The 4th International Conference on Sustainability Agriculture and Biosystem IOP Publishing

IOP Conf. Series: Earth and Environmental Science 1059 (2022) 012032 doi:10.1088/1755-1315/1059/1/012032

COD Removal of Batik Wastewater using Microbial

Immobilization System for Agricultural Purposes

Yuniati M D 1, Rachmawati V1, Nurjayati R1, Marganingrum D1, Lisdiana A1,

Noviardi R R1W Purwoko W1
1
Research Center for Geotechnology, Research Organization of Earth Sciences -
National Research and Innovation Agency (BRIN), Kompleks BRIN, Jl. Sangkuriang
Bandung 40135

Corresponding author’s email address: muti001@brin.go.id

Abstract. Batik is made through a series of processes that requires large amount of water. One
of these processes is dying the fabric using chemical dyes. Many batik producers in Yogyakarta
dispose the wastewater straight into the river or on the ground. Since batik is recognized as
UNESCO's Intangible Cultural Heritage of Indonesia, batik industries have been grown rapidly.
As a result, batik wastewater became a huge contributor to environmental pollution, especially
land and water resources. In the present study, application of microorganisms (bioremediation)
to remove or degrade pollutant of batik wastewater was investigated. Microorganisms of Bacillus
licheniformis were immobilized in bentonite mineral. This microbial immobilized system then
applied in aerobic reactor of continuous process. The concentration of Chemical Oxygen
Demand (COD) was calculated as daily parameter. Although the final COD concentration still
doesn't meet the government standard, this microbial immobilization system shows promising
COD removal results. The highest COD removal is 75%. This experiment is still in the early
stages and further phases of the experiment are required to reach safe chemical parameters of
batik wastewater for agricultural purposes as regulated by the Indonesian government.
Keywords:, Bacillus licheniformis, Bioremediation, Batch process, Continuous process

1. Introduction
In 2009, UNESCO (The United Nations Educational, Scientific and Cultural Organization) recognized
Indonesian batik as a Masterpiece of Oral and Intangible Heritage of Humanity. Since then, a blast of
batik industries has occurred with rapid progress. Therefore, besides being a symbolic icon to
Indonesians, batik is also an essential source of income for the local communities. It means the batik
industry has a significant role in supporting the national economy [1,2].
Batik is made through a series of processes with a considerable amount of water. One of these is
dying the fabric that consumes approximately 50 liters of water to make one piece of batik. Many batik
producers dispose the wastewater straight into the river or on the ground. As a result, extensive water
pollution is unavoidable and has more significant effects on the environment. One of the affected is the
agricultural field.
Many farmers in developing countries grow crops, especially vegetables using this contaminated
water to irrigate their crops. They may not be aware of the pollution level of the water they use.
Evenmore they have no choice but to use this water to irrigate their crops because it is the only water
available. To avoid or lowering the risk in using batik wastewater, easy and suitable treatment is needed.

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The 4th International Conference on Sustainability Agriculture and Biosystem IOP Publishing
IOP Conf. Series: Earth and Environmental Science 1059 (2022) 012032 doi:10.1088/1755-1315/1059/1/012032

Batik wastewater specifically contains paraffin, dyes (natural and synthetic dye), starch, and caustic
soda. Those components make wastewater characteristic with a high load of COD and pH [3]. Besides,
these effluents are also known to be one of most difficult treat due to the recalcitrant nature of dyes [4].
The dyed water also causing various health problems such as jaundice, vomiting and increased heart
rate [5]. Based on government regulation number 82 of 2001 regarding water quality management and
water pollution control, the standard pH for crop irrigation is 5-9, while the standard COD is 100 mg/L
[6].
Many methods for treating batik wastewater have been investigated. The most common method is
coagulation-flocculation [7,8,9]. However, this method still generates a high amount of toxic sludge [3].
Other techniques are the use of nanofiltration membrane [4], photocatalytic for decolorization and
demineralization [5], TiO2 nanoparticles [10], phytoremediation [11], integrated upflow anaerobic filter
(UAF)-Constructed Wetlands (CWs) [3], etc. Those techniques are relatively costly and simply not
affordable for individual home-based batik artisans.
It was previously reported that wastewater from textile and sugar industry in West Java could be
treated by using preservated microorganism material (MPMO) [12,13]. MPMO is one of microbial
immobilization systems that use Bacillus licheniformis as immobilized microorganism in bentonite
mineral. A major objective of the present study was to determine whether MPMO can be used to treat
batik wastewater by lowering its COD concentration to reach safe chemical parameters of wastewater
as regulated by the Indonesian government.

