Extended Abstract : Environmental Science and Ecology

Faculty of Science, Thaksin University
Phatthalung (August 26-29, 2022)
Environmental Science and Ecology

12th SCiUS Forum

12th SCiUS Forum
August 26 – 29, 2022
at Thaksin University

organized by
Ministry of Higher Education, Science, Research and Innovation

and Thaksin University

12th SCiUS Forum

คำนำ

คณะกรรมการบริหารโครงการห้องเรียนวิทยาศาสตร์ในโรงเรียน โดยการกำกับดูแลของมหาวิทยาลัย
(โครงการ วมว.) เห็นชอบให้มีการจัดกิจกรรม 12th SCiUS Forum ในระหว่างวันท่ี 26 – 29 สิงหาคม 2565
เพ่ือให้นักเรียนโครงการ วมว. ระยะท่ี 2 ชั้นมัธยมศึกษาปีที่ 5 ประจำปีการศึกษา 2564 ได้นำเสนอผลงานโครงงาน
วิทยาศาสตร์และแลกเปล่ียนองค์ความรู้กับเพ่ือนๆ นักเรียนต่างโรงเรียนในโครงการ วมว. จำนวน 19 แหง่ ได้แก่

1. โรงเรยี นสาธิตมหาวิทยาลยั เชยี งใหม่ – มหาวทิ ยาลัยเชียงใหม่
2. โรงเรียนมธั ยมสาธติ มหาวิทยาลยั นเรศวร – มหาวิทยาลัยนเรศวร
3. โรงเรยี นราชสีมาวิทยาลัย – มหาวิทยาลัยเทคโนโลยีสรุ นารี
4. โรงเรียนสาธิตมหาวทิ ยาลยั ขอนแกน่ ฝา่ ยมธั ยมศึกษา (ศกึ ษาศาสตร์) – มหาวทิ ยาลยั ขอนแกน่
5. โรงเรียนสาธติ มหาวิทยาลยั มหาสารคาม (ฝ่ายมัธยม) – มหาวทิ ยาลยั มหาสารคาม
6. โรงเรียนดรุณสกิ ขาลัย – มหาวิทยาลัยเทคโนโลยพี ระจอมเกล้าธนบุรี
7. โรงเรียนสาธิตแห่งมหาวิทยาลัยเกษตรศาสตร์ วิทยาเขตกำแพงแสน ศูนย์วิจัยและพัฒนาการศึกษา –

มหาวิทยาลยั เกษตรศาสตร์ วิทยาเขตกำแพงแสน
8. โรงเรยี นสาธิต "พบิ ูลบำเพ็ญ" มหาวทิ ยาลัยบรู พา – มหาวิทยาลยั บรู พา
9. โรงเรยี น มอ.วิทยานสุ รณ์ – มหาวทิ ยาลยั สงขลานครนิ ทร์ วทิ ยาเขตหาดใหญ่
10. โรงเรียนสาธติ มหาวิทยาลยั สงขลานครินทร์ – มหาวทิ ยาลัยสงขลานครนิ ทร์ วิทยาเขตปตั ตานี
11. โรงเรียนป่าพะยอมพทิ ยาคม – มหาวิทยาลัยทักษณิ
12. โรงเรียนสาธติ มหาวิทยาลัยพะเยา – มหาวทิ ยาลยั พะเยา
13. โรงเรียนลอื คำหาญวารินชำราบ – มหาวิทยาลัยอบุ ลราชธานี
14. โรงเรยี นสิรนิ ธรราชวิทยาลยั – มหาวิทยาลัยศิลปากร
15. โรงเรียนสวนกหุ ลาบวิทยาลยั รงั สิต – มหาวทิ ยาลยั ธรรมศาสตร์
16. โรงเรยี นสาธติ มหาวทิ ยาลัยขอนแกน่ ฝา่ ยมธั ยมศึกษา (มอดนิ แดง) – มหาวทิ ยาลัยขอนแก่น
17. โรงเรยี น มอ.วิทยานสุ รณ์ สุราษฎรธ์ านี – มหาวทิ ยาลยั สงขลานครินทร์ วิทยาเขตสรุ าษฎรธ์ านี
18. โรงเรยี นสรุ ววิ ัฒน์ – มหาวทิ ยาลัยเทคโนโลยีสรุ นารี
19. โรงเรยี นสาธิตอิสลามศึกษาฯ – มหาวทิ ยาลยั สงขลานครนิ ทร์ วทิ ยาเขตปัตตานี

กิจกรรม 12th SCiUS Forum ดำเนินการภายใต้มาตรการป้องกันการระบาดของโรคติดเช้ือไวรัส
โคโรนา 2019 (COVID-19) และแนวปฏิบัติในการเข้าร่วมกิจกรรม 12th SCiUS Forum ซึ่งในการนำเสนอ
ผลงานประกอบด้วยการนำเสนอโครงงานประเภท Oral presentation และ Poster presentation จำแนก
สาขาวิชาโครงงานวิทยาศาสตรอ์ อกเป็น 7 สาขา ได้แก่ สาขาวิชาเคมี สาขาวชิ าชีววิทยาและความหลากหลาย
ทางชีวภาพ สาขาวิชาฟิสิกส์และดาราศาสตร์ สาขาวิชาวิทยาศาสตร์สิ่งแวดล้อมและนิเวศวิทยา สาขาวิชา
คณิตศาสตร์และสถิติ สาขาวิชาเทคโนโลยีและคอมพิวเตอร์ และสาขาวิชาสะเต็มและนวัตกรรม สำหรับ
เอกสารเล่มนี้เป็นการรวบรวม Extended Abstract ของโครงงานวิทยาศาสตร์ประเภท Oral
presentation สาขาวิชาวทิ ยาศาสตรส์ งิ่ แวดลอ้ มและนิเวศวทิ ยา

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12th SCiUS Forum

คณะผู้จัดทำหวังเป็นอย่างย่ิงว่า เอกสารฉบับน้ีจะเป็นประโยชน์ต่อนักเรียน ครู คณะกรรมการตัดสิน
ผู้เข้าร่วมกิจกรรม รวมถึงคณะทำงานจากทุกหน่วยงานที่เกี่ยวข้อง และขอขอบพระคุณผู้เก่ียวข้องทุกท่านท่ีได้
ให้ความรว่ มมือสนบั สนุนการจดั กิจกรรม 12th SCiUS Forum ในครัง้ น้ี

คณะผ้จู ดั ทำ
กรกฎาคม 2565

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สารบัญ 12th SCiUS Forum

คำนำ หน้า
สารบญั ก
รายชอ่ื โครงงานวิทยาศาสตร์ ค
ประเภท Oral presentation สาขาวิทยาศาสตรส์ ง่ิ แวดลอ้ มและนเิ วศวทิ ยา
1
กล่มุ ท่ี 1 วันท่ี 27 สงิ หาคม 2565 55
กล่มุ ที่ 2 วนั ที่ 27 สงิ หาคม 2565

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12th SCiUS Forum

List of Science Projects 12th SCiUS Forum
Oral presentation

Environmental Science and Ecology Group 1
Saturday August 27, 2022

No. Code Title Author School
Islamic Science
1 OE1_19_02 Effect of seaweed on water Mr. Peerapat Madyusoh Demonstration School

treatment for mud crab Mr. Hanis Mutichun Piboonbumpen
Demonstration School,
fattening in the recirculating Burapha University

condo system Demonstration School,
University of Phayao
2 OE1_13_02 Microplastics contamination Miss Kanyawee Sodapukd
Kasetsart University
in marine animals at the Miss Natchanan Khantee Laboratory School
Kamphaeng Saen
coastal area of Chonburi Miss Paphada Jantraphinun Campus Educational
Research and
province Development Center
Naresuan University
3 OE1_02_01 Chromosomal Aberration of Miss Sutipa Tangparitklu Secondary
Demonstration School
Swamp eel (Monopterus Miss Jinatta Nojak
Rajsima Witthayalai
albus) and Nile tilapia School

(Oreochromis niloticus) in Naresuan University
Secondary
Huai Tap Chang, University Demonstration School

of Phayao

4 OE1_10_01 Preliminary finding on the Miss Chutimon Promkittyanon

present and anatomical Mr. Nathachanok Chaiwiriyapong

distribution of microplastics Miss Yanisa Chaikaew

in giant freshwater prawns in

cultured ponds

5 OE1_03_02 Active surveillance of Covid- Miss Sarintorn Saranporn

19 spreading in school and Miss Pranpriya Thongkam

surrounding community by

monitoring of SARS-CoV-2

in wastewater

6 OE1_06_02 Investigation of the elemental Mr. Muangthai Thongnok

composition of different Mr. Thanadech Saengrueangrak

types of cigarette and particle Mr. Phasin Ponvisetsit

exposure from smoking

7 OE1_03_03 The value added products and Miss Warintorn Sangwattanarat

life cycle greenhouse gas Miss Wanrada Namwong

emissions and cost of the

products from pickled ginger

residue.

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12th SCiUS Forum

No. Code Title Author School
Chiang Mai University
8 OE1_01_01 The effects of forest fires on Miss Phimpapond Pongtiwapan Demonstration School

seed germination of some Miss Thitichaya Teamtanomp Demonstration School
Prince of Songkla
deciduous trees Miss Suthinee Roeksupha University, Pattani
Campus
9 OE1_16_01 Environmentally Friendly Miss Nantachaporn Sudjai Mahasarakham
University
Waterproof and Anti-fungal Mr. Raffel Niyomdecha Demonstration School
(Secondary)
Paint from Natural Rubber Paphayomphittayakom
School
Latex
Piboonbumpen
10 OE1_07_01 Soil carbon contents of Miss Tanjira Punyasarn Demonstration School,
Burapha University
Northeastern’s Home Garden Miss Naruedee Wongyai
Kasetsart University
and Monoculture Plantation Laboratory School
Kamphaeng Saen
11 OE1_14_01 Reducing methane emissions Miss Jarupa Chuanklin Campus Educational
Research and
from rice fields by Miss Phawida Duangkhwan Development Center

methanotrophic consortium

augmentation for the

environmental friendly rice

production process

12 OE1_13_01 The contamination of Mr. Krithiran Vayakornvichitr

microplastics in beach Miss Natnaree Ruangdech

sediment in Chonburi province Miss Napatploy Pichetjindakul

in different monsoons

13 OE1_10_02 Microplastic residues in the Miss Chayakan

commercial fish off the coast Rassameecharoenchai

of Samut Songkhram Miss Pakpicha Moonjit

Province Miss

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Title : Effect of seaweed on water treatment 12th SCiUS Forum
for mud crab fattening
in the recirculating condo system OE1_19_02

Field : Environmental Science and Ecology
Author : Mr.Peerapat Madyusoh

Mr.Hanis Mutichun
School : Islamic Sciences Demonstration School

Prince of Songkla University
Adviser : Assoc.Prof.Dr.Sukree Hajisamae

Abstract

Management of water quality is the most important limiting factor for the mud crab (Scylla
paramamosain) fattening process in recirculating condo system (RCS). The effluent especially ammonia
circulating in the system affects the survival and growth of the crabs. The purpose of this study was to assess the
efficiency of different types of seaweeds in reducing ammonia concentration in the RCS. Two experiments were
conducted; (1) a preliminary study in the experiment tank without crabs and (2) in the RCS system with crabs fed
regularly with fresh fish. Four species of seaweed were used in the experiment 1 for 5 days, namely, Gracilaria
fisheri, Chaetomorpha linum, Ulva rigida, and Caulerpa lentillifera. Result showed different filtering rates of
ammonia between these five seaweed species (P<0.01) and indicated that G. fisheri and C. linum were the most
efficient seaweeds in removing ammonia in water. In the experiment 2, these two seaweed species were used in
the RCS system together with control. It was again confirmed that seaweeds can effectively remove ammonia from
the RCS system (P<0.05) and G. fisheri was the most efficient compared to C. linum. Apart from that other water
parameters indicated that using seaweeds in the RCS system can improve water quality by resulting in higher
dissolved oxygen value and lower pH value. Therefore, it is suggested that using seaweeds, especially G. fisheri
is essential to remove ammonia in water and help the healthy status of the system leading to healthy crab production
and reducing toxic to nature. Additionally, it is observed that apart from reducing ammonia, C. linum also helps
in trapping sediment in the RCS, thus a combination of using both seaweed species is recommended.

Keywords : Seaweed, Recirculating condo system, Mud crab, Aquaculture technology, Wastewater

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12th SCiUS Forum

Introduction

Mud crab (Scylla paramamosain) is an economically important aquatic species due to its prominence and
high demand in the market. Crabs can be raised in a variety of ways. Crab farming technique in recirculating condo
systems is relatively new and becomes more popular as it saves space and is simple to maintain. It is a method of
rearing crabs in a high-density environment. The circulating water system is a closed system that uses water and
does not require water transfer. They do, however, usually have problems with water quality conditions especially
ammonia from excess feed and waste from crab’s excretion (Weihrauch et al., 2004). As far as an environmental
problem is the main concern for the current farming system, reducing all waste as much as possible is the most
important way for modern aquaculture. Moreover, as BCG model (Biological, circulating, and green technology)
is the international and national policy, applying the model in all agricultural sectors is important. Seaweeds have
been used for wastewater treatment in many areas. This is due to the low cost compared to other methods. Seaweed
offers a wide range of applications, such as industry, use to maintain a balanced environment, fertilizer extraction,
and biological water treatment. Seaweed can absorb nutrients or pollutants such as metals, nitrogen, and
phosphorus (Arumugam et al., 2018). However, there is no report so far on the application of using seaweeds in
crab fattening in the recirculating condo system (RCS). The purpose of this study was to assess the efficiency of
different types of seaweeds in reducing ammonia concentration for fattening mud crabs in the RCS. Results from
this study can help crab’s farmers to have an alternative and environmentally friendly method to apply on their
farms.

