EAFTA 2023, On-site/online, May 17th-20th, 2023, Morioka, Japan

Prospective Plasma Contribution to Fourth Agricultural Revolution
Masaharu Shiratani1,21, Center of Plasma Nano-Interface Engineering, Kyushu University, Japan
2, Institute of Advanced Study
The third agricultural revolution or the Green Revolution is the period when agrichemicals
had been intensively employed and genetically modified crops began to be used, leading togreater output of the products. In spite of the success, the Green Revolution has left our lands
over-cultivated and over-used, making them unfit for use in the future and negatively impacting
the ecological balance. Industrial nitrogen fixation, for instance, has increased exponentiallysince the 1940s, and human activity has doubled the amount of global nitrogen fixation [1]. In
agricultural systems, fertilizers are used extensively to increase plant production, but unused
nitrogen, usually in the form of nitrate, can leach out of the soil, enter streams and rivers, andultimately make its way into our drinking water [2]. Such large changes in the availability ofnitrogen can lead to severe alterations of the nitrogen cycle in both aquatic and terrestrial
ecosystems. Hence, we need the fourth agricultural revolution, which supports the use oftechnology in order to realize sustainable farming. Plasma technology can contribute to solve
such issues through the novel and effective ways of agricultural productivity enhancement in an
ecofriendly manner [3-6].
[1] P. M. Vitousek, et al.. Ecological Applications, 7 (1997) 737.
[2] A. Bernhard, Nature Education Knowledge, 3 (2010) 25.
[3] I. Adamovich, et al., J. Phys. D, 55 (2022) 373001.
[4] T. Okumura, et al., Sci. Rep., 12 (2022) 12525.
[5] D. Suriyasak, et al., ACS Agr. Sci. Technol., 1 (2021) 5.
[6] P. Attri, et al., Processes, 8 (2020) 1002.

EAFTA 2023, On-site/online, May 17th-20th, 2023, Morioka, Japan

Everything You Need to Know to Make Sushi and Finding Fascinating
Japanese Foods Products
All-Japan Sushi Association, Director,
Masayoshi Kazato
sakaezushi8126@gmail.com

Today, with the world's population increasing and people becoming more health-conscious, there is a
growing demand for seafood in countries around the globe. Raw seafood dishes, such as sashimi and sushi,
from Japan's food culture are quickly gaining popularity in Europe, the US, and China, among other places.
Japan's seafood quality is highly praised globally due to the careful handling of the catch and advanced coldchain technology that ensures the fresh supply of safe and trustworthy raw seafood. As of 2013, more than
55,000 Japanese restaurants were open worldwide, and this number continues to grow year by year. However,in many countries where the food culture of eating raw fish has not taken root, sushi is often adapted to suitthe local market. Unfortunately, accidents related to raw fish occur frequently, and restaurants that prioritize
food safety still feel uneasy about this situation. The basic principle of sushi preparation is hygiene. To
alleviate such concerns and ensure safe consumption of raw seafood, methods for making sushi rice to prevent
bacteria growth caused by raw fish, techniques for preparing and processing fish (including marinading, salttreating, vinegar-treating, blanching, searing, and washing), and fish preservation methods (such as
refrigeration, freezing, thawing, and maintaining freshness) will be introduced.

Keywords: Sushi, Sashimi, Nigiri, gygiene, food safety

EAFTA 2023, On-site/online, May 17th-20th, 2023, Morioka, Japan

To boost rice productivity under a changing climate
Hiroyuki Shimono1,2,
1, Faculty of Agriculture, Iwate University
2, Agri-Innovation Center, Iwate University

Global food demand has been rising to support future population growth under changing
climates. My research interest is to increase crop productivity (per unit land area) in response toenvironmental changes, genotype by environmental interaction, using methods of field
phenotyping, mathematical simulation model, QTL and genome-wide association studies(GWAS) for mainly rice (Oryza sativa L.), a staple food for the more than half of the world's
population. I want to share three topics of my current works, (1) Innovative technology: Early–
winter direct–sowing cultivation of paddy rice, (2) Breeding new rice cultivars adapted to futureelevated atmospheric CO2, (3) Big data analysis for high yielding rice breeding.
Sustainable food production and supply is important issue in the direction of SDGs. I will
introduce activities of Agri-Innovation Center, Iwate University, which was reinitiated from 2022.
Keywords: Atmospheric CO2, Big data, Direct-seeding, Dormancy, Productivity, Rice, SDGs

EAFTA 2023, On-site/online, May 17th-20th, 2023, Morioka, Japan

Thermal Gelation Mechanisms of Fish Meat Paste - How Does Myosin
Molecule Participate?
Shugo Watabe
School of Marine Bioscience, Kitasato University, 1-15-1 Kitasato, Minami, Sagamihara,
Kanagawa 252-0373, Japan*Correspondence: swatabe@kitasato-u.ac.jp

We analyzed changes in the rheological properties of white croaker (Pennahia
argentata) meat paste made from surimi added with 0.5 M NaCl during the following
two-step heating procedure: first preheating at 0-70 °C with 5 or 10 °C intervals andsubsequent secondary main heating at 85 °C. Changes in the breaking strength werehardly observed for both preheating and main-heating gels in the preheating temperature
range of 0–30 °C. In the range of 35-45 °C, however, a marked increase was observed
for the two gels with the increase of preheating temperature where the protein solubility
in the SDS-urea solution and myosin heavy chain monomer content therein were rapidly
decreased in a preheating temperature-dependent manner. Changes in viscoelastic
properties such as storage modulus (G’), loss modulus (G”) and tangent delta (G’/G”)
during preheating in the range of 35-45 °C and subsequent main heating were also
markedly different from those during preheating in other temperature ranges andsubsequent main heating. Interestingly, white croaker meat paste also showed prominentchanges of viscoelastic properties at 35-45 °C in the temperature sweep analysis. We
previously reported that the myosin molecule of white croaker is unfolded mainly at 34.8
and 44.1 °C due to the disruption of alpha helix and/or coiled-coil structure1. The rod
part of the myosin molecule has a unique coiled-coil structure with heptad repeats ofamino acid residues where hydrophobic residues are buried inside the coiled-coilstructure of two alpha-helices. Thus, it is likely that the unfolding of the myosin
molecule at 35-45 °C, especially in the rod part, is important to rearrange myosin
molecules in order to provide elastic thermal gels.
Reference:
1. Hideto Fukushima, Yoshie Satoh, Misako Nakaya, Shoichiro Ishizaki, and Shugo
Watabe: Thermal effects on fast skeletal myosins from walleye pollack, whitecroaker and rabbit in relation to gel formation. J. Food Sci., 68(5), 1573-1577 (2003).

Keywords: fish, myosin, surimi, thermal gelation, thermal unfolding

EAFTA 2023, On-site/online, May 17th-20th, 2023, Morioka, JapanThe Current State and Prospect of Recirculating Aquaculture Systems in
China○Ying Liu1*
1, College of Biosystems Engineering and Food Science, Zhejiang University, 866
Yuhangtang Rd, Hangzhou 310058, P.R. China*Corresponding author: liuyingzju@zju.edu.cn

Recirculating Aquaculture System (RAS) in China has developed
rapidly in the past decade, and has also made great contributions to theprovision of high-quality animal protein. This report covers the current
status of RAS in China, developing opportunities of RAS, main production
modes of RAS in China, technical level analyses of Chinese RAS, andperspective. It is hoped that this report will explore ways to upgrade theChina’s facility aquaculture engineering equipment to promote sustainable
development of aquaculture, and provide a responsible development
approach to world food security and economic growth.
Keywords: Recirculating Aquaculture System, Aquaculture engineering,
Current state, Prospect

EAFTA 2023, On-site/online, May 17th-20th, 2023, Morioka, Japan

Studies on Measuring of Fruit and Vegetable Losses Throughout theSupply Chain and Awareness-Raising Campaign “Save Your Food” inTürkiyeRuhşan Özdemir Çifçi
Faculty of Agriculture, Ege University, 35100 İzmir, Türkiye
Ministry of Agriculture and Forestry Konak District Directorate, Umurbey Mah. İşçiler Cad.
No:139 Konak, Izmir, Türkiye
ruhsanozdemir@gmail.com,ruhsan.ozdemircifci@tarimorman.gov.tr

A study conducted between 2015-2017 on Measuring of Fruit and Vegetable Losses
Throughout the Supply Chain in İzmir / Türkiye. The aim of the study was to determine
the causes and extent of pre-harvest, harvest and post-harvest losses in agricultural
production and to propose solutions that could be effective in reducing losses. Other
purposes of the research are listed below.
- Identify the causes of food losses in the fruit and vegetable supply chain in the Worldand Türkiye,- Determine the extent of food losses in the fruit and vegetable supply chain in the Worldand Türkiye.
After the studies for measuring food losses, the national “Save Your Food Campaign” isconstructed. “Save Your Food Campaign” aims to create awareness and improveunderstanding of the causes and impacts of food losses and waste and to drive behavior
change in individuals and communities in Türkiye. With this campaign, the Ministry ofAgriculture and Forestry and FAO aims to combat food loss and waste both at the national
and international levels, raise awareness of the issue, and make Türkiye a role model for
efforts in this regard in the international arena.

Keywords: Food losses, food waste, fruit and vegetables, supply chain, save your food,Türkiye

EAFTA 2023, On-site/online, May 17th-20th, 2023, Morioka, Japan

Plant polyphenols as natural compounds to modulate muscle food quality
attributes
Youling L. Xiong
University of Kentucky, USA
ylxiong@uky.edu
Polyphenol-rich herbs and spices are common food ingredients added in fish and meat
processing to modify product flavor and appearance. When extracted plant phenolic
compounds are mixed into minced muscle or proteins, their roles are beyond flavor andscavenging free radicals; they are found to modify product texture through theinteraction with myosin or actomyosin. The ability of polyphenols to modulate gelationand emulsification of muscle protein, especially under oxidative conditions, explains
their dose-dependent role in affecting the textural attributes of processed fish, poultry,
and red meat products. In-depth studies have shown that phenolic compounds modify
muscle proteins in a structure-dependent manner. For example, the variation in hydroxyl
group distribution in flavonoid B-ring between mono-, di-, and tri-phenol compounds is
an important factor affecting phenolic efficacy. The texture-modifying activity of plant
phenolic compounds in muscle foods underscores their multi-functionality nature in fishand meat products.

