Extended Abstract : Biology and Biodiversity

12th SCiUS Forum

Currently, the diagnosis of H. pylori infection can be detected by a variety of processes, including Urea
Breath Test, Stool Antigen Test, RUT, Polymerase Chain Reaction, SEM and NBI.

However, some patients underwenta gastrointestinal NBI and a primary detection of H. pylori infection
by the RUT was negative while H. pylori infection was detected by scanning electron microscopy resulting in
the patient unable to receive treatment. SEM can solve the problem by visualizing the damage of the gastric
surface and reducing the false negative from the RUT. Therefore, the purpose of this research is to evaluate the
relationship between RUT and SEM in patients with H. pylori related gastritis.

Methodology

1. Preparing biopsy samples to study pathology.

1.1 Collected biopsy from the operating room.
1.2 Fix the biopsy in glutaraldehyde and phosphate buffer overnight at 4 °C.
1.3 Wash the biopsy with a PBS buffer for 20 minutes, 3 times.
1.4 Stain biopsy in Osmium tetroxide for 2 hours and cover with a box to protect biopsy from light.
1.5 Rinse the biopsy with distilled water for 20 minutes, 3 times. After that, wrap it in filter paper.
1.6 Prepare 20%, 40%, 60% and 80% concentrations of acetone.
1.7 Dehydrate the water from the biopsy with acetone in each concentration for 20 minutes.
1.8 Dehydrate the water from the biopsy with 100% concentration Acetone for 20 minutes, 3 times.
1.9 Take the biopsy into the Critical point dryer machine, then add acetone until it covers the sample.
1.10 Set up the Critical point dryer machine, attach the CO2 to the machine and start working.

1.11Coated the dried sample with carbon paint before observing under a SEM.
2. Analysis data by using SPSS.

In this experiment, thirty patients with gastric disease diagnosed with H. pylori infection with the
RUT and NBI at Suranaree University Hospital of Technology were categorized as 5 types. All statistical
analyses were conducted using SPSS version 25. Differences in RUT result, NBI result and SEM result were
assessed using chi-square test.

OB5_18_06/2

394

12th SCiUS Forum

Results
1. Pathology of H. pylori infection

Coccoid form of H. pylori
The figure shows H. pylori in the coccoid form, in which H. pylori usually appears in the spiral
form. However, when conditions are not suitable for the growth of H. pylori, it will adapt by transform into a
coccoid form to survive. In coccoid form, very few reactions occur within cells and when the conditions return
to be suitable for growth again, H. pylori changes its shape back to spiral-shape again, so when patients with
infection are taking the drug, in the beginning, it seemed to disappear. but later found the same infection. It’s
also known as false negative

2. Relationship of experimental results
H. pylori infection was detected in 19 patients (63.33%) using RUT and NBI. In SEM, 26 cases

were detected (86.67%).

Table 1 Correlations between the diagnosis of RUT and the detection of infection using NBI

Table 2 Correlations between the diagnosis of infection using RUT and detection by SEM

OB5_18_06/3

395

12th SCiUS Forum

The results from the chi-square test showed that the RUT results were related to the detection of
infection by using endoscopy with P=0.000 and RUT results were also associated with the infection detected
using scanning electron microscopy, with P=0.005.

Discussion
The results of RUT diagnosis were significantly related to NBI and SEM, and the detection of

pathogens. H. pylori coccoid morphology is associated with a false negative in RUT diagnosis. SEM can be
used to identify H. pylori infection that may not be detected by RUT and high-resolution endoscopic images.

Conclusion
1. RUT results were associated with NBI and SEM.
2. Present of coccoid forms were associated with negative RUT.
3. SEM is an alternative method for diagnosing H. pylori infection.

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. Zhao J, Xu S, Gao Y, Lei Y, Zou B, Zhou M, et al. Accuracy of Endoscopic Diagnosis of Helicobacter
pylori Based on the Kyoto Classification of Gastritis. Frontiers inOncology 2020;10: 599218.
2. Calvet X, Delgado J, Montserrat A, Lario S, Lazaro M, Quesada M, et al. Accuracy of Diagnostic Tests for
Helicobacter pylori: A Reappraisal. Clinical Infectious Diseases 2009;10: 1385–1391.
3. Alzoubi H, The Use of 13C-Urea Breath Test for Non-Invasive Diagnosis of Helicobacter pylori Infection
in Comparison to Endoscopy and Stool Antigen Test. Diagnostics 2020;10: 448.

4. ทวศี ักด์ิ ทองทว.ี ผลของ Probiotics ในการรักษากระเพาะอาหารอักเสบจากการตดิ เชือ้ H. pylori. มหาวทิ ยาลัยเทคโนโลยีสุรนาร;ี 2558.
5. ทนพญ.วราภรณU ฟกW โพธ.์ิ การทดสอบเพือ่ การวินิจฉัยการตดิ เชอื้ Helicobacter pylori จากเลือด. มหาวทิ ยาลัยธรรมศาสตร;U 2561.

