Extended Abstract : Environmental Science and Ecology

12th SCiUS Forum

Fig. 6. The chart shows Methane production rate
The graph gradually diminishes for the rest of the day. Because bacteria use methane as a carbon source,
the amount of methane in each bucket decreases, with the maximum methane in the fourth bucket on day two and
the lowest on day 4 . This could be because organic fertilizers have been added to the soil. Organic fertilizers
produce more methane than chemical fertilizers.

Conclusion
Group of methanotrophs. A formula has been developed to benefit regional farmer communities and

reduce methane. Field trials were conducted in paddy fields under conventional cultivation methods to reduce
methane emissions using clusters. The results show the use of groups of methanotrophs in the early stages of rice
emergence. The addition of 20% bacteria to the paddy fields reduced the average methane production to 0.26 ml
from 0.31 ml in the control experiment. However, when 20% bacteria and compost were added to the paddy fields,
the average methane production increased to 0.73. mL may be due to microorganisms in the methane-producing
compost. At the same time, the addition of chemical fertilizer produced 0.18 ml of methane, the lowest among all
experimental kits. This may be because chemical fertilizers affect the function and growth of microorganisms in
rice fields. Methanotrophs combined with organic and chemical fertilizers to continuously reduce methane and
by-products from the oxidation of carbon dioxide. This will ultimately reduce the greenhouse gas emissions of rice cultivation.

Acknowledgement
This project was supported by Science Classroom in University Affiliated School (SCiUS). The funding of SCiUS

is provided by Ministry of Higher Education, Science, Research and Innovation. This extended abstract is not for citation.

References
มูรนี เวาะเด็ง. การคดั แยก และการคดั เลอื กแบคทีเรียย่อยสลายมีเทน ในสภาวะตอ้ งการออกซิเจนจากตวั อยา่ งดินนาขา้ ว. 2561.

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Title: The contamination of microplastics in beach sediment OE1_13_01
Field:
Author: in Chonburi province in different monsoons

School: Environmental Science and Ecology
Advisor:
Mr. Krithiran Vayakornvichitr

Ms. Natnaree Ruangdech

Ms. Napatploy Pichetjindakul

Piboonbumpen Demonstration School, Burapha University

Asst. Prof. Dr. Anukul Buranapratheprat, Department of Aquatic Science,

Faculty of Science, Burapha University

Abstract
Microplastics are plastic particles smaller than 5 millimeters that are from decomposed large plastic

waste, or microbeads. They contaminate the coastal environments such as Beach sediment, and seawater that
considerably impact the marine ecosystem. Chonburi province where the coasts adjoin the eastern Gulf of
Thailand is important to the country's economy in terms of tourism, fishing, and industrial sites of Thailand.
The purposes of this study were to investigate the contamination of microplastics in the Beach sediment along
the coast of Chonburi province in the southwest and the northeast monsoon. The samples were collected at a
depth of 5 centimeters by using a core sampler at 5 stations consisting of Bang Saen Beach, Wonnapha Beach,
Bangphra Beach, Bangphra Jetty, and Ko Loi in July and November 2021. Microplastics were filtered from
the clear brine after it was mixed with the sediment sample and left. Six shapes of microplastics, which are
fiber, fragment, film, pellet, granule, and foam were identified from photographs taken from a
stereomicroscope. The highest and the lowest numbers of microplastics were found in the southwest monsoon
at Wonnapha Beach (6,749 pieces/Kg dry weight), and in the northeast monsoons at Bangphra Jetty (2,525
pieces/Kg dry weight), respectively. The results also showed that the number of microplastics was higher in
the southwest monsoon than in the northeast monsoon at Wonnapha Beach, Bangphra beach, and Ko Loi.
Only Bang Saen Beach and Bangphra Jetty were found to have more microplastics in the northeast monsoon
than in the southwest monsoon. The variation of microplastic did not relate to the grain size (p-value > 0.05).

Keywords: Chonburi province, coastal pollution, microplastic, the inner Gulf of Thailand

Introduction
Nowadays, Thailand produces a large amount of plastic waste and Thailand is the 6th country in the

world for releasing waste into the sea (3). The large plastic waste would decompose into microplastics that are
smaller than 5 millimeters. Some of these microplastics were distributed to beaches by the current and would
contaminate the beach sediment which is the habitat of benthic animals (4). They could take the microplastics
into their body. That could affect seafood consumers in the future.

Chonburi province where the coasts adjoin the eastern Gulf of Thailand is important to the country's
economy in terms of tourism, fishing, and industrial sites of Thailand. These activities could increase plastic
waste.

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Therefore, we did this study for investigating the contamination of microplastics in the beach
sediment along the coast of Chonburi province in the southwest and the northeast monsoon. We had chosen 5
stations consisting of Bang Saen Beach, Wonnapha Beach, Bangphra Beach, Bangphra Jetty, and Ko Loi to
study.

Methodology
Microplastics sampling
1. Use quadrat (50 x 50 cm.) to take the random areas along the high tide line.
2. Collect the samples at a depth of 5 centimeters by using a core sampler (7 cm. diameter)
3. Collect the samples from 5 stations consisting of Bang Saen Beach, Wonnapha Beach,
Bangphra Beach, Bangphra Jetty, and Ko Loi in July and November 2021.
Microplastics extraction and observation (2)
1. Fill the beaker with 20 g. of the sample.
2. Mix the sediment sample with 100 ml. of 250 g/L NaCl solution and leave it for 6 hours.
3. Filter microplastics from the sediment sample through a glass microfiber filter (GF/C).
4. Bake the filtered GF/C at the temperature of 60 oC for 3 days.
5. Investigate and identify shapes of microplastics from photographs taken from a
stereomicroscope.
Grain size classification
1. Sift 20 g. of sediment sample through 1000, 425, 250, 125, and 63 µm sieve by wet sifting.
2. Bake the sieved sediment sample at a temperature of 60 oC for 3 days.
3. Weigh the sample and calculate to percent by using the following formula

Sediment quantity (%) = Sediment dry weight of each size (g) 100
Total sediment dry weight (g)

Statistical analysis
Check the data distribution and compare the southwest and northeast monsoons using the

Minitab program. If the data has a normal distribution, use One Way ANOVA to analyze. If the data
has a non-normal distribution, use the Kruskal-Wallis test.

Results and Discussion
The result shows six shapes of microplastics which are fiber, fragment, film, pellet, granule, and foam

(Fig. 1 – Fig.6).

Fig. 1 Fiber Fig. 2 Fragment Fig. 3 Film Fig. 4 Foam Fig. 5 Granule Fig. 6 Pellet

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Fig. 7 Box and Whisker Plots of the number of microplastics at each station in southwest
and northeast monsoon

The numbers of microplastics found in the southwest monsoon at Bang Saen Beach, Wonnapha
Beach, Bangphra Jetty, Bangphra Beach, and Ko Loi were 1,791 6,749 2,498 3,228, and 5,161 pieces/Kg dry
weight, respectively. Fragments were found the most at Bang Saen Beach (653 pieces/Kg), Bangphra Jetty
(1,308 pieces/Kg), and Ko Loi (2,028 pieces/Kg). Fibers were found the most at Bangphra beach (1,455
pieces/Kg). Wonnapha Beach had the same number of fragments and fibers (2,339 pieces/Kg). The numbers
of microplastics found in the northeast monsoons at Bang Saen Beach, Wonnapha Beach, Bangphra Jetty,
Bangphra Beach, and Ko Loi were 2,265 2,013 2,525 1,172 and 1,862 pieces/Kg dry weight, respectively.
Fibers were found the most at every station (939 1,510 1,839 553 and 889, respectively).

100%
80%
60%
40%
20%
0%

BS/NE BS/SW W/NE W/SW BP1/NE BP1/SW BP2/NE BP2/SW KL/NE KL/SW
425 µm 250 µm 125 µm 63 µm < 63 µm

Fig. 8 Proportion of grain size of sediment at each station in southwest and northeast monsoon
In the southwest monsoon, the main constituents of sediment were grave and very coarse sand ( > 1

mm) at Wonnapha beach and Bang Phra beach, there was medium-coarse sand ( 250 – 425 µ m) at Ko Loi,
and fine sand ( 125 – 250 µ m) at Bang Saen beach and Bang Phra jetty. In the northeast monsoon, the main
constituents of sediment were grave and very coarse sand ( > 1 mm) at Bang Phra beach and Bang Phra jetty,
there was medium-coarse sand ( 250 – 425 µ m) at Wonnapha beach, and fine sand ( 125 – 250 µ m) at Bang
Saen Beach and Ko Loi.

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The variable analysis result showed that the numbers of microplastics had no significant difference
between stations (p-value > 0.05) but they had a significant difference between monsoons (p-value < 0.05)
caused by the current in the Gulf of Thailand. The current flow is anticlockwise and clockwise in the southwest
and the northeast monsoon, respectively (1). Thus, the microplastics were distributed and contaminated in the
beach sediment in the southwest monsoon more than northeast monsoon. In terms of the grain size, it had no
significant difference between both stations and monsoons (p-value > 0.05).

Conclusion
There were six shapes of microplastics encountered. Most of them were fibers and fragments. The

different monsoons affected the number of microplastics (p-value < 0.05). Most of the microplastic found in
the southwest monsoon was more than northeast monsoon but Bang Saen Beach and Bangphra jetty had
microplastics in the northeast monsoon more than southwest monsoon. That might be caused by the variable
current and wind. However, the difference in stations did not affect the number of microplastics (p-value >
0.05). Both different stations and monsoons did not affect the grain size (p-value > 0.05).

Acknowledgments
This project was supported by Science Classroom in University Affiliated School (SCiUS) under

Piboonbumpen Demonstration School, Burapha University. The funding of SCiUS is provided by the Ministry
of Higher Education, Science, Research, and Innovation. We also thank you to Asst. Prof. Dr. Anukul
Buranapratheprat for the advisement and thanks to Mr. Tontrakarn Obrom, Ms. Benjawan Khotchasanee, Ms.
Pattinee Kongpradit, and Mr. Nattapong Satja for the facilitation and supporting laboratory. This extended
abstract is not for citation.

References
1 Buranapratheprat, A., Yanagi, T., and Sawangwong, P. Seasonal variations in circulation and salinity

distributions in the upper Gulf of Thailand: modeling approach. (2002). La mer 40, 147-155.
2 Institute for Research and Development of Marine Resources and Mangrove Forests, and Faculty of Marine

Technology, Burapha University. Survey and classification of microplastic marine debris. (2014).
3 Jambeck J. R., Geyer, R., Wilcox, C., Siegler, T. R., Perryman, M., Andrady, A. and Law, K. L. Plastic

waste inputs from land into the ocean. (2015). Science 347:768-771.
4 Praditsap. N., Panichphol. A., Inkong. S., and Tantawanich, T. Population structure of marine

microbenthic animals at the sandy beach of Koh Sichang, Chonburi Province. National Budget Subsidy
Research Report 2008. (2008).

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Title : Microplastic residues in the commercial fish OE1_10_02
Field : off the coast of Samut Songkhram Province
Author :
Environmental Science and Ecology
School :
Adviser : Miss Pakpicha Moonjit

Miss Chayakan Rassameecharoenchai

Miss Phatcharaporn Jittirach

Kasetsart University Laboratory School Kamphaeng Saen Campus Educational Research
and Development Center

Dr. Varangkana Jitchum

Assist. Prof. Dr. Siraprapha Premcharoen

Abstract

Thailand's fishing industry is worth more than 80 billion baht which is one of the countries with the
highest fishery productivity in the world, producing more than 1.6 million tons per year. Fisheries provide a
source of income make a living for many households. It is also an important source of food for the people in
this country. The purpose of this study is to investigate microplastic contamination in greenback mullet
(Planiliza subviridis) and narrow-barred Spanish mackerel (Scomberomorus commerson), of those taken from
local fisheries at Ban Bang Bo Lang, Samut Songkhram Province. The study's findings have revealed
polystyrene (PS) and polyethylene terephthalate (PET) microplastics in greenblack mullet and narrow-barred
Spanish mackerel. These results must be reflected in our consumption. PS is likely to decompose from food
packaging such as foam boxes. and microplastics PET which are likely to degrade from clear drinking water
bottles. From the results, it also highlights the practice of dumping plastic waste into bodies of water without
proper disposal which causes another serious environment problem. Those improper disposal waste have
eroded by wave and sunshine, resulting in shreds which from polymer chain degradation. The microplastic
shreds do not dissolve in water and can absorb toxins found in the sea, which can affect not only fish and other
aquatic animals, but also humans who are at the bottom of the food chain.

Keywords : microplastic; commercial fish; environment

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Introduction

Microplastics have become a major source of pollution in marine and coastal ecosystems all over the
world. This makes them difficult to dispose of and ensures that their properties remain constant. Microplastics
residues in the environment are caused by a variety of factors, including those caused by large, broken, or
weathered plastics. The synthetic fibers clothes, tire parts, plastic bags, or plastic covers for drinking water
bottles which are deteriorating microplastics can easily contaminate and leave the residues in the environment
when water is drained into them. Because microplastics can absorb toxins in the sea, the longer they stay in
the water, the more toxic they become. Microplastics have been found in water, soil sediments, seas, and
oceans all over the world. When the marine life consumes microplastics, they accumulate in the food chain
and are passed down the food consumption hierarchy in the ecosystem. Due to the smaller microplastics can
pass through cell walls then they could cause the effects on organisms' health and livelihoods. For example,
the destruction of vascular tissue, the organisms at the end of the food chain, such as humans, may be exposed
to toxic residues. It influences the heart and contains toxins that can lead to cancer. Drinking alcohol can cause
microplastics to enter the body. The researchers, about water bottles, have also speculated that these
microscopic plastic particles could float in the air and enter the lungs of living organisms. There also have
been reported the effects of microplastics on a variety of aquatic organisms, such as fish from the Northeast
Atlantic Sea, according to Barboza et al. (2020), The microplastics have been found in most gastrointestinal
areas, according to studies. Polyethylene and polyesters are two common types of polymers which have been
found. According to the report by Jabeen et al. (2018), the goldfishes can get microplastics into their bodies
after these microplastics have been exposed to them. In addition, the fish has lost a significant amount of
weight. Microplastics are also accumulating in the gums and gastrointestinal tract, as evidenced by fibers.
Inflammation of the liver and intestines is caused. Microplastics in small pieces and granules are chewed and
excreted from the body rather than ingested. The jaw and jaw muscles are damaged because of polymer
textures. Microplastics are found in a wide range of the important commercial aquatic animals in various
marine environments around the world. This research is interested in investigating the microplastic residues
in commercial fish on the coast of Ban Bang Bo Lang Bang Kaeo Subdistrict, Mueang District, Samut
Songkhram Province. By using the chemical technique to analyze the microplastics in gastrointestinal tract of
the different feeding habit fish.