EAFTA 2023, On-site/online, May 17th-20th, 2023, Morioka, Japan

Effective Utilization of Oyster Processing Byproduct○Min-Jie Cao, Ling Weng, Yan Wang, Meng-Ya Ji, Qian Zhang
College of Ocean Food and Biological Engineering, Jimei University, Xiamen, 361021,
China)

China is the major oyster production country in the world, with a production of 5.82
million tons in 2021. Shell is the major byproduct during oyster processing which
occupies nearly 70% of the whole weight. Discarded oyster shell caused serious
environmental problems and thus effective utilization of oyster shell is essential.
On the other hand, because of excessive application of chemical fertilizers and acid rain,soil acidification in China is quite serious, especially in southern east provinces.
Acidified soil will cause low production and low quality of the agricultural products and
impede sustainable development. Calcinated oyster shell mainly contains CaO, CaCO3
as well as other metal ions and can be utilized as soil conditioner to neutralize acidified
soil. We have applied this soil conditioner to more than 60 crops in 20 different counties
in Fujian province. The results showed that application of oyster shell soil conditioner
could significantly neutralize soil acidification, increase the content of soil organic
matter and exchangeable calcium, and effectively improve the yield and quality of thecrops while no side effect was observed.

EAFTA 2023, On-site/online, May 17th-20th, 2023, Morioka, Japan

Overview of Vietnam's aquatic sector during the period 2015-2022
and vision to 2045
Hanh Tran Thi My* and Duy Nguyen Xuan
Faculty of Food Technology, Nha Trang University, Khanh Hoa Province, Vietnam*Corresponding author: myhanhtt@ntu.edu.vn
Vietnam is a country in Southeast Asia with a land area of 329,560 km2 and a coastline of
3,260 km, an exclusive economic zone of 1 million km2. In 2022, total fishery output was 9
million tonnes including 3.86 million tons from exploitation and 5.19 million tons fromaquaculture. The export value of the fisheries sector in 2022 was $11 billion. The workforce
is more than four million people. Fisheries are defined as the country's leading economic
sector, comprising about 4-5% of GDP, 9-10% of the country's total exports, ranking fifth inexport value. In 2015 Vietnam's fisheries output increased 38% from 6.56 (2015) to 9.05
million tonnes (2022). Of these, aquaculture accounted for 57% and exploitation was 43%.
Fisheries exports during 1998-2022 increased 13 times from $817 million (1998) to $11
billion (2022). The fisheries development objectives of the nation to 2030 and vision of 2045
are defined as follows: a 3.4-4% annual growth rate, total export output of 9.8 million tons,
export value $14-16 billion and employment of more than 3.5 million workers.
Keywords: Aquatic sector, export, Vietnam, vision

EAFTA 2023, On-site/online, May 17th-20th, 2023, Morioka, Japan

Rethinking on food system for sustainable world○Noriyuki Tanaka1,2
1, Food Society of Modern International Lifestyle Education(FSMILE)
2, Faculty of Agriculture, Iwate University, Ueda 3-18-8, Morioka, Iwate 020-8550, Japan*Corresponding author: norit22999@hotmail.com

This presentation provides you a part of research outcome from coordinated study on
bio-production (food production), ecosystems and humanity under the Integrated
Research systems of Sustainability Science (IR3S), in which the presenter deeply
involved. In my presentation,
Our environmental impact through food production, process, transportation, andconsumption, is no longer trivial on earth. I try to convince you that food system scienceand education may play an important role for social and economical transformation of
our society toward sustainability of the earth.

Keywords: Sustainability, Environment, Food System Science , Education

EAFTA 2023, On-site/online, May 17th-20th, 2023, Morioka, Japan

Food literacy behavior change toward post-pandemic society○Kun Qian1
1, Institute for Asian and Oceanian Studies, Kyushu University, Motooka 744, Nishi-ku,
Fukuoka 819-0395, Japan*Corresponding author: qk@kyudai.jp
The 2019 coronavirus (COVID-19) pandemic caused various changes in people’s
behaviors, particularly those related to eating and diet. Changes in eating behaviors
caused by the pandemic have been investigated in diverse cultural and social contexts. In
this presentation, the author will introduce several studies mainly conducted by his
research team, focusing on the general food literacy behavior, including food waste, foodconsumption, cooking ability and preference. These studies were based on a series oflongitudinal questionnaire surveys started from April, 2020. More than 30,000 samples
were collected in these online surveys, from nationwide respondents in Japan. Based on
the statistical analyses on and investigations into the dataset, we proposed a structuralmodel of eating literacy behavior change during the pandemic. The model defined in our
research provides informative suggestions for household food management during the
COVID-19 pandemic, and similar situations in the future.
Keywords: SARS-CoV-2, cooking behavior, food waste, food consumption, stress, selfefficacy, communal eating

EAFTA 2023, On-site/online, May 17th-20th, 2023, Morioka, Japan

Functionality of natural pigments in food products○Hayato Maeda *1,2
1, Faculty of Agriculture and Life Science, Hirosaki University, 3 Bunkyo-cho, Hirosaki,
Aomori 036-8561, Japan
2, United Graduate School of Agricultural Sciences-Iwate University, 3-18 Ueda, Morioka
city, Iwate 020-8550, Japan*Corresponding author: hayatosp@hirosaki-u.ac.jp

Foods contain various natural pigment components such as anthocyanins andcarotenoids. These pigment components show various physiological functions such as
anti-oxidation, anti-cancer, and anti-obesity activity. In this presentation, I introduce thefunctionality about new apple cultivars rich in anthocyanins and paprika rich incarotenoids.
Kurenai no Yume, literally “crimson dream”, is a red flesh apple cultivar developed
by Hirosaki University. This apple is sweet, mildly tart, rich in anthocyanin, anddelicious fresh or cooked. Its natural red color remains distinct even after cooking,making it excellent for the creation of richly colored apple products. It showed high antioxidant activity. Further, this apple extracts showed inhibitory effect of α-glucosidase
and ameliorated blood glucose up-regulation in animal experiment1).
Paprika (Capsicum annuum) contain various carotenoids such as capsanthin, capsorubin,
cryptocapsin cucurbitaxanthin A, β-cryptoxanthin, capsanthin epoxide, zeaxanthin, andβ-carotene. Especially, capsanthin and capsorubin are characteristic carotenoid in
paprika. They show strong anti-oxidative and anti-cancer promoting effects. Furthermore,these carotenoids show preventive effects of obesity related diseases2,3). Paprika
carotenoids promoted adiponectin secretion by promoting differentiation of small size
adipocyte cells. Moreover, Paprika carotenoids ameliorated chronic inflammation in
obesity condition adipocyte cells.

Keywords: anthocyanin, carotenoid, apple, paprika, anti-obesity, diabetes, adipocyte
1) Matsumoto K et al. ISHS Acta Horticulturae, 1127:29-33(2016). 2) Maeda H et al.
ISRN Inflammation, 2013:763758(2013). 3) Maeda H et al. Biosynthetic and Bio
functional Approaches Chapter 26, Springer Singapore, 285-293(2021)

Biological functions of phenethyl isothiocyanate
for glucose and lipid metabolisms
Yoshiaki Ito1,2, Moe Nagami1, Maiko Chiba1 and Takashi Nagasawa1
1

Biological Chemistry and Food Science, Faculty of Agriculture, Iwate University, Morioka, Iwate, Japan
2

Agri-Innovation Center, Iwate University, Morioka, Iwate, Japan

Keywords: phenethyl isothiocyanate, adipocyte, skeletal muscle, insulin resistance, Akt
Phenethyl isothiocyanate (PEITC), a natural dietary isothiocyanate, has been shown to havebeneficial effects in improving cellular defense activities against oxidative stress throughactivation of nuclear factor erythroid-2 related factor 2 (Nrf2) pathway. However, little evidence
exists if the antioxidative activity has beneficial effects on glucose metabolism. Here, we
examined the preventive potential of PEITC for impaired insulin induced glucose uptake byoxidative stress in 3T3-L1 adipocytes. Treatment with PEITC increased the expression ofantioxidative enzymes regulated by Nrf2 such as γ-glutamylcysteine-synthetase, heme oxygenase
1, NAD(P)H:quinone oxidoreductase 1 and glutathione S-transferase, and reduced oxidative
stress induced by H2O2. Furthermore, PEITC restored impaired insulin-stimulated glucose uptake,translocation of glucose transporter 4 (Glut4) and insulin signaling by H2O2. These results
indicate that PEITC protected insulin-regulated glucose metabolism impaired by oxidative stressthrough the antioxidative activity in 3T3-L1 adipocytes. Next, we tested whether PEITC directly
promotes glucose utilization in mouse skeletal muscle cells, C2C12 myotubes. PEITC induced
glucose uptake, Glut4 translocation to the plasma membrane, and activation of Akt and ERK in
C2C12 cells. Inhibition of Akt suppressed PEITC induced Glut4 translocation and glucose uptake,whereas ERK inhibition did not. These results indicate that PEITC promotes glucose utilization
through Akt pathway in C2C12 myotubes. Taken together, PEITC may serve as a dietary
constituent with beneficial effects on the carbohydrate metabolism.

EAFTA 2023, On-site/online, May 17th-20th, 2023, Morioka, Japan

Anthocyanins food innovation: from laboratory bench to healthy dining table
De-Xing HOU 1,2
1, Department of Food Science and Biotechnology, Faculty of Agriculture,
2, The United Graduate School of Agricultural Sciences, Kagoshima University, Japan*Corresponding author: hou@agri.kagoshima-u.ac.jp
Abstract
Anthocyanins are naturally occurring polyphenolic compounds that give the intense color to
many fruits and vegetables such as berries, red grapes, red rice, purple sweetpotato and red
cabbages. There are of great nutritional interest because their intakes have been estimated to
be up to 9-fold higher than that of other dietary flavonoids. Thus, we have been working on
chemical properties, bioactive functions and molecular mechanisms of anthocyanins-richfoods for almost two decades. In this talk, I will summarize some key results from ourlaboratory studies and our practices to innovate anthocyanin-rich foods for healthy dining
table. First, we found that a clear chemical-bioactivity relationship is presented in kinds ofanthocyanins. Thus, we clarified the quantity and quality of anthocyanins from a variety offruits and vegetables, and provide chemical-bioactivity information to plant breedingscientists to breed fruit or vegetable that contains higher functional anthocyanins. Second, ouranimal experimental data demonstrated that anthocyanin-rich foods could ameliorate high fat
diet-induced obesity, inflammation and dysbiosis of gut microbiome. Molecular analysis
further revealed that active anthocyanins could upregulate Nrf2-mediated antioxidantsignaling pathways and downregulated NFb-mediated inflammatory pathways. These data
provide the evidence and underlying mechanisms for the health-promoting properties ofanthocyanin-rich foods. Third, we also observed that anthocyanins are easily oxidized andbecome pro-oxidants. The factors including light, pH and metals affect anthocyanin color andfunctions. Thus, we have notified these factors to keep anthocyanin color and functions infood processing. Our final goal is to make health-promoting properties of anthocyanin-richfoods into our dining table, thus, we are innovating the healthy menu from anthocyanin-richfoods to dining table, defined “KUROZEN”, basing on the scientific knowledges on chemical
properties and functions of anthocyanins-rich foods, and nutrition-balance with other nutrients.
Keywords: Anthocyanin; Functions, Mechanisms; Processing, Healthy dining table
References
Hou et al., International Journal of Oncology,23,705 (2003). Hou et al., Current Molecular
Medicine 3,149 (2003). Hou et al., Carcinogenesis, 25,29 (2004). Hou et al., Archive ofBiochemistry and Biophysics,418,99 (2005).
Hou et al., Biochemical Pharmacology, 70,417(2005). Chen et al., Nutrition and Cancer, 60,43 (2008). Hou et al., Antioxidant andRedox Signaling, 13, 691 (2010). Sogo et al., Planta Medica, 81,26 (2015). Sogo et al.,Biofactors, 41,58 (2015). Wu et al., Journal of Functional Foods, 17: 514 (2015). Wu etal., Molecular Nutrition and Food Research, 61, 887 (2017).
Wu et al., Journal of
Agricultural and Food Chemistry, 65,5133 (2017).
Wu et al., Molecules. 23,12 (2018).
Nakano et al., Nutrients, 12, 3252(2020).

