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Final Project Report
Rehabilitation of Ex-Mining Quarry Area with
Phytoremediation Plants and Biological Fertilizer
Application
Project Coordinator:
Nisa Rachmania Mubarik1*
Other Researchers:
Hirmas Fuady Putra1
Dini Fitriyanti1
Marini Adani1
Idin Abidin2
1Department of Biology, Faculty of Mathematics and Natural
Sciences, Bogor Agricultural University, Bogor 16680, INDONESIA
2Gunung Ceremai National Park, Kuningan, INDONESIA
September 2016
INDONESIA
mailto:[email protected]
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1 Contestant profile
Contestant name Dr Nisa Rachmania Mubarik
Contestant occupation Lecturer
University/Organization Bogor Agricultural University
E-mail
Phone
Number of people in your team Four person
2 Project overview
Title Rehabilitation of Ex-Mining Quarry Area
with Phytoremediation Plants and
Biological Fertilizer Application
Contest INDONESIA
Quarry name Hambalang
Prize category Education and Raising Awareness
Habitat and Species Research
Biodiversity Management
Student Project
Beyond Quarry Borders
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ABSTRACT
Several phosphate (P) and sodium (K) solubilizing bacteria,
nitrogen fixing bacteria, and indole-3-
acetic acid (IAA) producing bacteria isolates have been isolated
from soil in mining limestone Cirebon.
These bacteria were selected, blended, and packaged together
into biological fertilizer product. The use
of biological fertilizers on seedling plants could replace
synthetic chemical fertilizers on seedling plants.
Phosphate and potassium solubilizing bacteria have the ability
to produce a substance that dissolves the
compound P and K bound contained in the soil, so that it becomes
available to plants and can replace
synthetic fertilizers (chemical) P and K. Nitrogen-fixing
bacteria ( N) can reduce the use of urea fertilizer.
Several bacteria also produce indole-3-acetic acid known as
auxin, one of growth promoting hormone.
The basic philosophy underlying this research project is to
rehabilitate sandy clay at ex-mining area
based on new technology and low capital expenditure. The
experiments conducted were (1) use of
bacterial formulation consist of P and K solubilizing bacteria,
N fixing bacteria, IAA producing bacteria as
biofertilizer for seedling plant that planted in ex-mining land
in Hambalang, (2) re-vegetation on ex-mining
area by using phytoremediation plants such as leucaena (Leucaena
leucocephala), albizia (Albizia
saman), and pterocarpus (Pterocarpus indicus) application with
compost and consortium of bacteria on
ripping and without ripping soil treatments, and (3) evaluation
of ecological condition on ex-mining area.
Preparation of bacterial formulation was done in Department of
Biology, Faculty of Mathematics and
Natural Sciences, Bogor Agricultural University. The field
experiments were initially with determined the
location of the research in the reclaimation area near the lake
in Hambalang. A 100 m transect line
stretched for ecological observation and vegetation prior to
planting. Observation area were divided into
three locations corresponding elevation of the land starting
from higher land area (zero point) to lower
land area (100 m from zero point). The re-vegetation experiments
has been done on higher land area,
center land area, and lower land area. Each location is divided
into six plots with a size of 6 mx 4 m per
plot, consist of planting treatment with leucaena, albizia, and
pterocarpus (3 plots) and without planting (3
plots). There were 3 soil treatments i.e. ripping, ripping with
soil enrichment with compost and bacterial
consortium), and control without treatments.
The conclusion of the research project that soil treatments with
ripping and application with compost
before planting could rehabilitate of ex-mining Hambalang area.
The bacterial consortium from similar
quarry condition could use as biofertilizer for seedling plants
which planted at the area and making plant
growth better.