Methodology

1. Materials

1.1 Equipment and chemicals related to field visits

1.1.1 Refrigeration temperature 0 degrees Celsius
1.2 Equipment and chemicals involved in laboratory experiments

1.2.1 General glassware
1.2.2 Wooden stand for measuring the length of the fish

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1.2.3 Clear plastic zip-lock bag, size 5×7 cm.
1.2.4 Deionized water
1.2.5 Crucible
1.2.6 Desiccator
1.2.7 Hot air oven
1.2.8 Fourier Transform Infrared Spectrometer (FTIR), Perkin Elmer, Spectrum II
1.2.9 Acetone AR Grade
1.2.10 Ethanol AR Grade
2. Experimental Method
2.1 Fish Sampling
The fish samples were collected with local floating nets and then frozen for further analysis
in the laboratory. The 8-12 greenback mullet and narrow-barred Spanish mackerel samples were
selected to measure the standard length (SL), the total length (TL) and weight. Then the results
were recorded for comparison.
2.2 Plastics/microplastics Collecting from fish samples
The collected fish samples are caesarean section to separate the fish's stomach. The stomach
is then weighed and dissected to collect a liquid sample. To remove the contents of the stomach,
the deionized water was poured into the sample. The liquid from the sample was filled in a zip-
lock bag. And then, the 4.0 mL of liquid sample was placed in crucible and incubated at 100°C
overnight before being transferred to desiccator. Then, the residual sample was collected and
stored in the desiccator for further experiment.
2.3 Chemical Structure Analysis Using Fourier Transform Infrared Spectrometer (FTIR)
The functional group of residual samples were analyzed using FTIR technique. The area of
the FTIR was cleaned with ethanol and acetone. The background was scanned and collected
before testing the sample . The microplastic samples were placed onto the machine by using the
spatula stainless. After analysis, the samples were returned to the zip-lock bag. The machine was
cleaned with ethanol and acetone.

Results, Discussion and Conclusions
From the study of microplastics residues in commercial fish off the coast of Samut Songkhram

Province, polystyrene (PS) and polyethylene terephthalate (PET) have been found in both greenback mullet
and narrow-barred Spanish mackerel. These polymers can be related to human habit directly. PS is commonly
found in food packaging such as styrofoam, and PET is used for clear drinking water bottles.

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Samples Number Feeding habit Weight Standar Wave number (cm-1) and Type of
of fish (g) d length Chemical bonding plastics
that
have (cm)
ingested
plastic

Greenback 12 Bottom/ 75.7 16.8 3024 Aromatic C-H stretching PS
mullet
midwater ± 9.46 ± 0.7 2928 C-H stretching

1625 Aromatic ring stretching PET

1458 CH2 bending

1400 Aromatic ring stretching

1200 C-C-O ester group

1050 Aromatic CH bending

840 para substituted benzene ring

698 Aromatic CH out-of-plane bending

mono substituted benzene ring

517 Aromatic ring out-of-plane bending

Narrow- 8 Water surface/ 133.2 21.9 3024 Aromatic C-H stretching PS
barred
midwater ± 71.88 ± 4.0 2928 C-H stretching

Spanish 1625 Aromatic ring stretching PET

mackerel 1458 CH2 bending

1400 Aromatic ring stretching

1200 C-C-O ester group

1050 Aromatic CH bending

840 para substituted benzene ring

698 Aromatic CH out-of-plane bending

mono substituted benzene ring

517 Aromatic ring out-of-plane bending

The presence of the micro-polymers in all fish samples would be implied that the use of compostable
plastics might not be safe for living organisms. There are no researches indicating that the metabolism in
human body will be changed the chemical structure of these microplastic substances to make them more toxic
or not. The health effects of plastic/microplastics entering the human body are more important to understand
and beyond a reasonable doubt.

Acknowledgement

This project was supported by Science Classroom in University Affiliated School (SCiUS). The
funding of SCiUS is provided by Ministry of Higher Education, Science, Research and Innovation. This
extended abstract is not for citation.

References

1. Jung MR, Horgen FD, Orski SV, Rodriguez C V, Beers KL, Balazs GH, et al. Validation of ATR FT-IR to

identify polymers of plastic marine debris, including those ingested by marine organisms. Mar Pollut Bull
[Internet]. 2018;127:704–16.

2. Barboza LGA, Lopes C, Oliveira P, Bessa F, Otero V, Henriques B, et al. Microplastics in wild fish from

North East Atlantic Ocean and its potential for causing neurotoxic effects, lipid oxidative damage, and

human health risks associated with ingestion exposure. Sci Total Environ [Internet].

2020;717(134625):134625.

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List of Science Projects 12th SCiUS Forum
Oral presentation

Environmental Science and Ecology Group 2
Saturday August 27, 2022

No. Code Title Author School
Demonstration School
1 OE2_04_01 Elimination of toxic Miss Yosita Wijarnarong of Khon Kaen
University
cyanobacterium Microcystis Miss Nichaboon Budsarakam
Rajsima Witthayalai
aeruginosa using biological School

approach Surawiwat School,
Suranaree University of
2 OE2_06_01 Synthesis of pigments from Miss Pattaraporn Kowasupat Technology
Suankularbwittayalai
automotive industrial waste Mr. Natchanon Aimesungnern Rangsit School

for using in ceramic glaze Miss Thanaporn Wanthong Engineering Science
Classrooms
systems (Darunsikkhalai School)

3 OE2_18_01 The study of chicken feathers Miss Papawarin Maikla PSU Wittayanusorn
Surat Thani School
waste degradation by soil Mr. Kongpop Thummapimuk
Rajsima Witthayalai
bacteria Miss Gulisara Inpaeng School

4 OE2_11_01 Effects of Crude Extract from Miss Pawarisa Bunyakalumpa Islamic Science
Demonstration School
Sea Holly on Growth of Miss Jitsupa Arunpirom

Colletotrichum Miss Phinapat Poldej

gloeosporioides Causing

Chili Anthracnose Disease

5 OE2_09_02 Preparation of Coffee Waste- Miss Trakoonkaew Ketkan

to-Briquette for Producing a Miss Wanitcha Tunnikorn

Potential Alternative Solid Miss Pimphatcha Rethanu

Fuel

6 OE2_17_02 Development of biomass Miss Lakkhika Sukphan

pellet fuel using torrefied oil Miss Jidapa Theppanich

palm fronds with used

bleaching earth from palm oil

industry

7 OE2_06_03 Utilization of waste-derived Miss Napat Tangcharoen

biodiesel in a compression Miss Jedpreeya Siriprasertsilp

ignition engine Miss Nicha Seehanavee

8 OE2_19_01 Production of liquid organic Mr. Zuhairee Ramong

fertilizer from fishmeal Miss Thanaporn Kaewmak

factory by-product and study

on its efficiency of lettuce

growth

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No. Code Title Author School
Lukhamhanwarin
9 OE2_08_01 Tapioca Starch-Based Miss Thunchanok Nacha chamrab School

Biodegradable Film: Effect of Miss Piyamat Janthong Surawiwat School,
Suranaree University of
Glycerol and Silica Extracted Miss Nichapa Ruangkitwanit Technology

from Sugarcane Bagasse Ash PSU Wittayanusorn
Surat Thani School
10 OE2_18_02 Removal of cyanide from Mr. Wongsatorn Prommeechai
Engineering Science
wastewater of auto parts Mr. Patiphat Ratchaponsaen Classrooms
(Darunsikkhalai School)
manufacturing industry by Mr. Korakrit Boonprasatsuk
Naresuan University
electrocoagulation process Secondary
Demonstration School
11 OE2_17_01 Wood plastic composite from Mr. Krittapat Thongfua

mangosteen peel and plastic Mr. Nitipoom Phramsrichai

bottle wastes

12 OE2_09_01 Practices for Environmental Mr. Phoowasit Vipaschewin

Management of Waste from Mr. Phongsakorn

MICE Industry: Data Chimchoeysuwan

Compilation and Preparation Mr. Panat Lekpittaya

13 OE2_03_01 Compostable Colostomy bag Miss Sutheekan Pokajao

Miss Pemika Kangwanlertpanya

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Title: Compostable Colostomy bag OE2_03_01
Field: Environmental Science and Ecology
Author:
Miss Pemika Kangwanlertpanya
School:
Advisor: Miss Sutheekan Pokajao

University Naresuan University Secondary Demonstration School , Naresuan University

Asst.Prof.Supinda Sirilak M.D., Faculty of Medicine, Naresuan University

Assoc.Prof.Dr.Sukunya Ross, Department of Chemistry, Faculty of Science, Naresuan

Abstract:
Colorectal cancer is the one of most common cancer around the world. The primary treatment for this

cancer is surgery which aims to remove cancerous tissue. Some patients may need to insert a stoma and
colostomy for temporary or permanent drainage up to their conditions. Therefore, colostomy bags (plastic bags)
are necessary for them in everyday use. In terms of environment-friendly products and collaborating with the
Department of Chemistry, Faculty of Science, Naresuan University, we decided to develop the bio-degradable
(BD) colostomy bag from bioplastic. We created the sample of BD1 to BD8 which were different components
of bioplastic and run laboratory testing with mechanical testing, Scanning Electron Microscope (SEM), and
Optical Contact Angle (OCA). We found that BD1 to BD3 (Polylactic acid-based bioplastic) was not flexible
enough. About BD7 and BD8 (Polylactic acid and Polycaprolactone-based bioplastic), their texture was not
soft enough and irritated. BD4 to BD6 consisted of Polybutylene succinate-based bioplastic which was flexible
and non-irritating compared to others, but BD6 was not as flexible as BD4 and BD5. Together with
questionnaires of user satisfaction, BD5 was the final sample of our project. In conclusion, BD5 was suitable
for developing BD colostomy bags in the future medical industry to help patients and our world.

Keyword: Biodegradable, Bioplastic, Colorectal cancer, Colostomy, Colostomy bag

Introduction
Today, the world is facing the problem of colorectal cancer. In some cases, a colostomy bag is needed

to support the excretion of waste from the ostomy. According to the data of Naresuan University Hospital, a
survey as of February 14 2022, found that the price of colostomy bags that are sold in the hospital has 2 models,
namely B.Braun, priced at 170 baht per bag, and Proxima 2, priced at 68 baht per bag. If using social security
and gold card privileges, only 1 set per month will be received. Therefore, low-income patients have to reuse
the colostomy bag. This increases the risk of infection rates even higher. In addition, colostomy bags are made
from non-biodegradable plastic. Therefore, when the patient has finished using it, it will take time to
decompose, unlike the general plastic waste that the world is currently a problem with. The organizers,
therefore, thought to develop a colostomy bag that is sold in the general market. From non-degradable plastic
to biodegradable plastic from the Department of Chemistry, Faculty of Science Naresuan University It must
also have a price that is accessible to patients.

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Initially, the authors will develop a colostomy bug using a prototype from an economical colostomy bag.
assistance program patients with prosthesis Chongkonnee Nithi Foundation Chulalongkorn Hospital, Thai Red
Cross Society.

Methodology
To test bioplastics obtained from the Department of Chemistry, Faculty of Science, Naresuan

University, divided into 3 tests.
1. Tensile test
2. Contact angle
3. Color test
4. Stratification Questionnaire

Results, Discussion, and Conclusion
In the study of samples of bioplastics with properties that are flexible, strong, waterproof, opaque,

biodegradable, Cheap price that makes it accessible to all Thai people and does not irritate the patient's skin,
which affects the to be used in the further production of colostomy bags. All tests concluded that the BD4 and
BD5 samples were closest to the required properties mentioned above and could be further developed in the
biomedical industry.

BD4

20

15

10

5

0

-5 1
62
123
184
245
306
367
428
489
550
611
672
733
794
855
916
977
1038
1099
1160
1221
1282
1343
1404
1465
1526
1587
1648
1709
1770
1831
1892
1953
2014

Tensile stress (MPa) Tensile stress (MPa) Tensile stress (MPa)
Tensile stress (MPa) Tensile stress (MPa) Tensile stress (MPa)
Tensile stress (MPa) Tensile stress (MPa) Tensile stress (MPa)
Tensile stress (MPa)

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BD5

14
12

10

8

6
4
2
0

-2 1
31
61
91
121
151
181
211
241
271
301
331
361
391
421
451
481
511
541
571
601
631
661
691
721
751
781
811
841
871
901
931
961
991
1021
1051
1081
1111

Tensile stress (MPa) Tensile stress (MPa) Tensile stress (MPa)
Tensile stress (MPa) Tensile stress (MPa) Tensile stress (MPa)
Tensile stress (MPa) Tensile stress (MPa) Tensile stress (MPa)
Tensile stress (MPa)

Acknowledgment
This project was supported by Science Classroom in University Affiliated School (SCiUS). The

funding of SCiUS is provided by the Ministry of Higher Education, Science, Research, and Innovation. This
extended abstract is not for citation.