EAFTA 2023, On-site/online, May 17th-20th, 2023, Morioka, Japan

Polysaccharides from brown algae: relationship between structure andfunction○Yuya Kumagai*1, Hiroki Kojima1, Junpei BaBa1, Hideki Kishimura1
1, Faculty of Fisheries Science, Hokkaido University, Minatocho 3-1-1, Hakodate, Hokkaido
041-8611, Japan*Corresponding author: yuyakumagai@fish.hokudai.ac.jp
Kombu is an important ingredient used in the dashi soup of Washoku. Although theumami component is attracting attention as a feature of Kombu, it also contains fucoidan
as a functional polysaccharide. Fucoidan is a sulfated polysaccharide with L-fucose as
the main chain, and the structure differs sugar compositions, glycosidic bonds, andsulfate group content depending on the seaweed species and collection date. Fucoidan
has been reported to have functions such as anti-inflammatory and antioxidant activity.
Although the relationship between fucoidan structure and function has been pointed out,
the detailed relationship remains unclear. In this study, we investigated the structuralcharacteristics of Japanese Kombu (Saccharina japonica) depending on the collection
date. Fucoidan was obtained from Kombu powder by hot water extraction. The
supernatant was added calcium chloride to remove alginic acid, and the supernatant waslyophilized to obtain crude fucoidan after dialysis. Crude fucoidan was separated byanion-exchange column chromatography. Fucoidan structure was confirmed by FT-IR.
The amount of crude fucoidan in Kombu from March to June was stable. Crude fucoidan
was separated into three peaks by chromatography (P1, P2, and P3). Among them, thelatest peak (P3) had a large number of sulfate groups compared to P2 and P1. We now
plan to clarify the relationship between sulfate groups and functionality.
Keywords: Kombu, fucoidan, FT-IR, Polysaccharides

EAFTA 2023, On-site/online, May 17th-20th, 2023, Morioka, Japan

Make Your Idea a Cuisine: How Innovation is Made on Foods○Weng Ling
College of Ocean Food and Biological Engineering, Jimei University, 43 Yindou Road,
Xiamen 361021, China
Email: weng1201@jmu.edu.cn

Foods are the most essential to our livelihood and shall not be defined to what satisfy
taste buds, and quell hunger. Making foods is a nouveau life style and a fountain inspiring
talents. How shall we make ordinary ingredients ascend into a masterpiece appealing to themass, as well as being innovative to outcompete others in major events? This lecture aims toelaborate the winning design on their original idea, concept and points of innovation based onrecent innovative food design competitions for university students held by Chinese Institute ofFood Science and Technology (CIFST). Opening discussions on the topic of designinginnovative foods are welcomed.

Keywords: Foods, Innovation, Food design competition, Chinese Institute of Food Scienceand Technology

EAFTA 2023, On-site/online, May 17th-20th, 2023, Morioka, Japan

Oxidation and reduction mechanisms of fish myoglobins
Yoshihiro Ochiai
Faculty of Agriculture, Tohoku University, Aramaki, Sendai, Miyagi 980-8572, Japan
E-mail: yochiai@tohoku.ac.jp

Myoglobin (Mb) is an oxygen-binding heme protein involved in intracellular oxygenstorage and oxygen transport from the plasma membrane to the mitochondria. Mb forms
various derivatives depending on its redox state. Especially, desirable bright red meat color isdue to oxygenated form (oxyMb), while accumulation of oxidized form (metMb) results inbrowning or discoloration of meat. Due to the high contents of Mb in tuna meat as well as
beef, undesirable discoloration proceeds very fast during postmortem storage. In addition,
mitochondria involved in chemical energy production in live muscle could affect the redox
state of Mb. In this presentation, molecular dynamics simulation of Mb oxidation mechanism
will be discussed. The role of mitochondria in the reduction of metMb will also be referred to.
The simulation revealed the interaction between oxyMb and oxygen molecules. The
number of water molecules in the heme pocket decreased under lower partial oxygen pressure,while higher oxygen pressure tended to reduce metMb formation. On the other hand, themitochondrial fraction from tuna dark muscle reduced metMb. The involved enzyme issupposed to be cytochrome b5 reductase located in the outer membrane of mitochondria. No
significant change in the reduction activity was observed in the acidic pH range (5.7-6.8).
NADH favored metMb reduction, while exhaustion of NADH resulted in metMb formation,suggesting that maintenance of NADH level is important for keeping the fresh meat color
during chilled storage.

Keywords: myoglobin, oxidation, reduction, discoloration

EAFTA 2023, On-site/online, May 17th-20th, 2023, Morioka, Japan

Application of plant extracts as natural additives for the preservation offishery products○A. K. M. Azad Shah*

Department of Fisheries Technology, Faculty of Fisheries, Bangabandhu Sheikh Mujibur
Rahman Agricultural University, Gazipur 1706, Bangladesh*
E-mail: azad@bsmrau.edu.bd

Fish and fishery products have become increasingly popular due to their high nutritional
value owing to the presence of high-quality protein, n-3 polyunsaturated fatty acids,
vitamins, and minerals. However, fish are recognized as being highly perishable, having
a relatively short shelf life and being highly susceptible to enzymatic, microbiological,
and chemical deterioration. To prevent these chemical deteriorations and delay the
microbial growth in fish, various preservation methods are used. Among them, lowering
the temperature of fish and using of synthetic preservatives are renowned; however, they
do not completely inhibit the chemical deterioration and microbial growth in fish.
Moreover, excessive use of synthetic preservatives has proven to be carcinogenic andpotential health risks for humans. Therefore, there is an increasing interest in natural
additives with potent antioxidant and antimicrobial properties to replace the synthetic
preservatives in seafood preservation. Plant extracts with rich phenolic compounds fromseveral plant sources, such as seaweed, guava leaves, water lily, and stevia leaves havedemonstrated notable in-vitro antioxidant and antibacterial activity. Due to the presenceof phenolic compounds, plant extracts retard bacterial growth as well as inhibit theformation of ammonia and other primary and secondary lipid oxidation products, which
extends the shelf life, retains the quality, texture, and flavor of the fish fillets. Therefore,
these plant extracts can be used as natural additives in the seafood industry for thepreservation of fish and fishery products.

Keywords: Plant extract, Synthetic preservatives, Phenolic compounds, Antioxidant andantibacterial activity, Quality, Shelf life.

EAFTA 2023, On-site/online, May 17th-20th, 2023, Morioka, JapanThe study on marine natural product a new isoindolone FGFCI for
fibrinolysis○Wenhui Wu1, Chunli Gao1, Haixing Zhang1, Bin Bao*1, 2, 3
1, Collage of Food Science and Technology, Shanghai Ocean University, Huchenghuan Road
999, Pudong New Area, Shanghai 201306, China
2, Marine Biomedical Science and Technology Innovation Platform of Lin-gang Special Area,
Haiji Sixth Road 218-4, Shanghai 201306, China
3, Putuo Branch of International Combined Research Canter for Marine Biological Sciences,
Zhongke Road 2, Putuo District, Zhoushan City 316104, Zhejiang Province, China*Corresponding author: bbao@shou.edu.cn

Fungi fibrinolytic compound 1 (FGFC1) is a rare pyran-isoindolone derivative withfibrinolytic activity. The aim of this study was to further determine the effect of FGFC1
on fibrin clots lysis in vitro. We constructed a fibrinolytic system containing single-chain
urokinase-type plasminogen activator (scu-PA) and plasminogen to measure thefibrinolytic activity of FGFC1 using the chromogenic substrate method. After FITCfibrin was incubated with increasing concentrations of FGFC1, the changes in the
fluorescence intensity and D-dimer in the lysate were measured using a fluorescencemicroplate reader. The fibrin clot structure induced by FGFC1 was observed andanalysed using a scanning electron microscope and laser confocal microscope. We foundthat the chromogenic reaction rate of the mixture system increased from (15.9 ± 1.51) ×
10−3 min−1 in the control group to (29.7 ± 1.25) × 10−3 min−1 for 12.8 μM FGFC1 (p <0.01). FGFC1 also significantly increased the fluorescence intensity and D-dimer
concentration in FITC fibrin lysate. Image analysis showed that FGFC1 significantly
reduced the fiber density and increased the fiber diameter and the distance between
protofibrils. The inhibition of fibrinolytic activity of FGFC1 by 6-aminohexanoic acid
(EACA) and tranexamic acid (TXA) together with the docking results revealed that thelysine-binding sites (LBSs) play a crucial role in the process of FGFC1 binding toplasminogen. The action mechanism of FGFC1 binding to plasminogen was inferred,and FGFC1 was able to induce plasminogen to exhibit an open conformation by binding
through the LBSs. The molecular docking results showed that docking of ligands (EACA,
FGFC1) with receptors (KR1–KR5) mainly occurred through hydrophilic andhydrophobic interactions. In addition, the binding affinity values of EACA to KR1–KR5
were −5.2, −4.3, −3.7, −4.5, and −4.3 kcal/moL, respectively, and those of FGFC1 to
KR1–KR5 were −7.4, −9.0, −6.3, −8.3, and −6.7 kcal/moL, respectively. The findings

EAFTA 2023, On-site/online, May 17th-20th, 2023, Morioka, Japandemonstrate that both EACA and FGFC1 bound to KR1–KR5 with moderately highaffinity. These results show that FGFC1 can effectively promote fibrin lysis in vitro andmay represent a novel candidate agent for thrombolytic therapy.