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INTRODUCTION
The topsoil is the layer that provides elements of macro and
micro essential for the plant. The
existence of microbes in the soil layer plays a role in the
provision of nutrients, litter decomposition, and
improve soil structure. The open mining land and excessive use
of synthetic fertilizer can lead to
decrease of soil quality and environmental pollution. The
previous research project around Cirebon
limestone quarry, we have found 55 isolates of phosphate
solubilizing bacteria and 12 isolates of
potassium (K) solubilizing from around of limestone Cirebon
quarry (Mubarik et al. 2014). In addition, one
of the K solubilizing bacteria, Burkholderia cepacia KQC.5C.5,
was able to release soluble K from soil
after ten days incubation (Anggraeni et al. 2016). Other strain
of Burkholderia sp QC3.a.1 was capable of
solubilizing the highest P from Pikovskaya media (Mursyida et
al. 2015). Dwiana (2015) reported that
Bacillus megaterium QC 7B41 and QC 5C32 isolated from Cirebon
quarry could produce indole-3-acetic
acid (IAA or auxin) and N fixed. .Application the B. megaterium
showed positive effect on the growth
primary root and lateral root number of seedling plant of
leucaena in green house experiment (Yandra
2015). Serratia marcescens QC5C32 has potency as P and K
solubilized, indole-3-acetic acid (IAA)
production, and could convert N2 to NH3 by using nitrogenase
enzyme activity. The use of S. marcescens
as biofertilizer on seedling plant of leucaena showed a better
effect on the dry weight of roots.
The project was aimed to rehabilitate of ex-mining area by
planting of phytoremediation plants.
The category of phytoremediation in the project as
Phytostimulation or rhizodegradation which use of
rhizospheric associations between plants and symbiotic soil
microbes to degrade contaminants from
quarry area. The bacterial consortium was used as biofertilizer
for seedling plant which planted on
quarry. The impact of the research results generated
technologies for the rehabilitation of ex-mining area
by using compost with consortium of soil bacteria and ecological
treatments such as soil ripping and
revegetation.
OBJECTIVES
The research project was conducted to (1) use of bacterial
formulation consist of P and K
solubilizing bacteria, N fixing bacteria, IAA producing bacteria
as biofertilizer for seedling plant that
planted in ex-mining land in Hambalang, (2) re-vegetation on
ex-mining area by using phytoremediation
plants such as leucaena (Leucaena leucocephala), albizia
(Albizia saman), and pterocarpus
(Pterocarpus indicus) application with compost and consortium of
bacteria on ripping and without ripping
soil treatments, and (3) evaluation of ecological condition on
ex-mining area.
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BACKGROUND INFORMATIONS
Location of clay quarry in Hambalang at District Citereup and
Gunung Putri, Bogor Region, West
Java Province, Indonesia. The topographic looks like elongated
hill and sloping hill with an altitude
between 175-630 meters above sea level and Hambalang Hill are
rear of the quarry. Several locations
are still in the reclamation area, namely Parumpung, Pondok
Manggu, and Pasir Gadung. There are two
main rivers around the mining area, namely Cileungsi and
Ciherang. Both of the river flow to the
springhead namely Citeureup river. Surface water from the the
upper area of quarry is collected in the
pond at reclamation activities. Hambalang quarry is located a
bit far from the main highway and only a
local road that connects from the village area within 10 km.
There are several quarry of other company
near PT Indocement Tunggal Prakarsa TBK (Figure 1). The site
area for research project were located
around ex-mining area of sandy clay in Hambalang, Citeureup
Bogor. The project has been done since
May until July 2016. The GPS of location and environment data at
the project area as shown on Table 1.