References
1. วรวิทย์ วาณิชยส์ ุวรรณ และคณะ. นวตั กรรมผลติ ภณั ฑท์ างการแพทยจ์ ากพอลิเมอร์เชิงคลีนิกมหาวทิ ยาลยั สงขลานครินทร์. 2562 [เขา้ ถงึ เมือ่ 5 กมุ ภาพนั ธ์ 2565] เขา้ ถงึ
ไดจ้ าก http://www.thailandplus.tv/archives/319109
2. นฤทธ์ิ ฝ่ ายบุตร. ผลของความขรุขระเชิงผิวต่อความสามารถในการเปี ยกน้าของฟิ ล์มบางคาร์บอนคลา้ ยเพชร. [วิทยานิพนธ์ปริญญาวิทยาศาสตร์มหาบณั ฑิต].
ขอนแก่น: มหาวิทยาลยั ขอนแก่น; 2556.

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Title : Elimination of toxic cyanobacterium OE2_04_01
Microcystis aeruginosa using biological approach
Field :
Authors : Environmental Science and Ecology

School : Miss Nichaboon Budsarakum
Advisor :
Miss Yosita Wijarnarong

Demonstration School of Khon Kaen University, Khon Kaen University

Assoc. Prof. Dr. Theerasak Somdee

Department of Microbiology, Faculty of Science, Khon Kaen University

Abstract
Water resource, one of crucial ecosystems for all organisms, has been suffered from water pollutants.

One of the most common natural water problems is cyanobacterial blooms, resulting in cyanotoxin
contamination in water, which is potential adverse effects on the animal and human health. Previous studies
revealed that physical and chemical eliminations of toxic cyanobacteria resulted in cell lysis, causing the
release of the toxins into water sources. Therefore, biological approach, which is a safe and natural treatment
for eliminating toxic cyanobacteria is a promising solution. Actinomycetes, a group of bacteria, are known as
a source of biological active compounds and have ability to inhibit other organisms. The aims of this study
were to isolate actinomycetes for elimination of toxic cyanobacterium Microcystis aeruginosa, the most
common toxic cyanobacterial bloom in eutrophic fresh water and select the most effective isolate of bacterium
for elimination of the cyanobacterium. Actinomycetes were isolated from soil in six different areas of Khon
Kaen University, Khon Kaen, Thailand. Thirty-three actinomycete isolates were isolated and screened for anti-
cyanobacterial activity against M. aeruginosa using co-culture assay. The result demonstrated that twenty-two
isolates were capable of inhibiting M. aeruginosa. The strain KKU-A7 exhibited the strongest anti-M.
aeruginosa with removal efficiency of 99.25%. Hence, this is promising to utilize KKU-A7 to eliminate
harmful cyanobacterial blooms in natural water.

Keywords : Elimination, Biological approach, Microcystis aeruginosa, Actinomycetes

Introduction
Eutrophication is one of the most important aquatic problems. A part of the problems is cyanobacteria

blooms, which can cause adverse effect to aquatic organisms by contaminated cyanotoxins. Microcystis
aeruginosa is a cyanobacterial bloom-forming species that is the most common in eutrophic lakes and
reservoirs throughout the world and can produce neurotoxins and peptide hepatotoxins called as microcystins
which are toxic to animals and humans. There are three common approaches of cyanobacterial removal
including physical, chemical and biological methods. However, physical and chemical methods can cause cell
lysis, which can increase a release of toxins in water resources. Therefore, the biological method is the most
suitable. Actinomycetes which are known to produce a wide variety of industrially and medically relevant
compounds are interesting alternatives for eliminating cyanobacteria. Therefore, the present study has a
hypothesis whether actinomycetes, which are isolated from soil samples from areas of Khon Kaen university,
can inhibit Microcystis aeruginosa or not.

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Methodology
Part 1 : Isolation of actinomycetes

Soil samples were collected from 6 different areas of Khon Kaen University, Khon Kaen, Thailand;
A) Central main stadium, B) agriculture area, C) Bueng Si Than reservoir, D) Romklao Kallapruek Park, E)
Agricultural Technology Park and F) Kaen Kallapapruek Building. Samples were air-dried for 1 week, crushed,
and sieved. Each sieved sample was treated with CaCO3 at a ratio of 10 to 1 and incubated at 37°C for 1 week.
The incubated samples were diluted down to 10-3 dilution, then each dilution was spread-plated on starch
casein agar plates in triplicate and incubated at 30°C for 1 week. Then actinomycetes were selected to be
streak-plated and incubated at 30°C for 1 week. Each isolate was stocked up in agar slant for further studies
and cultured in a 100-mL flask containing 45 mL starch casein broth for 1 week.

Part 2 : Screening of anti-Microcystis bacteria
Culture of Microcystis aeruginosa was grown in MLA medium for 1 week. For an experiment group,

each culture broth (part 1) was co-cultured with culture of M. aeruginosa at ratio 1 to 9 in duplicate and
cultured for 1 week. For a control group, cultures of M. aeruginosa were added starch casein broth at ratio 9
to 1 in duplicate and cultured for 1 week. After a week ago, the experiment group was compared with the
control group to determine an anti-Microcystis activity of each isolate and to find the most effective isolates
in the anti-Microcystis activity for further studies.

Part 3 : Testing of anti-Microcystis activity
Selected actinomycete isolates (part 2) were streak-plated on starch casein agar (part 1) and cultured

for 1 week. Spores of each actinomycete isolate were collected by scraping surface of cultured starch agar
casein, diluted down to 10-7 dilution, stocked up in microtubes and some of them were frozen for further
studies. Cultures of selected isolates were grown from 10-7 dilution of their spores in starch casein broth for 1
week. For a testing group, one-week cultured of M. aeruginosa was co-culture with cultured of selected
isolates at ratio 9 to 1. For a control group, one-week cultured of M. aeruginosa was added starch casein broth
at same ratio as the testing group. Both groups were cultured and shaken at 150 rpm for 1 week. On the first,
third, fifth, and seventh day of co-culture, 5 mL of each co-cultured medium was collected and frozen in
centrifuge tube.

Part 4 : Extraction of chlorophyll a
Frozen co-cultured medium was melted and centrifuged at 4000 rpm for 10 minutes. A liquid part in

each centrifuge tube was poured out, then 90% methanol was added in every centrifuge tubes. Methanol-added
centrifuge tubes were soaked in water bath at 70°C for 20 minutes and cooled down before they were
centrifuged at 4000 rpm for 10 minutes.

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Part 5 : Calculation of removal efficiency
A chlorophyll-a concentration was measured at 630, 645, 665 and 750 nm with a spectrophotometer

and calculated with the following equation:
Chlorophyll a (mg/L) = [11.6 x (A665-A750) - 1.31 x (A645-A750) - 0.14 x (A630-A750)] x F

When A630 A645 A665 and A750 are values that were measured at 630, 645, 665 and 750 nm in order
and F is value of volume of methanol (mL) x volume of sample liquid (L) x (1/cuvette width (cm)). Then
calculate the removal efficiency (%) with the following equation:

Removal efficiency (%) = [ 1 – (Ct/C0)] x 100
When Ct is a chlorophyll-a concentration of actinomycetes testing group and C0 is a chlorophyll-a
concentration of control group.

Results
Thirty-three different isolates of actinomycetes were isolated from soil samples that were collected

from 6 different areas. Sample was isolated for 7 isolates from central main stadium, 10 isolates form farming
area, 9 isolates from Bueng Si Than, 2 isolates from Romklao Kallapruek, 3 isolates from Agricultural
Technology Park and 2 isolates from Kaen Kallapapruek Building. Screening of anti-Microcystis bacteria
revealed that 22 isolates had anti-Microcystis ability. Each isolate was compared and showed that the strain
KKU-A7, KKU-B8 and KKU-B10 were possessed the anti-Microcystis activity. Removal efficiency of KKU-
A7, KKU-B8 and KKU-B10 was shown in Figure 1, 2, 3 and 4.

Figure 1 : Removal efficiency (%) of co-culture day 1. Figure 2 : Removal efficiency (%) of co-culture day 3.

Figure 3 : Removal efficiency (%) of co-culture day 5. Figure 4 : Removal efficiency (%) of co-culture day 7.

According to Figure 4, it demonstrated that strain KKU-A7 had anti-M. aeruginosa ability with
removal efficiency of 99.25%, strain KKU-B8 had anti-M. aeruginosa ability with removal efficiency of
0.00%, and strain KKU-B10 had anti-M. aeruginosa ability with removal efficiency of 87.29%.

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Conclusion
From this study, it found that 33 actinomycete isolates were isolated from soil of 6 areas and screened

for anti-cyanobacterial activity against M. aeruginosa using co-culture assay. The result demonstrated that 22
isolates have ability to inhibit M. aeruginosa. From calculating and comparing the removal efficiency, the
result showed that the strain KKU-A7 was the strongest anti-M. aeruginosa with removal efficiency of 99.25%.
So, KKU-A7 tended to eliminate harmful cyanobacterial blooms in natural water.
Acknowledgements

This project was encouraged by our advisor, Assoc. Prof. Dr. Theerasak Somdee. This project was
supported by Science Classroom in University Affiliated School (SCiUS). The funding of SCiUS is provided
by Ministry of Higher Education, Science, Research and Innovation. This extended abstract is not for citation.
References
1. Phankhajon K, Somdee A, Somdee T. Algicidal activity of an actinomycete strain, Streptomyces rameus,
against Microcystis aeruginosa. Water Science and Technology 2016;74(6):1398-1408.
2. Somdee T, Sumalai N, Somdee A. A novel actinomycete Streptomyces aurantiogriseus with algicidal
activity against the toxic cyanobacterium Microcystis aeruginosa. J Appl Phycol 2013;25:1587-1594.

3. ณัฐวฒุ ิ หวงั สมนกึ และ จรี พร เพกเกาะ. การควบคมุ ไซยาโนแบคทเี รียและย่อยสลายสารพิษไมโครซสิ ตินโดยแบคทเี รยี
จากแหล่งนา้ บางแหง่ ในจงั หวัดเชยี งใหม่. วารสารวจิ ยั และพัฒนา มจธ. 2558;38:155-66.

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Title : Synthesis of pigments from automotive industrial waste OE2_06_01

for using in ceramic glaze system
Field : Environmental Science and Ecology

Author : Natchanon Aimesungnern , Thanaporn Wanthong, Pattaraporn Kowasupat

School : Rajsima Wittayalai School, SCiUS-Suranaree University of Technology

Advisor : Asst. Prof. Dr. Siriwan Chokkha, Insititute of Engineering, Suranaree University of Technology

Abstract
The waste produced from Thailand’s automotive industry has alarmingly increased to approximately 70-100

tons per year. The cheaper way is landfill but this waste will corrode soil and this has caused considerable concerns
on the environmental impacts. The rich source of Fe-iron and Fe2O3 in this waste suggest another possibility of
using it as starting materials for producing compounds such as a pigment. The automobile waste in the form of
powder was first cleaned and mixed with other starting raw materials, including ZnO, Cr2O3 and AlCl3.6H2O in
various proportions. The homogeneous powder mixture was heated at 1250oC in an electric furnace, to form a new
formula ceramic pigment. The 6% by weight of the prepared ceramic pigment was then added into a mixture of
glazing components before applied onto a biscuit tile. Then, the sample was subjected to thermal treatment at
1200oC in an electric furnace. Finally, CIE L*a*b* color scale and structural phase of pigments were analyzed using
a HunterLab spectrophotometer and X-ray diffraction (XRD), respectively. The results suggested that the
automotive industrial waste can be utilized to synthesize a pigment with a variety of shades in Fe-Cr-Al-Zn system.
This leads to the production of ceramic workpieces to be more distinctive and more diverse. More importantly, the
pigment produced from the automobile industrial waste from this work could help decrease the disposal costs, and
efficiently reducing discharged wastes and environmental impacts.

Keywords: automotive industrial waste, ceramic pigments, clay-based ceramic, ceramic glazing

Introduction
The automobile is an important factor for everyday activities in facilitating human transportation and consumer demand for

automobiles has increased (Preeti S. et al., 2016). As a result, Thailand’s automotive industry is growing rapidly with a high
production rate, leading to an increase in the level of automotive industrial waste. the automotive parts are manufactured by metal
casting for use as engine components such as bolts and screws, etc. This production causes automotive industry waste in large
amounts of 70 -100 tons per year per small company. There are at least three ways for automotive waste management, which is 3
Rs. The first R is to recycle by using pyro-metallurgical processing. The product from recycle is high grade alloys but this is
expensive method and need high skilled worker and advanced technology. Another approach is to reform using chemical

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processing. Since the waste riches in Fe which is transition elements. Therefore, the products obtained can be coordination
compounds, catalysts and pigments.Final way is relocation as Landfill may be the cheapest way with waste disposal costs of 2,000
baht per ton. However, the consequence is that chemical reactions in corrosion can occur, leading to toxins contaminating in soil
and posing a danger to the environment (Geoffrey H., 2003).

Ceramic color pigments are important additive materials used for the decorating of ceramic products.The color or shade of
ceramic pigment are always derived by chemical compounds of transition elements, such asvanadium, iron, cobalt, manganese,
nickel, copper, chromium,praseodymium, etc.The starting composition are mixed and fired at the appropriate temperature to form
a new crystalline structure of ceramic pigment, depending on chemical composition and ratio of starting precursor(Deraz, N.et al.,
2013; Medeiros, P. et al., 2015; Yüngevis, H. 2013). Therefore, the aim of the present research work is interested in helping our
environment by study on the possibility of utilizing Fe-waste from automotive industry as a starting raw material for pigment
synthesis and use the pigment as additive in ceramic glazing system.