Methodology

The experiment was conducted in four parts, as follows:

Part 1: Study of ammonia removal efficiency in plastic tanks (without mud crab)

The experimental design was CRD (Completely Randomized Design) with five
treatments (3 replications), namely control, G. fisheri, C. linum, U. rigida, and
C. lentillifera which were placed in separate tanks containing (area 0.14 m2) with
a density of 3 kg/m2 and a salinity of 28 ppt. The experiment was conducted for
5 days. The findings from the first experimental trials were carried over to
Experiment 2

Part 2: Study of ammonia removal efficiency in the RCS

As G. fisheri and C. linum were the most efficient seaweeds to reduce ammonia in the experiment 1, they were
chosen as treatments together with control (without seaweed) to test
their filtration capacity. To test the biofiltration capacity, G. fisheri
and C. linum were placed in a wastewater tank. This tank density was
determined by an experiment conducted by Abreu et al. (2011), who
discovered that 3 kg/m2 is the best density for achieving a high
production level of seaweed. The crabs were fed daily with fresh fish
at about 5% body weight at about 1700-1800. The left-over feed was
removed the next morning. The experiment was conducted for 5 days
with circulating water for 5 hours/day.

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Part 3: Measurement of water quality
Salinity, Dissolved Oxygen (DO), and pH was measured using a multi-probe, and ammonia was analyzed

using the Phenate Method (APHA, AWWA, ANDWEF,1995). Water quality was assessed on days 0, 3, and 5.
Part 4: Data analysis

The data was displayed by the mean ± standard deviation. Data analysis was performed using Microsoft
Excel software. One-way ANOVA was used to assess the difference in ammonia reduction by different types of
seaweeds from the experiments 1 and 2.

Results Figure 1: Efficiency removal of ammonia in the tank
Experiment 1 (without mud crab)
The result is shown in Figure 1. After 5 days, the ammonia

concentration of each treatment was reduced. ANOVA indicated
different reductions between treatments (P<0.01), with the control
unit had the lowest value of reduction. The graph shows that G.
fisheri and C. linum have the highest reduction rate compared to
others. Thus, G. fisheri and C. linum were selected for the RCS.

Experiment 2
The results level of ammonia showed in Figure 2. After 5days, the ammonia of each treatment was
reduced. The control unit had the lowest reduction of ammonia and different filtering rates of ammonia between
them (P<0.05). The graph shows that G. fisheri has the highest reduction rate. The salinity value of each
treatment is higher because the water has evaporated (Figure 3). Moreover, the higher DO and lower pH value
(Figure 4 and Figure 5) were found in the RCS with seaweeds compared with the control. This is a good
environmental condition for crabs in the RCS system.

Figure 2: Efficiency removal of ammonia in Figure 3: Salinity in the RCS systems
the RCS systems

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Figure 4: Dissolved Oxygen (DO) in the Figure 5: pH value in the RCS systems
RCS systems

Conclusion

Seaweed can absorb ammonia, with G. fisheri and C.linum being the best choices for use in The RCS
systems compared to other seaweeds species. Gracilaria fisheri and C. linum were best suited for use in the RCS
system due to their higher rate of ammonia absorption capability. Therefore, it is suggested that using seaweeds,
especially G. fisheri is essential to remove ammonia in water and help the healthy status of the system leading to
healthy crab production and reduce toxic to nature. Additionally, it is observed that apart from reducing ammonia,
C.linum also helps in trapping sediment in the RCS, thus a combination of using both seaweed species is
recommended.

Acknowledgments

This project was supported by Science Classroom in University Affiliated School (SCiUS) under Prince of
songkla University and Islamic Sciences Demonstration School. The funding of SCiUS is provided by the Ministry
of Higher Education, Science, Research and Innovation, which is highly appreciated. This extended abstract is not
for citation.

Reference

Abreu, M.H., Pereira, R., Yarish, C., Buschmann, A.H., Sousa-Pinto, I., 2011. IMTA with gracilaria
vermiculophylla: productivity and nutrient removal performance of the seaweed in a land-based pilot
scale system. Aquaculture, 312 (1–4), 77–87.

APHA AWWA WEF. (2017). Standard Methods for the Examination of Water and Waste water (23rd Edition).
American Public Health Association, Washington DC, 10(4), 114-122.

Nithiya, A., Shreeshivadasan, C., Hesam, K., Sathiabama, T., Norazli, O., and Noor, S. N., 2018. Treatment of
Wastewater Using Seaweed: A Review. International Journal of Environmental Research and Public
Health, 15(12), 1-17.

Dirk, W.,Steve, M., and David, W., T., 2004. Ammonia excretion in aquatic and terrestrial crabs.The Journal of
Experimental Biology 207, 4491-4504.

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Title: Microplastics contamination in marine animals OE1_13_02
Field:
Author: at the coastal area of Chonburi province

School: Environmental Science and Ecology
Advisor:
Ms. Kanyawee Sodapukd

Ms. Natchanan Khantee

Ms. Paphada Jantraphinun

Piboonbumpen Demonstration School, Burapha University

Dr. Vichaya Gunbua, Department of Aquatic Science, Faculty of Science, Burapha

University

Abstract
The study of microplastics contamination in marine animals at the coastal area of Chonburi province

was investigated. The samples were collected twice (November and December 2021) from 6 stations (Koh
Loy, Bangphra, Wonnapha beach, Khao Sammuk, Angsila, and Phli port) to investigate the aquaculture and
the Phli port was also represented the urban area while the Wonnapha beach represented the tourism site. The
manta net, submersed pump and grab sampler were used to collect the microplastics at the surface, 1.5 meter
under water and sediment sampling, respectively. In addition, microplastics contamination in marine animals
was investigated from pelagic fish (Rastrelliger brachysoma), benthic animal (Crassostrea gigas) and infauna
animal (Meretrix meretrix), respectively. The 5 compositions of microplastics found were fragments, films,
granules, filaments and foams while the filament was the most abundance. The results also indicated that the
total microplastics density in November was higher than in December but no statistical significance in pelagic
sediment and marine animal within 2 months of study.

Keywords: Chonburi province, Contamination, Marine animals, Microplastics

Introduction
Nowadays, the environmental problem because of the plastic waste in the seas has increased. The

plastic waste may accumulate in organisms which are naturally unable to digest. The data from Plastic Waste
Management Sub-committee in 2018 has estimated the number of microplastics as 5-10% of the total amount
of plastic waste in 2017, in Thailand, the total number of microplastics is approximately 39,570.4 - 79,140.8
tons per year that entering to the environment. So, this plastic waste will accumulate and increase all the time,
and may reduce in size and become the secondary microplastics.

However, Thailand has several researches about microplastics in the environment, mostly involved
with the sea. Some examples include the contamination of microplastics in bivalve at Chaolao and Kungwiman
beach Chanthaburi province. The results of the research showed that microplastic contamination in Danax sp.
in Chaolao beach was quite related to Kungwiman beach which was 3.13±2.75 particle/individual and
2.98±3.12 particle/individual (2) . Ingestion of microplastics by some commercial fishes in the lower Gulf of
Thailand: A preliminary approach to ocean conservation. The result of the research highlighted the ingestion
of plastics in 54.29% of samples. The ingested plastics were microplastics (27.27%; <5 mm), mesoplastics
(69.88%; 5-25 mm) and macroplastic, (2.85%; >25 mm) (1) . From the results of these microplastic studies can

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conclude that in many aquatic animals there was quite a bit of microplastic contamination. As time passed this
invisible plastic will accumulate a lot of bane from the ocean before entering the last consumer in the food
chain: a human who consumes the seafood.

Therefore, we have objectives which study in appearance, quantity and type of microplastics that
accumulated in economic aquatic animals of Chonburi province: pelagic fish (Rastrelliger brachysoma),
benthic animals (Crassostrea gigas) and infaunal animals (Meretrix meretrix). We had selected the 3 stations:
Phli port, Wonnapha beach and Koh Loy to represent the contamination of microplastics in Chonburi,
including the analysis related to the plastic wastes. This approach could lead to find the way to solve the plastic
waste problem.

Methodology
The samples were collected twice in November and December 2021 once a month, from 6 stations

which are Koh Loy (The buoy aquaculture), Bangphra (The hanging oyster farming), Wonnapha beach (The
coastal fishery and tourism site), Khao Sammuk (The staking oyster farming), Ang sila (The hanging oyster
farming and coastal fishery), and Phli port (The urban area and scallop and clam fishery). Each station
represents different water activities and approximately 10 km apart.

Manta net was dragged with flow meter attached and Submersed pump was used with filter bag which
mesh of both are 300 µm to collect microplastics at the water surface and under water at 1.5 m depth,
respectively. Each way collects 1,000 L of water samples.

At the bottom of the sea, Grab sampler was used to collect sediment samples. Then, 250 g/L sodium
chloride solution was added to 500 g of dried sediment samples, stir and leave to settling for 24 hr.

For investigated marine animal, mackerel (Rastrelliger brachysoma), only gills and digestive organs
were used while in clam (Meretrix meretrix) and oyster (Crassostrea gigas), only the meat of them were
included. The 10 g each of them was digested by adding 30% H2O2 solution at 60 °C. Then, 250 g/L sodium
chloride solution was added when they were all dried. Then, all were stirred and left to settle for 24 hr.

All samples which include water samples obtained by both Manta net and Submerse pump, sediment,
and marine animal samples were filtered through GF/C filter papers. The clear part especially at the surface
will be used for sediment and marine animal samples. Then, placed them at 60 °C for 24 hr. After that, the
classification of the microplastics in term of type, color and size were investigated by a Stereo microscope.

Results and Discussion
The results of the study found the microplastics composition in water and sediment in 5 types of

different colors which were Foam (white), size 169-596 μm, Filament (purple, blue, red, black, green and
orange), size 153-6,666 μm, Fragment (grey, purple, blue, red and orange) size 100-1,548 μm, colorless film,
size 650- 1,211 μm, and Bead (blue, green and white) size 122-412 μm. In economic aquatic animals, the
results were also found 5 types of microplastics which were mackerel: Filament (black, blue, red, green and
orange) size 436-1,623 μm, Fragment (red, blue and black) size 151-1,312 μm, and colorless transparent film
2,566 μm. In scallops, 4 types were found: Foam (white) with size 978μm, Filament (green, black, red-purple
and blue) with size 167-1,743 μm, Fragment (red, blue, green) with size 1,208-2,005 μm, and Bead (white)
318μm and in oyster. Found 2 types: Filament (green-black and red-blue), size 108-1,911 μm, and Fragment

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(red), size 768-1503 μm. Moreover, the Filament and Fragment microplastics were found in all aquatic
samples.

Fig.1 Foam Fig.2 Filament Fig.3 Fragment Fig.4 Film Fig.5 Bead

Density of microplastics in water samples collected by manta net in November and December 2021
were found to be in the range of 14–49 and 19-42 pieces/ m3, according to Sequential and microplastics
samples collected by submerse pump method showed microplastics in the range of 9-43 and 4-16 pcs/ m3,
respectively, and microplastics in the sediment samples. Microplastics were found in the range of 5-31 and 4-
17 pieces/500 g (dry weight), respectively, and the microplastics densities found in the 3 aquatic species
showed the highest microplastics densities in mackerel. found in the average of 9.17 pieces per head followed
by oysters, which were found in the average of 6.25 pieces per fish, and microplastics in clams were found in
the average of 6.00 pieces per fish, with the most filament microplastics found and could be found in all

economic fish samples.

Fig.6 Density of microplastics Fig.7 Density of microplastics Fig.8 Density of microplastics
by manta net in November 21. by submersed pump in by Grab sampling in

Fig.9 Density of microplastics Fig.10 Density of microplastics Fig.11 Density of microplastics
by manta net in December 21. by Grab sampling in December
by submersed pump in
December 21.

Fig.12 Density of microplastics in aquatic species

The results of this study were consistent with “Contamination of microplastic in bivalve at Chaolao
and Kungwiman beach Chanthaburi province” (2) including in accordance with “Microplastic Contamination
in Marine Animals Used as Seafood” (5) which the main type of microplastics is found, Filament is also the

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same. In addition, the research results of a study to study Contamination of microplastic in bivalve at Chaolao
and Kungwiman beach Chanthaburi province Microplastic densities were found in 2.98±3.12 pieces of
scallops each in Khung Wiman Beach. and Chao Lao Beach were found 3.13±2.75 pieces/body (2), which had
a density similar to that of the microplastics found in oysters and clams in this study.

Conclusion
The microplastic contamination at the coastal area in Chonburi province was found from the seawater

and sediment in the total of 5 types: Foam, Filament, Fragment, Film and Bead. The 5 types of microplastics
in marine animals were also found and split into 3 types in Rastrelliger brachysoma: Filament, Fragment and
Film, 4 types in Meretrix meretrix: Foam, Filament, Fragment and Bead, and 2 types in Crassostrea gigas:
Filament and Fragment. From the results, the Filament is represented the majority. The density of microplastics
in seawater was around 4-49 particles per cubic meter and sediment was around 4-31 particles per 500 grams
(dry weight). The average densities in Rastrelliger brachysoma, Crassostrea gigas and Meretrix meretrix were
9.17, 6.25 and 6.00 particles per individual, respectively. Filament type was the microplastics that was found
the most in marine animals.

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 Dr.Vichaya Gunbua for the
advisement. This extended abstract is not for citation.