2. Materials and Methods

2.1 Materials
The waste used in this study was liquid waste from the intake of wastewater treatment plant of batik
wastewater storage in Yogyakarta City. For bioremediation process, the dominant type of Bacillus
licheniformis were packaged in bentonite-Ca mineral as preservation material (MPMO). The MPMO
act as a microbial immobilization system. Nutrients for growth of microorganisms that used in the
seeding and acclimatization process before continuous process were tapioca (1.6 gr/L), ammonium
sulfate (1.5 gr/L), potassium di-hydro phosphate (1 gr/L), calcium chloride (1 gr/L), magnesium chloride
(0.5 gr/L), iron (II) chloride (0.05 gr/L), and broth nutrient (0.5 gr/L). The concentration of nutrients in
this study was adjusted to the chemical oxygen demand (COD) concentration of batik wastewater used
in the experiment.

2.2 Methods
Laboratory-scale experiment was conducted in two stages processes. There are batch process and
continuous process. Even though, only continuous process data will be discuss in this article.
The processes of batik wastewater treatment were using small scale wastewater treatment plant (mini
IPAL) consist of 5 units, namely equalization, sedimentation I, biological, sedimentation II, and effluent
reservoir tank (Figure 1).
Seeding and acclimatization of Bacillus licheniformis using synthetic wastewater were done in batch
process prior to the continuous process. These processes could make Bacillus licheniformis and other
microorganism consortium in batik wastewater adjust to a new environment containing new wastewater.
As a result, they can improve their degradation ability.
Sampling were done in the inlet of the equalization tank (as influent point) and the outlet of effluent
reservoir unit (as effluent point). The concern parameter in this study is COD.

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The 4th International Conference on Sustainability Agriculture and Biosystem IOP Publishing
IOP Conf. Series: Earth and Environmental Science 1059 (2022) 012032 doi:10.1088/1755-1315/1059/1/012032

1 2 3 4 5

Figure 1. The mini IPAL used in laboratory-scale experiment of batik wastewater (1. Equalization
tank; 2. Sedimentation I tank; 3. Biological processing tank; 4. Sedimentation II tank; and 5.
Effluent reservoir tank)
The removal efficiency was determined by comparing the value of COD before and after the
processing process with the following equations:
𝐶𝑂𝐷𝐼 −𝐶𝑂𝐷𝐸
𝐸𝑅 = ( )𝑥 100 (1)
𝐶𝑂𝐷𝐼

ER is removal efficiency (%), CODI is COD influent, i.e., the result of COD measurement in the inlet
of the equalization unit (mg/L), CODE is COD effluent, i.e., the result of COD measurement in the outlet
of effluent reservoir unit (mg/L).
The retrieval and measurement of COD influent and effluent were carried out as much as ten times
running, meaning ten times the flow process through the five mini IPAL processing tanks. All
experimental work was carried out at the Research Center for Geotechnology (RCG) of National
Research and Innovation Agency (BRIN).

3. Results and Discussion

3.1 Characteristic of batik wastewater

Wastewater sample that used in this study come from production process of batik that consist of dyeing,
waxing, bleaching, and washing. Because of these processes, the wastewater of batik contain organic
and inorganic contaminant. The colour of batik wastewater is pitch black that come from accumulation
of colour substances. The COD concentration is above 1145 mg/L.

3.2 The efficiency of chemical oxygen demand (COD) removal

Chemical Oxygen Demand (COD) test is the best and rapid method for estimation of organic matter
present in the wastewater sample. Before continuous process, we should ensure that there is a sufficient
quantity of active biomass available to consume the applied quantity of organic pollutant at any time.
This could be reach by Mixed Liquor Suspended Solid (MLSS) value. MLSS is the concentration of
suspended solids in an biological tank which occurs during the treatment of wastewater. MLSS consists
mostly of microorganisms and non-biodegradable suspended matter.
In the batch process, MLSS values obtained during the seeding and acclimatization process reached
the range of 790 to 2656 mg/L. Those values produce high COD removal efficiencies during batch
process. Following batch process, the ten-days study measured the COD of influent and effluent through