Methodology
The waste powder was washed by using the water to remove oil and the magnetic was used to separate a contaminant from

the grinding process. Then, the waste slurry was dried in the oven at 100ºC for 24 hours to create a dark brown powder. The
chemical composition and structural phase of the automotive industrial waste were analyzed by X-ray fluorescence (XRF)and X-
ray diffraction (XRD). After confirming a pure phase of Fe-waste powder, it was sieved through a 325 mesh before used as a
precursor for synthesis of a new Fe-pigment. The cleaned Fe-waste was used as a precursor by mixing with Cr2O3, ZnO and
AlCl3.6H2O in various ratio.Each composition ratio was weighted balance before mixing and firing at 1250ºC for 1 hour, to form
a specific formula of ceramic pigment. Finally, the glaze slurry was prepared by mixing glaze powder and 6% by weight of an
additive pigment in the medium as water.Then glaze slurry was coated on the ceramic body by using Dip coating technique.The
specimen was fired at 1200ºC for 1 hr., to transform soft mineral into hard glazing ceramic and high density.The pigment color
characteristics were quantified by HunterLab spectrophotometer with CIE L*a*b*color scale.The structural phase of the new Fe-
pigment composition was identified by X-ray diffraction.

Results, Discussion and Conclusion
From Fig. 1, the elemental composition of automotive industrial waste was analyzed by XRF and shown in the rich source

of 97 wt%of iron (Fe)and small amounts of others including Cr, Al, Niand Mn.The XRD pattern of the dried automotive industrial
waste powder is shown in the forms of Fe-metal and Fe2O3 with a high purity of 97 wt% similar to a laboratory grade of chemical
compound, indicating the possibility of using an automotive waste as a starting material for ceramic pigment synthesis.

For the 4-axial diagram based on ceramic theory, six shades in Fe-Cr-Al-Zn system such a white, green, orange, ovaltine,
light brown and dark brown can be observed by the naked eyes and simulated in Fig. 2. The digital pictures and CIE L*a*b*
standardized color scale of example work pieces in each shade are clearly different. For example, all workpieces showed in a high

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L* value, indicating that the stain color was white and perfect reflecting diffuses. However, the difference in color was primarily
caused by differences in the values of a* and b* axes, which can be explained as follows: For the white no. 36, a* and b* were the
lowest values near zero. Therefore, it can be seen the color as white. For the green no. 32, the value of a* was low near zero and b*
was so high. From this result, it should actually see as yellow. But because the precursor contains Fe-waste which gave a* green
color at the testing temperature, so that the glazing appeared green. For the Orange no. 3, Ovaltine no. 30, Light Brown no. 2 and
Dark Brown no. 15, a* and b* axis are the highest values. From this result, the glaze was a mixture of red from a* axis and yellow
from b*axis, so, the glaze color range was orange-brown, which various shades depended on the different amount of precursor that
used to produce the pigment.

Fig. 1 Fig. 2

Fig. 1 The chemical composition by XRF and XRD and Fig. 2 The digital pictures and CIE L*a*b* color scale

The analytical XRD patterns of all specific Fe-waste pigment compositions are shown in the Fig. 3(a) – 3(f). All the XRD
results showed new structural phase of chemical products and some remaining precursor. For white no. 36 from Fig. 3(a), it was

found that the starting raw materials reacted and converted to a new structure of ZnAl2O4, and the excess ZnO was the remaining
precursor from the reaction. For green no. 32 from Fig. 3(b), Zn(Al1.8Fe0.2)O4 was found as a new structure formed by precursor
reaction and Fe2O3 was the remaining precursor. For orange no. 3 from Fig. 3(c), precursors reacted completely and forming in a
new of ZnCr2O4 and (Fe0.6Cr0.4)2O3. For ovaltine no.30 from Fig. 3(d), Zn(Al1.8Cr0.2)O4 was a new structure formed by precursor
reaction and ZnO was remaining precursor. For light brown no.2 from Fig.3(e), all precursors reacted completely and forming in a
new compound of ZnCr2O4 and (Cr1.3Fe0.7)O3.And finally for dark brown no.15 from Figure 7(f), all precursors also reacted
completely and forming in a new compound of ZnCr2O4, (Fe0.6Cr0.4)2O3 and Zn(AlFe)O4. It can be seen that the starting materials

mixed in different ratios lead to different reactions which produced specific products with different phases and different quantities
that gave a wide variety of color-shades.

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3(a) White Pigment No.36 3(b) Green Pigment No.32 3(c) Orange Pigment No.3

Intensity (a.u.) Intensity (a.u.) Intensity (a.u.) ZnCr O PDF 82-1048
24
ZnAl2O4 PDF 82-1043 Zn(Al Fe )O PDF 82-1038
ZnO PDF 36-1451 1.8 0.2 4 (Fe Cr ) O PDF 34-0412
Fe O PDF 39-1346 0.6 0.4 2 3
23

10 20 30 40 50 60 70 80 10 20 30 40 50 60 70 80 10 20 30 40 50 60 70 80
2-Theta (degree) 2-Theta (degree) 2-Theta (degree)
3(d) 3(e) 3(f)
Ovaltine Pigment No.30 Light Brown Pigment No. 2 Dark Brown Pigment No.15

Intensity (a.u.) Intensity (a.u.) Intensity (a.u.) ZnCr2O4 PDF 87-0028
(Fe Cr ) O PDF 34-0412
Zn(Al Cr )O PDF 82-1039 ZnCr O PDF 01-1123
1.8 0.2 4 24 0.6 0.4 2 3
ZnO PDF 70-2551
Cr Fe O PDF 35-1112 Zn(AlFe)O PDF 82-1047
1.3 0.7 3 4

10 20 30 40 50 60 70 80 10 20 30 40 50 60 70 80 10 20 30 40 50 60 70 80

2-Theta (degree) 2-Theta (degree) 2-Theta (degree)

Fig. 3(a) – 3(f) The XRD patterns of Fe- waste based pigment

The confirmed results from this study which illustrated the reformation of Fe waste into more valuable
pigments, does not only provide waste management’s benefit for automotive industry but also provide pigments
with a variety color-shades for ceramic industry as well.

Acknowledgements
This project was supported by Science Classroom in University Affiliated School (SCiUS) under Suranaree

University of Technology and Ratchasima Wittayalai School. The funding of SCiUS provided by Ministry of
Higher Education, Science, Research and Innovation. This extended abstract is not for citation.

References
1. Liu Y., Liu Y., Chen J.(2014).“The Impact of the Chinese AutomotiveIndustry:Scenariosbasedon the NationalEnvironmental

Goals”.Journal of Cleaner Production.
2. Manojit G., Arkajit G.and Avinava R.(2019).“Renewable and Sustainable Materials in Automotive Industry”. Encyclopedia of

Renewable and Sustainable Materials.
3. Preeti S., Aksha S., Ajay S. and Preeti S. (2016). “Automobile Waste and Its Management”. Research Journal of Chemical and

Environmental Sciences.Vol 4 [2].
4. Deraz, N., Abd-Elkader, O., (2013).“Investigation of magnesium ferrite spinel solid solution with iron-rich composition”, Int.J.

Electrochem.Sci.8; 9071–9081.
5. Medeiros, P., Gomes, Y., Bomio, M., Santos, I., Silva, M.(2015).“Influence of variables on the synthesis of CoFe2O4 pigment

by the complex polymerization method”, J.Adv.Ceram.4135–141.

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Title : The Study of Chicken Feathers Waste Degradation by OE2_18_01
Field : Soil Bacteria
Environmental sciences

Author : Mr. Kongpop Thummapimuk

Ms. Gulisara Inpaeng

Ms. Papawarin Maikla

School : Surawiwat School, Suranaree University of Technology

Advisor : Dr. Sakesit Chumnarnsilpa, School of Chemistry, Institute of Science, Suranaree University of

Technology
Dr. Monrawat Rauytanapanit, Surawiwat School

Abstract
In 2016, 3,000 tons of chicken feathers wastes were discarded which are hard to be destroyed. The

usual treatment of chicken feathers by using burying or burning processes which generate acidic soil and pollute
air. In this project, we dispose of chicken feathers by using bacteria from soil. The aim of the study was to
identify bacteria which is extracted from soil and to study the degradation process in each bacteria. There are 5
bacteria - Bacillus subtilis, Bacillus siamensis, Escherichia coli, unknown bacteria1 (FD1) and unknown
bacteria2 (FD2). As a result, every bacteria can degrade chicken feathers. After the identification of FD1 and
FD2 based on 16s rDNA sequences analysis revealed that FD1 has homology to Lysinibacillus capsici and FD2
has homology to Bacillus safensis. The analytic results revealed that pH values illustrate the alkaline nature of
feathers degradation and optical density (O.D.) values indicated the rapid rate of feathers degradation. FD2 can
degrade chicken feathers within 8 days, while others have some rachis left.

:Keywords Chicken feathers, Soil bacteria, Bacillus, Escherichia, Lysinibacillus

Introduction
Chicken feathers are a waste from the poultry industry which is hard to be destroyed. Chicken feathers

waste is about 3,000 tons per week. Nowadays, the poultry industry degrades chicken feathers waste by burying
and increasing heat and pressure which pollute soil and air due to sulfur dioxide in chicken feathers structure.
Chicken feathers consist of keratin over 90%. Keratin is a very strong structure formed by several types of
amino acids. The most common type is cysteine. In cysteine there is Thiol which is a function group that can
form disulfide bonds. The strength of keratin depends on the amount of disulfide bond. There are 2 types of
keratin - alpha-helix which is stronger and the second one is beta-sheet. Chicken feathers are beta keratin.
The rachis resembles a sandwich-structured composite: a dense keratin cortex surrounded by a spongy keratin.

According to previous research, chicken feathers can be degraded by soil bacteria which use keratin

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in feathers as a carbon and nitrogen source. Soil bacteria can produce keratinase and decompose feathers
by destroying disulfide bonds. Then the structure is changing from network structure to
beta-sheet structure.

The present study is aimed to identify bacteria which is extracted from soil and to study the white
chicken feathers degradation process in each bacteria. Keratinolytic bacteria are economically and
environmentally friendly. We expected keratin as a product from degradation process and keratin can be used
as various objectives for example culture media and fuel source.

Methodology
Materials white feather, distilled water, centrifuge tube, salt, Dipotassium phosphate, culture medium, Bacteria 3
species Escherichia coli , Bacillus siamensis, Bacillus subtilis and 2 Unknown bacteria are Unknown bacteria
1 (FD1), Unknown bacteria 2 (FD2), Crystal violet, Iodine solution, Safranin, Ethylalcohol 95%, Shaker, Hot air
oven, Centrifuge, Autoclave, UV-VIS Spectrophotometer, Universalindicator
Bacteria identification According to the previous researcher, they have already identified 3 bacteria which
are Bacillus subtilis, Bacillus siamensis, Escherichia coli and then these bacteria were stored in the fridge (1).
So we continue identifying other bacteria - FD1 and FD2 by 16s rDNA sequences analysis.
Bacteria culture we did a streak plate for every bacteria to purify bacteria then selected the single colonies
to culture in liquid broth. After that stain Gram and observed colony for FD1 and FD2.
Feathers preparation We defatted chicken feathers by washing it with 150g of detergent per 200g of white
feathers in 5 liters of water. Then the defatted feathers were dried in the oven for 48 hours. The dried feathers
were stored in the closed container.
Feathers degradation We degraded chicken feathers in the tube, in each tube containing 2 ml of culture
medium and bacteria, 0.2 g of chicken feathers, 0.01 g of sodium chloride, and potassium phosphate buffer
solution. The total solution was 20 ml per tube. The controlled group was the same as the experimental group
without bacteria. The bacteria were as follows: Bacillus subtilis, Bacillus. siamensis, Escherichia coli, FD1,
FD2 and the controlled group were repeated in a total of 3 experiments. The feathers were degraded in a shaker
at 150 rpm at 37 °C. The pH and absorbance value (Optical Density: OD) at 550 nm was measured by universal
indicator and UV-visible spectrophotometer continuously throughout the experiment.

Results
1.Bacteria identification After staining Gram and colony observation, Both FD1 and FD2 are Gram
positive and the shape is a short cylinder. As well as the colony observation results, FD1 and FD2 were
irregular on the top, flat from the side and the outer rim was undulate.

Picture 1 shows colony of FD1 Picture 2 shows Gram staining of FD1

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Picture 3 shows colony of FD2 Picture 4 shows Gram staining of FD2

Identification of FD1 and FD2 based on 16s rDNA sequence analysis revealed that FD1 has
homology similar to Lysinibacillus capsici at 100% identity and FD2 has homology similar to Bacillus safensis
at 99.92% identity.

2. Results of chicken feathers degradation After feathers were degraded for 8 days, There was colloid

solution in every tube and Bacillus safensis can totally degrade chicken feathers within 8 days while others
have some rachis left.

Table 1 feathers degradation at day 1, 2, 3 and 8

Bacillus safensis

Lysinibacillus capsici

Bacillus siamensis
Bacillus subtilis
Controlled group

Escherichia coli

Graph 1 O.D. values everyday during the degradation Graph 2 pH values everyday during the degradation

Graph 1 indicated that O.D. values tend to increase wherewith Bacillus safensis has the highest O.D.
value and totally degraded feathers the fastest, followed by L. capsici, B. siamensis, E. coli, B. subtilis, controlled
group, respectively. Graph 2 indicated that pH at day 8 in 5 bacteria and controlled groups is higher than the
beginning of the process. pH value at day 8 is around 7-8.

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Conclusion & Discussion
The result showed that all bacterias (Escherichia coli , Bacillus siamensis, Bacillus subtilis, Bacillus

safensis, Lysinibacillus capsici) have the ability to degrade chicken feathers within 8 days. On day 8th
B.safensis lost more than 50% of chicken feathers weight. However, they still have some rachis left, because
rachis arrangements are more complicated than chicken barbs (4).

Trichophyton mentagrophytes and Trichophyton terrestre which are fungi but they can produce
keratinase as same as the present bacteria (3). Within 7 days T.mentagrophytes and T.terrestre lost chicken
feathers weight 25% and 20%, respectively. After 21 days T.mentagrophytes and T.terrestre only lost 69.7%
and 59.6% of feathers respectively. As a result, B.safensis degraded chicken feathers faster thanT.mentagrophytes.