References
1. Azad, S. M. O., Towatana, P., Pradit, S., Patricia, B. G., and Hue, H. T.. (2018). Ingestion of microplastics
by some commercial fishes in the lower Gulf of Thailand: A preliminary approach to ocean conservation.
International Journal of Agricultural Technology 2018, 14(7), 1017-1032.
2. ปิ ตพิ งษ์ ธาระมนต,์ สุหทยั ไพรสานฑก์ ุล และนภาพร เลียดประถม. (2559). การปนเป้ื อนของไมโครพลาสตกิ ในหอยสองฝา บริเวณชายหาดเจา้ หลาวและ
ชายหาดคุง้ วมิ าน จงั หวดั จนั ทบุรี. แก่นเกษตร 44 (1) (พเิ ศษ), 738-744.
3. มิถิลา ปรานศิลป์ , สิรินาถ ชยั ศรี และพชั รภา สวา่ งวงศ.์ (2562). ไมโครพลาสติกในตะกอนดินในพ้นื ท่อี ่าวไทยฝั่งตะวนั ออก. กรมทรัพยากรทางทะเลและ
ชายฝ่ัง.
4. Alysse Mathalon, Paul Hill. (2014). Microplastic fibers in the intertidal ecosystem surrounding Halifax
Harbor, Nova Scotia. Marine Pollution Bulletin, 81(1), 69-79.
5. พนั ธุ์ทิพย์ วเิ ศษพงษพ์ นั ธ์, อภิญญา ชาติทวสี ุข และ วชิระ ใจงาม. (2563). การปนเป้ื อนของไมโครพลาสตกิ ในสัตวท์ ะเลทเี่ ป็นอาหาร. การประชุมทางวชิ าการ
ของมหาวทิ ยาลยั เกษตรศาสตร์ คร้ังท่ี 58.

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Title : Chromosomal Aberration of Swamp Eel (Monopterus OE1_02_01

Field : albus) in Huai Tap Chang, University of Phayao
Biology and Biodiversity

Author : Miss Jinatta Nojak
Miss Sutipa Tangparukul

School : Demonstration School University of Phayao
Advisor : Asst. Prof. Dr. Kriengkrai Seetapan, School of Agriculture and Natural Resources, University

of Phayao

Abstract
Huai Tap Chang is a reservoir in the University of Phayao that obtained wastewater from several

activities within the university. It might be the cause of the water pollution. This study aimed to investigate the
heavy metal contaminations in water, sediment, and swamp eel (Monopterus albus) to compare with the
Thailand’s standard limits. Not only did the heavy metal concentrations be measured but the swamp eel
chromosomes were also assessed in M. albus from Huai Tap Chang (affected area) compared with from the
Fisheries Training Area, School of Agriculture and Natural Resources, University of Phayao (unaffected area).
The samples (water, sediment, and fish) were collected from affected and unaffected areas. All samples were
analyzed the heavy metal concentrations (As, Cr, Cu, Pb, Ni and Zn) by inductively coupled plasma optical
emission spectrometry (ICP-OES). Furthermore, five eels were collected for each site to assess the
chromosomal aberration. The results showed that, in the sediment sample, the contrations of Zinc (Zn) and
Arsenic (As) were higher than those from the standard limits. Additionally, 10 types of chromosomal aberration
were observed in Huai Tap Chang including centric fusion (CF), centric fragment (C Fr.), single chromatid gap
(SCG), ring chromosome (Ring C), dicentric chromosome (DC), sister chromatid gap (SSCG), deletion (D),
single chromatid decomposition (SCD), fragmentation (F) and polyploid (P). Notedly, the normal
chromosomes were found from unaffected area. These findings revealed that the Huai Tap Chang be
contraminated with the heavy metals until them can cause the chromosomal mutation in swamp eel.

Keywords: Huai Tap Chang, wastewater, heavy metal, chromosomal aberration, Monopterus albus

Introduction
Huai Tap Chang is the wastewater rest point in the University of Phayao near student’s dormitories

that gathers the water usage from several buildings around this area. Relatively, the water is low quality, bad
smell, and many dead fishes found floating on pond. As previous reports, several sources of metals from the
domestic water including food, tap water, detergents, cosmetics, laundry, and bath (Aonghusa and Gray, 2002).
The metals have soluble properties that can be turned into a complex compound. It can precipitate or become
colloidal particles, and it is a stable substance that cannot decompose in natural processes (Tengjaroenkul et
al., 2 0 1 8 ) . Therefore, the metals can accumulate in the sediment, water as well as aquatic animals, causing
their genetic abnormalities (Yadav and Trivedi, 2009). Some cytogenetic abnormality demonstrated that a high
level of arsenic (As) contraminated in water and sediment from gold mine can cause the chromosomal

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aberration in Asian swamp eel (Monopterus albus) (Neeratanaphan et al., 2019). Additionally, previous
research investigated the contamination of several heavy metals in water, sediment and nile tilapia
(Oreochromis niloticus) and the chromosomal aberrations in the domestic wastewater canal. The results
showed that the heavy metals concentrations were not higher than Thailand’s standard limits, except Ni
concentration in the water. However, the domestic wastewater resulted in the chromosomal abnormality in nile
tilapia (Sriuttha et al., 2017)

In the Huai Tap Chang, there are many species of aquatic organisms. This reservoir receives the
weastwater from many areas in the university, especially the weastwater from the doministries that might be
the cause of polluted water and aquatic animal abnormality. To assese these problems, we measure the
concentration of heavy metals in the water, sediment, and eels. Additionally, the cytogenetic abnormality was
observed in the swamp eel (M. albus) compared to the unaffected area.

Methodology
All samples (water, sediment, and eels) were collected from 2 areas including Huai Tap Chang

(affected area; yellow star) and the Fisheries Training Area, School of Agriculture and Natural Resources
(unaffected area; blue star), University of Phayao, Phayao, Thailand (Figure 1). Furthermore, these samples
were analyzed by Central Laboratory (Thailand) Co., Ltd. (Chiang Mai).

Figure 1: Overview of the Huai Tap Chang and unaffected area.

To investigrate the chromosomal abnormality, five eels were collected from affected and
unaffected areas and carried to the fishery laboratory. The eels were intramuscularly injected with colchicine
solution at 0.7ml/ 100g body weight. After 1 hour and 45 minutes, the injected eels were knocked on ice and
then the kidneys were dissected into small pieces. To induce the cell swelling, the tissues were suspended into
the hypotonic solution (0.075 M KCl) and incubated at room temperature for 30 minutes. After incubation, the
samples were centrifuged at 1,000 rpm for 10 minutes and the supernatant were discarded. The pellets were
gentally fixed and resuspended into a cool fixative solution (3 methanol: 1 acetic acid). The fixation process
was repeated 3 times and the prepared samples were kept in -20 oC for further study. To investigrate the
chromosome, the cells were stained with Giemsa’s solution and dropped onto glass slide. Then the slides were
observed and photographed under a light microscope. The chromosomal aberrations were assessed by
examining the percentage of 50 metaphase cells per sample. The sample preparation in this study was approved

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by the Animal Ethics Committee of University of Phayao based on the Ethics of Animal Experimentation of
National Research Council of Thailand UP-AE62-01-04-019.

Results
The concentration of heavy metals that contaminated in the sediment, water and eels from Huai Tap

Chang and unaffected areas are shown in Table 1. Interestingly, we found 2 contents of heavy metal including
As and Zn were higher that the standard limit (Thailand Pollution Control Department, 2018)

Table 1: Heavy metal concentrations in the sediment, water, and Monopterus albus from Huai Tap Chang and

unaffected area.

Heavy metals Huai Tap Chang area unaffected area

Sediment Water M. albus Sediment Water M. albus

(mg/kg) (mg/L) (mg/kg) (mg/kg) (mg/L) (mg/kg)

1. Arsenic (As) 10.8 0.0023 <0.13 ND ND ND

2. Chromium (Cr) 14.86 ND ND ND ND ND

3. Copper (Cu) 18.14 <0.0050 ND ND ND ND

4. Lead (Pb) 16.9 ND ND ND ND ND

5. Nickel (Ni) 11.68 ND <0.12 ND ND ND

6. Zinc (Zn) 97.78 ND 8.5 ND ND ND

Remarks: ND = Not detected

At unaffected area, we found that the diploid chromosomes of M. albus was 2n = 24. On the other
hand, at Huai Tap Chang, we discovered 10 types of chromosomal aberations including centric fusion (CF),
centric fragment (C Fr.), single chromatid gap (SCG), ring chromosome (Ring C), dicentric chromosome (DC),
sister chromatid gap (SSCG), deletion (D), single chromatid decomposition (SCD), fragmentation (F) and
polyploid (P) (Figure 2). Notedly, the most common chromosomal aberration in the affected area was CF.

Figure 2: Different types of chromosomal aberrations in the metaphase cells of Monopterus albus, showing a centric
fusion (CF), centric fragment (C Fr.), single chromatid gap (SCG), ring chromosome (Ring C), dicentric
chromosome (DC), sister chromatid gap (SSCG), deletion (D), single chromatid decomposition (SCD),
fragmentation (F) and polyploid (P) (A: unaffected area, B–H: Huai Tap Chang area).

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Conclusion
At Huai Tap Chang, the concentration of As and Zn in the sediment were exceed the Thailand

standard limits. Additionally, the most common of chromosomal aberrations was CF, D, SCG, Ring C, and F.
Nonetheless, the chromosomal aberration was not found at unaffected area.

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, Rescarch and Innovation, Jointed by
Demonstration School and School of Science, University of Phayao. This extended abstract is not for citation.

References
1. Aonghusa, C. N., & Gray, N. F. (2002). Laundry detergents as a source of heavy metals in Irish domestic

wastewater. Journal of Environmental Science and Health, Part A, 37(1), 1-6.
2. Neeratanaphan, L., Kanjanakunti, A., Intamat, S., and Tengjaroenkul, B. (2019). Analysis of

chromosome abnormalities in the Asian swamp eel (Monopterus albus) affected by arsenic
contamination near a gold mine area. International Journal of Environmental Studies. DOI:
10.1080/00207233.2019.1704087
3. Sriuttha, M., Khammanichanh, A., Patawang, I., Tanomtong, A., Tengjaroenkul, B., amd Neeratanaphan,
L. (2017). Cytotoxic assessment of Nile tilapia (Oreochromis niloticus) from a domestic wastewater
canal with heavy metal contamination. Cytologia, 82(1), 41-50.
4. Tengjaroenkul, B., Intamat, S., Thanomsangad, P., Phoonaploy, U., & Neeratanaphan, L. (2018).
Cytotoxic effect of sodium arsenite on Nile tilapia (Oreochromis niloticus) in vivo. International Journal
of Environmental Studies, 75(4), 580-591.
5. Thailand Pollution Control Department (TPCD). (2018). The Standard Levels of Heavy Metals in
Surface Water Quality. Available: https://www.pcd.go.th/wp-content/uploads/2020/05/pcdnew-2020-05-
25_04-11-22_154407.pdf. Accessed Day 29, 2022.
6. Yadav, K. K., & Trivedi, S. P. (2009). Sublethal exposure of heavy metals induces micronuclei in fish,
Channa punctata. Chemosphere, 77(11), 1495-1500.

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Title: Preliminary finding on the present and anatomical distribution of microplastics in giant
Field:
Author: freshwater prawns in cultured ponds

School: Environmental Science and Ecology OE1_10_01
Advisors: Mr. Nathachanok Chaiwiriyapong

Miss Chutimon Promkittiyanon

Miss Yanisa Chaikaew

Kasetsart University laboratory School Kamphaeng Saen Campus

Educational Research and Development Center

Assoc. Prof. Dr. Taeng On Prommi

Abstract:

Despite the fact that microplastic (MP) pollution is a global problem in aquatic environments, research on MPs

in commercial freshwater prawns is very limited. This study reports the shapes, sizes, types, and counts of

microplastics (MPs) found in 90 Macrostemum rosenbergii collected from three cultured ponds in central

Thailand. The study focused on four anatomical compartments (intestinal tract, hepatopancreas, muscle, and

exoskeleton), each of which was studied separately. MPs were found in all the investigated individuals, with
an average of about 79.3±2.92 MPs/individual. MPs were found in the intestinal tract, which had an average
of about 26.82± 0.92 MPs/individual, the hepatopancreas, which had an average of about 21.09± 1.11
MPs/individual, the muscle, which had an average of about 16.70± 0.63 MPs/individual, and the exoskeleton,
which had an average of about 14.69± 0.65 MPs/individual. Fibers were predominant over fragments and
spheres with an average 54.27-69.23%, 30.77-45.70%, and 0.40-0.56%. The majority of MPs were in the 200–
250 µm and 250–500 µm height ranges. The most common polymers were polyethylene terephthalate (PET),

polyvinyl acetate (PVAc), and poly(acrylonitrile-co-butadiene), accounting for approximately 61 percent of all

MPs discovered. A correlation (r = 0.85, p = 0.000 and r = 0.054, p = 0.002) was discovered between the

weights of male and female prawn organs and the number of microplastics identified in each organ. In terms

of the size of the microplastics identified in different organs, there was no difference between male and female

prawns in microplastics > 500 µm in all organs (p>0.05). The data presented here helps us better understand

the potential dangers associated with human consumption of various tissues from contaminated prawns.

Keywords: Microplastics, Macrobrachium rosenbergii, anatomical distribution, pond, FT-IR

1. Introduction
The presence of microplastics (less than 5 mm) in freshwater and marine organisms is a major source of

concern for biodiversity and poses a risk to human consumers of seafood and aquaculture
products. Microplastics have been found in a variety of marine and freshwater biota around the world,
including prawns and shrimp, bivalves, birds, and fish. Many laboratory investigations have also revealed that
microplastics and any related chemical pollutants attached to these microplastics are ingested by a variety of
organisms from various trophic levels. The primary uptake pathway of microplastics in aquatic species is
thought to be by ingestion. They are expected to remain in the digestive tract (gut) after ingestion and ultimately
be eliminated. Physical damage occurs after ingestion, including internal and/or external abrasions, ulcers, and
digestive tract obstructions. While some ingested pollutants can accumulate in tissues and cause internal
exposure, microplastic exposure is more temporary. Gills were also identified as a key pathway for collecting
microplastics in fish in several laboratory studies, despite the fact that this has rarely been documented in the
field. Due to an exponential population increase, aquaculture has become an increasingly essential source of
food. Due to an exponential population increase, aquaculture has become an increasingly essential source of
food. Macrobrachium species are among the most widely cultivated freshwater prawns on the planet (NEW et
al., 2010). In fact, Macrobrachium rosenbergii has been the most cultivated species on a commercial scale,
with production representing an excellent alternative to agribusiness due to its biological characteristics such
as growth, omnivory, high fertility, and resistance to disease. Furthermore, due to its soft texture and low
environmental impact, it is widely accepted on the market in all countries (Gupta et al., 2007; NEW et al.,
2010). The goal of this study was to provide the basis for future investigations into the presence of MPs in
distinct anatomical compartments and/or edible sections of M. rosenbergii from three farmed ponds in central
Thailand. The GI tract, hepatopancreas (HEP), muscle (MU), and exoskeleton were chosen as potential
contamination regions. MPs were chemically analyzed and classified as fragments or fibers. The size and sex
of the collected specimens were also considered when analyzing possible differences in MP distribution.