Fig. Confocal laser microscopy images of fibrin networks based on fibrinolysis by FGFC1

Keywords: Fibrinolytic compound, Marine natural product, Protofibrils, Moleculardocking,

EAFTA 2023, On-site/online, May 17th-20th, 2023, Morioka, JapanK -value, a freshness index of fish.
Its application for modern demand*Kunihiko Konno1,2
1, Faculty of Agriculture, Iwate University, Ueda 3-18-8, Morioka, Iwate 020-8550, Japan2, Hokkaido University, Hakodate, Hokkaido 041-0843, Japan*Corresponding author: konnokabuki2022@outlook.jp
One of the most important factors that determines the quality of fish is its freshness.
Sensory methods are eventually used to judge the freshness. They are convinient but not
quantitative. Among scientific freshness indices, the most popularly used one is K-value,
which was proposed in 1960s and still valuable to show the freshness of fish. The value isbased on ATP metabolism in fish muscle after its death.
ATP in fish muscle is sequentially degraded into much simper compounds after its death.
The degradation of IMP into HxR is the slowest among the steps, and the relative content of
HxR and Hx derived from IMP is defined as K-value. As the IMP degradation is enzyme
catalyzed reaction, the process is determind by the the storage temperature and period,
namely storage history. The index has been accepted by people. The reasons are;
1. A complete comsumption of ATP leads ”Rigor-mortis”, traditional freshness index.
2. IMP gives gsynergitic effect with Soy sauce on the taste of fish meat (Sashimi).
3. K-value increase and freshness decrease judged by traditional methods fit very well.
Analysis technology was remarkably improved from ion exchange chromatography to
HPLC. Although the separation mechanism of HPLC differs from original one, the sample
preparation method designed for original method was used until now. The hardest part in theoriginal method was pH adjustment to pH 7. As HPLC analysis does not require the sample
to be pH 7, pH was adjusted to 3, where ATP was kept stable. Adjusting pH to 3 is simply
achieved by adding a fixed volume of KOH solution. The simplified method make it possibe
to analyze a large number of samples in short time even in night time by equipping
autosampler.
K-value is can be applicable not only to fresh fish but also to frozen stored and cooked
seafood products. In the latter cases, the value gives the freshness of the raw material (fish)
before the treatments.
Established K-value measurement protocol can garantee the freshness of any seafoods
distributed in the world scientifically.
Keywords: ATP, IMP, freshness, frozen fish, HPLC, Rigor-mortis,

EAFTA 2023, On-site/online, May 17th-20th, 2023, Morioka, Japan

Molecular interaction and properties characterization of anthocyaninbased film and its application in freshness monitoring and preservation
Yaqin Hu1*
1, College of Food Science and Technology, Hainan Tropical Ocean University, Sanya,
572022*Corresponding author: 1004346262@qq.com
The increasing demand of food quality and safety has given rise to the intelligent and active
packaging. The natural colorants with eco-friendly properties are integrated into packaging
films. Among which, anthocyanins have been widely applied as the functional components
due to their wide sources and multiple functions.
It is believed that various molecular interactions contained in the film matrix play the
crucial role in the film properties. Hence, the kinds of molecular interactions between the
bio-polymers and anthocyanins were studied based on our previous studies. Results
demonstrated that molecular interactions could not only affect film physical properties by
the formation of structural network, but also affect film functional properties (including
pH-responsive, active and barrier characteristics) by controlling of anthocyanin release.
Furthermore, the anthocyanin-based films with desirable functions were proved to show
potent abilities in real-time freshness monitoring and efficient food preservation (such as
better sensory profiles, lower TVB-N, TVC and k values, etc).
Our series of studies provided the basic insight into the molecular interactions, properties
and applications of anthocyanin-based films. Nevertheless, further studies focusing on
anthocyanin stability enhancement and preservation mechanism investigation ofanthocyanin-based film are critically needed.

Keywords: Anthocyanin, Food packaging, Molecular interactions, Film properties,
Real-time freshness monitoring, Food preservation

EAFTA 2023, On-site/online, May 17th-20th, 2023, Morioka, Japan

Changes in the K-value in several fishes and the benefits of applying Kvalue in the distribution of fisheries products○Yuko Murata1
1, Fisheries Technology Institute, Japan Fisheries Research and Education Agency, Fukuura,
Kanazawa-ku 2-12-4, Yokohama, Kanagawa 236-8648, Japan*Corresponding author: murata_yuko56@fra.go.jp
K-value is used as an index to determine the freshness of fish. It is especially suitable fordetermining the freshness of raw fish to be eaten such as sashimi and sushi. Our researchso far determined that K-value is not only an indicator of freshness but may also be
useful in quality control and determining the handling history of fish from catch to
distribution. In this study, we present cases of the application of K-value in the
distribution of fisheries products. In the case of red sea bream, Pagrus major, andrainbow trout, Oncorhynchus mykiss, it was found that the handling history of fish may
be determined after the catch and during storage. It was also found that the time required
to develop the best taste can be estimated in the case of yellowtail, Seriola
quinqueradiata, and silver salmon, Oncorhynchus kisutch. The study finally presents theapplication of K-value from catch to consumption.
This research was supported by the research program on development of innovativetechnology grants from the Project of the Bio-oriented Technology Research
Advancement Institution (BRAIN), and research and development projects forapplication in the new policy of Agriculture, Forestry and Fisheries from the Ministry ofAgriculture, Forestry, and Fisheries, Japan. Also funded by Nippon Kaiji Kentei Kyokai.
Keywords: K-value, freshness, quality control, handling history, distribution, catch,storage

EAFTA 2023, On-site/online, May 17th-20th, 2023, Morioka, Japan

Validation of a simplified method for determination of K-value
by interlaboratory study.
Takeya Yoshioka
Hokkaido Industrial Technology Center, Kikyo 379, Hakodate, Hokkaido 041-0801, Japan*Corresponding author: yoshioka@techakodate.or.jp[Background] K-value is determined in many research institutes and testing laboratories
as a fish freshness indicator. However, it was difficult to compare the K-value data fromdifferent laboratories because each laboratory used different determination procedure,
reagents and apparatus. In addition, the conventional K-value measurement method wascomplicated and required skilled persons to perform accurate measurement. Therefore,
in order to standardize the K-value determination method, simplification and validation
were examined.
[Methods] To simplify the measurement method, the well-known conventional
procedure was modified. An interlaboratory study to validate a simplified new method
was designed based on the IUPAC protocol. As the test samples, frozen fish meat
powder prepared from white muscle was packed in plastic bottles and distributed to eachlaboratory. The fish species selected considering white and red fish, natural and farmed
fish, etc. According to a provided method, eleven laboratories quantified ATP-related
compounds in the 10 test samples (5 pairs of blind duplicates) and calculated K-values.
[Results] The neutralization process of the perchloric acid extract was simplified by a
two-step pH adjustment using alkaline solution and neutral phosphate buffer. In theinterlaboratory study, specified statistical indices were calculated from K-value datareported from each interlaboratory. The discernible range of K-value (4sR) calculated
from the reproducibility standard deviation (sR) was approximately 1% for highfreshness samples (K-value 6-10%), approximately 2% for medium freshness samples(K-value 25-40%), and approximately 5% for low freshness samples (K-value 84%).
These results were approved by a project committee composed of academic experts andothers.

Based on this method, a draft standard was submitted to the Ministry ofAgriculture, Forestry and Fisheries (Japan), and "Test Method for Freshness (K-value) of
Fish - High Performance Liquid Chromatography" was established as a Japanese
Agricultural Standard in March 2022.
Keywords: K-value, standardization, interlaboratory study

EAFTA 2023, On-site/online, May 17th-20th, 2023, Morioka, Japan

Effect of Freshness before Freezing on the Quality of Thawed Meat inJapanese Pilchard Landed at Ports in Eastern Hokkaido○Keisuke Moriya*1, Akiko Miyazaki2, Hiroyuki Kodama3, Koji Ebitani3
1, Food Processing Research Center, Hokkaido Research Organization, 589-4 Midori-machi,
Ebetsu, Hokkaido, 069-0836, Japan2, Abashiri Fisheries Research Institute, Hokkaido Research Organization, 7-8-5, Minato-cho,Mombetsu, Hokkaido 094-0011, Japan3, Kushiro Fisheries Research Institute, Hokkaido Research Organization, 4-25, Nakahamacho, Kushiro, Hokkaido 085-0027, Japan*Corresponding author: moriya-keisuke@hro.or.jp

Fish with good freshness and quality for sashimi (sliced raw fish) are highly valued
because Japanese consumers prefer sashimi. It is well known that freshness of Japanesepilchard (Sardinops melanostictus) that is migratory red-flesh fish deteriorates rapidly.
The port in eastern Hokkaido is close to the fishing ground of Japanese pilchard, whichis advantageous for maintaining the freshness of Japanese pilchard. The purpose of thisstudy was to clarify the freshness of Japanese pilchard just after landing on eastern
Hokkaido and the effect of ice storage period prior to freezing on the quality of thawed
meat. Japanese pilchard which was landed at ports in eastern Hokkaido stored in ice fordifferent periods (0, 5, 24 h) and then, these fishes were frozen in a liquid freezer. Eachfrozen pilchard was kept at -20 °C or -40 °C until analysis for 0 or 2 months and then,thawed for 15 h at 2 °C. Japanese pilchard just after landing was considered highly fresh
due to K value of ≤4 % and some of which remained adenosine triphosphate (ATP). The
K value of frozen meat of 5 h of ice storage after landing was about ≤5 %. The maximum
strength as a physical property of the thawed meat decreased with ice storage period
before freezing. The b*/a* value of thawed dark meat after keeping at -20 °C for 2
months with 0 h of ice storage, or after keeping -40 °C for 2 months with 0-24 h of ice
storage were not changed during frozen storage. The pilchards, that were rapid-frozen
just after landing or that were rapid-frozen within 5 h after landing and then stored at 40 °C, were suitable for consumption as an ingredient in frozen product for sashimi as
they demonstrated good physical properties and insignificant color deterioration postthawing.