Optical density (O.D) measurement of bacteria cultures in graph 1 showed the growth curve of
bacteria. The early stage of the graph is the exponential curve, which is the growth stage. After that bacteria
lack of food because there the number of food is insufficient for bacteria. As the result, some bacteria are dead
andthen looping inall experiments. Although we cleaned chicken feathers by detergent it was still contaminated
and graph 1 showed the control group has the same curve as other bacteria.

pH measurement in each tube showed the result in graph 2. After day 8th pH of chicken feathers is 8
with B.safensis.pH demonstrated the alkaline nature of the experiments that release amino groups from chicken
feathers (2).Since pH of our experiments have a difference between day 1st and 8th less than Prasanthi’s
research. We might extrapolate that there are more left keratin than previous research (3). Since B.safensis is
not pathogenic bacteria it is a good choice to use B.safensis in industry.

Acknowledgements
This project was supported by Science Classroom in University Affiliated School (SCiUS) under

Suranaree University of Technology and Surawiwat school. The funding of SCiUS is provided by the Ministry
of Higher Education, Science ,Research and Innovation. This extended abstract is not for citation.

References
1. Chaowatthanaphanit, T., Meesawat, P., Yoovanichanon, C. E-Poster SCiUs Forum #11 @ BUU

[Internet].Screening of feather hydrolytic bacteria for agriculture and microbial industry applications. 2021
[cited 10 October 2021]. Available from: https://fliphtml5.com/bookcase/jmmvb

2. Chilakamarry C, Mahmood S, Saffe S, Arifin M, Gupta A, Sikkandar M et al. Extraction and application
of keratin from natural resources: a review. 3 Biotech. 2021;11(5).

3. Prasanthi, N., Bhargavi, S., Machiraju, P.V.S. Chicken feather waste - a threat to the environment.
International Journal of Innovative Research in Science, Engineering and Technology. 2016;5:16759-16764.

4. Wang B, Yang W, McKittrick J, Meyers M. Keratin: Structure, mechanical properties, occurrence in
biological organisms, and efforts at bioinspiration. Progress in Materials Science. 2016;76:229-318.

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Title: Effects of Crude Extract from Sea Holly on Growth of OE2_11_01
Colletotrichum gloeosporioides Causing Chili Anthracnose Disease

Field: Environmental Science and Ecology

Author: Miss Pawarisa Bunyakalumpa

Miss Jitsupa Arunpirom

Miss Phinapat Poldej

School: Suankularbwittayalai Rangsit School, Thammasat University

Advisor: Dr. Varundhorn Chuaboonmee and Dr. Tiwtawat Napiroon, Department of Biotechnology,

Faculty of Science and Technology, Thammasat University

Abstract
Bio-pesticide is one of the environmental friendly pest control. Chili which is one of importance economic

plants are greatly damaged by anthracnose disease fungi called Colletotrichum gloeosporioides. In this study
extraction, identification and anti-fungal activity analysis of Sea Holly plant were investigated. The experiments
were divided into three parts. The first experiment, crude extract of leaf and stem of Sea holly were extracted using
methanol with and without sonication methods. The second experiment, crude extract were identified using Thin
Layer Chromatography (TLC) and Gas Chromatography-Mass Spectrometry (GC- MS) technique. The last

experiment, effect of crude extract on inhibiting Colletotrichum gloeosporioides growth were investigated using
agar diffusion technique. The results showed that the highest yield (1.606 g) of crude extract were obtained from
leaf extraction by using methanol with sonication for 120 minutes. The results of phytochemical screening of crude
extracts by Thin-Layer Chromatography (TLC) were not shown positive test of alkaloids and terpenoid. However,
the results of GC-MS analysis showed that crude extracts from leaves contained n-Hexadecanoic acid as the major
compound, while the crude extracts from stem barks contained Tricyclo [4.1.0.0(2,7)] hept-3-ene as the major
compound. The effect of crude extracts on Colletotrichum gloeosporioides growth showed that crude extracts from
stem barks tended to inhibit Colletotrichum gloeosporioides growth.
 
Keywords: Sea Holly, Crude extract, Anthracnose disease, Colletotrichum gloeosporioides
 
Introduction 

Colletotrichum gloeosporioides is a cause of anthracnose disease which damage so many plants,
especially chili. The infected chili develop juicy blisters and expand around the fruit and fall off before harvesting.

According to famous methods, synthetic chemical pesticides called mancozeb and benomyl were used for get rid
of the fungi. However, these chemicals cause human health and environmental problems. Therefore, another
method for eliminating this fungi is the use of bioactive compounds from plants as bio- pesticides.

In this study leaves and stem barks of Sea Holly which is a medicinal plant were extracted and tested for
the anti- fungal activity against Colletotrichum gloeosporioides.

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Methodology 
1. Plant extraction

Sea Holly’s leaves and stem barks were dried at room temperature and kept in dry container. Leaves
and stem barks were then ground using pounder machine. The fine powder was extracted in two steps including
maceration in 98.8% methanol and sonication at difference times. First, the mixtures were shaken in rotary
shaker for 3 days. After that the mixtures were sonicated at 0, 30, 60 and 120 min, respectively depending on the
condition. The extracts were filtered through filter paper and then dried by evaporating method. The dry crude
extract were filled methanol until to 500,000 ppm. The extracts were kept at 4° C temperature.
2. TLC and GC-MS analysis of bioactive substances from Sea holly

TLC plates were prepared of the appropriate size (10x10 cm) and the crude extracts were spotted at
baseline using a capillary tube at 30 μg/mL concentration (20 drops/spot). The TLC was developed in
different solvent systems which arrange by polarity and then the Rf values were calculated to be 8 cm. After
development, the TLC plates were air-dried and observed under the UV light at 254 and 365 nm. Two different
specific reagents including Dragendorff’s reagent for alkaloids detaction and Anisaldehyde sulfuric acid
reagents for terpenoids detection were sprayed on to TLC plates. In addition, a test tube screening of alkaloids
detection were also examined. Regarding GC-MS analysis, the crude extracts of interesting sample were
submitted for analyzing MS at the National Nanotechnology center, Science Park, Pathum Thani province.
3. Anti-fungi analysis against Colletotrichum gloeosporioides

The 100 µL of 100,000 ppm crude extracts were spread on PDA plate. Colletotrichum gloeosporioides
culturing on agar plate were cut by cork borer and placed on the PDA plate covering with crude extracts. Methanol
was used as negative control and Mancozeb was used as positive control. The colony size of fungi on agar pate
were measured and recorded.
 
Results  
1. Crude Extracts Yield

The crude extract extraction of leaf and stem of Sea holly using methanol with and without sonication methods
were conducted. The results showed that the highest yield (1.606 g) of crude extract was obtained from leaf
extraction by using methanol with sonication for 120 minutes as shown in Table 1. 

Table 1 Yield of dry-crude extract

Sample Code Dry weight Sample Code Dry weight
(g) (g)
Leaf-Non sonication-0 L-NS-0 S-NS-0
minute L-S-30 0.793 ± Stem bark- Non sonication -0 S-S-30 1.368 ±
L-S-60 0.268 minutes S-S-60 0.068
Leaf-Sonication-30 L-S-120 S-S-120
minutes 1.323 ± Stem bark- Sonication -30 1.225 ±
0.018 minutes 0.419
Leaf- Sonication -60
minutes 1.446 ± stem bark- Sonication -60 0.947 ±
0.077 minutes 0.052
Leaf- Sonication -120
minutes 1.606 ± Stem bark- Sonication -120 1.299 ±
0.122 minutes 0.200

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2. TLC and GC-MS analysis of bioactive substances from Sea holly
The results of TLC plate of leaf crude showed that absorbing bands were found at wavelength 365 nm (Rf

value = 0.14 and 0.31) as shown in Figure 1. When sprayed with Anisaldehyde sulfuric acid, the resulting
substance was green-gray color.

Figure 1 TLC chromatograms of 1.1 leaf extracts from Sea Holly

The TLC plate of stem bark extracts was observed under UV light with wavelengths of 365 nm and 254

nm, the total absorbance bands were found at the baseline and when sprayed with Dragendorff, there was no
color on the TLC plate.  When dragendorff's reagent was added to the crude extracts for alkaloid detection, all
extracts were not shown a positive test of alkaloids (orange color and sediments). 

The results obtained from the GC- MS analysis showed that leaves crude extract contained n-
Hexadecanoic acid as the main component and the stem bark crude extract S- NS- 0- 3 contained Tricyclo
[4.1.0.0(2,7)] hept-3-ene as the main component. Some details of compounds were shown in Table 2.

Table 2 Main bioactive compounds from crude extract of leaf and stem bark

Explant Name of Compound Some Biological Activity
Leaf Hexadecanoic acid -Inhibit the fungi A.hydrophila P.fiuorescens
-Suppresses Candida albicans
Stem bark Tricyclo[4.1.0.0(2,7)]hept-3-ene -Used as an insect repellent

3. Anti-fungi analysis against Colletotrichum gloeosporioides

The effect of crude extracts on growth of Colletotrichum gloeosporioides inhibition were test by agar
plate diffusion method. The colony size of fungi on agar pate were measured and recorded. The results showed
that sample S-NS-0 showed highest inhibition activity. The details were shown in Table 3 and Figure 2.

Table 3 Colony size of Colletotrichum gloeosporioides after incubate with the PDA plate covering with crude

extract

Sample Diameter (cm)
L-NS-0 8.1 ± 0.1

L-S-30 7.9 ± 0.1
S-NS-0 7.8 ± 0

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S-S-30 7.73 ± 0.03
Methanol (Negative control) 8.5 ± 0
Mancozeb (Positive control)
7.83 ± 0.06

*The results were obtained from 3 replications of the experiment

Figure 2 Colony size of Colletotrichum gloeosporioides after incubate with the PDA plate covering with Sea
Holly’s crude extracts A = Methanol (Negative control) /B = Mancozeb (Positive control)/
C = L-NS-0/ D= L-S-30/ E = S-NS-0/ F = S-S-30

 
Conclusions

The highest yield (1.606 g) of crude extract were obtained from leaf extraction by using methanol with
sonication for 120 minutes. The results of GC- MS analysis showed that crude extracts from leaves contained n-
Hexadecanoic acid as the major compound, while the crude extracts from stem barks  contained Tricyclo
[ 4. 1. 0. 0( 2,7) ] hept- 3- ene as the major compound. In addition, crude extract from stem barks tended to inhibit
Colletotrichum gloeosporioides growth. 

Acknowledgments 
This project was supported by Science Classroom in University Affiliated School ( SCiUS) under

Thammasat University and Suankularb Wittayalai Rangsit School. The funding of SCiUS is provided by Ministry
of Higher Education, Science, Research and Innovation, which is highly appreciated. This extended abstract is
not for citation.
 
References 
1. พิกลุ นชุ นวลรตั น,์ อจั ฉรา บญุ โรจน.์ ผลของสารเคมี Prochloraz, Benomyl, Carbendazim, Azoxystrobin, Mancozeb และ Copper

oxychloride ตอ่ การควบคุมโรคแอนแทรคโนสของแกว้ มงั กร. วารสารวิจยั ราไพพรรณี 2558;2:15-20.
2. วิชชดุ า สุขปานพรหม, “การทดสอบประสิทธิภาพสารป้องกนั กาจดั เช้อื รา 3 ชนิดในการยบั ย้งั การเจริญของเส้นใยและการงอกของรา Collettotrichum

gloeosporioides และ Colletotrichum Capsici.” 2540

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Title : Preparation of Coffee Waste-to-Briquette OE2_09_02
for Producing a Potential Alternative Solid Fuel

Field : Thaksin University, Phatthalung

Author : Trakoonkaew Ketkan

Pimphatcha Rethanu

Wanitcha Tunnikorn

School : Darunsikkhalai Science School

King Mongkut'S University of Technology Thonburi.

Advisor : Dr.Trairat Muangthong-on (King Mongkut's University of Technology Thonburi).
Mr.Thiti Jarangdet (Darunsikkhalai Science School).

Abstract

In recent years circular economy of waste use have been proposed from various purposes by the

government and private sectors. Huge amount of spent coffee ground is generated around the world. However, the

practical use of spent coffee ground is needed to perform by addressing the model in circular economy. We have

proposed the preliminary study to prepare briquette from spent coffee ground by using different organic materials

as binders. Three types of binder comprised cassava starch, corn starch and glutinous rice flour. In this work, the

briquettes were proposed to use three different ratios of binder to spent coffee ground by 1:10, 2:10 and 3:10. The

ignition efficiency of each briquette was conducted by home experiment.

Keywords : Spent Coffee Ground, Briquette, Biomass, Waste, Carbonization

Introduction
From the statistics in 2020, the average Thais coffee consumption rate is 300 cups per person

per year. That is a reason why we have a lot of spent coffee grounds. As mentioned above, the huge amount of
spent coffee ground has a negative effect, that is waste from coffee grounds. The most common method of disposal
is landfill. Disposing of it as waste is very dangerous because it will turn into carbon dioxide and methane. These
gases cause global warming. Therefore, reducing or modifying them helps reduce greenhouse gas emissions.

There are many methods for waste disposal. One of them is using waste to generate energy which
is used to make fuel. From the research of Miss. Neeranoot (2016), the coffee ground has a calorific value of 17-
20 MJ/Kg, which is a high value compared to the calorific value of other biomasses. Its high calorific value
indicates that it is a good quality fuel. Burning coffee grounds does not cause odor pollution like other biomasses.
This is the reason why we choose this method to dispose of spent coffee grounds.

From the research of Mr. Rung-roj (2010), there was a study on the production of char from
coconut shells and char from cassava rhizome which is the biomass. Therefore, our project has been extended by
using coffee grounds to experiment in the same way. Energy from the spent coffee ground can be generated in a
short time and it is renewable energy. It also reduces agricultural waste for a benefit before landfills.