2. Methodology
1. Sampling and sample preparation

A modified seine net was used to gather fresh specimens of both female and male giant freshwater prawns
(Macrobrachium rosenbergii) from three local prawn farmed ponds in Thailand's central area in January 2022.

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A total of 90 giant freshwater prawn specimens (45 females and 45 males) were collected and kept cool in an
icebox before being transported to the laboratory.

Fig. 1 2. Extraction of microplastics from giant freshwater prawn
The shrimp were defrosted at room temperature before being
dissected. To remove any potential contamination from the plastic
freezer bags in which the shrimp were kept, the exteriors of each
shrimp were rinsed twice with deionized water (DI). A digital
weighing scale was used to weigh each shrimp, and a ruler was
used to measure the total length. Individually dissecting the
specimens on metal trays and removing their gastrointestinal tracts
(GI), hepatopancreas (HEP), muscle (MU), and exoskeleton (EX)
were done with metal forceps and metal scissors, which were
cleaned with DI after dissection (Fig. 1).

Each tissue organ was separately transferred and weighted into a 100-mL conical flask. After that, 20
mL of a hydrogen peroxide solution (30% H2O2) was added into individual flask to remove the organic matter.
All flasks were then covered with aluminum foil and and heated at 60 °C in a shaken water bath at 150 rpm for
3 hours, or until all of the organic matter was digested. The blanks were inspected for the presence of MPs
while running parallel to the soft tissue breakdown. In the blanks, no MP particles were found. Each sample
was transferred to a glass separation funnel to which 12 g potassium formate (99% HCO2K) was added. The
solution was drained and filtered onto nylon membrane filters (pore size 0.45 µm; diameter 47 mm) using a
vacuum filtration. The filters were placed in small aluminum bowls, covered them with aluminum foil and
placed them in a drying cabinet (50°C) for 2 d.

3. Visual observation of the microplastics
The microplastics on each filtered paper were observed and photographed using a stereomicroscope (Leica
EZ4E). A visual assessment was also used to identify suspected microplastics according to their morphological
characteristics such as color and shape (Hidalgo-Ruz et al. 2012). The shapes for microplastic particle
classification were fiber, sphere, film (thin and small layer) and fragment (part of a larger plastic item) (Su et
al. 2016, 2018).
4. FT-IR analyses of microplastics found in giant freshwater prawn
Microplastic particles were selected and identified using a Spectrum-Fourier transform infrared
spectrometer (FT-IR). Polymer type analysis and the characterization of functional groups were compared with
spectra database and instrument libraries, and the characteristic peaks of the functional groups were combined
to determine the polymer type (Bergmann et al. 2017).
5.Statistical analyses
The MP type, size, and color were analyzed and measured for each prawn tissue organs. One-way ANOVA
in combination with Tukey’s (HSD) post hoc pairwise comparisons was performed to determine significant
differences in the abundance of the MPs in the prawn tissue organs using SPSS software version 20.0 (IBM,
Armonk, NY, USA). Further, to generate graphs, Microsoft Excel 2013 (Microsoft Corp., Redmond, WA,
USA).

3. Results Number of MPs (item)
1. Abundance of microplastics
The mean weights of 30 male and female prawns in each pond revealed that prawns in pond 1 weighed

29.21±0.744 g/ind. and 19.93±1.06 g/ind., in pond 2 it was 33.44±0.42 g/ind. and 19.98±1.18 g/ind., and in
pond 3 it was 28.67±1.23 g/ind. and 17.99±1.69 g/ind., respectively. There were 2,412 microplastic items in
the intestinal tract of 90 prawn individuals from three cultured ponds; 1,898 items in the liver, 1,493 items in
muscle, and 1,323 items in the exoskeleton (Fig. 2, Table 1).

M_intes F_intes M_hep F_hep M_mus F_mus M_exo F_exo
300

250

200

150

100

50

0
1 3 5 7 9 11 13 15 17 19 21 23 25 27 29 31 33 35 37 39 41 43 45
Number of individual

Fig. 2 Example of microplastics type, size, and color in the intestinal tract of the freshwater giant prawns.

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A total of 7126 microplastic items were found in 90 prawn individuals. All prawns were contaminated with

microplastics. The mean was 79.3±2.92 item/ind. When considering microplastic contamination in prawn

organ tissue, the gastrointestinal tract was contained 26.82±0.92 item/ ind., the hepatopancreas contained

21.09±1.11 item/ind., the muscle contained 16.70±0.63 item/ind., and the exoskeleton contained 14.69±0.65

item/ind., respectively. In the 45 male prawns, 3,880 microplastic items were found, and 3,246 microplastic

items were found in the 45 female prawns. The lowest number of microplastics in the prawn was 89 and the

highest was 271. The number of microplastics discovered in each organ was found to be correlated (r = 0.85,

p = 0.000 and r = 0.054, p = 0.002) in an examination of the individual organ weights of male and female

prawns in three cultured ponds.

Table 1 Mean values and standard deviation of microplastic particles in each part of prawn.

Pond 1 Male (n=15) MPs (item) Female (n=15) MPs (item)

Intestine (g) 1.5736±0.5882 25.33±16.08 0.0558±0.0282 24.73±8.78

Hepatopancreas (g) 4.5799±0.8192 28.26±14.01 2.4486±0.9038 20.53±11.43

Muscle (g) 14.5057±1.9871 18.46±7.27 9.1032±1.1761 19.60±7.89

Exoskeleton (g) 6.5676±0.44 20.20±6.77 4.1991±0.5611 13.00±8.29

Pond 2 Male (n=15) MPs (item) Female (n=15) MPs (item)

Intestine (g) 1.6625±0.3561 36.80±14.49 0.0668±0.0173 29.40±15.23

Hepatopancreas (g) 5.5824±0.6179 30.06±19.91 2.2068±0.5447 17.53±5.26

Muscle (g) 16.3911±0.6768 20.60±8.33 9.5358±1.0821 17.53±10.58

Exoskeleton (g) 7.7098±0.6038 18.00±9.92 4.8714±0.8792 15.40±11.12

Pond 3 Male (n=15) MPs (item) Female (n=15) MPs (item)

Intestine (g) 1.4097±0.2002 22.73±9.64 0.5851±0.4989 21.80±9.30

Hepatopancreas (g) 3.3861±0.3379 16.26±5.27 2.7621±0.2769 13.40±6.12

Muscle (g) 14.0405±1.2774 11.60±2.22 7.4244±0.9630 12.40±5.47

Exoskeleton (g) 7.1259±0.2378 10.40±3.43 3.5744±0.4079 11.13±5.57

2. Color and type of microplastics
Detected MPs were mostly fiber (54.27–69.23%), fragment (30.77–45.70% %), and spher (30.77–45.70%)

in a total of 7,126 microplastics in 90 prawns (Table 2)

Six different colors were found in tissue organ of prawn, and the dominant color was blue (49.41-64.20%),
followed by white (transparent) (26.41-49.30%), red (1.29–11.54%), violet (1.17-4.63%), green (0.09-0.54%),

and yellow microplastics (0.15%), respectively (Table 2).

Table 2. Microplastic type and color in both the female and male of the giant freshwater prawn.

Category of microplastics Pond 1 Pond 2 Pond 3
Female Male Female Male Female Male

Type (%) Fiber 58.19 56.61 69.23 60.47 54.27 54.30

Fragment 41.41 43.39 30.77 39.53 45.17 45.70

Sphere 0.40 0 0 0 0.56 0

Color (%) Blue 63.29 59.45 57.18 64.20 55.02 49.41

Violet 1.17 4.63 1.21 1.49 0 0

Red 9.13 7.81 8.25 4.36 11.54 1.29

Transparent 26.41 28.01 33.36 29.94 32.75 49.30

Green 0 0.09 0 0 0.54 0

Yellow 0 0 0 0 0.15 0

3. Size of microplastics
Identified MPs were classified into four different sizes, i.e. <100 µm, 200˗250 µm, 250˗500 µm, and > 500
µm (Table 3). Microplastics with diameters of 200–250 µm and 250–500 µm are the most common, while

microplastics with diameters of 100 µm and > 500 µm are less common. In comparison to the size of

microplastics detected in various organs of male and female prawns, there was no difference in microplastic

size > 500 µm (p > 0.05).

Table 3. The size of microplastics in different organs of prawns.

Organ <100 µm 200 - 250 µm 250 - 500 µm > 500 µm
male female male female male female male female

I (n=45) 7.3±5.9 6.1±5.8 7.4±6.4 7.4±6.4 8.8±5.4 8.0±4.6 4.8±3.8 3.4±5.4

H (n=45) 5.4±6.6 4.8±4.6 8.7±6.1 8.7±6.1 4.8±3.2 4.8±3.2 3.2±2.6 2.1±2.0

M (n=45) 2.0±3.6 2.4±4.0 6.0±4.0 6.0±4.0 4.6±4.9 6.4±4.4 4.2±2.5 2.5±2.7

E (n=45) 1.4±2.7 2.3±4.6 5.0±4.5 5.0±4.5 6.2±3.0 4.2±3.5 3.9±2.2 2.6±1.9

Chi-square 37.752 20.994 9.590 18.566 22.134 21.644 6.104 2.239

df 3333 3 3 3 3

Asymp. Sig. 0.000 0.000 0.022 0.000 0.000 0.000 0.107 0.524

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4.Identification of polymers by FT-IR
A selected 727 plastic-like particles (about 10.2% of total MPs) were identified by a Fourier Transformed
Infrared Spectroscope (FT-IR). Of 695 MP particles (95.59%) were identified as microplastics, 4.41% as non-
microplastics. Some particles were confirmed as polyethylene terephthalate (PET), polyvinyl acetate (PVAc,
PVA), poly(acrylonitrile-co-butadiene), poly(ethylene glycol) methyl ether (mPEG), poly(propylene glycol)
methacrylate (PPGMA), polydiallyl phthalate (PDAP), polyethylenimine (PEI), poly(styrene-co-
divinylbenzene) (P(S-DVB)), and nylon. The spectral characteristics of these polymers are shown in Fig. 3

Poly(styrene-co-divinylbenzene) Nylon, 1, 0% None-microplastic, 32, 4%
(P(S-DVB)), 22, 3%
Polyethylene
Polyethylenimine (P… terephthalate (PE…

Polyvinyl
acetate (PV…

Polydiallyl
phthalate
(PDAP),
55, 8%

Poly(acrylonitrile- Poly(propylene Poly(ethylene
co-butadiene), 76, glycol) glycol) methyl…
11% methacrylate, 66,
9%

Fig. 3 FT-IR analysis and photos of the most common forms of microplastics detected in samples.

4. Discussion
In a study by Reunura and Prommi (2022), microplastic pieces were found in the gastrointestinal tracts of

female and male prawns. However, this is the first study to compare the presence of microplastics in different
regions of the shrimp pond system's anatomical Macrobrachium rosenbergii. This exploratory study can
provide important information on food security because it contains the edible portion (muscles and liver). It
emphasized the importance of sharing knowledge about the presence of microplastics in edible aquatic
organisms, with a special focus on seafood products.

Some of the plastic polymers discovered in M. rosenbergii are ubiquitous components of plastic debris
found in aquatic bodies. Packaged foods, plastic bags, bottles, and other kitchen plates or bowls are examples
(Strafella et al., 2019). The key components are PET (polyethylene terephthalate), which accounts for 37.13
percent of all plastic objects, and PVAc (polyvinyl acetate), which is used to make chewing gum. It's used in
latex glue, plastic water-based paint, and as a precursor to the creation of polyvinyl alcohol (PVOH), which is
used to make fibers instead of cotton, accounting for 13.34% of all plastic waste. They made up more than half
of the total number of M. rosenbergii members found in this study. It could be a piece of plastic mixed in the
shrimp pellet feed, fishing gear, lime to be sown to adjust the water conditions, and household waste or solid
waste that may be blown into the pond. Polymers related to synthetic fabrics, such as nylon, are found only in
small amounts in M. rosenbergii, possibly due to the washing process of synthetic garments. Washing machines
are one of the main sources of microplastic pollution in freshwater ecosystems, resulting from improper
management (Napper and Thompson, 2016; De Falco et al., 2019). In addition, atmospheric fiber can also
increase fiber contamination in freshwater cultured ponds (Dris et al., 2017).

The four organs (intestines, hepatopancrease, muscle, and exoskeleton) of the M. rosenbergii studied in this
study revealed that the intestine and hepatopancreas were the tissues most contaminated with microplastics,
followed by muscle and exoskeleton. The hepatopancrease plays an important role in various metabolisms and
serves as a storage area for fats and carbohydrates (Stathopoulou et al., 2017). In general, hepatopancreas
alterations are regarded as a useful technique for determining the impact of pollution on shrimp (Batista de
Jesus et al., 2020).

Based on the findings, we confirmed the presence of microplastics in all sections of the male and female
prawn tissue organs investigated. This prawn is frequently sold in the country's fresh markets and for export.
Despite the fact that microplastics are well-known contaminants in freshwater environments, their existence in
cultured agricultural systems has not been documented. This study is the first to report the discovery of
microplastics in lobsters in freshwater shrimp farming systems.