Figure 1 Location of Hambalang quarry at Bogor Region, West Java
Province, Indonesia (https://www.google.co.jp/maps)
Table 1 Environment data at project area at Hambalang
reclamation area on May 2016
No Area Location area with Global Positioning
System (GPS)
Light intensity (lux)
Humidity (%)
Ambient temperature
( 0C)
Wind velocity (Km/h)
1 Higher area (0 m)
S 06°31.737'
E 106°54.750'
1964 39 42.5 9.3
2 Centre area S 06°31.737'
E 106°54.720'
1965 46.5 37.3 8.8
3 Lower area (100 m from 0 m)
S 06°31.737'
E 106°54.696'
1825 46 36.7 7.7
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METHODS
Soil Sampling from Hambalang Quarry and Measurement of
Microclimate
Soil was obtained from mining quarry area and around .An amount
of 1 Kg soil surface (0-10 cm) from
each soil sampling was taken in polyethylene bag. Whole soil
samples were taken with the permission of
PT Indocement Tunggal Prakarsa to analyzed at Bogor Agricultural
University.
Microclimate of the project area (temperature and humidity) is
measured by sling psychrometer and light
intensity by using lux meter.
Total plate count of bacteria
In amount of 3.0 g of soil samples is diluted in 27 mL of NaCl
0.85% and it was agitated 120 rpm in
shaking incubator, room temperature (25 oC) for 4 hours. Then,
the culture was done with serial dilution
from 10-1
to 10-5
in NaCl 0.85 %. Suspension was spread on Nutrent Agar (NA),
Pikovskaya agar with
composition of 1 L medium: 10 g glucose, 5 g Ca3(PO4)2, 0.5 g
(NH4)2SO4, 0 .1 g MgSO4.7H2O, 0.5 g
yeast extract, 0.2 g KCl, 0.2 g NaCl, 0.002 g FeSO4.7H2O, 0.002
g MnSO4.7H2O, and 15 % bacto agar)
for phosphate solubilizing bacteria isolation (Nautiyal 1999),
and Nitrogen free base medium consist of
media Nitrogen Fixation Bacteria (NFB) consist of 5.0 g/L,
K2HPO4 0.5 g/L, MgSO4.7H2O 0.2 g/L, NaCl
0.1 g/L, CaCl2.2H2O 0.02 g/L, indicator of bromthymol blue (BTB)
0.5% in KOH 0.2 M 2 mL and vitamin B
1 mL micronutrient solution 2 mL (CuSO4.5H2O 40 mg/L, ZnSO4.7H2O
0.12 g/L, H2BO3 1.4 g/L,
Na2MoO4.2H2O 1.0 g/L, MnSO4.H2O 1.175 g/L, FeEDTA 1.64% 4 mL,
KOH 4.5 g/L. pH 6.8 and added
with 1.8 g/L agar (Okon et al. 1977). The cultures were
incubated at room temperature for 48-72 hours.
Consortium of Bacterial Formulation
The carrier material used in this study was talcum with the
composition and mixing process of the
material as followed: 200 mL suspension of selected isolate (108
cells/mL), 100 g of talcum, 1 g carboxy
methyl cellulose, and 1 g of CaCO3 (Suryadi et al. 2013).
Control treatment was prepared only with sterille
talcum (without suspension bacteria). The viability test of the
isolate was routinely done every weeks..
Revegetation of Ex-mining Area
Before revegetation of field experiment on at sandy clay of
ex-mining at Hambalang area, data of
ecological condition was measured by by using line transect from
upper land area (0 m, zeo spot) to
lower area (100 m) (Putra 2013). The area + 2000 m2 was
excavated by using tractor and divided of six
plots (6 m x 4 m per plot) at three regions (upper, centre, and
lower) There were 3 soil treatments by
using ripping, ripping with soil enrichment with compost, and
control without treatments. Ripping was
done by making 7 line mounds on ripping plots. Compost treatment
was used one week before
revegetation treatment on rippling plot and without ripping
(non-ripping) (Figure 2).
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A half meter until one meter of plant of pterocarpus
(Pterocarpus indicus) and albizia (Albizia
saman) are prepared by researcher from Gunung Ciremai National
Park, Kuningan, West Java, Indonesia
and leucaena (Leucaena leucocephala) seedling plants (< 30
cm) are found from Hambalang area. Two
plants of each species are planted randomly on ripping and
non-ripping area, and on the treatment with
and without compost application. Bacterial consortium was
applied after planted at composting treatments
(Figure 3). Data of plant growth of assisted and spontaneus
growing plants are measured and collected
every two weeks from each plant of three region from June until
last July 2016.