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Methodology
1) Dry the coffee grounds to repel moisture until the weight is constant which means it is completely dry
2) Mix the coffee grounds with the binder ( glutinous rice flour, corn starch, and tapioca starch). The mixing

ratios of coffee grounds and binder are 1:10 2:10 3:10 respectively.
3) Take the mixture into a PVC pipe extruder. The compression will give the shape and density of the fuel rods.

The size and shape will depend on the purpose of use. The easiest way is to use your hands to form and press the
mixture into bars.

4) Dry and dehumidified the coffee grounds in sunlight for approximately 7 days. The standard moisture content
has to be less than 8% w/w. Time duration can be changed according to the appropriate weather conditions.

5) After drying, the briquette is complete. Then, test the properties of the briquette. When ignited, there must be
no sparks, less smoke, and no erupt. The property of the briquette can be divided into two parts: the ignition speed
and the length of time the fuel stick can ignite.

6) Burn the briquette in the brazier.
7) Start a timer from the ignition until the briquette can ignite by itself. observing the red dot. If the briquette can
ignite by itself then, there will turn red. When you see that, stop the timer. Then record the time in a flammability
timetable.
8) Perform the experiment in step 7) with all types of the binder. Then record the results in the flammability
timetable until the briquette can ignite by itself.
9) Start a timer when the briquette can ignite by itself until the fire goes out. This can be noticed by the red dot
on the briquette. The red dot will disappear and the briquette will turn gray ash. If the briquette has gone out, stop
the timer and record the time in a timetable.
10) Perform the experiment in step 9) with all types of binder then record the results in a timetable.
11) Bring the result in 10) to compare. Each briquette has a different weight therefore, it cannot be compared.
Weigh each briquette to 20 grams. Record the results of the experiment in an ignition timetable.
12) All the experimental results are displayed in graphs to show the trends and compare which binder in
which ratio is the best.

Result
Briquette from spent coffee grounds when it ignited, the results are as follows: When the

Briquette ignited, it will be no spark and no eruption. The smoke does not have a bad smell. Most of the smoke is
the smell of coffee grounds. Briquette from coffee grounds takes less time to ignite and can be ignited for a long
time. This project recorded the time of the Briquette from the three different binders and recorded the results in
the experimental timetable to compare the ignition efficiency. The timer is divided into two parts: The time from
the start of the fire until the Briquette can ignite by itself. The time from the moment the Briquette is self-igniting
until the moment the fire goes out. The results of the above-mentioned experiments were as a bar chart.

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Flammability Time

150

Time (s) 100

50

0

cassav a starch corn starch glutinous rice
flour

Binder

1:10 2:10 3:10

Ignition Time

Time (s) 1400
1200
1000

800
600

400
200

0

cassav a starch corn starch glutinous rice
flour

Binder

1:10 2:10 3:10

Conclusion
The experimental results were conducted to compare briquettes by using different binders in

three ratios to find the best one. In conclusion, the best quality binder is cassava starch in a ratio of 3:10. It tends
to ignite the briquettes the fastest and can ignite longer than the others.

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Flammability Time

150

Time (s) 100

50

0

01:10 02:10 03:10

Ratio of binder : spent coffee grounds

cassav a starch corn starch glutinous rice flour

Ignition Time

1500

Time (s) 1000

500

0 02:10 03:10
01:10

Ratio of binder : spent coffee grounds

cassav a starch corn starch glutinous rice flour

Acknowledgements
This project was supported by Science Classroom in University Affiliated School (SCiUS).

The funding of SCiUS is provided by Ministry of Higher Education, Science, Research and Innovation. This
extended abstract is not for citation.

References
Neeranoot W. Studying of heating value for tillandsia usneoides L. as biofuel[dissertation].

Bangkok: Songkla university; 2016.
Rung-roj P. The prodution of charcoal briquette by coconut shell and cassava

rhizome[dissertation]. Bangkok: Srinakharinwirot university; 2010.

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Title: Development of biomass pellet fuel using torrefied OE2_17_02
oil palm fronds with used bleaching earth from palm

oil industry

Field: Environmental and Ecology

Authors: Ms. Jidapa Theppanich

Ms. Lakkhika Sukphan

School: PSU. Wittayanusorn Surat Thani School

Prince of Songkla University, Surat Thani Campus

Advisors: Asst. Prof. Dr. Saysunee Jumrat (Prince of Songkla University, Surat Thani Campus)

Asst. Prof. Dr. Yutthapong Pianroj (Prince of Songkla University, Surat Thani Campus)

Mr. Thanet Kunamaspakorn (PSU. Wittayanusorn Surat Thani School)

Abstract
This research examined the biomass: oil palm fronds (OPF) and waste from industry: used bleaching

earth (UBE) for processing biomass pellet fuel. The OPF was taken to the torrefaction process that fixed
temperature at 300 °C with various times 20, 40, and 60 min. The OPF, UBE, and the torrified oil palm frond
(TOPF) were analyzed for heating properties: calorific value, ash content, and moisture content. Therefore,
the torrified condition with the highest calorific value at 5,495.53 cal/g (300 °C with 40 min) was chose and
blended with UBE to produce the biomass pellet fuel, which varied mixing ratios of TOPF:UBE (30:70, 50:50,
and 70:30) and OPF:UBE (30:70, 50:50, and 70:30). The biomass pellet fuels were characterized by proximate
analysis and net calorific value. The results were found that the biomass pellet fuel with the ratio of TOPF:UBE
as 70:30 depicted the most properties of net calorific value, moisture content, ash content, volatile matter,
fixed carbon, bulk density, length, and diameter were 4,394.07 cal/g, 0.25%, 40.74%, 1.52%, 57.49%, 0.82
g/cm3, 6-25 mm, and 4-12 mm, respectively in accordance with Solid biofuels – Fuel specifications and classes
– part 8: Graded thermally treated and densified biomass fuels (ISO/TS 17225-8:2016). It shows that the pellet
fuel using torrefied OPF with UBE could be practical and environmentally friendly due to utilization of
residues and wastes.

Keywords: Oil palm fronds (OPF), Used bleaching earth (UBE), Pellet fuel, Torrefaction

Introduction
Now a day, the high energy demand of human activities and the environmental concern are the most

important crisis of mankind. Every country tries to increase energy production but reduce greenhouse gases
emission. The solution to this crisis is increasing renewable energy usage. Biomass is a source of renewable
energy like Thailand which is an agricultural country. Each year, Thailand has a lot of agricultural waste from
various processing. Especially, the southern part of Thailand has a lot of oil palm industries. One of the most
potentials is oil palm fronds, which is a large amount of agricultural waste from the oil palm plantation around
52 million tons per year (Office of agricultural economics, 2014). However, the disadvantages of direct
utilization of biomass are large volume, difficult storage, difficult transportation, and low heating value.

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The way to overcome these problems is the dense pellet fuel, thus it is a solid fuel conversion from biomass
by compression to make a high density and a small cylinder rod solid fuel. Not only pure biomass is produced
a pellet fuel, but also a binder material is an important one, which holds a pellet to form solid durability. The
potential one is waste from the palm oil refining industry named used bleaching earth (UBE). It has capacity
144,000 tons per year (Ministry of industry, 2010). Thus, they are possible to produce pellet fuel, they can
analyze the production feasibility.

Torrefaction is a properties improvement process for upgrading biomass as a solid fuel by using the
thermal process in an absence oxygen condition at 200-300 ℃. (Chokchai, 2019). Therefore, torrefaction of
oil palm fronds would be a good way to decrease moisture and increase net calorific value, it makes properties
of pellet fuel efficiency better than oil palm fronds that are not torrefied to produce pellet fuel.

Therefore, this project studied the torrefied conditions of oil palm fronds by selecting the highest net
calorific value, which was characterized by a bomb calorimeter, then the best one was blended with UBE to
produce the pellet solid fuel and analyze the heating value, also the important pellet properties. Finally, cost
estimation and production feasibility of the pellet solid fuel were reported.

Methodology

The experimental process consists of 4 parts.

Part 1: The preparing of oil palm fronds fiber

Oil palm fronds (OPF) were cut to 50 cm of

length and dried with the sun to reduce moisture a) Cut to 50 cm of length b) Dried with the sun
content as shown in Fig.1 a) and b) respectively. Then, to reduce moisture

size reducing of them were performed with a length

less than 1.5 mm and decreased moisture in the oven

about 8-10% and called them as oil palm fronds fiber c) Oil palm fronds fiber (OPFF)
(OPFF) as shown in Fig.1 c). Fig. 1 Preparation of oil palm fronds (OPF)
Part 2: The study of torrefied condition of OPFF

Duran bottle was used as a reactor, OPFF were

put inside it. Then, took this reactor in microwave

system (Fig 2). The torrefied consisted of 3 conditions

at 300 °C with various time of 20, 40, and 60 min. After

microwave torrefaction, the torrefied oil palm frond

fiber (TOPF) were analyzed thermal properties (net

calorific value, moisture content, and ash content) for

Fig. 2 Microwave torrefied system. selected the optimum condition to produce the biomass
pellet fuel with used bleaching earth (UBE).

Part 3: The study of the influence of the proportion of torrefied oil palm fronds and used bleaching

earth on the properties of biomass pellet fuel

The TOPF with the optimum torrefied condition/ OPFF were mixed with UBE to produce pellet fuel

through a pellet machine at 100 °C. The ratio of TOPF/OPFF and UBE were studied at 30:70, 50:50, and 70:30

as shown in table 1.

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Table 1 Mixed proportion of biomass pellet fuel with TOPF/OPF

and UBES.ample Percent (by weight) Then, the biomass pellet fuel was tested fuel
properties such as net calorific value, proximate
OPFF 30: UBE 70 OPFF TOPF UBE analysis (moisture content, ash content, fixed carbon
OPFF 50: UBE 50 content, volatile matter), length and diameter, and bulk
OPFF 70: UBE 30 30 0 70 density in accordance to ISO 17225-8:2016.
TOPF 30: UBE 70
TOPF 50: UBE 50 50 0 50
TOPF 70: UBE 30
70 0 30

0 30 70

0 50 50

0 70 30

After properties testing in Part 3, the biomass pellet fuel with the optimum ratio of TOPF:UBE as

70:30 (focused on net calorific value) were calculated the production cost to analyze the feasibility for

industrial manufacturing.

Results and Discussion

Part 1: Condition of torrefied oil palm fronds (TOPF)

The properties of UBE, OPFF, and TOPF at 300 °C with various times (20, 40, and 60 min) were

shown in Table 2 and Fig. 3.

Table 2 Properties of UBE, OPFF, and TOPF at various conditions

Sample Image Color 6000.00

UBE Black Net calorific value (cal/g) 5000.00
OPFF White 4000.00
3000.00

TOPF (300°C, 20 min) White, brown, and black 2000.00
TOPF (300°C, 40 min) Brown and black 1000.00

0.00

TOPF (300°C, 60 min) Brown, Gray, and back Fig. 3 net calorific value of UBE, OPFF, TOPF
at various times

Part 2: Influence of the proportion of torrefied oil palm fronds and used bleaching earth
The result of properties testing of biomass pellet fuel using OPF and UBE (Fig. 4 a) found that the

net calorific value of biomass pellet fuel. The ratio of TOPF: UBE at 70:30 was the highest and passed the
biomass pellet standard (ISO17225-8:2016). Table 3 shows the result of properties: moisture content, volatile
matter, ash content, and fixed carbon. Moisture content every ratio met the pellet fuel standard and the ratio
TOPF:UBE at 70:30 had the lowest moisture content. The lowest ratio that gave volatile was 50:50 of TOPF:
UBC and the highest was 30:70 of OPFF: UBE. This property value was compared with other research because
the biomass pellet fuel standard was not specified. Ash content was not passed the biomass pellet fuel standard.
Fixed carbon results, the highest ratio that gives fixed carbon was OPFF: UBE at 30:70 and the lowest was
TOPF: UBE at 50:50. Lastly was bulk density (Fig. 4 b), every ratio was passed the biomass standard and the
ratio of TOPF:UBE at 70:30 was the highest of bulk density

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Table 3 Properties of biomass pellet fuel using TOPF/OPFF and UBE at various ratio

Sample Color Properties 5500.00 ISO17225-8:2016
Moisture Ash Volatile Fixed 5000.00
content (%) content (%) matter (%) carbon (%) Net calorific value (cal/g) 4500.00
4000.00
OPFF 30: UBE 70 0.55 37.73 2.71 59.01 3500.00
3000.00
OPFF 50: UBE 50 0.49 41.17 2.56 55.78 2500.00
2000.00
1500.00
1000.00

500.00
0.00

OPFF 70: UBE 30 0.47 42.48 1.02 56.02 Bulk density (g/cm3) 0.85 a) Net calorific value
TOPF 30: UBE 70 0.34 42.04 1.14 56.48 0.80
TOPF 50: UBE 50 0.30 44.72 0.62 54.36 0.75 ISO17225-8:2016
0.70
0.65
0.60
0.55
0.50

TOPF 70: UBE 30 0.25 40.74 1.52 57.49 b) Bulk density

ISO17225-8:2016 - ≤10 ≤20 - Fig. 4) Net calorific value and bulk density of
- biomass pellet fuel using OPFF, TOPF, and UBE

Production cost of pellet fuel using torrefied OPFF (TOPF) with UBE is very expensive, is more

expensive than the commercial pellet fuel (4702.02 bath/ton). This is because using microwave torrefied

system need N2 gas and electricity, although OPF and UBE are residue materials for free and cheap price,

respectively.

Conclusion
The study of preparing torrefied oil palm frond consists of the fixed temperature at 300°C with

various times 20, 40, and 60 min. The torrified oil palm fronds at 300°C with 40 min give the highest net
calorific value and the ratio of torrefied oil palm fronds and used bleaching earth at 70:30 depicted the best
result of proximate analysis, ultimate analysis, and net calorific value according to ISO 17225-8:2016
standard. Besides, the biomass pellet fuel in this ratio was the most suitable for production feasibility for the
industry.