Keywords: Freezing, Freshness, K value, Sashimi, Quality

EAFTA 2023, On-site/online, May 17th-20th, 2023, Morioka, Japan

Introduction to Standardization in ISO
Yoshiaki Yamano*
Standard Consulting Unit, Japan Standard Association, Mita MT Bldg., 3-13-12 Mita,
Minato-ku, Tokyo, 108-0073, Japan*Corresponding author: yamano@jsa.or.jp
This presentation is a general explanation of the standardization in ISO (International
Organization for Standardization). International standards play an important role inworld trade, import and export. They bring many kinds of benefits to our activities, suchas food safety, environmental and quality management, etc. We also explain theacceptance criteria with a new work item proposal for the standardization in ISO.
Keywords: International standardization, ISO, Roles in world trade, Acceptance criteria,
New work item proposal

EAFTA 2023, On-site/online, May 17th-20th, 2023, Morioka, Japan

Optimisation of Virgin Coconut Oil Extraction of Natural Pigment fromMicroalgae by Response Surface Methodology (RSM)○Eko Susanto*1, Febriansyah Hanik Dwi saputri1, Putut Har Riyadi1, Lukita Purnamayati1
1, Department of Fish Products Technology, Faculty of Fisheries and Marine Sciences,
Universitas Diponegoro, Jl. Prof. Jacub Rais, Tembalang Semarang 50275*Corresponding author: eko.susanto@live.undip.ac.id

The present study reports on the extraction of natural pigment from three different
speciesofmicroalgae(Arthrospiraplatensis,

Nannochloropsis

oculata,andThalassiosira sp). Response surface methodology (RSM) was used to investigate theeffect of process variables on the extraction using virgin coconut oil (VCO). Two
independent variables, temperature (50–60 °C) and extraction time (60–120 minutes),
were investigated. According to the results, the optimal extraction condition with VCO
were microalgae specific. For Arthrosphora platensis and Thalassiosira sp, the ideal
conditions were achieved at a temperature of 60 °C and an extraction period of 90
minutes; whereas, for Nannochloorpsis oculata, the optimal conditions were achieved at
a temperature of 59.83 oC for 88.09 minutes.The experimental results under ideal
conditions were in good consistency with the predicted values, which showed that theextraction of pigments from microalgae using VCO will enhance the functionality of
VCO itself and will be adaptable into products for functional foods.

Keywords: Virgin coconut oils, Microalgae, Carotenoids, Chlorophylls, RSM

EAFTA 2023, On-site/online, May 17th-20th, 2023, Morioka, Japan

Enhancement of anti-inflammatory activity of fish myofibrillar protein
by Maillard-type glycation and its molecular mechanism○Wenzhao Li1, Ga-Hyun Joe1, Yutaka Shimizu1, Hiroki Saeki1
Laboratory of Marine Food Science and Technology, Faculty of Fisheries Sciences, Hokkaido
University, Minato 3, Hakodate, Hokkaido 041-8611, Japan*lee19930206@eis.hokudai.ac.jp[Objective] Myofibrillar protein (Mf) from spawned-out chum salmon

gained strong

anti-inflammatory activity in vitro and in vivo by the glycation with alginate
oligosaccharide (AO) through the Maillard reaction. Considering that alginate
oligosaccharide is a copolymer linked by two types of uronic acid possessing one
carboxyl group in each molecule, the role of carboxyl group of the attached sugars inenhancing the anti-inflammatory activity of Mf was investigated in this study.
[Methods] The lyophilized Mf was glycated with monosaccharides and their oxidized
derivatives (uronic acid) at 60 °C and 35% relative humidity through the Maillard
reaction. The glycated Mfs were digested by pepsin-trypsin and evaluated the antiinflammatory activity by measuring the secretions of inflammatory mediators inlipopolysaccharide (LPS)-stimulated RAW 264.7 macrophages. In addition, the effect on
gene expression related to LPS-stimulated signaling pathways was also examined to
discuss the molecular mechanism of the enhanced anti-inflammatory activity.
[Results] The anti-inflammatory activity of Mf was not affected by glucose andgalactose, whereas carboxyl-containing glucuronic acid and galacturonic acid conferredstrong anti-inflammatory activity upon addition to Mf, as the same as alginate
oligosaccharide. These results indicate that the presence of carboxyl group in reducing
sugar is an important factor enhancing the anti-inflammatory activity of Mf in the
Maillard type glycation. In the gene expression analysis, the uronic acid-glycation
suppressed LPS-stimulated inflammation by inhibiting the ability of CD14 recognizing
LPS, thereby enhancing the suppressive effect of Mf in TLR4-MyD88-dependent
inflammatory signaling pathway. In conclusion, the uronic acid-glycation, which
modulates cell signaling and enhances anti-inflammatory function, would be a useful
method for developing food-functional materials from the Mf.
Keywords: Fish myofibrillar protein, Anti-inflammatory activity, Maillard reaction, Uronic
acid, Carboxyl group, Macrophage

EAFTA 2023, Online/onsite, May 17th -20th, 2023, Morioka, Japan

Fresh-keeping effect of 3D printed surimi improved by Ca2+-nano starchlutein and its freshness indicator mechanism based on printed
anthocyanin
Gaoshang Li1, 2, Yaqin Hu2, *1
Institute of Food Engineering, College of Biosystems Engineering and Food Science, Zhejiang University,
Hangzhou, Zhejiang, China, 310058
2College of Food Science and Technology, Hainan Tropical Ocean University, Sanya 572022, PR China*1004346262@qq.com[Objective]
Protein-based surimi is suitable for 3D printing to produce customed foods. However, the insufficientbonding ability and weakened function (such as short shelf) become the weak points of 3D printed
functional surimi foods. To facilitate its printing effects and fresh-keeping, a surimi added with a
combination of Ca2+ and nano starch-lutein (NS-L) was used. Additionally, though the freshness of somematerials changed during storage period, it could not be perceived due to the color of material itself.
Hence, it is meaningful to research the printable freshenss indicator materials.

[Methods]
The surimi was added to NS-L and Ca2+, printing properties (texture, gel strength, rheological properties)
and freshness indexes of the materials were determined. Additionally, printable color-changing material-anthocyanin-hydroxypropyl methyl cellulose-xanthan gum-carrageenan (AHXK) was prepared. Its
printing effects and discoloration mechanism (response to ph and ammonia) were determined.
Furthermore, the real-time monitoring of freshness on printed surimi was explored.

[Results]
1.

The NS-L and Ca2+ increased gel strength, hardness, τ, viscosity of surimi by 1.2-3 times, forming
denser structure and strengthening supporting and bonding ability to enhance printing effects.

2.

They also inhibited bacterial growth and spoilage bacteria of printed functional surimi, as well asamino acids decomposition of surimi was decreased by 12%, which limited the ATP decomposition(decreased K) and production of ammonia (decreased TVB-N).

3.

The AHX with 5% K had suitable material properties (gel strength, texture, rheology) for printing.

4.

The HXK with 0.75% A was sensitive to pH and ammonia, exhibiting significant color-changing on
freshness monitoring of printed surimi during refrigeration.

[Conlusion]
Printed AHXK-functional surimi could achieve the aim of printing effect enhancement, fresh-keeping
and freshness monitoring simultaneously, which could promote the application of 3D printing technology
on food process.
Keywords: Printability; Functional surimi; Preservation; Color-changing; Freshness monitoringEAFTA 2023, On-site/online, May 17th-20th, 2023, Morioka, Japan

Preservation and bacteriostasis functions of citral nanoemulsion on
golden pomfret during 4 degrees C storage
Zhiheng Hu12, Yaqin Hu*2

1, College of Biosystems Engineering and Food Science, Zhejiang University, Hangzhou
3100582, College of Food Science and Technology, Hainan Tropical Ocean University, Sanya
572022*Corresponding author: 1004346262@qq.com
Golden pomfret is an economical marine fish specie which is easily prone tospoilage during process and storage. The spoilage in aquatic products is normally
attributed to the oxidation, action of endogenous enzymes and the pollution of spoilage
microorganisms. Hence, effective methods of preservation and bacteriostasis are vital forthe fresh keeping of golden pomfret.
In our previous study, a citral nanoemlusion has been prepared, while its inhibition
effects to Shewanella putrefaciens, one of the SSOs in golden pomfret, has also beenclarified. In this study, the practical fresh keeping functions of citral nanoemulsion on
golden pomfret were investigated. During storage, the citral nanoemulsion could
significantly reduce the TVC value in fish. The SSOs of golden pomfret were mainly
Shewanella sp. and Pseudomonas sp., among which Shewanella putrefaciens occupied
the dominant status. The citral nanoemulsion could affect the distribution of microbial
composition, which was mainly reflected by the stronger bacteriostasis on Shewanella
sp.. Meanwhile, the nanoemulsion treatment effectively delayed the increase of freshness
deterioration indexes such as TVB-N, TBARS and K value, following with higher
texture parameters and sensory scores.
Overall, our study verified the ideal preservation and bacteriostasis functions of
citral nanoemulsion on golden pomfret, the citral nanoemulsion could be applied as apotential fresh keeping agent in practical aquatic products.
Keywords: Golden pomfret, Citral nanoemulsion, Preservation, Bacteriostasis

EAFTA 2023, On-site/online, May 17th-20th, 2023, Morioka, Japan

Sensory Evaluation and Lipid Analysis of Marlin Kajikia audax for Sashimi
Kanako Hashimoto*1, Tokifusa Kawashima2, Yuko Murata1, Takuya Seko1, Shintaro
Imamura1, Kenji Ishihara1
1Fisheries Technology Institute, Japan Fisheries Research and Education Agency, 2-12-4
Fukuura, Kanazawa, Yokoyama, Kanagawa 236-8648, Japan
2
Chiba Prefectural Fisheries Research Center, 2492 Hiraiso, Chikuramachi, Minamiboso,
Chiba 295-0024, Japan*Corresponding author: hashimoto_kanako05@fra.go.jp
Identifying the sensory properties of fish consumers is important for providing theirpreferred seafood. The sensory preference for fish is affected by odor, flavor, texture,
and appearance. Moreover, several compounds are involved with sensory preference. In
this study, we focused on fats that enhance the intensity of umami and examined the lipid
content preference in the marlin Kajikia audax sashimi. First, we compared the lipid
contents and fatty acid compositions of marlin by different individual size and body parts.
Furthermore, the preference for total, taste, texture, odor, and lipid content in marlin
sashimi taken from different parts of the body with different amounts of lipid wasinvestigated by the paired-preference method. We also evaluated the relationshipbetween the lipid content preference and total, taste, texture, and odor preferences by thechi-square test. The lipid contents in the bigger-sized marlin were higher than that in thesmall-sized. In bigger-sized marlin dorsal parts, the lipid contents in the anterior parts
were the higher than that in the posterior parts. No differences were observed in the fatty
acid composition of different size and body parts. The lipid content preference for theanterior dorsal part (high lipid part) was higher than that for the posterior dorsal part(low lipid part). The lipid content preference was associated with total and taste
preferences. Therefore, the lipid content was found to be associated with the preference
for marlin sashimi.
Keywords: sensory preference, sashimi, lipid content