Statistical Analysis
The data were to analyzed by using analysis of variance with
Microsoft Excell 2010 and SAS 16.0.8
(Statistical Analysis System) at P
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Table 2 Chemical composition of soil sample from ex mining
Hambalang area
Soil Region
pH 1:5 H
2O
C Org (%)
N Total (%)
P (ppm)
K (cmol /Kg)
CEC (cmol/Kg)
Texture (%)
Sand Silt Clay
Higher area
7.99 0.46 0.09 10.5 0/19 7.92 34.16
52.13 13.71
Centre area
8.01 0.69 0.12 12.4 0.24 9.51 30.99 54.48 14.53
Lower area 8.13 0.69 0.12 9.8 0.22 12.28 36.42 45.78 17.80
CEC: Cation exchange capacity
Abundance of Soil Bacteria
The abundance of soil bacteria was measured before land
treatment and after plant treatments. There
are increasing of soil bacteria after one month after
revegetation as much as log 1.17 (Table 3).
Enhancement of total bacteria after revegetation can be caused
by plant produce exudates or
metabolites that used as nutrient for bacteria and soil
microbes. Rhizobacteria which grow around roots
plant also produce chemical signal to invite the bacteria of the
same species to create a consortium near
roots plant (Madigan et al.. 2008). The bacteria could
categorize as phosphate solubilizing bacteria,
nitrogen fixing bacteria, plant growth promoter that benefit to
plants (Table 4). Treatment with biofertilizer
on plant seedling could increase the population of bacteria on
soil. Previous measurement of total
bacteria which contained in compost is log 7.80.
Consortium of bacterial population which used as biofertilizer
consist of Bacillus sp. QC7B41 and
Bacillus QC5C32. Both bacteria are categorized as phosphate
solubilizing bacteria, nitrogen fixing
bacteria, and plant growth promoter (IAA producing bacteria).
The bacterial consortium of both bacteria
was stable on talcum as carrier substance up to 6 weeks
incubation with log number of bacteria 6.94
(Mulyani 2016).
Table 3 The abundance of soil bacteria before land treatment and
after revegetation
No Region Total of bacteria (log) Increasing of log number of
bacteria
Before land treatment After revegetation
1 Higher area 3.94 6.70 2.76
2 Centre area 4.85 7.11 2.26
3 Lower area 5.43 6.6 1.17
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Table 4 Total plate count of phosphate solubilizing and N fixing
bacteria after plant treatments
No Region Total of bacteria (log)
Phosphate solubilized N fixed
1 Higher area 5.77 6.47
2 Centre area 5.30 6.27
3 Lower area 4.77 5.53
Evaluation of Ecological Condition on Ex-Mining Area
Data of field treatments was analyzed using Excell and SAS
programs at P 0.05 or no significant between treatments. This is
presumably because the data is taken
in a short time (two months) and the plants used are not the
same on the age and plant height. However,
there was a tendency that the treatment with ripping and adding
with compost and bacteria showed
higher than control (non-ripping or without ripping) for plant
assisted (Figure 3). The use of bacteria and
compost on condition ripping treatment has no effect on plants
that grow spontaneously in the ex-mining
area at Hambalang.
Figure 3 Comparison of plant productivity between vegetation in
various soil conditions
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Comparison of plant productivity among species showed the
tendency that the treatment with
ripping and adding with compost and bacteria showed higher than
ripping treatment and control (non-
ripping) for pterocarpus and leucaena (Figure 4). Enhancement of
leucaena plant cover showed that it
was higher than other plants. It was suggested that all leucaena
which used were seedling plants (
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formulation Rp 500,000 per Kg. It is required 5 Kg/ha with cost
Rp 2,500,000. The total of cost of Rp
33.850,000 approximately 2507.4 USD (1 USD = Rp13,500).