Acknowledgments
This project was supported by Science Classroom in University Affiliated School (SCiUS). The

funding of SCiUS is provided by Ministry of Higher Education, Science, Research and Innovation. This
extended abstract is not for citation.

References
1. Chokchai, M, Panadda, I. (2019). Plackett-Burman Design Screening the Factors Affecting Torrefaction of

Palm Empty Fruit Bunches by Using Plackett-Burman Design. Journal19-4: October-December 2019. Khon
Kaen University, Khon Kaen.
2. Office of agricultural economics. 2014. Oil palm fronds. (online). available from https://www.oae.go.th/
3. Ministry of Industry. 2010. Manual Requesting permission to bring waste or materials Do not use and leave
the industrial area. (online) available from https://www.google.co.th/url?webintra.diw.go.

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OE2_06_03Title : Utilization of waste-derived biodiesel in a compression ignition engine

Field : Environmental Science and Ecology

Author : Jedpreeya Siriprasertsilp, Nicha Seehanavee and Napat Tangcharoen

School : Ratchasima Witthayalai School, SCiUS-Suranaree University of Technology

Advisor : Asst. Prof. Dr. Ekarong Sukjit, Suranaree University of Technology

Abstract

The purpose of this project was to study biodiesel from different feedstock as an alternative fuel in
diesel engine in order to reduce the amount of emission and particulate matters (PM) that was originated from
the combustion in diesel engine. Waste derived biodiesels, WCOME and CKOME were prepared from waste
cooking oil and waste chicken oil, respectively by transesterification process. Palm oil-derived biodiesel was
also prepared and used as reference. Fatty acid profiles of prepared biodiesel and the biodiesel properties
were analyzed. The engine performance, combustion characteristics and exhaust gas emissions of waste-
derived biodiesel were compared with palm oil biodiesel and diesel fuel. In addition, the oxidation
temperature of particulate matters was analyzed by a Simultaneous Thermal Analyzer (STA). The results of
the study revealed that biodiesel derived from waste cooking oil and waste chicken oil can be considered as
a potential candidate to replace the biodiesel derived from palm oil as main feedstock of biodiesel production
in Thailand without any penalty in exhaust emissions.

Keywords: biodiesel, diesel engine, waste cooking oil, waste chicken oil, emission from combustion

Introduction
Biodiesel is a renewable, less toxic and more environmental friendly alternative to petro-diesel fuel.

Biodiesel can be produced from several renewable feedstocks such as vegetable oils, animal fats, used
cooking oils and algae by using transesterification reaction which triglycerides react with alcohol (methanol
or ethanol. However, biodiesel made from non-edible feedstock is more attractive to avoid the competition
between food and fuel. Biodiesel derived from waste feedstocks, waste cooking oil and waste chicken oil,
was therefore selected in this study to investigate its effect on fuel properties, engine performance,
combustion characteristics and exhaust emissions. In addition, the characteristic of particulate matter
emissions produced by the biodiesel combustion was also reported. Diesel fuel and palm biodiesel were used
as reference fuels so that the use of waste-derived biodiesel in a diesel engine was evaluated.

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Method and Experimental Details
Table 1 Conditions for biodiesel production from different feedstock.

Raw materials Molar Catalyst Reaction Time Reaction
Palm Oil ratio (%wt) (min) Temperature (oC)
12:1 60
Waste Cooking Oil 1.5% KOH 60
Waste Chicken Oil 6:1 65
1.0% KOH 65
6:1 60
0.5% KOH 60

a. Biodiesel production

The transesterification was used for biodiesel production, The conditions used to the transesterification of
palm oil, waste cooking oil and waste chicken oil are shown in Table 1.

b. Determination of fuel properties

The fuel properties were measured according to American Society for Testing and Materials (ASTM) standard
which include kinematic viscosity, specific gravity, flash point, cetane index, gross calorific value and
distillation temperature. The fatty acid profiles of biodiesel derived from palm oil and waste feedstocks were
characterized by a Gas Chromatography–Mass Spectrometer (GC–MS).

c. Engine test and PM analysis

The engine test was carried out on a single cylinder diesel engine without any modification at the engine
speed of 1,500 rpm with full engine load.

PM was collected using a PM collector, as equipped with exhaust pipe of the diesel engine. The Simultaneous
Thermal Analysis (NETZSCH, STA 449 F3 Jupiter) was applied to characterize soot as composition of PM.

Result and Discussion
a. Fuel characteristics

The fatty acid profiles of biodiesel derived from palm oil (POME), waste cooking oil (WCOME) and waste
chicken oil (CKOME) had quite in similar characteristic. The physical and chemical properties of biodiesel
produced from different feedstocks showed kinematic viscosity, specific gravity, gross calorific value and
cetane index of biodiesels above the limit prescribed by the diesel fuel standard.

b. Engine performance, Combustion Characteristics and Emissions

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Comparison of the brake specific fuel consumption (BSFC) and brake thermal efficiency (BTE) obtained for
biodiesel from different feedstocks with diesel fuel at full engine load is shown in Figure1. It can be seen that
the BSFC was found to be higher in all biodiesel than diesel fuel due to the effects of lower calorific value
of biodiesel. While the BTE of all biodiesel was observed to be lower than that of diesel fuel. It is very
attractive that the combustion of waste-derived biodiesel (WCOME and CKOME) produces similar engine
performance compared to POME.

*

Figure 1. Engine performance Figure 2. Combustion characteristics

The in-cylinder pressure (ICP) and rate of heat release (RoHR) against crank angle diagram for test fuels at
full engine load is shown in Figure 2. ICP of all biodiesels were obtained to be higher than diesel fuel due to

high oxygen content in the biodiesel molecules leading to increase rate of combustion, peak temperature and
pressure . Moreover, the higher oxygen content in chemical composition of biodiesel has decreased ignition

delay for the use of POME, WCOME and CKOME resulted in shorter ignition delay when compared with
diesel fuel . In the case of the exhaust gas emissions obtained for biodiesel from different feedstocks and
diesel fuel at full engine load is shown in Figure 3. This study showed that the concentrations of CO, HC and

smoke emissions from all biodiesels were higher than those of diesel fuel while NOx emissions were lower
when compared to diesel fuel. Among biodiesels, the combustion of WCOME produced lower CO, HC and

smoke emissions with respect to POME while the combustion of CKOME produced similar CO, HC and
smoke emissions with more benefit in NOx emissions compared to POME

Diesel POME WCOME CKOME

1.0 Oxidation of Carbonaceous Soot
Oxidation of Volatile Organic Compounds

0.5

0.0

DTG (% /min) -0.5 Fuel TSO,max
-1.0 Diesel 484.0
-1.5 POME 475.9

WCOME 476.4
CKOME 472.0

-2.0

-2.5

-3.0

-3.5 TSO,max = Temperature for Maximum Soot Oxidation
100 160 220 280 340 400 460 520 580 640 700
-4.0
40 Temperature (oC)

Figure 3. Exhaust emissions Figure 4. TGA analysis

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c. PM emissions characteristics

Particulate matter (PM) emissions consists mostly of volatile organic compounds (VOC) and carbonaceous
soot and it is one of the major harmful emissions produced by diesel engines. The first derivative of weight
loss (DTG) according to the heating program of TGA under oxygen atmosphere for diesel fuel and biodiesels
is shown in Figure 4. The first peak of DTG represents the oxidation of volatile organic compounds and the
second peak represents the oxidation of carbonaceous soot. Soot emissions caused by the combustion of
biodiesel derived from waste cooking oil and waste chicken oil seem to oxidize easier compared to diesel
fuel.

Conclusion
• The biodiesel derived from waste cooking oil and waste chicken oil shows the potential to replace

the biodiesel derived from palm oil as main feedstocks of biodiesel production in Thailand.

• The lower oxidation temperature of soot from waste-derived biodiesel indicates the lower energy
to oxidize soot emission. This is beneficial in prolongation of the lifetime for diesel exhaust filter.

• The possibility of utilizing biodiesel derived from waste feedstocks as blend component in diesel
fuel to form the commercial diesel fuel complying the national standard under the supervision of the
Department of Energy Business, Ministry of Energy, Thailand can be considered as future works.

Acknowledgment
This project was supported by Science Classroom in University-Affiliated School (SCiUS) under Suranaree
University of Technology and Rajsima Witthayalai School. The funding of SCiUS provided by the Ministry
of Higher Education, Science, Research and innovation, is highly appreciated. This extended abstract is not
for citation.

References
[1] Katekaew, S., Suiuay, C., Senawong, K., Seithtanabutara, V., Intravised, K., Laloon, K. (2021). Optimization
of performance and exhaust emissions of single-cylinder diesel engines fueled by blending diesel-like fuel
from Yang-hard resin with waste cooking oil biodiesel via response surface methodology. Fuel, 304, 121434.

[2] Sayyed, S., Das, R.K., Kulkarni, K. (2022). Experimental investigation for evaluating the performance and
emission characteristics of DICI engine fueled with dual biodiesel-diesel blends of Jatropha, Karanja, Mahua,
and Neem. Energy, 238, 121787.

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Title : Production of liquid organic fertilizer from fish meal factory OE2_19_01

by-product and study on its efficiency of lettuce growth
Field : Environmental Science and Ecology
Author : Mr. Zuhairee Ramong

Mrs. Thanaporn Kaewmak
School : Islamic Sciences Demonstration School , Prince of Songkla University
Advisor : Dr. Natthakorn Woraathasin (Department of Agricultural Technology, Faculty of Science and

Technology, Prince of Songkla University, Pattani Campus)

Abstract :
The liquid waste from fish meal is a prime byproduct contained the nutrient sources for the growth of

various microorganisms including the anaerobic fermentation. When the fermentation process completed, the
various elements were released. These soluble elements consisting of macroelements and microelements are
crucial molecules for plant growth. Consequently, this study aimed to exploit the liquid waste from fish meal
factory for liquid organic fertilizer (LOF) to plant compared to that of chemical fertilizer with the formula 15-15-
15. LOF firstly generated from the liquid waste fermentation for 2 months was explored for the comprising
elements. The LOF carried the pH value of 4 and consisted of various elements such as nitrogen, phosphorus,
potassium, calcium, magnesium and sulfur for 2.52, 1.15, 2.09, 1.29, 0.18, and 0.15 percentage, respectively. The
plant, Green Cos lettuce, was divided into 3 groups composed of 3 treatments, LOF, chemical fertilizer and
LOF+chemical fertilizer. All treatments were recorded the growth every 3 days for 35 days of planting. The factors
of growth measurement, canopy width, height and fresh weight, were observed a great value in the treatment of
LOF. However, all treatments showed no significant difference in root length and dry weight of Green Cos lettuce.
Moreover, the disease incidence and severity in Green Cos lettuce were also investigated. LOF and LOF+chemical
fertilizer treatments showed the similar result that possessed 9 6 % of disease incidence index with low severity.
Whereas, a complete disease incidence index with moderate severity was observed in the treatment of chemical
fertilizer. In addition, the total chlorophyll content was also analyzed. The result showed all treatments were no
significant difference. LOF fermented from the factory liquid waste was therefore sufficient macroelements and
microelements for Green Cos lettuce growth according to the FERTILIZER ACT B.E. 2518 and FERTILIZER
ACT B.E. 2518 AMENDED BY FERTILIZER ACT (NO. 2) B.E. 2550.

Keywords : Liquid organic fertilizer , Liquid waste from fish meal, Lettuce

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Introduction
Fish meal is an important raw material for the aquaculture and livestock industry as protein nutrients

essential for the growth of all types of animals. The production of fishmeal produces waste products such as water
from cooked process, and there are also fish bones, shells, and ink cores.

Most of the wastewater from fishmeal plants comes from cooked process, fish washing water. If the
wastewater is not treated with a standard, it may affect the houses close to the factory. And although the waste
from fishmeal factories is properly disposed of at present, it cannot be used to create value.

Therefore, this study focuses on using the waste from fishmeal factories as the main raw material for
liquid fertilizer production to promote plant growth and able to resist plant diseases. It is the use of waste material
from seafood processing plants to create economic value and reduce the disposal of waste that may pollute the
local area. Moreover, the use of water from cooked process of fishmeal factories to produce liquid organic
fertilizers is consistent with the BCG Economy (Bio-Circular-Green Economy) principle and reduced the use of
chemical fertilizers in agricultural production and to promote the policy of expanding the production of organic
agricultural products in Thailand as well.

Methodology
Part 1 Liquid organic fertilizer production process: Shell, fish bone, and squid core with 0.5 kg were mixed
and followed by water from fish boiled process 1 kg, molasses 0.3 kg, 5 liters of water, and PD2. The mixer was
fermented for 2 months in anaerobic conditions. When the fermentation process completed, the various elements
were analyzed.
Part 2 Experimental design: The experimental plan was to use the Completely Randomized Design (CRD)
experimental plan to study the growth of Green cos from 3 different fertilizer types, 25 plants of each type were
planted for a total of 75 plants as follows:

Treatment 1: Liquid Organic Fertilizer (LOF) from fishmeal factory.
Treatment 2: Chemical Fertilizer Formula 15-15-15.
Treatment 3: Liquid organic fertilizer from fishmeal factory and chemical fertilizer formula 15-15-15,

ratio 1:1.
Part 3 Study on the growth effect of lettuce: After 14 days of lettuce seedlings grown in pit trays, they were
transplanted into a plastic bag. The potting soil must have a pH in the range of 6-7. Then the treatments were
treated according to the formulas mentioned in the experimental method above. The growth parameters of Green
cos were recorded for 18 days.

Results
When the fermentation process completed, the various elements were released. These soluble elements

consisting of macroelements and microelements are crucial molecules for plant growth. The LOF carried the pH
value of 4 and consisted of various elements such as nitrogen, phosphorus, potassium, calcium, magnesium and

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sulfur for 2.52, 1.15, 2.09, 1.29, 0.18, and 0.15 percentage, respectively (Table 1), which would indicate that the
elements content was in accordance with the Fertilizer ACT.