EAFTA 2023, On-site/online, May 17th-20th, 2023, Morioka, Japan

Improvement Added Value of Fishery Byproduct as Halal Gelatin○Mala Nurilmala*1,3, Agoes M Jacoeb1, Noviyan Darmawan2,3, Tun Tedja Irawadi2,3,
Yoshihiro Ochiai4
1, Aquatic Product Technology Department, Faculty of Fisheries and Marine Sciences, IPB
University, 16680, Bogor, Indonesia
2, Department of Chemistry, IPB University, 16680, Bogor, Indonesia
3, Halal Science Center, IPB University, 16129, Bogor, Indonesia
4, Graduate School of Agricultural Science, Tohoku University, Aramaki, Aoba, Sendai,
Miyagi 981-8572, Japan*Corresponding author: mnurilmala@apps.ipb.ac.id

Fishery industries leave waste such as heads, bones, skin, scales, swim bladder, andviscera. The amount of these byproducts is in the range of 30-50%. However, theutilization of them is still limited, then need efforts to increase its valorization. Recently,
we have been developing gelatin from these byproducts to increase its added value andmeet the requirement for ‘halal’ food as well as non-food such as cosmetics andpharmacy, since the common gelatin come from source of skin and bone of porcine. It
has been known gelatin has been widely used as a multifunctional ingredient for manyindustries. Here, we use skin, scale, and swim bladder to produce halal gelatin by acidic
method. The characteristic resulted gelatin were in accordance with those standard ofIndonesian gelatin and GMIA (Gelatin Manufacturer Institute of America)

Keywords: Fishery byproducts, Gelatin, Halal, Valorization

EAFTA 2023, On-site/online, May 17th-20th, 2023, Morioka, Japan

Contribution of structural rearrangement and molecular interactions on
quality improvement of frozen fish fillets○Qingqing Jiang1*, Shiyu Huang1, Xichang Wang1, Yaqin Hu2, Emiko Okazak3
1, College of Food Science and Technology, Shanghai Ocean University, Shanghai 201306,
China
2, College of Food Science and Technology, Hainan Tropical Ocean University, Sanya
572022, China
3, Department of Food Science and Technology, Tokyo University of Marine Science andTechnology, Tokyo 108-8477, Japan*Corresponding author: qqjiang@shou.edu.cn

The present study evaluated the effects and underlying mechanisms of light salting on quality
properties of fish fillets during repeated freezing-thawing. Light salting was found to improvewater-holding capacity and decelerated texture softening in fillets. Instead of tissue distortionand heterogeneous aggregates in control groups, light salting promoted myofibril disassemblyand formation of an ordered protein network with the solubilized myofibrillar proteins. The
myofibrils presented an overall amorphous appearance with the loss of M-lines, removing the
restraints to myofibril swelling and solubilization from A-binds in salted groups during
repeated freezing-thawing. The structural rearrangement caused by light salting facilitated theenlargement of water-holding space, transformation of tissue water, and tissue recoverability,
improving water-holding capacity and texture properties of fillets during freezing-thawing.
The finding provided novel insight into the improvement of quality properties of fish fillets bylight salting when subjected to drastic temperature fluctuations.

Keywords: Structural rearrangement; Molecular interactions; Water transformation; Lightsalting; Freeze-thaw stability; Fish fillets

EAFTA 2023, On-site/online, May 17th-20th, 2023, Morioka, Japan

Involvement of Endogenous Proteases in Abalone Muscle Softening○Chen Yulei1,2, Zhang Minghui1, Li Wanyu1, Sun Lechang1,2, Cao Minjie*1,2
1, College of Ocean Food and Biological Engineering, Jimei University, 43 Yindou Road,
Xiamen 361021, China
2, National & Local Joint Engineering Research Center of Deep Processing Technology for
Aquatic Products, 43 Yindou Road, Xiamen 361021, China*Corresponding author: mjcao@jmu.edu.cn

The extracellular matrix (ECM) is greatly degraded by endogenous proteases in aquatic
animals after death, thus leading to muscle softening. We revealed collagenases to participate
in postmortem tissue self-degradation of Haliotis discus hannai by degrading type I collagen.
Hence, recombinant matrix metalloproteinase 1 and 14 (rMMP1 and rMMP14) with highpurity and enzymatic activities were expressed using a prokaryotic expression system.
rMMP1 and rMMP14 effectively degraded type I collagen into small fragments and peptides.
Tissue inhibitor of metalloproteinase (TIMP), an endogenous inhibitor of MMP, was also
expressed using HEK 293F cells. Recombinant TIMP (rTIMP) showed great inhibitory
activity toward rMMP1, but not rMMP14. Hence, it can significantly inhibit rMMP1’s
degradation activity toward collagen. Inhibition kinetics analyses revealed rTIMP to be acompetitive inhibitor of rMMP1. Biolayer interferometry revealed rTIMP can effectively bindwith rMMP1, with an equilibrium dissociation constant value of 263 nM. Furthermore, wecloned the full-length cDNA sequence of prolyl endopeptidase (PEP) from abalone.
Recombinant PEP (rPEP) was expressed and characterized in detail. We for the first time
determined the 1.5 Å crystal structure of rPEP. Using collagen peptides as substrates, HPLCESI-MS analysis confirmed that rPEP specifically cleaved at the carboxyl side of proline
residues, suggesting its role in the degradation of collagen peptides. These results elucidate
the possible mechanism of abalone muscle softening in the aspect of endogenous proteases.

Keywords: Abalone, Muscle softening, Matrix metalloproteinase 1, Prolyl endopeptidase,
Collagen degradation

EAFTA 2023, On-site/online, May 17th-20th, 2023, Morioka, Japan

Microstructure and physicochemical properties: Effects of different
pretreatment combined with dual-frequency ultrasound on quality oflarge yellow croaker (Pseudosciaena crocea) during cold storage
Weiqing LAN1,2,3 ○Peiling YAN1 Xinyu ZHAO1 Jing XIE1,2,3*

1, College of Food Science and Technology, Shanghai Ocean University, Shanghai
201306, China
2, Shanghai Aquatic Products Processing and Storage Engineering Technology
Research Center, Shanghai 201306, China
3, National Experimental Teaching Demonstration Center for Food Science andEngineering (Shanghai Ocean University), Shanghai 201306, China*Corresponding author: jxie@shou.edu.cn

Abstract: The quality changes of cold-stored (4 ºC) large yellow croaker(Pseudosciaena crocea) were studied by using stable chlorine dioxide combined withdual-frequency ultrasound assisted-slightly acidic electrolyzed water (CUSS). The
results of microbial indicators demonstrated that CUSS treatment inhibited microbial
growth. Moreover, CUSS treatment had lower values of pH, total volatile basic
nitrogen (TVB-N) and thiobarbituric acid (TBA), and higher immobilized watercontent. CUSS treatment also inhibited the degradation of inosine monophosphate andthe accumulation of hypoxanthine riboside and hypoxanthine. Through the results oftexture profile analysis (TPA), protein degradation, and scanning electron microscopy(SEM), CUSS treatment could effectively protect the protein structure and maintain
good texture characteristics of Pseudosciaena crocea. According to microbial indexes,
K-value and sensory analysis, compared with dual-frequency ultrasound treatment,
CUSS treatment could prolong the shelf-life of Pseudosciaena crocea for another 6days. Therefore, CUSS treatment is a potential method to preserve Pseudosciaena
crocea and improve its protein characteristics.
Key words: Stable chlorine dioxide, dual-frequency ultrasound, slightly acidic
electrolytic water, Pseudosciaena crocea, quality1Condition-dependent Adenosine Monophosphate Decomposition Pathways by Endogenous
Enzyme in Striated Adductor Muscle from Japanese scallop (Patinopecten yessoensis)

Huamao Wei1, Wenge Yang1, Tao Huang1, Chunhong Yuan2(1. College of Food and Pharmaceutical Sciences, Ningbo University, 315211; 2. Faculty of Agriculture, Iwate
University, Ueda 3-18-8, Morioka, Iwate 020-8550, Japan)[Objective] The post-mortem ATP and related compounds have been extensively studied for
quality evaluation of fish or shellfish for over 60 years. "K-value" is based on the ATP breakdown
and the subsequent formation of inosine (HxR) and hypoxanthine (Hx). It is well-known that IMP
is the main nucleotide present in most fish species such as mackerel, whereas AMP remains the
major nucleotide in crustaceans and mollusks. It is controversial whether IMP in scallops
generates. Therefore, the aim of this study was to investigate the AMP decomposition pathway inscallop adductor muscle and its influencing factors, such as EDTA or EGTA addition, heating,ions metal concentration change, et al.
[Methods] Crude enzyme solution was made from scallop striated muscle after chopping andhomogenization at 4°C, and divided into three groups: control group, with EDTA or EGTA
addition. Then, AMP, IMP and AdR were added into the crude enzyme solution as the substrate todetect the decomposition rate at at 25°C. In addition, the crude enzyme solution was dialyzed toremove metal ions, and the deionized crude enzyme solution was divided into two groups: thecontrol group (C) and the addition of 0.1% chloramphenicol (CP). Subsequently, AMP was added
and changes in its ATP-related compounds was measured by HPLC at 25°C.
[Results] The results showed that IMP accumulated due to AMP decomposition via endogenous
enzymes in scallops stored at both 4°C and 20°C. The AMP decomposition rate was highest in the
supernatant of homogenized scallop adductor muscle, followed by the suspended solution andprecipitate, while IMP could not be decomposed in scallop. AdR decomposed rapidly since highactivity of AdR deaminase. The IMP generation rate increased dramatically in scallop crude
enzyme solution containing 5 mM EDTA. Moreover, small amount of IMP also generated rapidly
in scallop crude enzyme solution containing 5 mM EGTA within 2 h. Moreover, K+ was
confirmed to promote the decomposition of AMP to IMP, Mg2+ promoted the decomposition of
AMP to AdR, and Ca2+ slightly inhibit the decomposition of AMP. Meanwhile, the protein
content and protein composition were also measure during dialysis and incubation. The results
demonstrated that, the protein in the scallop enzyme solution of control group gradually denatured
due to microbial activity during incubation, while CP group did not. Adenosine deaminase andadenosine kinase were detected in denatured protein fraction (40 kDa) by LC-MS/MS.
[Conclusion]. The decomposition path of AMP in scallops could be changed when concentrationof metal ions change. In addition, the production of IMP in scallop was confirmed to be produced
by endogenous enzymes via decomposition of AMP, while the generation of HxR and Hx iscaused by endogenous enzymes and microbial activities.
Key words: AMP; endogenous enzyme; EDTA; IMP