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 .

Reference
Reunura, T. & Prommi, T. 2022. Detection of microplastics in Litopenaeus vannamei (Penaeidae) and

Macrobrachium rosenbergii (Palaemonidae) in cultured pond. PeerJ 10:e12916

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Title : Active surveillance of Covid-19 spreading in school OE1_03_02
and surrounding community by monitoring of SARS-

CoV-2 in wastewater

Field : Environmental Science and Ecology

Author : Miss Pranpriya Thongkam

Miss Sarintorn Saranporn

School : Naresuan University Secondary Demonstration School, Naresuan University

Advisor : Asst. Prof. Tanapon Phenrat (Naresuan University)

Abstract
Presently, schools in Thailand are still suffering from the COVID-19 pandemic and have a high

risk of forming new clusters, which causes the schools to be shut inevitably. Shutting down schools affects the
quality of the students’ lives in both the short and long term. Therefore, this project aimed to apply wastewater-
based epidemiology techniques for the N-gene of SARS-CoV-2 identification in wastewater derived from
buildings in Naresuan University Secondary Demonstration School and the nearby community (samples from
Pumping station 4) in case there were any infected individuals with or without symptoms before receiving the
3-14 days individual RT-qPCR test for surveillance and early warning of COVID-19 pandemic, which would
reduce the risk of cluster leading to shutting down the schools.

In the analysis of wastewater samples from Pumping station 4 6 times from December 2021 until
February 2022, the N-gene of SARS-CoV-2 was found in every sample, which could earlier warn about the
spread twice by warning 5 and 7 days before the spread happening. We reported the data to the Phitsanulok
Provincial Public Health Office to proceed with the proactive measures for preventing the spread. On the other
hand, the analysis of samples in Naresuan University Secondary Demonstration School 11 times, the N-gene
of SARS-CoV-2 wasn’t found in any wastewater sample. But, 1 out of 190 students was reported confirmed
infection on 2 nd January 2022 and 3 out of 900 students were also reported during 7-11 th February 2022.
However, the cases were not a cluster. Thus, the SARS-CoV-2 identification in wastewater for early warning
of the cluster still succeeded.

Keywords : COVID-19, Wastewater-based epidemiology, Surveillance, SARS-CoV-2

Introduction
Presently, schools in Thailand are still suffering from the COVID-19 pandemic and have a high

risk of forming new clusters, causes the schools to be shut inevitably. Shutting down schools affects the quality
of the students’ lives in both the short and long term. The research stated that online learning due to the shut
down of schools will decrease the students’ lifetime earning power by 3%. For this reason, there should be
surveillance and early warning in schools. A method that has gained attention is Wastewater-based
epidemiology (WBE). In wastewater can detect SARS-CoV-2 and also detect pepper mild mottle virus
(PMMoV). The research shows using as an internal reference for quantification of SARS-CoV-2. The ratio of
SARS-CoV-2 to PMMoV varies with the number of infected people.

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In addition, wastewater data captures all COVID-19 cases including symptomatic and
asymptomatic. So, we will be able to notify the spread of covid-19 3-14 days in advance. This project aimed
to identify the N-gene of SARS-CoV-2 in wastewater derived from buildings in Naresuan University
Secondary Demonstration School and the nearby community (samples from Pumping station 4) by collecting
wastewater from communities in District 1 and District 2, Phitsanulok Municipality, Phitsanulok for
surveillance and early warning of the COVID-19 pandemic, which would reduce the risk of cluster leading to
shutting down the schools.

Methodology
Equipment : Bottle volume 250 ml, Water pump, Funnel, RT- PCR Machine
The experiments were divided into 2 parts,
Part 1 : SARS-CoV-2 detection

1.1. Collect wastewater samples from a septic tank at Naresuan University Secondary
Demonstration School and Pumping station 4 by using water pump and keep them in bottle.

1.2. Capture concentration of Total nucleic acids. This method allows rapid purification and
concentration of Total nucleic acids from large volumes of wastewater samples.

1.3. Extract Total Nucleic Acid and clean-up.
1.4. Use a nanodrop machine to identify concentration of Total Nucleic Acid.
1.5. Analyze the expression of SARS-CoV-2 related genes with RT-qPCR.
Part 2 : Surveillance and early warning
If the N-gene of SARS-CoV-2 is found in wastewater sample, we will analyze and interpret the
results by calculating the ratio of SARS-CoV-2 to PMMoV for predicting infected people trends 2 weeks in
advance and write a report to the Phitsanulok Provincial Public Health Office and inform to Naresuan
University Secondary Demonstration School’s director.

Results
Collecting wastewater samples from Pumping station 4 6 times from November 26 th 2021 until

February 18 th 2022, the N-gene of SARS-CoV-2 was found in every sample.

This bar and area chart shows the number of 7-day average infected people in Phitsanulok Muncipality in
Districts 1 and 2, detected by RT-qPCR and the ratio of N-gene to PMMoV in wastewater.

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In the analysis of data, on January 5th and February 4th, we could earlier warn about 5 days, 7 days
respectively before the spread happening. The lowest percentage of infected people per population that we
detect in wastewater is 0.24.

Collecting wastewater samples from Naresuan University Secondary Demonstration School from
December 10 th 2021 until February 25 th 2022, the N-gene of SARS-CoV-2 wasn’t found in any samples.

The line and area chart shows the number of 7-day average infected people in Naresuan University Secondary
Demonstration School, detected by RT-qPCR and the N-gene of SARS-CoV-2. We didn’t find the N-genes of
SARS-CoV-2 in wastewater. Although on January 2nd, there was one infected person from one-hundred ninety
people in a male dormitory representing 0 . 5 3 % , and during February 7-11th, there were three infected people
from nine hundred people in the school representing 0.33%.

Discussion
At Naresuan University Secondary Demonstration School, the percentage of infected people per

population is higher than the lowest percentage of infected people per population that we detect in wastewater
from Pumping station 4, but we couldn’t find the N-gene of SARS-CoV-2. It was probably a result of infected
people may not have defecated at the school or there were a few infected people which released less virus,
diluting virus in wastewater. On the other hand, Pumping station 4 gathers wastewater from Phitsanulok
Muncipality in Districts 1 and 2, have many infected people and a high virus concentration which makes it has
more chance to detect virus in wastewater than at the school.

Conclusion
In the analysis of wastewater samples from Pumping station 4, the N-gene of SARS-CoV-2 was

found in every sample, which could earlier warn about the spread twice by warning 5 and 7 days before the
spread happening. We reported the data to the Phitsanulok Provincial Public Health Office to proceed with the
proactive measures for preventing the spread. On the other hand, the analysis of wastewater samples from
Naresuan University Secondary Demonstration School, the N-gene of SARS-CoV-2 wasn’t found in any
samples. But there were a few infected people in the school. However, the identified cases were not a cluster.
Thus, the SARS-CoV-2 identification in wastewater for early warning of the cluster still succeeded.

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Acknowledgements

This project was supported by Science Classroom in University Affiliated School (SCiUS) under
Naresuan University and University Secondary Demonstration 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. K. Hill et al. Future perspectives of wastewater-based epidemiology: Monitoringinfectious disease
spread and resistance to the community level. Water Quality Research Journal, 2020;56(2):57-67.

2. M Kitajima et al. Pepper mild mottle virus as a water quality indicator. npj Clean Water,
2018;19(1): 1-9.

3. Voorn and de Jong. COVID. Multidisciplinary Digital Publishing Institute, 2021;1(6): 39-70.

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Title : Investigation of the elemental composition of different OE1_06_02

Field : types of cigarette and particle exposure from smoking
Environmental Science and Ecology

:Author Thanadech Saengrueangrak, Phasin Ponvisetsit, Muangthai Thongnok

:School Rajsima Wittayalai School, SCiUS Suranaree University of Technology

:Advisor Asst.Prof.Dr. Kiattisak batsungnoen, Institute of Public Health,

Suranaree University of Technology

Abstract
This project aims to develop an apparatus setup for the determination of elemental composition, as

well as the size and particle emission during smoking and after smoking. The apparatus setup experiment to
investigate the elemental composition and particle emission was performed by constructing the apparatus
chamber simulated human smoking. Three types of cigarette were investigated: 1. Thai cigarette 2. imported
cigarette 3. hand-made cigarette. Scanning electron microscope-energy dispersive X-ray spectroscopy (SEM-
EDS). Showed that hand-made cigarette had the highest amount of C, or soot (71.17 wt.%), whereas Thai
cigarettes and imported cigarettes contained 66.62 and 64.25 wt.%, respectively. It was found that size and
amount of particles emission during smoking consisted of various size and very high levels of particulate
matter. There were also some left behind after smoking which can be accumulated into high and exceed the
safety standard limit.

:Keywords tobacco smoking, elemental composition, particle emission, particulate matter (PM)

Introduction

In present-day society, smoking in Thailand is seen as prevalent among people ranging

from teenagers to the elderly, even though they already know that it is harmful to health for both the smokers
themselves and the people around them [1].

It was the aim of this project to investigate emission during and after smoking of three different
types of cigarettes.

In the experimental part, the elemental content was determined by connecting a cigarette to a smoker
detector which has been assembled with a trap filter inside for analysis in an elemental analyzer and also
detecting the particulate matter in the environment during and after smoking by using Portable Laser Aerosol
Spectrometer Model Mini-LAS 11-E.

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Methodology PLC Draw- out Cylinder
I. Apparatus setup and cigarettes preparation (exhale)
To setup an apparatus for investigating the elemental Linear
Draw- in
composition and particulate matter. The apparatus was conducted in (inhale)Actuator
several parts. The apparatus parts carried out in this study are following;
A. Control Part: Programmable logic controller (PLC) was used as a Filter
controller for the apparatus to start-stop the system and determine puff Cassette
volume by controlling the range of Linear Actuators to draw in and out.

B. Convert Part: To inhale air Linear Actuators was used to pump the

air to simulate the work of the human lungs by being controlled by

PLC. Figure 1: Smoking chamber simulated to human lung

C. Accumulate Part: Cylinder used as a tank to store cigarette smoke in the tank and then pumped out by

Linear Actuators.

The chamber simulating human smoking with a smoking simulator to smoke inhaled air at
55 mL ± 0.3 mL which is equivalent to human lungs were assembled, as shown in Figure 1 [2]. In this project,
three types of cigarettes were used as Thai cigarette, imported cigarette and hand-made cigarette

II. Experimental parts for analyzing elemental composition, size and amount of particulate matter

determination
The experiment started by preparing human smoking with a smoking simulator to smoke inhaled air. The
filter used was a 37 mm cassette filter with cellulose ester (MCE). The particulate
matter on the filter was further analyzed by using an electron microscope (FESEM -
EDS JEOL Model JSM 7800F, Japan) by scanning electron microscope energy
dispersive X-ray spectroscopy (SEM-EDS) [3] (Figure 3).

In the second part of the experiment to determine the Figure 3: Electron microscope (FESEM -
size and amount of particles produced when smoking Thai EDS JEOL Model JSM 7800F, Japan)
cigarette. Volunteer smokers were recruited to smoke four

rolls of cigarette using Thai cigarette and installed Aerosol Spectrometer (Portable Laser Aerosol

Spectrometer Model Mini-LAS 11-E) (Figure 4) in the smoking area and divided the experiment into three

periods.

1. Average background (measure surrounding particles before smoking) for 3 minutes.

2. Average during smoking (measure surrounding particles while smoking).

3.Average after smoking (measure surrounding (a) (b)
particles after the smoking session) for 5 minutes and

the measured values were graphed for further data

analysis.

Figure 4: (a) Guard volunteer, (b) Portable laser Aerosol Spectrometer

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Results, Discussion and Conclusion
I. Elemental Composition
The results from analyzing elemental composition showed that most of the elements found were

carbon (C) or soot. It’s a black, odorless solid that can be absorbed into the body through inhalation, in contact
with the eyes, causing irritation to the respiratory system and eyes and can also cause lymphoma. The elemental
composition from the emission of different types of cigarette was shown in Table 1. It was found that emissions
from Hand-made cigarettes contained soot as high as 71.17% which is higher than Thai cigarette (66.62%) and

Table 1:imported cigarette (64.25%). The particles were shown in Figure 5.
Elemental composition of smoking emission from cigarette.

Electron C O Electron C O Electron C O

Cl K N Cl K N CK N

Figure 5: X-ray map of smoke. (Right hand-made cigarette, Middle Thai cigarette, Left imported cigarette).
II. Size and amount of particulate matter
Each size and amount of PM measurement was performed repeatedly 4 times and averaged. The

results were shown in graphs of particle concentration (g/m3) versus time (s) (Figures 6, 7, 8, 9 and Table 2)
The total average suspended particulate (TSP) of all sizes produced during smoking was 1143.25 g/m3. The
average PM of each size produced during smoking was in the order: PM 10 (1,101.32 g/m3) PM 2.5 (980.22
g/m3) PM 1 In the experiment the average particle concentration during smoking, PM2.5 was 980.22 g/m3
whilst PM 10 was 1,101.32 g/m3 which higher than 65 time and 24 time of exposure standard respectively.
The world health organization (WHO) recommended for PM2.5 in 24hr exposure was 15 g/m3and PM 10
was 45g/m3 [4].

Table 2: Size and amount of particulate matter

Average Back-ground Average During Average After Smoking
[g/m3] Smoking [g /m3] [g /m3]

TSP 72.81 1143.25 266.64
PM10 238.26
PM2.5 58.61 1,101.32 192.48
PM1 122.41
43.96 980.22

40.39 465.99

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1. The developed smoking chamber with trapped filter in this project was successfully used for the
determination elemental composition with size and concentration of particulate matter.

2. The carbon (soot) was the main component in emission and hand-made cigarette produced the
highest amount of soot than the others.