OB5_18_06/4

396

12th SCiUS Forum

Title : Development of edible coating combined with bitter gourd

Field : extract on fruit peel
Biology

:Author Mr. Phummiphat Promnoi

:School Naresuan University Secondary Demonstration School

:Advisor Assistant Profressor Dr. Maliwan Nakkuntod

Abstract
Anthracnose is a plant disease that causing damage to produce both in quantity and quality. The

important cause of anthracnose is in the genus Colletotrichum causing losses to fungal crops. This disease is
found all over the world, especially in the humid tropics. There will be severe outbreaks. The authors therefore
studied the amount of concentration ratio between bitter gourd and chitosan coating that is most effective in
slowing the spread. Rotten fruit and slow down the development of fungal diseases. Colletotrichum spp. that
causes anthracnose in plants and to develop an edible coating. Mixing bitter gourd on fruit skin can slow down
anthracnose incidence from the results of development experiments. Bitter gourd edible coating: 5 kg of bitter
gourd can be used to make the coating, 6.74 g, with a percentage yield of 0.1348%, which can be used to make
four coatings and add more batches. The other four trials were the control, bitter gourd coating, Chitosan
coating 0.5%, Chitosan coating 1%, Chitosan coating 2%. Chitosan coating agent 0.5% with bitter melon,
Chitosan coating 1% with bitter melon. and 2% chitosan coating with bitter gourd Applying the coating for 8
days, the percentage change of the mean fresh weight of all 3 results per 1 experimental set. The size of the
lesions was assessed by dividing scores of 1-5 points. The absence of lesions in most trials was 3 - 4 points,
but for the bitter gourd and antibacterial coatings. chitosan coating 2% will have a score of 2-3 points and the
number of withered fruit. The number of bipolar lesions in the control unit and 2% chitosan coating were found
to be high. 1% bitter gourd mixed with the best performance in coating of Nam Dok Mai mango was based on
the percentage change of mean fresh weight, lesion size, wilt number and number of fruits with terminal rot.

:Keywords Chitosan, Bitter gourd

Introduction
Many kinds of fruits, when fully ripe, will have silt on the surface of the fruit as a waxy coating to

prevent loss of water, and after harvesting, the “soft” of the fruit will gradually disappear, causing the fruit.
This causes water loss and begins to wilt faster. Food coating is one of the popular methods of prolonging life.

1

397

12th SCiUS Forum

Bitter gourd is a herb that has been used for thousands of years. Currently, there have been research
and experimental properties of bitter melon, which has been found to have many benefits and importantly,
bitter gourd can enhance the benefits of both medicinal properties. It improves the benefits of all aspects of
fruit, prolong life and prevent fungal infection.

Anthracnose is a plant disease that causes damage to both quantity and quality of crops. The most
important fungi that cause anthracnose are in the genus. Colletotrichum causing losses to economic crops

Therefore, the project organizer would like to develop an edible coating mixed with bitter gourd
on the surface of the fruit. It can inhibit the expression of anthracnose so that it can be applied to various fruits
to help reduce water loss, can slow down the ripening of the fruit and reduce the expression of anthracnose as
well.

Methodology
The experiments were divied into 2 parts as follows,

Part 1: Preparation of bitter gourd extract
1. Take 5 kg of bitter gourd, cut it and remove the pulp and seeds inside. Then cut into small pieces.
2. Put the bitter gourd in a tray, spread evenly and not too thick, put in a hot air dryer at a
temperature of 50+-2 °C.
3. Then take the bitter gourd after baking. Go to a blender until it is like powder.
4. Bring the powdered bitter gourd to be divided into beakers 25 grams each.
5. Put the ethanol into the beakers that put the powdered bitter gourd as well. Ratio 1: 4 (25 grams
of bitter gourd per 100 ml of ethanol)
6. Mix the bitter gourd with ethanol together and cover the lid with aluminum sheet tightly and
leave to ferment for at least 5-7 days.
7. Take the strand between bitter gourd and ethanol to filter out the sediment.
8. Take the solution after filtering and put it in a vacuum at 45 °C to evacuate the ethanol from the
solution. The remaining viscous liquid in the beaker
9. Take the remaining substance to freeze dry to get 6.74 grams of bitter gourd extract.

Part 2: Preparation of edible coating
The experiment was divided into 8 sets. It is divided as follows: Control unit, bitter gourd coating,

0.5% chitosan coating, 1% chitosan coating, 2% chitosan coating, 0.5% chitosan coating with bitter gourd, 1%
chitosan coating with bitter gourd and 2% chitosan coating with bitter gourd. The Nam Dok Mai mango applied
for 8 days, the percentage change of mean fresh weight of all 3 outcomes per experimental set was obtained.
The size of the lesion was assessed by dividing scores from 1-5 points. Then storage mangos in incubator at
25°C.

Results, Discussion and Conclusion
5 kg of bitter gourd was used as a coating mixture for 6.74 g, accounting for 0.1348% of the yield, and

was coated for 8 days to obtain the percentage change of the average fresh weight of all 3 results per 1 experimental
set as follows: 1.0975, 1.1625, 0.98125, 0.98375, 1.335, 0.99875, 0.90375, and 1.14375, respectively. The size of
the lesion was assessed by 1-5 score division, with the absence of lesions in most trials having a score of 3 - 4. Scored

2

398

12th SCiUS Forum

Bar graph: Percent fresh weight lost
only in the part of the bitter gourd and chitosan coating. 2% will have a
score of 2 - 3 points and the number of withered fruit The number of
outcomes for terminal gangrene was high at the control and 2% chitosan
coating.

From the experiments, it was found that the most
effective coating for the Nam Dok Mai mango was chitosan coating.
1% was mixed with bitter gourd due to the percentage change in mean
fresh weight of the chitosan coating. 1% mixed with bitter gourd was the least. The lesion size was 2-3 points
and the number of wilts 1 result and the number of terminal rot disease 2 results showed that the chitosan coating
1% bitter gourd mixture is effective in preventing water loss and slowing the incidence of anthracnose, as well
as preventing wilt and terminal rot.

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. Nanthawan Hadthamard. Hot water dipping combined with chitosan coating to maintain quality of bird

chili [disssertation]. [place unknown]: Rajamangala University of Technology Suvarnabhumi; 2020

3

399

12th SCiUS Forum
ORGANIZED BY

คณะวทิ ยาศาสตร มหาวทิ ยาลัยทกั ษิณ
Faculty of Science, Thaksin University