EAFTA 2023, On-site/online, May 17th-20th, 2023, Morioka, Japan

Shape Analysis and Vitality Estimation of pacific oyster (Crassostrea gigas)
Using Three-Dimensional Morphometric Techniques○Xin Lu3, Shiliang Dong1, Ryosuke Bizen2, Sota Hayashi2, Koichi Takaki3,4,
Kouichi Konno3, Kouichiro Kobayashi3, Katsuyuki Takahashi3, Chunhong Yuan2,4*
1 United Graduate School of Agricultural Sciences, Iwate University,
2 Faculty of Agriculture, Iwate University
3 Faculty of Science and Engineering, Iwate University
4 Agri-Innovation Center, Iwate University*Corresponding author:chyuan@iwate-u.ac.jp
Pacific oysters (Crassostrea gigas) are widely cultivated as an important marine fisheryresource. Despite their production steadily rising over the years, their short survival andpreservation period make the selection and maintenance of highly vital oysters a prime
concern. While previous studies have explored the correlation between the shape of an oyster
and its quality, this research proposes a novel, non-invasive, and immediate method forgrading the freshness of Pacific oysters intended for brand sales. By exclusively applying 3D
morphometric techniques to analyze oyster's external shapes, this method can replace theinconvenient but highly accurate chemical freshness assessment with a more streamlined
approach..
In this study, the authors accurately describe the unique oyster shapes, including aspects
like mean flattening ratio, the difference in this ratio at the front and back, the center axis
curvature, and its orientation angle. Along with the shell height to length ratio, these fivefeatures form the external shape features to obtain detailed biochemical data about oyster
vitality. They then use principal component analysis (PCA), a multi-variable technique, tosimplify this five-dimensional feature space to one, and create a formula for classifying
oysters as round or flat. Using this formula, they first separate the oysters into these twocategories. Next, they arrange the oysters based on the A.E.C. value, a freshness and vitality
indicator, separating the round (higher A.E.C.) and flat (lower A.E.C.) oysters again. Finally,
they compare these classification results to calculate their accuracy.
The study used 90 oysters for the experiment, including 40 from Toyama-ken, purified indeep-sea water, and 50 from Yamada-machi, Iwate-ken. The proposed method accurately
classified 70.0% of the round and flat Toyama oysters (20 each) and 76.0% of the round andflat Yamada Town oysters (25 each). As the two different species were collected at different
attachment and growth periods in two major macaque production areas in different marine
areas, the effectiveness of the classification method proposed in this study was demonstrated.
The experimental results aslo prove that there are disparities in vitality maintenance between
round and flat oysters defined by the proposed method. Applying this study in practicalsettings could enhance oyster branding and quality during distribution, such as at wholesalemarkets or during transportation.
Keywords: Pacific oyster, Crassostrea gigas, 3D morphometric techniques, vitality, shapefeatures, flattening ratio, curvature, center axis, A.E.C. value, principal component analysis

EAFTA 2023, On-site/online, May 17th-20th, 2023, Morioka, Japan

Enzyme-assisted extraction and characterization of collagen from Basa fish
(Pangasius bocourti) skin
○Quyen T.H. Tran*, Tang V. Nguyen, Thinh V. Phan
Faculty of Food Technology, Nha Trang University, 02 Nguyen Dinh Chieu, Nha Trang,
Khanh Hoa 57000, Vietnam*Corresponding author: quyentth@ntu.edu.vn
It’s generally accepted that type I collagen is a fibrillar structure collagen, which plays animportant role as the essential structural composition and mechanical scaffold of several
tissues. The objective of this study is to extract type I collagen from Basa fish skin using
acetic acid as the extraction solvent with the assistance of pepsin. The small pieces ofBasa fish skin were immersed in 0.2 M sodium hydroxide solution under stirring in 24 h
to eliminate lipids. After that, the skin was washed to neutral pH with distilled water. In
the next step, the skin pieces were dipped and stirred in 0.003 M citric acid in 30 min toremove minerals. The skin pieces were immersed in 1% hydrogen peroxide solution for
3 h for colorant and odorant elimination. The collagen sample was obtained with 0.5 M
acetic acid solution in addition to pepsin in 24 h. The extract filtration was performed
through filter paper. The protein precipitation was done by adding dropwise sodium
chloride solution in 24 h to the extract so that the final sodium chloride concentration
was 2.5 M. The precipitate was collected after centrifugation at 4oC and 3000 rpm in 20
min. The extraction process was followed by dialysis in phosphate buffer with pH 7.8.
Finally, the sample was lyophilized and stored in sealed containers until use at -20oC.
The obtained collagen was characterized by quantitative and qualitative methods. The
denaturation temperature of collagen samples was determined based on the change inviscosity of collagen solution as increasing temperature. The purity of collagen proteins
was examined by ultraviolet-visible spectroscopy (UV-Vis). The molecular weight (Mw)of collagen subunits was determined by sodium dodecyl sulfate-polyacrylamide gelelectrophoresis (SDS-PAGE). The amino acids composition was analysed by highperformance liquid chromatography (HPLC). The surface morphology of collagen wasobserved by scanning electron microscope (SEM) images. The chemical groups were
determined by fourier transform infrared spectroscopy (FTIR). The results showed thatthe collagen extraction yield was 520.5 mg/g of fresh fish skin on the basis of lyophilized
dry weight. The denaturation temperature (Td) was 34.8oC by measuring viscosity. UVVis spectrum with one peak at the wavelength of 230 nm confirmed the purity of theEAFTA 2023, On-site/online, May 17th-20th, 2023, Morioka, Japan

collagen. Based on SDS-PAGE, Mw of collagen α1, α2, and β subunits wereapproximately 130, 118, and above 200 kDa, respectively. By HPLC, 17 proteinogenic
amino acids were found in the collagen sample, in which the hydroxyproline content was
68.3 mg/g. SEM images confirmed the fibril structure of collagen. FTIR spectrum
indicated characteristic bands according to the presence of amide A, B, I, II, and III
bonds in collagen chemical structure. It is concluded that the preparation of collagenfrom Basa fish skin (Pangasius bocourti) was done. Therefore, the purified collagenobtained from this study can be further used in various fields of application.
Keywords: Basa fish skin, type I collagen, enzymatic extraction, proteinogenic aminoacids, hydroxyproline

EAFTA 2023, On-site/online, May 17th-20th, 2023, Morioka, Japan

Factors influencing the nutritional composition, quality and safety of dried
fishery products○Md. Golam Rasul*1, Chunhong Yuan2, Kefeng Yu3, Koichi Takaki4, A.K.M. Azad Shah1
1, Department of Fisheries Technology, Bangabandhu Sheikh Mujibur Rahman Agricultural
University, Gazipur -1706, Bangladesh
2, Department of Food Production and Environmental Management, Faculty of Agriculture,
Iwate University, 3-18-8 Ueda, Morioka, Japan
3, Sanriku Fisheries Research Center, Iwate University, Heta 3-75-1, Kamaishi, Japan4, Faculty of Science and Engineering, Iwate University, Ueda 4-3-5, Morioka, Japan*Corresponding author: rasul@bsmrau.edu.bd

Dried fishery products are popular processed food items throughout the world. However,there is a frequent complain about the quality and safety of dried products. To reduce
quality loss of dried fishery products, excellent quality raw materials with vacuum
drying, oven drying, solar tunnel drying with different pre-treatments of chlorinatedwash, bleeding, gutting, blanching, ultrasound, and treatment with various plant extracts
have a broad potential application for the improvement of quality and safety of the driedproducts. Moreover, various chemical preservatives have also been found to be effective
in increasing better organoleptic scores of the products although they have potential
health hazards. Drying temperature and relative humidity should below 60°C and 1043%, respectively to prevent loss of nutrients, quality decrease and consumer
acceptability. Multilayer plastics, polyethylene films of high gauge, poly vinyl alcohol
film containing 2% green tea extract along with vacuum packaging and oxygen
scavenger packaging ensure the best quality protective effectiveness to prevent the dried
fishery products from absorbing moisture and oxidizing of lipid. Recommended storage
conditions for dried products are <60% relative humidity and temperature from 0-10°C;
low oxygen levels of 0.5%. Low dose gamma irradiation (<5 kGy) reduces microbial
loads, extends the shelf life and improves the taste and texture of dried fish. Ultraviolet
light treatment, 10 mins cold oxygen plasma treatment, 3 mins atmospheric plasma
treatment, corona discharge plasma (0-3 mins) and high hydrostatic pressure effectively
inactivate the microbial contaminants from dried fishery products with better sensory
properties. These findings will be helpful to develop an effective quality control for
producing safe fishery products for domestic consumption and earning foreign currency.

Keywords: Dried fishery products, Factors, Nutritional composition, Quality, Safety

EAFTA 2023, On-site/online, May 17th-20th, 2023, Morioka, Japan

Effects of dry storage on mitochondrial structure and metabolic pathway ofMizuhopecten yessoensis○Zhongzhuan Yin1,2, Man Li1, Yan Cai1 , Lin Qi , Chunhong Yuan3, Yuanyong Tian1*
1, College of Food Science and Engineering, Dalian Ocean University, Dalian, 116023,
Liaoning, China
2, Department of Food Science and Technology, Tokyo University of Marine Science andTechnology, 4-5-7, Konan, Minato-ku, Tokyo 108-8477, Japan3, Faculty of Agriculture, Iwate University, Morioka, 020-88550, Japan*Corresponding author: tianyuanyong@foxmail.com

Postharvest dry storage was simulated by storing scallops at 4 °C for different periods.
The quantification of 40 energy metabolites related to the pathways of glycolysis,
tricarboxylic acid cycle (TCA) and oxidative phosphorylation in the scallop adductor
muscle was performed by LC‒MS/MS. The results showed that during the early stage,
the TCA cycle was maintained at a low level. Meanwhile, the aspartate pathway andopine pathway were opened, and alanine and opines were the early metabolites during
this stage. During the middle and late stages of dry storage, muscle pH decreased.
Phosphoenolpyruvate carboxy kinase (PEPCK) was activated, leading to the competitive
inhibition of pyruvate synthesis by PEPCK. Phosphoenolpyruvate enters the TCA
reverse cycle via the phosphoenolpyruvate → malate pathway to produce succinate.
Changes in metabolic pathways led to a change in the NADH level and mitochondrial
membrane potential. The change of energy pathway is earlier than mitochondrial
morphology change. The mitochondria just began to swell, the outer membrane wasbroken, and the matrix was cloudy after 12 h of dry storage. After 36h, mitochondria
were basically completely broken. In summary, during the dry storage of scallops, themitochondrial morphology was gradually destroyed, and the energy metabolism pathway
was changed to maintain the ATP levels in the scallop adductor muscle.
Keywords: Dry storage; Mizuhopecten yessoensis; Energy metabolism, Mitochondrial
morphology