3. Very high levels of particulate matter were emitted in this order:
PM 10  PM 2.5  PM 1

Inhalation of soot and PM from smoking can irritate respiratory and
lymphatic system and especially can cause lung cancer [5]. Figure 6: Total suspended particulate matter (TSP)

Figure 7: Particulate matter 10 Figure 8: Particulate matter 1 (PM1) Figure 9: Particulate matter 2.5
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
[1] T. Boonma, B. Sawnagwong, and W. Noichan, “Attitudes of the students in University towards
smoking electroniccigarettes and cigarettes.”
[2] ISO, “ISO 3308:2012.” International Organization for Standardization, 2012. Accessed: Feb. 17, 2022.
[Online]. Available: https://www.iso.org/cms/render/live/en/sites/isoorg/contents/data/standard/06/04/60404.html
[3] I. Guseva Canu, K. Batsungnoen, A. Maynard, and N. B. Hopf, “State of knowledge on the
occupational exposure to carbon nanotubes,” Int J Hyg Environ Health, vol. 225, p. 113472, Apr. 2020, doi:
10.1016/j.ijheh.2020.113472.
[4] WHO, “Ambient (outdoor) air pollution.” World Health Organization, 2021. Accessed: Feb. 11, 2022.
[Online]. Available: https://www.who.int/news-room/fact-sheets/detail/ambient-(outdoor)-air-quality-and-health
[5] O. US EPA, “Health and Environmental Effects of Particulate Matter (PM),” Apr. 26, 2016.
https://www.epa.gov/pm-pollution/health-and-environmental-effects-particulate-matter-pm (accessed Feb. 20,
2022).

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Title : The value added products and life cycle greenhouse gas OE1_03_03
emissions and cost of the products from pickled ginger

residue.

Field : Environmental science and Ecology

Author : Miss Wanrada Namwong

Miss Warintorn Sangwattanarat

School : Naresuan University Secondary Demonstration School, Naresuan University

Advisor : Asst. Prof. Dr. Suchada Ukaew (Industrial Engineering, Naresuan University)

Abstract
The objective of this research is to produce the value added products from pickled ginger residue,

and evaluate the greenhouse gas emissions (GHG) and costs of the three flavor pickled ginger and pickled
ginger salad products. In addition, the amount of water consumption for washing pickled ginger residue, by
single rinsing and counter current rinsing were carried out. The salt content in pickled ginger residue before
washing, after washing and in rinsing water were determined by the titration method. From the results, the
water consumption for the single rinsing and the counter current rinsing were 4 and 2 liters/250 g of pickled
ginger residue, respectively. The salt content in the single rinsing was 247.33 ml/liter. The salt content in the
first and last counter current rinsing were 889.46 and 199.71 ml/l, respectively. The system boundary of the
GHG emissions of two products is cradle to grave, covers from ingredient preparation, production and
packaging, transportation and waste disposal. The functional unit is one package of product (250 g of pickled
ginger). The GHG emissions of three flavors pickled ginger and pickled ginger salad were 0.24 and 0.21 kg
CO2 eq/package, respectively. The ingredient preparation stage of three flavors pickled ginger and pickled
ginger salad produced the highest GHG emission, accounting for 65.11% and 90.87%, respectively. The system
boundary of the production cost of the two products is cradle to gate, covers from ingredient preparation,
production and packaging, and transportation. The production cost of the three flavors pickled ginger and
pickled ginger salad were 23.96 and 20.46 Baht/package, respectively. Cost of the production mostly come
from workers accounting for 52.71% and 57.92%, respectively.

Keywords : Pickled ginger residue, GHG emissions, Cost of production, Life cycle assessment (LCA)

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Introduction
Thailand has processed and exported pickled ginger about 6,000 tons of pickled ginger per year.

These pickled gingers are decorated to get a beautiful shape by cutting off the unattractive parts, which can
create ginger residue about 1,800 tons per year, and they have to be landfill. Landfill creates methane (CH4)
about 45-60%, and carbon dioxide (CO2) about 40-60%, that are the main cause of global warming. Therefore,
this project is interested in increasing the value of the leftover pickled ginger residue. In addition, the project
evaluates the GHG emission and cost that used in the production of products made from pickled ginger residue.

Methodology
The experiments were divided into 3 parts as follows,

Part 1: Determination of salt content

1.1 Determine the amount of salt in pickled ginger residue before and after washing with water

Dry the pickled ginger at 105 Degree celsius, then take them to blend by the blender. After blending,
take 1 g of blended ginger, and add 10 ml of distilled water to make the blended ginger solution. After that,
prepare KCrO3, NaCl and AgNO3 solution. Then, standardize the AgNO3 solution by used 10 ml of NaCl
solution. Bring 10 ml of the blended ginger solution before and after washing with water, start titrating until
their color change into reddish brown. After that, calculate the salt content by the equation [1]

%Salt (NaCl) = (V)(N)(5.84) [1]

WSample

1.2 Determine the amount of salt in waste water in liquid form.

Bring 10 ml of waste water, start titrating until its color change into reddish brown. After that,
calculate the salt content by the equation [2]

Cl = (A-B)(N)(35450) [2]

m

Part 2: Production of a value-added product from pickled ginger residue

2.1 Production of three flavors pickled ginger

Wash 250 g of pickled ginger residue by single rinsing and counter current rinsing methods to remove
salt. After that, put them into a jar. Bring the water to boil, then add vinegar, lime juice and sugar. Pour the
water into a jar, and pickled them for one week. Lastly, put them in plastic bag.

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2.2 Production of pickled ginger salad

Wash 250 g of pickled ginger residue by single rinsing and counter current rinsing methods to remove
salt. After that, put them into a plate. Prepare the ingredients, the sugar, lime juice and chili. Then, mix them
together. Lastly, put them in plastic bag.

Part 3: Life cycle assessment

3.1 Evaluation of greenhouse gas emissions

Calculate the greenhouse gas emission by the equation [3]

GHG = A x EF [3]

3.2 Estimation of the production cost

Calculate the cost of the production by the equation [4], [5] and [6]

scrap value = (current price)(scrap value rate) [4]
Equipment depreciation = current price – scrap value [5]

lifetime [6]
cost/year = (ingredient cost)(used amount)(production/year)

boughted amount

Results

It was found that the salt content in pickled ginger residues before washing was 1 1 1 .9 9 % , and after
washing was 7 4 . 7 0 % , or decrease by 3 7 . 2 9 % . The reason why there are more than 1 0 0 % of salt, maybe it
caused by the mistake during titration.

It was found that the salt content in single rinsing water was 247.33 ml/liter. The salt content in first
and last tanks of counter current rinsing water were 889.46 and 199.71ml/liter, respectively. This means the
counter current rinsing method was able to remove more salt and can safe more water than the single rinsing
method.

Figure 1: The salt content in pickled ginger Figure 2: The salt content in waste water

residues before and after washing with water from single and counter current rinsing

method

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It was found that three flavored pickled ginger had the GHG emissions of 0. 21 kg CO2eq/package.
The GHG emissions in each stage from high to low was ingredients preparation > production and packaging >
waste disposal > transportation. As for the pickled ginger salad, the GHG emissions was 0 . 2 1 kg CO2
eq/package. The amount of GHG emissions in each stage from high to low was ingredients preparation > waste
disposal > production and packaging > transportation.

The three flavors pickled ginger and pickled ginger salad had production costs of 23.96 and 20.46
baht/package, respectively. Which mostly come from variable costs (99.79%). The annual production cost of
these two values added products were 213,458.77 and 182,265.57 baht/year, respectively. Which the Cost of
the production mostly come from workers accounting for 52.71% and 57.92%, respectively.

Figure 3: The GHG emissions of the Figure 4: The cost of the
values added products production per package

Conclusion
The salt content in the single rinsing was 247.33 ml/liter. The salt content in the first and last

counter current rinsing were 889.46 and 199.71 ml/l, respectively. The GHG emissions of three flavors pickled
ginger and pickled ginger salad were 0.24 and 0.21 kgCO2 eq/package, respectively. The ingredient preparation
stage produced the highest GHG emission, accounting for 65.11% and 90.87%, respectively. The production
costs were 23.96 and 20.46 Baht/package, respectively.

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
1. Erica Rodrigues. Cost of Production: The Ultimate Guide [Internet]. [cited 2022 Jul 28]. Available from:

https://prodsmart.com/cost-of-production-the-ultimate-guide/
2. The Nature Conservancy. Calculate Your Carbon Footprint [Internet]. [cited 2022 Jul 28]. Available from:

https://www.nature.org/en-us/get-involved/how-to-help/carbon-footprint-calculator/

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Title : The effects of forest fires on seed germination of some deciduous trees
Field :
Author : Environmental Science and Ecology

School : Thitichaya Teamtanomp OE1_01_01
Advisor : Phimpapond Pongtiwapan

Suthinee Roeksupha

Chiang Mai University Demonstration School

Asst. Prof. Dr. Sutthathorn Chairuangsri (Chiang Mai University)

Kusol Raungprataungsuk (Chiang Mai University Demonstration School)

Abstract :
The frequent forest fires in the northern part of Thailand affect forest ecosystems in terms of both survival

rates and plant growth. These may affect the structure of plant society in the long term. The purpose of this research
was to study the effects of forest fires on germination and seed growth of 5 species of deciduous trees, namely
Afzelia xylocarpa, Tectona grandis, Xylia xylocarpa, Pterocarpus macrocarpus, and Dalbergia oliveri. The
factors from forest fires that were chosen to study the effects on tree seeds were heat and inorganic substances
from biomass combustion. Seeds from each species were divided into 4 groups 1) burned with leaf litter, 2) soaked
in ash solution, 3) burned with leaf litter and soaked in ash solution, and 4) control treatment. After that, five seeds
were randomly sampled from each treatment for seed viability testing with triphenyl tetrazolium chloride solution.
The treated seeds were planted to compare the germination rates of each species in all experiments. Heat reduced
seed germination and viability, with the exception of Tectona grandis, which was more viable than the control.
Seed germination and viability were altered by biomass combustion inorganic compounds, with higher
germination rates and viability greater than the control, except for Tectona grandis viability than the control and
Xylia xylocarpa with reduced germination rate. Seed germination and viability are affected by the heat and
inorganic chemicals produced by biomass burning, resulting in a lower germination rate and worse viability than
in the control.

Keywords : Forest fires, Biomass, Seed viability testing, Ash solution, and Germination rates.

Introduction
There are different types of forest fires, one of which is the most common type of wildfire in the forests

of northern Thailand, which is a surface forest fire. Although surface forest fires are not as deadly as other types,

they affect the forest ecosystem in terms of plant growth and survival and the community structure. A forest fire

can have both good and bad consequences, for example, it can destroy trees but it may induce sprouting and seed
germination in some species. The type of forest with the highest forest fire frequency in the north is deciduous
forests. The important factors that cause by forest fires and may affect forest plants were heating from the flame
and inorganic substances obtained from biomass combustion.

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The purpose of this research is to study the effects of forest fires
on seed germination in some deciduous forest species. The seeds from
deciduous tree species were treated with heat from flames, a solution of
inorganic substances from the combustion of biomass, and obtained both
heat and inorganic substances solution.

Methodology
Seeds of five deciduous tree species; Afzelia xylocarpa, Tectona grandis, Xylia xylocarpa, Pterocarpus
macrocarpus, and Dalbergia oliveri were selected for the study.
The experiments were divided into 4 parts as follows,

Part 1: Study the structure of the seed
The seed structures were studied. The seeds were cut vertically and horizontally and
photographs were taken under the stereomicroscope. The seed's husk thickness, the
position of the embryo, and the endosperm inside the seed were examined.
Part 2: Simulating forest fire conditions
The factors from forest fires that were chosen to study the effects on tree seeds were heat and inorganic substances
from biomass combustion. Seeds from each species were divided into 4 groups.
2.1 Burned with leaf litter
Thirty seeds of each species were prepared. The prepared seeds are burned with
500 g of leaf litter (the biomass combustion time was 136 seconds). After that, the
seeds are soaked in distilled water for 24 hours. Three replicate sets of the
experiment were done for each species.
2.2 Soaked in ash solution
Leaf litter was burnt and the ash was then dissolved in distilled water (the ratio of
ash to distilled water is 20 g : 1000 ml.). Thirty seeds were soaked in ash solution for 24 hours. Three replicate
sets of the experiment were done for each species.
2.3 Burned with leaf litter and soaked in ash solution
Thirty seeds are burned with 500 g of leaf litter, after which the seeds are soaked in ash solution for 24 hours (the
ratio of ash to distilled water is 20 g : 1000 ml.). Three replicate sets of the experiment were done for each species.
2.4 Control treatment
Thirty seeds were soaked in distilled water for 24 hours. Three replicates were done for each species.
Part 3: Seed viability testing with triphenyl tetrazolium chloride solution
Five seeds were randomly sampled from each treatment in Part 2, and the sampled seeds were tested for viability
with triphenyl tetrazolium chloride solution. The sampled seeds were cut in half lengthwise, then sampled seeds
were soaked in the 1% 2,3-5 Triphenyl tetrazolium chloride solution for 20 hours at 40 °C. In this TZ test, the
activity of the hydrogenase enzyme, which is specific to living cells was investigated. The part of seeds that has

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cellular respiration (living tissue) will turn to be red. After that photographs of seeds were taken under a
stereomicroscope. And the internal coloration of the seeds was recorded.

Part 4: Seed germination
The seeds from experiments simulating forest fire conditions and control
treatment (soaking in water for 24 hours) were germinated. The germination rate
from different treatments was compared to determine the effects of forest fires
on tree seeds. The seed trays were watered daily, and germination rates were
recorded every 3 days for 30 days.