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 The 4th International Conference on Sustainability Agriculture and Biosystem IOP Publishing
IOP Conf. Series: Earth and Environmental Science 1059 (2022) 012032 doi:10.1088/1755-1315/1059/1/012032

the continuous processes in a treatment plant (Figure 1). The feed in the form of batik wastewater entered
equalization tank and overflowed into sedimentation I tank. Wastewater degraded by microbes under
aerobic condition in biological processing tank. MPMO contained Bacillus licheniformis in powder form
suspended in the third tank as biological process occurs after equalization and sedimentation process.
The result come out into sedimentation II tank and the effluent collected in the effluent reservoir tank.
Based on observational data for ten days (Table 1) shows that MPMO has a good performance. In
the early observations on day 1, the COD reduced from 1145 mg/L in the inlet to 603 mg/L in the outlet.
After that, the removal rate was slightly slower and fluctuating until day 5 with a range of 593 mg/L -
683 mg/L. The lowest COD value obtained on day 9 was 442 mg/L, but the value is still higher than 100
mg/L which is required for irrigation water standard.
Table 1. The COD, TSS, Mass Loading, MLSS and Removal Efficiency of Batik Wastewater
Treatment

Data Standard Standard Efficiency Efficiency

Number CODI CODE COD TSSI TSSE TSS Mass Loading MLSS Removal Removal
(mg/L) (mg/L) (mg/L)1 (mg/L) (mg/L) (mg/L)1 (kg/m3.day) (mg/L) COD (%) TSS (%)
1 2656
1145 603 100 434 128 400 0.92 47 71
2 2357
1145 593 100 434 79 400 0.92 48 82
3 1986
1145 643 100 434 59 400 0.92 44 86
4 1308
1145 683 100 434 70 400 0.92 40 84
5 791
1145 633 100 434 71 400 0.92 45 84
6 1031
1145 432 100 434 55 400 0.92 62 87
7 1778
1145 412 100 434 37 400 0.92 64 91
8 1153
1145 467 100 434 148 400 0.92 59 66
9 1556
1748 442 100 471 64 400 1.4 75 86
10 1544
1748 643 100 471 71 400 1.4 63 85
1)
Water quality standard based on Government Regulation Number 82 of 2001 for class IV water quality [6]

4000 Influent Effluent Removal effiency 100

Removal Efficiency COD (%)

3500 90
COD measurements (mg/L)

75 80
3000
70
2500 60
2000 50
1500 40
30
1000
20
500 10
0 0
1 2 3 4 5 6 7 8 9 10
Data

Figure 2. Influent and effluent COD measurements that showing the removal
efficiency
The efficiency of COD removal is given in Figure 2. COD refers to the chemical decomposition of
organic substances and the result showed COD removal efficiency range from 40% to 75% after
biological processes using MPMO. These results show Bacillus licheniformis able to reduce COD value
and confirm the research conducted by Sonune et al. (2015) [14]. Microorganism can degrade the

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The 4th International Conference on Sustainability Agriculture and Biosystem IOP Publishing
IOP Conf. Series: Earth and Environmental Science 1059 (2022) 012032 doi:10.1088/1755-1315/1059/1/012032

complexity of chemical compounds with organic substances into smaller compounds such as CO2 and
water, as follows [15,16]:
organic substances + O2 CO2 + H2O (2)
microorganism

The maximum efficiency of COD removal reached up to 75%, indicating the potential microbial
immobilization system to treat batik wastewater for agricultural purposes.

3.3 COD and its related parameters

Besides MLSS and microorganism, COD also influenced by Total Suspended Solid (TSS). TSS is a
suspension caused by the presence of organic compounds in the water which relate with MLSS.
Inffluent TSS Effluent TSS
500 100
450 90

Removal Efficiency TSS (%)

400 80
350 70
TSS Measurements (mg/L)

300 60
250 50
200 40
150 30
100 20
50 10
0 0
1 2 3 4 5Data6 7 8 9 10

Figure 3. Influent and effluent of TSS measurements that showing the

removal efficiency
Figure 3 shows that the concentration of TSS in the effluent of batik waste treatment tends to
decrease. Still, there is a significant increase in TSS concentration in the 8 th data, with the TSS
concentration in the 7th data from 37 mg/L to 148 mg/L. While the value of TSS removal efficiency
tends to show an increasing value, the 8th data also experienced a significant decrease with the removal
efficiency value of 91% in the 7th data to 66% in the 8th data. The most significant TSS removal
efficiency value is 91%. It was obtained from the 7th data with the influent TSS concentration of 434
mg/L and decreased to 37 mg/L in the effluent. Thus, the overall concentration of TSS met the water
quality standard based on Government Regulation Number 82 of 2001 for class IV water quality. The
effluent still contains TSS even though the value has met the referenced quality standard. Suspended
solids contained in the effluent could come from detached biomass and suspended solids contained in
the influent but they were not degraded and deposited by precipitation or adsorption processes [17].