ACKNOWLEDGMENT
We thank and appreciate to Quarry Life Award Indonesia at PT
Indocement Tunggal Prakarsa Tbk.
Palimanan for start up financial support of this project, thank
to Head of Department of Biology, Faculty
of Mathematics and Natural Sciences, Bogor Agricultural
University and Head of Gunung Ciremai
National Park for research supporting. Special thank for
students i.e. Ahmad Suryadi, Kamila, Rina, Aris,
Abid, and Dijah who assisted in field experiments.
REFERENCES
Dwiana A. 2015. Isolation and characterization of Indole-3
acetic acid producing bacteria from limestone quarry [undergraduate
thesis] In Bahasa Indonesia. Bogor (ID): Bogor Agricultural
University.
Angraini E. 2015. The study of potassium solubilizing bacteria
from limestone mining area Palimanan,
Cirebonquarry [thesis]. In Bahasa Indonesia. Bogor (ID): Bogor
Agricultural University Hardjowigeno S. 1995. Soil Science. 2
nd Edition. In Bahsa Indonesia.. Jakarta (ID): Akademika
Presindo.
Madigan MT, Martinko JM, Dunlap PV, Clark DP. 2008. Brock
Biology of Microorganism. San Francisco
(US): Pearson Benjamin Cummings. Mubarik NR, Wibowo RH, Angraini
E, Mursyida E, Wahdi E. 2014. Exploration of bacterial diversity
at
Cirebon quarry [final report]. Bogor (ID):
www.quarrylifeaward.ae/ system/files/winners. Mulyani. 2016 Cells
viability and identification of bacteria that isolated from
post-mining limestone at
Cirebon. [undergraduate thesis] In Bahasa Indonesia. Bogor (ID):
Bogor Agricultural University. Mursyida E, Mubarik NR,
Tjahjoleksono A. 2015. Selection and identification of
phosphate-potassium
solubilizing bacteria from the area around the limestone mining
in Cirebon quarry. Research Journal of Microbiology 10 (6):
270-279. DOI: 10.3923/jm.2015.270.279.
Nautiyal CS. 1999. An efficient microbiological growth medium
for screening phosphate solubilizing
microorganisms. FEMS Microbiology Letter 170:265-270. Okon Y,
Albrecht SL, Burris RH. 1977. Methods for growing Spirilum
lipoferum and for counting it in pure
culture and in assosiation with plants. Applied of Environmental
of Microbiology 33:85-88. Putra HF. 2013. Evaluation of ecological
function and revegetation level of xx-mining of Lead at Air
Mungkul, Kabupaten Belitung [thesis]. In Bahasa Indonesia.
Bandung (ID): Institute Technology of Bandung.
.Suryadi D, Susilowati DN, Riana E, Mubarik NR. 2013. Management
of rice blast disease (Pyricularia
oryzae) using formulated bacterial consortium. Emirates Journal
of Food and Agriulture. 25: 349-357.
Yandra RF. 2015. The examination of potential bacteria that
producing Indole-3-acetic acid (IAA) from
limestone land at planting leucaena (Leucaena leucocephala). In
Bahasa Indonesia. Bogor (ID): Bogor Agricultural University.
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Appendix 1 Comparison of plant height increment between species
of plants in various soils conditions
Appendix 2 Comparison of the trunk diameter increment between
species of plants in various soils condition
1.667
1.2
4.15
1.61.417
2.483
1.2831.5
1.917
0
0.5
1
1.5
2
2.5
3
3.5
4
4.5
Tru
nk
dia
met
er in
crem
ent (
mm
)
Treatment
Pterocarpus Albizia Leucaena
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Appendix 3 Monitoring of reclamation model (July 2016):
Measuring of plant cover, biomass, rood trees diameter
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Appendix 4 Paper of previous research and used as reference
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Cover Final Project ReportQLA Project Report_Nisa R Mubarik et
al_Indonesia_2016