Results and Discussion
Tectona grandis, Afzelia xylocarpa, Xylia xylocarpa, Dalbergia oliveri, and Pterocarpus macrocarpus

had mean seed husk thicknesses of 4.34, 1.50, 0.449, 0.156, and 0.143 mm. respectively. The embryo is located
on the inner top of the seed, with the exception of Tectona grandis, which is located in the center, while the
endosperm inside the seed is located on the seed's inner flesh.

The result of the viability test using triphenyl tetrazolium chloride solution showed that the seeds of
Afzelia xylocarpa, Xylia xylocarpa, Dalbergia oliveri, and Pterocarpus macrocarpus treated with heat from
biomass burning and heat + ash solution had lower viability than the control experiment and the seeds that soaked
in ash solution. While for Tectona grandis, the seed viability test showed that the heat and inorganic chemicals
created by biomass combustion affect the viability of this species, the color of Tectona grandis in this treatment
compared with the control experiment was clearly less red than in other experiments. This may be due to the thick
seed husk that protects the embryo from heat and ash solution.

The result of the seed germination test was determined for one month, which may not enough for some
species to germinate (Table 1). Dalbergia oliveri, and Pterocarpus macrocarpus have very low germination rates
while non of Tectona grandis seed was germinate in all experiments.

Soaking Afzelia xylocarpa seeds in ash solution increased the germination rate of this species. However,
heat from burning biomass decreased its germination rate. The ash solution may soften the thick seed husk of this
species better than water so that water and air can penetrate into the seed and reach to embryo better resulting in a
higher germination rate.

The germination rate of Xylia xylocarpa seeds in the heat experiment and heat + ash solution experiment
was 0%. This indicated that heat could totally damage the seed of this species. Soaking the seed of Xylia xylocarpa
in ash solution also decreases its germination rate. This may be due to the size of the seed which is medium size
and its seed husk was thinner than Afzelia xylocarpa and Tectona grandis.

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Table 1. The seed germination rate of Tectona grandis, Afzelia xylocarpa, Xylia xylocarpa, Dalbergia oliveri and

Pterocarpus macrocarpus with different presowing treatments.

Species Burned with leaf Soaked in ash Burned with leaf Soaked in

litter solution litter and soaked distilled water

in ash solution (control)

Tectona grandis 0% 0% 0% 0%

Afzelia xylocarpa 27.28% 64.44% 20% 37.78%

Xylia xylocarpa 0% 31.11% 0% 47.78%

Dalbergia oliveri 1.11% 0% 0% 0%

Pterocarpus macrocarpus 0% 0% 0% 2.22%

Conclusion
Heat reduced seed germination and viability, with the exception of Tectona grandis, which was more

viable than the control. Seed germination and viability were altered by biomass combustion inorganic compounds,
with higher germination rates and viability greater than the control, except for Tectona grandis viability than the
control and Xylia xylocarpa with reduced germination rate. Seed germination and viability are affected by the heat
and inorganic chemicals produced by biomass burning, resulting in a lower germination rate and worse viability
than in the control.

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

Mai University and Chiang Mai University Demonstration School. The funding of SCiUS is provided by the
Ministry of Higher Education, Science, Research, and Innovation, which is highly appreciated. This extended
abstract is not for citation.

References
Fernandes, A.F., Oki, Y., Fernandes, G.W. et al. 2021. The effect of fire on seed germination of campo rupestre
species in the South American Cerrado. Plant Ecol 222, 45–55.

Kongwichai, Kaewsorn, and Chulaka. 2016. Viability Test by Tetrazolium Solution of Wax Gourd (Benincasa
hispida) Seed, Agricultural Sci. J. 47(2)(Suppl.): 337-340

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Title : Environmentally Friendly Waterproof and Anti- OE1_16_01
Field : Fungal Paint from Natural Rubber Latex
Ecology and Environment Science

:Author Ms. Nantachaporn Sudjai

School : Mr. Raffel Niyomdecha
Demonstration School Prince of Songkla University, Prince of Songkla University

Advisor : Dr. Nabil Hayeemasae, Department of Rubber Technology and Polymer Science, Faculty of
Science and Technology, Prince of Songkla University, Pattani Campus

Abstract

Waterproof paint is part of the waterproof system which consists of two important components:
pigment and binder. The binders that are available in the markets are mainly composed of harmful additives
such as heavy metals, volatile organic compounds, solvents, and so forth. In this research, we prepared
waterproof paint based on natural rubber latex that is environmentally friendly and meets TIS standard and
Green Label. Apart from that, the prepared paint is also antifungal, making it sustain its quality without microbe
living on it. The paint was prepared by mixing the pre-vulcanized natural rubber latex (NRL) compound with
acrylic emulsion (AE). When varying the ratios of NRL and AE, it was observed that the ratio of 70:30
(wt%/wt%), is proper for preparing paint. This ratio provided a homogenous mixture and suitable tensile
properties. Afterward, the paint was undergone certain testing and analysis to meet the requirements of TIS no.
1123-2555 (Masonry primer) and Green Label. The tests for TIS covered the film characteristics when dry,
fungal resistance, water resistance, and alkali resistance. Meanwhile, the Green Label contained an analysis of
volatile organic compounds and heavy metals (Pb, Cd, and Hg). It was observed that waterproof paint
successfully passed the TIS and Green Label requirements. It can be concluded that waterproof and antifungal
paint is successfully prepared, it also meets the Green Label requirement which can be claimed as
environmentally friendly paint.

Keywords : Pre-vulcanized Natural Rubber Latex, Volatile Organic Compounds, Heavy Metal

Introduction

Most commercial waterproofing paints use synthetic polymers as a binder such as polyurethane,
and acrylic emulsion, which contain substances that are harmful to the health of residents. In addition, general
waterproofing paint contains heavy metals and volatile organic compounds. If it accumulates in the body in
large quantities, it is harmful to the residents as well.1,3 Therefore, the research team is interested in developing
a waterproofing paint from natural rubber latex by using a binder based on Pre-vulcanized natural rubber latex

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and acrylic emulsion. Because natural rubber latex has outstanding properties. good adhesion water-resistant
and natural rubber latex is also low cost. By developing waterproof and anti-fungal paint that meets the
requirement of TIS no. 1123-2555 (Masonry Primer) and Green label criteria to reduce the amount of heavy
metals, and volatile organic compounds and has minimal impact on the environment and consumer health.

Methodology
Part 1: Preparation of Pre-vulcanized Natural Rubber Latex

The pre-vulcanized natural rubber latex was arranged for heating it at 60 °C and time variables of
30, 60, 90, 120, 150, and 180 min, and then prepared as a film. To be tested for the Degree of vulcanization by
using swelling test and chloroform test.
Part 2: Mix pre-vulcanized natural latex with acrylic emulsion

The prepared pre-vulcanized natural latex was mixed with the acrylic emulsion at the following
ratios of 60:40, 70:30, 80:20, and 90:10 (wt%/wt%). Then prepared polymer blend as a film. and used to test
mechanical properties such as modulus, tensile strength, and the Elongation at break by tensile test.
Part 3: Preparation and testing of waterproof and antifungal paint

Prepare the waterproof paint with a blend ratio between pre-vulcanized natural latex and acrylic
emulsion equal to 7 0 : 30 (wt%/wt%), and test for brush-applied properties and film properties when applied
dry. Fungal resistance, water resistance, and alkali resistance meet the requirement of TIS no. 1 1 2 3 - 2 5 5 5
(Masonry Primer), and test for the heavy metals content by ICP-AES and volatile organic compounds content
by TGA that meet the requirement of the Green Label.

Results, Discussion, and Conclusion
The experimental results from the preparation of pre-vulcanized natural latex by heating at 6 0 °C for

150 min will provide suitable properties to be mixed with acrylic emulsion. as a binder in paint Based on testing at
different ratios of pre-vulcanized natural rubber latex to acrylic emulsion 90:10, 80:20, 70:30, and 60:40 (wt%/wt%),
the study of mechanical properties by the tensile test shows that the 70:30 ratio has good mechanical properties and
the best characteristic quality compared to other ratios, the 7 0 :3 0 (wt%/wt%) ratio, therefore, used to mix into
waterproof paint as shown in Figure 1. and used to test its properties as a waterproof paint product, tested according
to TIS no. 1 1 2 3 - 2 5 5 5, testing the appearance of the film when dry, fungal resistance, water resistance, alkali
resistance. It was found that the waterproof paint from the research passed all test criteria. Also, when testing for

heavy metal substances No excess heavy metals were found in the prescribed amount, as shown in Table 1. And
testing for volatile organic compounds found that the value of 1.32% was within the criteria specified by the green

Stress (MPa)label (10%w/w), as shown in Figure 2. 60/40
70/30
35 80/20
30 90/10
25 100/0
20
15
10
5
0

0 200 400 600 800 1000 1200
Strain (%)

Figure 1. Stress-strain curves of NR/Acrylic blends

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105 1
100
95Weight % (%) 0
90 Derivative Weight % (%/min)
85 -1
80
75 -2
70
65 Thermogravimetric curve -3
60 (%) -4
55 -5
50
Derivative -6
30 thermogravimetry curve -7
(%/min) -8

-9
80 130 180 230 280 330 380 430 480 530 580

Temperature (°C)

Figure 2. Thermogravimetric analysis (TGA) curves showing the mass loss profile of VOCs

Table 1. Results of Heavy metal Determination Test by ICP-AES

Heavy metal Result (mg/kg)
Ni Not detected
Cd Not detected
Mn Not detected
Cr Not detected
Cu Not detected
Pb Not detected
Hg Not detected

Table 2. Results of testing according to TIS. 1123-2555 and Green Label

List Result
Application properties and film characteristics Pass
when dry
Fungal resistance test Pass
Water resistance test Pass
Alkali resistance test Pass
Heavy Metal Determination Test Pass
Pass
Volatile Organic Compound Determination Test

According to the results, it was found that incubation of pre-vulcanized natural rubber latex at
60°C for 150 min provided the most optimum conditions for the preparation of pre-vulcanized rubber latex,
compared to curing at other periods. By preparing pre-vulcanized natural latex and acrylic emulsion at different
ratios, it was found that the 70:30 (wt%/wt%) ratio had better mechanical properties than the different ratios.
And the preparation of waterproof and antifungal paint in this research meets the requirement of TIS no. 1123-
2555 (Masonry Primer) and the detection value of heavy metals and volatile organic compounds meet the
criteria of the Green Label, as shown in Table 2.

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Acknowledgments
First of all, we would like to pay our gratitude to our advisor, Asst. Prof. Dr. Nabil Hayeemasae

for warm guidance and lots of support to complete the project. Importantly, there would not have been this
project without the opportunity he had given to do this project. 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. Briffa J, Sinagra E, Blundell R. Heavy metal pollution in the environment and their toxicological effects on
humans. Heliyon. 2020;6(9):1-26.
2. El-Sherbiny S, Morsy FA, Atta AM, Ahmed SA. Preparation of Water Based Polymeric Binders for Paper
Surface Coating. Journal of Surface Engineered Materials and Advanced Technology. 2014;04(03):140-154.
3. Guo H, Lee S, Chan L, Li W. Risk assessment of exposure to volatile organic compounds in different indoor
environments. Environmental Research. 2004;94(1):57-66.
4. Ibrahim B, Helwani Z, Fadhillah I, Wiranata A, Miharyono J. Properties of Emulsion Paint with Modified
Natural Rubber Latex/Polyvinyl Acetate Blend Binder. Applied Sciences. 2021;12(1):1-296.

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Title : Soil carbon contents of Northeastern’s home garden OE1_07_01

Field : and monoculture plantation
Author :
School : Environmental Science and Ecology
Advisor :
Naruedee Wongyai, Tanjira Punyasarn

Mahasarakham University Demonstration School (Secondary), Mahasarakham University

Assistant Prof. Dr. Bhuvadol Gomontean (Department of biology, Faculty Science,

Mahasarakham University)

Abstract :
The investigation of soil organic carbon (SOC) stocks in different plantations is critical for

sustainable land management in the Northeast of Thailand. This study was conducted in Sahatsakhan and
Namon district of Kalasin Province in Thailand. The study was to evaluate soil organic carbon content and
storage in the different of plantation types e.g. home garden, teak plantation and cassava plantation. Soil
samples were collected at topsoil (0-30 cm depth) and subsoil (30-50 cm depth). Soil samples were analyzed
for physical and chemical properties such as soil texture, soil bulk density, soil moisture, soil pH, electrical
conductivity, organic matter, and soil organic carbon. The results showed that the soil texture of topsoil and
subsoil were sandy loam in all plantation types. The soil pH range was neutral in the home garden plantation,
but were moderately acidic in cassava and teak plantation soil. The electrical conductivity of all plantation
soils was classified as non-saline soil (0.010 to 0.025 dS/m). Bulk density in all plantation types soil showed
higher bulk density at subsoil than topsoil. The amount of organic matter and soil organic carbon content
between topsoil and subsoil of home garden and teak plantation were significantly difference (p<0.05),
except for cassava plantation. Among all plantation types, soil organic carbon content and storage in both
topsoil and subsoil were significantly difference (p<0.05) and the home garden showed the highest soil
organic content and storage than the other plantations (p<0.05). Moreover, at 0-50 cm soil depth, soil organic
carbon storage was significantly difference among all plantation types (p<0.05). Soil organic carbon had been
stored in home garden, teak plantation, and cassava plantation. (55.96±1.87, 25.43±0.96 and 8.71±1.14 tonha-
1, respectively). As the results home garden with higher plant diversity and less soil disturbance activities
representing a valuable option for storing of carbon and finally mitigating of climate change as climate
neutrality. Furthermore, reducing the risks of the loss of local biodiversity and be a viable option for
sustainable land management in Northeastern Thailand.