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The 4th International Conference on Sustainability Agriculture and Biosystem IOP Publishing
IOP Conf. Series: Earth and Environmental Science 1059 (2022) 012032 doi:10.1088/1755-1315/1059/1/012032

Removal Efficiency (%)

80 Mass Loading (kg/m3.day) 1,6
70 1,4
Removal Efficiency COD (%)

60 1,2

ML (kg/m3.day)
50 1,0
40 0,8
30 0,6
20 0,4
10 0,2
0 0,0
1 2 3 4 5Data 6 7 8 9 10

Figure 4. Concentration Effluent COD and The Mass Load

Figure 4 shows the relationship between COD removal and mass loading. Mass loading means the
total mass of one or more organic or inorganic effluent constituents delivered to the wastewater system
over a specified period.
𝑄 𝑥 𝑆𝑜
𝑀𝐿𝑅 =
𝑉
MLR is mass loading rate (Kg/m .day), Q is flow rate (m3/day), So is COD concentration (Kg/m3), and
3

V is volume (m3).
In the 9th data, there is an increase in the mass loading from 0.92 kg/m3.day to 1.4 kg/m3.day and
accompany by an increase in the value of removal efficiency from 59% to 75%. Thus, the removal
efficiency value in 9th data became the highest COD removal efficiency value during the processing
cycle. The increasing in mass loading value accompanied by increasing COD removal efficiency also
occurred in biological treatment research for sunflower oil from wastewater influents [18].
Another fact is the 9th data of COD removal efficiency as the highest removal efficiency related with
increasing MLSS concentration from 1153 mg/L to 1556 mg/L. The same thing happened with the rise
of MLSS concentration from 1031 mg/L to 1778 mg/L accompanied by an increase in TSS removal
efficiency, which became the highest efficiency value with a value reaching 91% in the 7th data. Based
on these results, it is shown that increasing the concentration of MLSS can increase the efficiency of
pollutant removal (Figure 5). These results are consistent with the research conducted on domestic waste
treatment at The Mahram Asia plant. The study stated that an increase in MLSS concentration resulted
in increasing COD and TSS removal efficiency, but an excessive rise in MLSS concentration would
decrease treatment efficiency [19].

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The 4th International Conference on Sustainability Agriculture and Biosystem IOP Publishing
IOP Conf. Series: Earth and Environmental Science 1059 (2022) 012032 doi:10.1088/1755-1315/1059/1/012032

MLSS Concentration Removal Efficiency TSS Removal Efficiency COD

100 3000

90
2500
80

MLSS Concentration (mg/L)

70
Removal Efficiency COD (%)

2000
60

50 1500

40
1000
30

20
500
10

0 0
1 2 3 4 5 6 7 8 9 10
Data
Figure 5. The effect of removal efficiency of COD and TSS at different value of
MLSS

4. Conclusions
The results show that biological processes using MPMO as microbial immobilization system can be
used to reduce the concentration of COD in batik wastewater. In addition, the bacteria (Bacillus
licheniformis) can grow well in bentonite-Ca while treating the wastewater. The levels of COD
concentrations in batik wastewater have reduced with maximum efficiency of COD removal reached up
to 75% in 10 days process, with COD value of 442 mg/L. This value is still higher than 100 mg/L which
is required for irrigation water based on water quality standard on Government Regulation Number 82
of 2001 Republic of Indonesia. This experiment is still in the early stages and further phases of the
experiment are required to reach safe chemical parameters of batik wastewater for agricultural purposes
as regulated by the Indonesian government.

Acknowledgments
The authors would like to acknowledge the Research Center for Geotechnology for their support in the
implementation of this experiment and analysis of samples.

