribution of contaminant/Polutant
o Urban or suburban area?
o Resident Area: Grid sampling 250 x 250 m
o Area near Septic tank: grid sampling 100 x 100 m
o Area near pollutant source: grid rapat
Geochemical mapping (cont.)
Collect Sample and subsampel
Collect sample from different soil depth (0-30, 30-60; depend
on pollutant type)
Sample weight is approx. 500-1000 g
Analysis:
o Atomic Adsorption Spectrometry: (Heavy) metal
o Gas Chromatography-Mass Spectrometry (GCMS):
Pesticida
o High Performance Liquid Chromatography (HPLC):
petroleum (polyaromatic hydrocarbon)
Expected map
Research Parameters
in contaminated soil bioremediation

Total Contaminant content and/or their derivates in soil

Contaminant Availability for plant uptake
Contaminant content in certain part of plant especially in edible
part
Essential nutrient uptake
Microbial population and compositin in rhizosphere
Soil biota population and composition
Plant parameter (shoot and root)
Bioremediaton Methods
In vitro
Microbial Growth media:
Physical & chemical
requirements to induce
microbial growth and
activities

Bioassay
Effect of microbial
innoculation and orrganic
matter on contaminant
content in accumulator
plants and plant growth.
Toxic level of contaminant
in plant and soil
microbes
Bioremediation Methods (cont.)
Pot research
in field or in
green house

Field research in
contaminated soil
due to anthropognic
activities (Do not
contaminate soil)
Control 0,5 % inokulan
Expected Results

Treatment Cd on shoot
(mg kg-1)
A = Control (wihtout 2,25 a
Azotobacter)
B = 50 mL Azotobacter (0,1 %) 2,31 a
C = 50 mL Azotobacter (0,5 %) 2,31 a
D = 50 mL Azotobacter (1 %) 2,50 a
E = 50 mL Azotobacter (1,5 %) 2,30 a
Real Case: Hg-contaminated gold mine tailing
Lahan dan Tanah Pertanian Buru

Tanah Aluvial (Fluvents)

Tanah Gleisol (Aquepts)

Tailing Characteristics
Parameter Satuan Nilai Kriteria
pH H2O (1:2,5) 9,17 Alkalis
C-Organik % 0,20 Sangat rendah
N-total % 0,04 Sangat rendah
C/N rasio 5 Rendah
P2O5 Olsen ppm P 3,16 Sangat rendah
P2O5 HCl 25% mg 100g-1 1,65 Sangat rendah
K2O mg 100g-1 7,93 Sangat rendah
KTK me/100g 7,66 Rendah
Tekstur tanah
Pasir % 67
Lempung
Debu % 22
Berpasir
Liat % 11
Kadar Hg Ppm 10,77 Tinggi
Physical Method:
Hg recovery by water leaching +
alkaline substances
Gold reextraction

Natural revegetation
Pedology and Biology Method