Keywords : soil, monoculture, home garden, carbon contents, Northeastern Thailand

Introduction
To date, the global major concerns are about food security, and the increase in greenhouse gas in the

atmosphere from human activities. Massive uses of the world's resources are associated with an increase in
the world's population in recent times. The increasing of population number is related to quantity of carbon
dioxide and other greenhouse gases in the atmosphere1. The demand of agricultural yields is couple with the
increasing of population and inevitably affect the earth's soil resources in various regions. Such concerns,
food security and atmospheric greenhouse gas, are linked to the soil quality which related to soil carbon and
dynamic. In agriculture ecosystems, soil organic carbon (SOC) is an effective index of soil quality and

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essential for sustaining agricultural productivity2. Declination or loss of soil organic carbon would be
decreasing of soil quality and crop yields3.

In addition, carbon reservoirs in the world besides the forest and wetland ecosystems agricultural
ecosystems are another important deposit source of carbon that should be given priority as a source or sink of
atmospheric carbon4. However, strategies and inappropriate land-uses practices in agricultural systems effect
to soil quality, carbon sequestration, and atmospheric carbon dioxide5. Although the increasing of carbon
dioxide emissions due to soil-related agricultural activities are not the primary cause compared to fossil fuel
combustion, agricultural ecosystems could be a climate mitigation solution if properly application of
management activities2. Carbon reservoirs in agricultural and degraded land areas can store up to 55-66 % of
past 42-78 Gt C loss. Generally, SOC accumulation rate depends on soil texture and soil structure, rainfall,
temperature, farm system and soil management. Enhancing the amount of carbon storage in the soil can be
achieved through soil remediation, regeneration of plant species, no-till farming, mulching, nutrient
management fertilization and soil improvement, water systems and mixed farming system6. Several studies
reported soil carbon storage in different farming systems and practices such as orchard plantation and
sugarcane plantation7, varieties of cropland and rangeland6, agricultural land-use and management practices8

From 105,554 Rai of total area in the northeast part of Thailand, there were 63,477 Rai of
agricultural area. There are various of farming system and practices those practices in Northeastern part of
Thailand especially in Kalasin Province such as monoculture, diverse cropping system, agroforestry
including home garden, and forest plantation. The researcher’s question of this study was how much of soil
carbon could be stores in the different of agricultural practices. The objective of this work was to assessed the
content of soil organic carbon in the different agricultural practices (home garden, forest plantation, and
monoculture). The results would be useful for agricultural management towards sustainability of soil quality
and climate mitigation.

Methodology
This study was conducted in agricultural areas with similarity of topographic factors in Kalasin

province, Thailand. They were classified into 3 types of plantation i.e. 1. Home garden plantation: HG (more
than ten year cultivation period) is represent for the traditional conservation farmland with the highest plant
diversity (58 species), 2. Teak plantation: TP (ten-year cultivation period) is represent for agroforestry
plantation with moderately plant diversity (12 species), and 3. Cassava (Manihot esculenta Crantz)
plantation: CP (ten-year cultivation period: rotation crop) is represent for monoculture area with highest
tillage frequency than the others (two times per month). Soil samples were collected from three plantations.
In each plantation type, two quadrats of 2×2 m were randomly laid down. A soil core and soil samples at 0–
30 cm depth (top soil) and 30-50 cm depth (sub soil) were collected. All soil samples were transferred to the
laboratory, for further processing and analyses (Table 1).

Statistical analysis comparison of soil organic carbon content, organic matter and SOC stock
between plantation types and between soil layers were analyzed using one-way ANOVA statistic with
DMRT (Duncan's Multiple Range Test) method.

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Table 1. Laboratory analysis methods

Analysis Method References
Organic matter Walkley-Black method Walkley and Black, 1934
Soil particle size Hydrometer method Soil Survey Staff, 1996
Soil reaction: pH Glass electrode 1:1 soil solution in water Soil Survey Staff, 1996
Soil bulk density Core method Black and Hartge, 1986
Electroconductivity: EC Electrical conductivity meter Jackson, 1958.
Soil moisture Gravimetric with oven drying Gardner, 1965

Result, Discussion and conclusion

The basic physical and chemical properties of the HG, TP and CP are shown in Table 2. Soil texture

All plantation types had the high sand particle proportion, and had the same soil texture as sandy loam. CP

showed highest soil moisture in both topsoil and subsoil layer while TP showed the lowest at the topsoil

layer. Soil bulk density were higher in subsoil of all plantation types with the lowest was at the topsoil of TP.

Table 2. Soil physical properties

Land use Depth Soil particle size (%) Soil texture Bulk density Soil
(cm) (g m-3) moisture
Home garden (HG) Sand Silt Clay sandy loam
Teak plantation (TP) 0-30 sandy loam 1.46 (%)
Cassava plantation (CP) 30-50 68.4 24.36 7.24 sandy loam 1.93 4.41
0-30 59.4 27.6 13.24 sandy loam 1.04 4.67
30-50 73.4 21.36 5.24 sandy loam 2.11 3.98
0-30 64.4 26.36 9.24 sandy loam 1.29 5.18
30-50 67.4 23.36 9.24 1.53 4.98
69.4 22.36 6.24 6.06

Soil reaction were neutral only at the HG site while the others were moderately acid. Soil electrical

conductivity (EC) value among all plantation types indicated that all soil samples were non-saline soil. In all

samplings, soil organic matter (OM) in the HG site was significantly highest while significantly lowest at the

CP site in both two depth. (Table 3).

Table 3. Soil chemical properties.

Land use Soil depth pH Soil reaction EC Salinity OM
Home garden (HG) (cm) (dS m-1) (g kg-1)
Teak plantation (TP) 0-30 7.03 neutral none 15.18±0.51Aa
Cassava plantation (CP) 30-50 6.73 neutral 0.025 none 7.77±0.26Bb*
0-30 5.64 moderately acid 0.016 none 7.95±0.50Ba*
30-50 5.59 moderately acid 0.013 none 4.52±0.77Cb*
0-30 5.73 moderately acid 0.011 none 2.17±0.51Da*
30-50 5.56 moderately acid 0.011 none 2.17±0.00031Da*
0.010

*Similar capital letters at among the plantation type in both top soil and sub soil indicates no significant treatment differences according
to least significant differences (Duncan) at P < 0.05. Similar small letters at among the dept in each plantation type indicates no
significant treatment differences according to least significant differences (Duncan) at P < 0.05.

Soil organic carbon: SOC
Soil organic carbon generally related and derived from soil organic matter4. The average soil carbon
content of 0-50 cm in HG, TP and CP soil was 13.31, and 7.23 g kg-1, respectively (Fig. 1). Soil carbon

content showed the same trend as OM in soil. The organic carbon content was significantly decreasing with

depth (P<0.05) in the HG (High plant diversity, least soil disturbance) and TP site (moderately plant

diversity), while there was not significantly difference between layers with a lowest carbon content in the CP

site (monoculture). The highest value was in the top soil layer of the HG site. The results correspond to the

report of Lecina-Diaz et al., 2018 that density and structural diversity had a strong positive effect on carbon

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stocks9. Moreover, soil with high tillage frequency reduced biological diversity and carbon content in soil10.
Acosta-Martinez et al. (2004) reported that continuous monoculture systems had a negative impact on soil
function and sustainability11.

Carbon content 15 4.51 2.62 1.26 13.31
(g/kg) 10 8.8 30-50 cm. 7.23
2.52
4.61
5 1.26 0-50 cm.

0
0-30 cm.

Home garden (HG) Teak plantation (TP) Cassava plantation (CP)

Fig. 1. Soil carbon content in different plantation types
As the results home garden with higher plant diversity and less soil disturbance activities
representing a valuable option for storing of carbon and finally mitigating of climate change as climate
neutrality. Furthermore, reducing the risks of the loss of local biodiversity and be a viable option for
sustainable land management in Northeastern Thailand.

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

Mahasarakham University and Mahasarakham University Demonstration School (Secondary). 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. Etheridge DM, Steele LP, Langenfelds RL, Francey RJ, Barnola JM, Morgan VI. Natural and
anthropogenic changes in atmospheric CO2 over the last 1000 years from air in Antarctic ice and
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Title : Reducing methane emissions from rice fields by OE1_14_01
methanotrophic consortium augmentation for the
Field : environmental friendly rice production process
Author :
Environmental Science and Ecology
School :
Advisor : Miss Jarupa Chuanklin

Miss Phawida Duangkhwan

Paphayom Phitthayakhom School, Thaksin University Phatthalung Campus

Assoc. Prof. Sompong O-thong (Thaksin University Phatthalung Campus)

Asst. Prof. Chaisit Niyasom (Thaksin University Phatthalung Campus)

Abstract

The food sector is one of the main drivers of climate change. Rice cultivation accounted for 0.62% (CH4
emissions) and field burning 0.62% of the total GHG emissions in Thailand agriculture. To achieve climate
neutrality of farms and farming systems, this work aims to use methanotrophs (Methane-oxidizing bacteria) to
reduce GHG emissions in flooded rice paddy systems by directly converting methane to methanol. Methanotrophic
bacteria were enriched from rice paddies soil, Phatthalung Province, Thailand, by cultivating on BA medium with
methane in the headspace of serum bottles. The enriched cultures were tested for methane consumption efficiency
and the inoculation size (10,20,30, and 40%) and applied to rice cultivation. The efficacy for methane reduction
of enriched bacteria from cattle farm effluent, swamp field sediments, ice field sediment, sludge from palm oil
factory, peat forest, and cattle farm soil was 97.9%, 95.5%, 92.0%, 79.5%, 75.6%, and 95.2%, respectively. The
enriched culture from cattle farm effluent had the most methane reduction efficiency. The best inoculum size was
20%. When applied to rice paddies, it can control methane emissions.

Keywords: Methanotroph, reducing methane, methane-oxidizing bacteria, flood rice paddy

Introduction

Thailand is mostly a farming country. Rice is the country's principal cash crop. However, Thai rice isn't
widely accepted outside of Thailand because the country's traditional rice production process has high GHG
emissions. Methane and carbon dioxide emissions intensify the greenhouse effect when discharged into the
atmosphere to exacerbate the greenhouse effect and cause global warming. Although methane from farming
accounts for only 10% of total methane emissions in Thailand's agricultural sector, rice fields constitute the largest
source of methane emissions. As a result, the issue of methane emissions in rice cultivation cannot be overlooked.

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The process of degrading organic materials by methanogenic archaea occurs during the growth of rice
plants that are inundated in rice fields. Methane-reducing archaea can be found in rice fields. Methanotrophic
bacteria produce energy by oxidizing methane with oxygen, also found in low numbers in rice fields. If farming
practices are established to limit methane emissions, the greenhouse effect on the atmosphere can be reduced.
Improve Thai rice production practices and address global warming issues

Methodology
The experiments were divided into 4 parts:
Part 1: Separating the Sangyod rice plants for planting
Bring the Sangyod rice plant from Paphayom District, Phatthalung Province, and the soil that has been

sown for three months in the individual bucket. Take measurements of soil & air moisture, pH, temperature, and
NPK in the soil during rice cultivation.

Fig. 1. Sangyod rice in each bucket. Fig. 2. Methanotrophic Bacteria Fig. 3. Absorb methane gas.
Cultivation Cultivation.

Part 2 : Methanotrophic Bacteria Cultivation

Bacteria were enriched from flooded soil samples obtained from 6 sites using a specific culture medium
for methane-degrading bacteria. The culture medium included NMS medium, trace element solution, phosphate
stock solution, vitamin stock. It was used for enriched methanotrophic bacteria. Methanotroph isolation as
indicated above, take the sample from the sampling location and place it in the prepared culture medium. Please
put it in a tube for serial dilution, then into a bottle with rubber and aluminium stoppers to seal it. Fill the bottle
with methane gas after that. In a dark environment, incubate the samples at 37.5 °C for 5-7 days. Examining the
finished product. Use Gas Chromatography to examine the final product. After that, choose the best bottle from
the best source. After that, the volume was raised by cultivating the bacteria in higher volumes of broth culture
medium, adding methane gas to the bottles, and incubating for 5-7 days at 37.5°C in a dark area.

Part 3: Methane consumption performance of enriched methanotrophic bacteria

Use a centrifuge was used to separate the methanotrophic bacteria. The precipitated cell was divided into
four bottles, ten percent, twenty percent, thirty percent, and forty per cent. The volume of each vial was adjusted to

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12th SCiUS Forum

equalize by adding ten per cent methane. Seal it to prevent outside air from entering. Then incubate for three days-
examination of the final product. Use gas chromatography, and the best % flask was chosen to enhance quantity
by cultivating them in higher volumes for adding to the bucket.

Part 4: Evaluation of methane removal efficiency in rice fields.

Add liquid (broth medium and methanotroph bacteria) as follows: The first bucket: do not add enriched
methanotrophic bacteria, the second bucket: add only cultivation liquid, the third bucket: add organic fertilizers,
the fourth bucket: add organic fertilizers and liquid, the fifth bucket: add chemical fertilizers, the sixth bucket:
add chemical fertilizers and liquid. Soil gas collection is used to capture soil methane. It is collected daily.
Then, use gas chromatography.

Results
The efficacy for methane reduction of enriched bacteria from cattle farm effluent, swamp field sediments,

ice field sediment, sludge from palm oil factory, peat forest, and cattle farm soil was 97.9%, 95.5%, 92.0%, 79.5%,
75.6%, and 95.2%, respectively. The enriched culture from cattle farm effluent had the most methane reduction
efficiency. Therefore, the researchers selected bacteria from the cattle farm effluent as the good methane
consumption bacteria for further experiments.

Fig. 4. The chart shows the percentage of methane Fig. 5. The amount of methane used by bacteria
used by the microorganisms in the system.

For the maximum efficiency and cost-effectiveness, The researchers tested the best bacterial
concentration for methane reduction of 20% per volume with the highest methane reduction value. Consequently,
we chose this concentration for use in field experiments. When methanotrophic bacteria were added to all 6 buckets
in Experiment 4.1, gas samples were taken daily for examination on days 1-2, and the methane gas content was

greater due to the methanotrophic bacteria processing. Early methane yield is caused by metabolism for growth,
and the graph steadily rises to its apex.

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