5. References
[1] Nurhasanah S, Sudrajat R L, Srinovita Y and Trisnawati E 2016 Optimization of batik fashion
based cultural heritage as a competitive advantage in the ASEAN Economic Anticipating
Commodity (AEC) in 2015. Proc. of the 29th Int. Conf. IRES vol 29 pp 18-23
[2] Mukimin A, Vistanty H, Zen N, Purwanto A and Wicaksono K A 2018 Performance of
bioequalization-electrocatalytic integrated method for pollutants removal of hand-drawn
batik wastewater. J. Water Process Eng. 21 77-83
[3] Handayani N I, Yuliasni R, Setianingsih N I and Budiarto A 2020 Full scale application of
integrated upflow anaerobic filter (UAF) - Constructed Wetland (CWs) in small scale batik
industry wastewater treatment J. ris. teknol. pencegah. pencemaran Ind. 11 27-35
[4] Rashidi H R, Sulaiman N M N, and Hashim N A 2012 Batik industry synthetic wastewater
treatment using nanofiltration membrane Procedia Eng. 44 2010-12. doi
10.1016/j.proeng.2012.09.025.

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The 4th International Conference on Sustainability Agriculture and Biosystem IOP Publishing
IOP Conf. Series: Earth and Environmental Science 1059 (2022) 012032 doi:10.1088/1755-1315/1059/1/012032

[5] Khalik W F, Ho L N, Ong S A, Wong Y S, Yusoff N A and Ridwan F 2015 Decolorization and
mineralization of batik wastewater through solar photocatalytic process Sains Malaysiana 4
607-12
[6] Indonesian Government Regulation No.82. Water Quality Management and Water Pollution
Control.
[7] Mehta P 2012 Treating textile effluents by coagulation - flocculation method using different
dosing composition Adv. Appl. Sci. Res. 3 (4) 2514-17
[8] Rodrigues C S D, Madeira L M and Boaventura R A R 2012 Treatment of textile dye
wastewaters using ferrous sulphate in a chemical coagulation/flocculation process. Environ.
Technol. 34 719-29. doi:10.1080/09593330.2012.715679
[9] López C L, Pascual J M, Díaz J C L, Toledo M V M, Muñío M M and Poyatos J M 2015
Combined treatment of textile wastewater by coagulation-flocculation and advanced
oxidation processes Desalin. Water Treat. 57 13987-94
[10] Sutisna, Wibowo E, Rokhmat M, Rahman D Y, Murniati R, Khairurrijal and Abdullah M 2017
Batik wastewater treatment using TiO2 nanoparticles coated on the surface of plastic sheet
Procedia Eng. 170 78-83
[11] Setiyono A and Gustaman R 2017 Pengendalian kromium (Cr) yang terdapat di limbah batik
dengan metode fitoremediasi Unnes J. Public Health 6(3) 155-60
[12] Sembiring H, Agustinus E T S, Estiati L M, Effendi, Gurharyanto, Dida E N and Saepuloh A
2013 MPMO (Material Preservasi Mikroorganisme) sebagai bahan aklimatisasi dalam
pengolahan limbah cair PT. Sivatex, Majalaya Prosiding Pemaparan Hasil Penelitian Puslit
Geoteknologi - LIPI 167-76
[13] Agustinus E T S, Sembiring H and Effendi 2014 An application of Preservated Microorganism
Material (MPMO) for organic wastewater processing in wastewater treatment plant Ris. Geo.
Tam. 24 65-76
[14] Sonune N A and Garode A M 2015 Bioremediation potential of bacterial isolates for municipal
wastewater treatment Curr World Environ. 10 (2) 619-25
[15] Joutey N T, Bahafid W, Sayel H, and El Ghachtouli N 2013 Biodegradation: involved
microorganisms and genetically engineered microorganisms Biodegradation - Life of Science
p 289
[16] Poznyak T I, Oria I C, and Poznyak A S 2019 Biodegradation: Ozonation and Biodegradation in
Environmental Engineering (Elsevier) p 353
[17] Pronk M, de Kreuk M K, Bruin B, Kamminga P, Kleerebezem R, and van Loosdrecht M C M
2015 Full scale performance of the aerobic granular sludge process for sewage treatment
Water Res. 84 207-17. doi:10.1016/j.watres.2015.07.011.
[18] Cisterna P 2017 Biological treatment by activated sludge with high biomass concentration at
laboratory scale for mixed inflow of sunflower oil and saccharose. Environments 4 69.
doi:10.3390/environments4040069
[19] Rafati M, Pazouki M, Ghadamian H, Hosseinnia A and Jalilzadeh A 2018 Effect of operating
parameters on the performance of wastewater treatment plant (Case study: The Southern
Tehran wastewater treatment) Adv. Environ. Technol. 4 211-21

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