Table 1: Contents of macroelements and microelements in liquid organic fertilizer

Element Element content (%)
Nitrogen (N) 2.52
Phosphorus (P) 1.15
Potassium (K) 2.09
Calcium (Ca) 1.29
Magnesium (Mg) 0.18
0.15
Sulfur (S)

To study the growth of lettuce after treated with the formulas, All treatments were recorded the growth
every 3 days for 35 days of planting. The results showed that canopy width, height and fresh weight, were observed
a great value in the treatment of LOF (Fig. 1A, 1B, 1D). However, all treatments showed no significant difference
in root length (Fig. 1E) and dry weight (Fig. 1C) of Green Cos lettuce. In addition, the total chlorophyll content
was also analyzed. The result showed all treatments were no significant difference (Table 2).

Figure 1: The growth of lettuce ; Canopy width (A), Height (B), Dry Weight (C), Fresh weight (D), Root length (E)

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Table 2 : The amount of chlorophyll A, chlorophyll B and total chlorophyll

Treatment kit Pigment quantity (micrograms per milliliter)

Liquid organic fertilizer (LOF) Chlorophyll A Chlorophyll B Total Chlorophyll
Chemical fertilizer (CF)
LOF and CF 5.52±1.49 2.53±1.88 8.05±3.35

5.78±1.36 2.46±0.62 8.25±1.94

6.81±0.22 3.34±1.36 10.15±1.56

The disease incidence and severity in Green Cos lettuce were also investigated. LOF and LOF+chemical
fertilizer treatments showed the similar result that possessed 9 6 % of disease incidence index with low severity.
Whereas, a complete disease incidence index with moderate severity was observed in the treatment of chemical
fertilizer (Table 3).

Table 3 : Disease incidence index and the severity of lettuce

Treatment kit Disease incidence (%) Severity level
Low
Liquid organic fertilizer (LOF) 96
Medium
Chemical fertilizer (CF) 100 Low

LOF and CF 96

Conclusion
Fishmeal factory can produce liquid organic fertilizers which contain macroelements and microelements

according to the FERTILIZER ACT B.E. 2518 and FERTILIZER ACT B.E. 2518 AMENDED BY FERTILIZER
ACT (NO. 2) B.E. 2550. Liquid organic fertilizer can be promoted the growth of Green cos, substitute chemical
fertilizers, able to reduce the disease index and severity of leaf spot disease in Green cos.

Acknowledgements
This project was supported by Science Classroom in University Affiliated School (SCiUS) under Islamic

Sciences Demonstration School and Prince of Songkla University. The funding of SCiUS is provided by Ministry
of Higher Education, Science, Research and Innovation. This extended abstract is not for citation.

References

กรมวิชาการเกษตร. ประกาศกรมวชิ าการเกษตร เร่อื ง กาหนดเกณฑ์ปยุ๋ อนิ ทรยี ์. 2557 [เขา้ ถึงเมือ่ 17 กันยายน 2564]. เขา้ ถึงได้จาก:

https://bit.ly/3CvJM8j

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Title: Tapioca Starch-Based Biodegradable Film: Effect of Glycerol and OE2_08_01

Silica Extracted from Sugarcane Bagasse Ash

Field: Environmental Science and Ecology

Author: Miss Thunchanok Nacha, Miss Nichapa Ruangkitwanit, and Miss Piyamat Janthong

School: Lukhamhan Warinchamrab School, SCiUS Project–Ubon Ratchathani University

Advisor: Asst.Prof.Dr.Saisamorn Lumlong, and Assoc.Prof.Dr.Pornpan Pungpo

Abstract
With the rise of food delivery, the growing concern about the amount of plastic packaging found in trash
requires the development of biodegradable film with low animal and human toxicity. Agricultural waste rich
in inorganic materials (such as silica) is often discarded, while it could be reused as a source of raw material.
Considering these points, silica extracted from sugarcane bagasse ash was utilized as a filler for starch
composite film. The starch-based biodegradable film was produced by solution casting and characterized by
mechanical properties (tensile strength and elongation at break), thermal properties, and biodegradation. The
purity of the silica was determined by XRF to be 80.5%, with a percentage yield of 46.34%. The starch-based
biodegradable films were prepared by dissolving tapioca starch in water and adding glycerol as a plasticizer.
The A1 sample, which contained 3.0% starch, and the C1 sample, which contained 3.0% starch and 7.5%
glycerol were the highest tensile strength of 28.50 and 28.12 MPa, respectively. On the other side, the C1
sample had the highest elongation at break values (4.69%). The tapioca starch-based biodegradable film was
decomposed in 7 days. In conclusion, tapioca starch was prepared from tapioca, a Thai industrial crop that can
be used as a raw material for a biodegradable film. Silica derived from renewable sources can be incorporated
into the starch-based film to produce biodegradable material, and silica improved the thermal properties of the
starch-based film.

Keywords: starch-based biodegradable film, sugarcane bagasse ash, silica, tapioca starch

Introduction
Plastic has been used in many applications such as food packaging, cosmetic packaging, furniture, and sports
equipment. According to the Thailand development research institute (TDRI), Thailand is the world’s tenth-
biggest dumper of plastic waste into the sea (2020). Thailand has an average of 1.03 tons of mismanaged waste
each year. The main issue with plastic is that it is complicated and time-consuming to decompose. It could
residual in the environment, such as microplastics, which can contaminate water sources and soil, as well as
return to the food chain and hide in raw materials in human foods. As a result, biodegradable films that are
biodegradable are an appealing solution to these issues. It can be recycled into nature, leaves no residue, and
is environmentally friendly. The purpose of this study was to extract silica from sugarcane bagasse ash, an
agricultural waste rich in inorganic materials, and to prepare biodegradable film from tapioca starch, a Thai
industrial crop grown in enormous volume in Ubon Ratchathani Province. The other purpose of this research
was to investigate silica as a filler for starch composite film in addition to improving characterized mechanical
properties (tensile strength and elongation at break), as well as thermal properties. Finally, the purpose of this
research is to encourage the development of zero-waste products.

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Methodology
2.1 Extraction of silica from sugarcane bagasse ash. Sugarcane bagasse ash was mixed with 3 M NaOH
(ratio 1:10 weight/volume) and refluxed at 90 °C for 24 hours. Then, the solution was filtered and crystallized
with 3M HCl. The crystallization was completed at 18 hours. The sample was washed with distilled water
until it reached a constant pH. Finally, the solution was dried at 80 °C for 2 hours. The chemical composition
of the obtained silica was determined using X-Ray Fluorescence (XRF).
2.2 Solution casting. The starch solution was prepared by dissolving tapioca starch 3.6 g in 120 mL water.
The amount of glycerol (0, 5, and 7.5% weight of starch) was added. The amount of silica (0, 1, 5, and 10%
weight of starch) was added. The solution was stirred and heated for 5 minutes at 60–80 °C. Then, the mixed
solution was poured onto rectangular Teflon plates and dried in an oven at 60 °C.
2.3 Characterization. Scanning electron microscope (SEM) images were recorded using a field emission
scanning electron microscope. Before the SEM analysis, The sample was fixed in the sample holder with
carbon tape and covered with platinum in the coater.
2.4 Mechanical properties. Specimens (size of 10×70 mm) were cut from each film and plastic bag
(Polypropylene, PP). The tensile strength and percent elongation at break were measured using a texture
analyzer and a 50 N load cell. The initial grip separation was 4 cm, and the cross-head speed was 50 mm/min.
2.5 Thermal analysis. Thermogravimetric analysis (TGA) was used to evaluate the thermal stability of
biodegradable films. TGA was performed by a thermogravimetric analyzer. The samples were analyzed under
a nitrogen atmosphere with a flow of 65 mL/min, using an alumina crucible, and heating at a temperature
range of 30 – 600 °C with a heating rate of 20 °C/min.
2.6 Biodegradability Assay. The biodegradable films and PP plastic bags were cut into 4×3 cm and weighed
as the initial dry weight (W0). Then, the samples were placed in cloth bags and buried in glasses of soil at a 5-
10 cm depth. Each sample contained 3 specimens and was branched into 6 groups (1, 3, 5, 7, 15, and 30 days).
The samples were watered on the starting planting day, 1, 3, 5, 7, and 15 days. When the deadline is over, dig
the samples up, wash them, then dried at 60 °C for 10 hours and weigh it as the final dry weight (Wd). The
decomposition rate was calculated by using equation 1.

Decomposition rate = W0-Wd ×100 (1)
W0

Where W0 is the initial dry weight and Wd is the final dry weight

Result
3.1 Silica extraction. The highest elemental products are SiO2 (80.5%), and other elements include Cl
(11.2%), Na2O (6.37%), and others. XRF determined the purity of the silica to be 80.5 percent, with a
percentage yield of 46.34 percent calculated from equation 2.

%Yield= Amount of product × 100 (2)
Amount of initial substance

3.2 Biodegradable films. Film-forming was chosen at a concentration of 3% starch due to the suitable

thickness of the films and the lack of air bubbles in the films. Glycerol concentrations of 0, 5, and 7.5%, and

silica concentrations of 0, 1, 5, and 10% were used for film preparation. There are 12 samples, and all the

conditions are described in table 1. Even though all samples' appearances are good, only 4 samples were

chosen for the testing; they are A1, C1, C2, and C4 because choosing a greater quantity of glycerol and silica

will show a greater difference in the analysis result.

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Table 1 Conditions of biodegradable film preparation.

3.3 Mechanical properties. The mechanical properties of Samples Amount of glycerol (% Amount of silica
w/w of starch) (% w/w of starch)
the film showed that the A1 and C1 samples have high
tensile strength of 28.5 MPa and 28.12 MPa, respectively, A1 0 0
A2 0 1
but when compared to polypropylene, which has a tensile A3 0 5
strength of 36.53 MPa, the tensile strength of the samples
was less than polypropylene. Meanwhile, the C1 sample has A4 0 10
the highest percent elongation at break, 4.69%. However, it B1 5 0
was less than polypropylene, which is 539.78%. The
B2 5 1
addition of glycerol does not affect the tensile strength of
the starch-based biodegradable films but increases percent B3 5 5
elongation at break, as shown in figure 1. On the other hand, B4 5 10
C1 7.5 0

C2 7.5 1

C3 7.5 5

C4 7.5 10

the addition of silica decreases the tensile strength and percent elongation at the break of films, as shown in
figure 1. As shown in figure 2, SEM images of biodegradable films revealed that adding more silica causes
silica to clump more. Because of the poor interaction between starch and silica, the gap between starch and
silica causes decreases in tensile strength and percent elongation at the break of films.

Figure 1 Mechanical properties of biodegradable films. c d

ab

Figure 2 SEM images of biodegradable films (x200): (a) C1, (b) C2, (c) C3, (d) C4

3.4 Thermal analysis. Thermogravimetric analysis was used to assess the thermal stability of the starch-based
biodegradable films. From the TG curves, the first weight

loss is between 30 and 275 °C for all samples except for the

C4 sample, where the first weight loss is between 30 and
240 °C. The first weight loss ranged from ~15 % to ~25 %
for all samples. The second weight loss, which is the onset

temperature of the films, is between 275 and 340 °C for all

Figure 3 Thermogravimetric (TG) curves of biodegradable films samples except for the C4 samples, where the second
weight loss is between 240 and 340 °C. A1 samples and C2
samples' second weight loss ranged from ~75 to ~85%, and

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C1 samples and C4 samples' second weight loss ranged from ~45 to ~60%. Therefore, the addition of glycerol
does not affect the thermal stability but the addition of silica decreases the onset temperature of the films and
the weight loss of samples, as shown in C4 in figure 3.
3.5 Biodegradability Assay. The C1 sample had more decomposition rate than the A1 sample. As a result,
adding glycerol to biodegradable films increased their decomposition rate. Furthermore, the C4 sample
decomposes at a lesser rate than the C1 sample, as shown in figure 4. As a result, adding silica decreases the

decomposition of biodegradable films. The decomposition
rate of polypropylene (PP, 1-2%) was lower than tapioca
starch-based biodegradable films. The tapioca starch-based
biodegradable films were decomposed in 7 days, which can be
regarded as an excellent degradation time when compared to
polypropylene.

Figure 4 The decomposition rate of the biodegradable films

Conclusions
The highest chemical component of silica is SiO2 (80.5%). Considering all the samples, A1, C1, C2, and C4
samples were chosen for testing. The biodegradable films containing 7.5% glycerol and 0% silica (C1) have
the highest percent elongation at break, 4.69%. Furthermore, the biodegradable films without glycerol and
silica (A1) and the biodegradable films containing 7.5% glycerol without silica (C1) showed the highest tensile
strength. For thermal stability, the biodegradable films containing 7.5% glycerol and 10% silica (C4) have
minimal weight change in each temperature range. In the biodegradability assay, the tapioca starch-based
biodegradable films were decomposed in 7 days. As a result of the tests, the addition of glycerol can improve
the mechanical properties of biodegradable films and increases the decomposition rate of biodegradable films.
The addition of silica does not improve mechanical properties but improves thermal properties of
biodegradable, and decreases the decomposition of biodegradable films. These biodegradable films can be
considered for use in biodegradable products in order to encourage the development of zero-waste products.

Acknowledgement
This project was supported by Science Classroom in University Affiliated School (SCiUS) under Ubon
Ratchathani University and Lukhamhan Warinchamrab School. The funding of SCiUS is provided by Ministry
of Higher Education, Science, Research, and Innovation, which is highly appreciated. This extended abstract
is not for citation.

References
Luciana C. de Azevedo, Suzimara Rovani, Jonnatan J. Santos, Djalma B. Dias, Sandi S. Nascimemto, Fábio
F. Oliveira, Leonardo G. A. Sliva, Denise A. Fungaro. 2020. Biodegradable films derived from corn and potato
starch and study of the effect of silicate extracted from sugarcane waste ash. Applied Polymer Materials, 20(2),
2160-2169.

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