EAFTA 2023, On-site/online, May 17th-20th, 2023, Morioka, Japan

Effects of stable chlorine dioxide combined with slightly acidic electrolyzed
water on quality of large yellow croaker (Pseudosciaen crocea) during cold
storage
Weiqing LAN1,2,3, ○Qi ZHOU1, Xinyu ZHAO1, Jing XIE*1,2,3
1, College of Food Science and Technology, Shanghai Ocean University, Shanghai 201306,
China
2, Shanghai Aquatic Products Processing and Storage Engineering Technology ResearchCenter, Shanghai 201306, China
3, National Experimental Teaching Demonstration Center for Food Science and Engineering(Shanghai Ocean University), Shanghai 201306, China*Corresponding author: jxie@shou.edu.cn

The quality changes of cold-stored large yellow croaker (Pseudosciaena crocea) werestudied by using stable chlorine dioxide (ClO2) combined with slightly acidic
electrolyzed water (SAEW). The results of microbial indicators demonstrated that ClO2
and SAEW treatment inhibited microbial growth. Moreover, ClO2+ SAEW treatment
had lower values of pH, total volatile basic nitrogen (TVB-N) and malondialdehyde(MDA) content, and higher water-holding capacity (WHC). Through the results of freeamino acids (FAAs) and trimethylamine (TMA), ClO2+ SAEW treatment could delay
the degradation of umami amino acids and the accumulation of bitter amino acids oflarge yellow croaker. According to microbial indexes, sensory analysis and K-value,compared with sterile distilled water treatment, ClO2+ SAEW treatment prolonged theshelf-life of large yellow croaker for another 4 days. Therefore, ClO2+ SAEW treatment
is a potential method to maintain the quality and flavor of large yellow croaker duringcold storage.
Keywords: Stable chlorine dioxide, Slightly acidic electrolytic water, Large yellow
croaker, Quality, Free amino acids

EAFTA 2023, On-site/online, May 17th-20th, 2023, Morioka, Japan

Insight into the antibacterial activity and mechanism of allicin andantioxidant of bamboo leaves against Shewanella putrefaciens
Weiqing LAN1,2,3, ○Zhe SHAO1, Jintao DU1, Jing XIE1,2,3,*
1, College of Food Science and Technology, Shanghai Ocean University, Shanghai 201306,
China
2, Shanghai Aquatic Products Processing and Storage Engineering Technology ResearchCenter, Shanghai 201306, China
3, National Experimental Teaching Demonstration Center for Food Science and Engineering(Shanghai Ocean University), Shanghai 201306, China*Corresponding author: jxie@shou.edu.cn

The antibacterial mechanism of allicin and antioxidant of bamboo leaves against
Shewanella putrefaciens (S. putrefaciens) was studied. The inactivation effect of allicin
combined with AOB against S. putrefaciens was evaluated by the growth curve. Thechanges in Na+ K+-ATPase activity, confocal laser scanning micrographs (CLSM),
nucleic acid and protein release were measured to evaluate the inactivation mechanism.
Scanning electron microscopy (SEM) was used to observe the morphology of bacterial
cells. Crystal violet biofilm was reflected the ability to inhibit the formation of biofilm.
The results showed that S. putrefaciens was effectively inactivated after single andcombined treatments of allicin and AOB, and the inactivation effect of combined
treatment was the best. The cell wall and cell membrane of bacteria were destroyed byallicin and AOB, resulting in the release of nucleic acid, protein. The results of Na+ K+ATPase activity showed that allicin and AOB could change cell membrane permeability
and destroy cell structural proteins. The results of SEM further proved the destructive
effects of allicin and AOB on the bacterial morphology. Therefore, the combination ofallicin and AOB can be used as a simple and effective method for the preservation of
aquatic products.

Keywords: Allicin, AOB, antibacterial mechanism, S.putrefaciens

EAFTA 2023, On-site/online, May 17th-20th, 2023, Morioka, Japan

Effects of seaweed extracts on the quality and shelf life of Nile tilapia(Oreochromis niloticus) fillets during frozen storage○Faria Afrin1,2, Md. Monirul Islam2, Md. Golam Rasul2, Chunhong Yuan3,4, A. K. M. Azad

Shah*2
1, United Graduate School of Agricultural Sciences, Iwate University, Ueda 3-18-8, Morioka,
Iwate 020-8550, Japan2, Department of Fisheries Technology, Bangabandhu Sheikh Mujibur Rahman Agricultural
University, Gazipur-1706, Bangladesh
3, Department of Food Production and Environmental Management, Faculty of Agriculture,
Iwate University, Ueda 3-18-8, Morioka, Iwate 020-8550, Japan4, Agri-Innovation Center, Iwate University, Morioka, Iwate 020-8550, Japan*Corresponding author: azad@bsmrau.edu.bd

This study investigated the effects of seaweed (Padina tetrastromatica, Sargassum muticum,and Spatoglossum asperum) extracts on the quality and shelf life of Nile tilapia(Oreochromis niloticus) fillets during frozen storage for a period of 20 weeks. Fish fillets
were dipped in seaweed ethanolic extracts solution (2%, w/v) separately for 10 min at 4°C.
The control and seaweed extracts-treated fillets were stored at −18±1°C, and chemical,
microbiological and sensory attributes were evaluated periodically. Results showed that P.
tetrastromatica extract-treated fillets significantly (P < 0.05) delayed the increment of pH,
total volatile basic-nitrogen, peroxide value, and thiobarbituric acid reactive substances inNile tilapia fillets than other seaweed extracts-treated and control fillets. The aerobic plate
count of fish fillets was within acceptable range during the storage period. Moreover, P.
tetrastromatica extract-treated fillets displayed comparatively better sensory attributes thanother seaweed extracts-treated fillets as well as control fillets. Results of this study indicatedthat ethanolic extract of P. tetrastromatica retains the quality and extends the shelf life offrozen Nile tilapia fillets for 8 weeks longer than control fillets. Thus, it can be concluded
that P. tetrastromatica extracts can be used as a natural preservative for Nile tilapia fillets.

Keywords: Nile tilapia, Seaweed extracts, Shelf life, Sensory evaluation, Frozen storage

EAFTA 2023, On-site/online, May 17th-20th, 2023, Morioka, Japan

Screening the tyrosinase inhibitory activity of red algae extracts –applying
in inhibition blackening of shrimp
Han The Nguyen, Quyen Le Vu, Huong Thi Thu Dang*
Faculty of Food Technology, Nha Trang University, 02 Nguyen Dinh Chieu, Nha Trang,
Vietnam*Corresponding author: dangthithuhuong@ntu.edu.vn

Tyrosinase (polyphenol oxidase -PPO) causes browning in fruits, vegetables, andblackening in seafood, especially shrimp. The activity of the tyrosinase can be inhibitedby bioactive substances which can be found in red algae. In the present study, PPO
inhibitory ability of the extract from 5 species of red algae (Gelidiella acerosa,
Gracilaria Salicornia, Acanthophora spicifera, Hypnea pannosa, Kappaphycus
alvarezii), collected in Nha Trang Vietnam were investigated. Screening was done under
the same conditions for 5 species (methanol 80% for 60 minutes, at 60C withmaterial/solvent (1:20). The highest inhibitory ability of PPO was found in Gelidiellaacerosa extract. The best extract was then fractioned through different polar solvents (nhexane, ethyl acetate, butanol, distilled water) to determine the segment with the highest
tyrosinase inhibitory ability. The optimum extraction conditions of Gelidiella acerosa
were found in distilled water during 60 minutes at 60C with material/solvent (1:40),
fractioned through ethyl acetate. Gelidiella acerosa extract increased shelf-life of shrimp
compared to the control sample.

Keywords: Red algae, extract, tyrosinase inhibitory ability, shrimp

EAFTA 2023, On-site/online, May 17th-20th, 2023, Morioka, Japan

Poster:Extracting biomaterials from coconut shells and plant disease control
applications
Duy Nguyen Xuan 1*, Khoi Nguyen Le Ngoc2 and Hanh Tran Thi My1
1

Faculty of Food Technology, Nha Trang University, Khanh Hoa Province, Vietnam
Faculty of Chemical Engineering, Vietnam National University Ho Chi Minh – University of
Science, Ho Chi Minh city, Vietnam*Corresponding authors: myhanhtt@ntu.edu.vn; duynx@ntu.edu.vn
2

The coconut is a popular tropical crop in Vietnam. The coconut can produce a wide variety of
products. Coconut shells are considered a by-product of the processing of coconut products.
The coconut shell contains many biologically active compounds. At present, however, they
are not used properly and effectively. The study was carried out to obtain valuable substancesfrom coconut shells and to find applications in the control of plant diseases. The pyrolytic
method at the appropriate temperature is used to extract biologically active substances fromthe coconut shells. Results of the study indicate that extracts obtained from coconut shells
have high antibacterial and antifungal activity. The typical class of substances found in
extracts of the organic acid and phenolic groups. GC-MS analysis found that 17 groups ofsubstances are phenolic derivatives. These are the principal substances that play an importantrole in the biology of the preparation. The results of early testing of preparations in theprevention and treatment of diseases on plants are quite good. These results open up the
potential for using biological preparations obtained from coconut shells in controlling disease
on plants.
Keywords: Biological preparations, coconut shells, clean agriculture, disease control

EAFTA 2023, On-site/online, May 17th-20th, 2023, Morioka, JapanThe water-soluble protein from walleye pollock attenuates obesity inducedby high-fat, high-sucrose diet.
○Shu Shimada1, Hiroki Saeki1, and Ga-hyun Joe 1*
1, Laboratory of Marine Food Science and Technology, Faculty of Fisheries Sciences,
Hokkaido University, Minato 3, Hakodate, Hokkaido 041-8611, Japan*Corresponding author: Gahyun@fish.hokudai.ac.jp[Objective] Obesity resulting from irregular eating habits induce chronic inflammationand various non-communicable diseases (NCDs). The waste effluent generated during
surimi manufacturing process contains a large amount of water-soluble protein (WSP)
from fish meat, but it is underutilized and mostly discarded. In this study, In this study,
we examined the impact of WSP on metabolic syndrome associated with obesity, todevelop advanced methods for utilizing WSP.
[Method] Waste effluent from surimi production was used to extract WSP. Frozen
walleye pollack muscle was suspended in 50 mM NaCl and pressed with a screw-press
to collect the effluent. WSP was then precipitated by shifting the pH from 6.8 to 4.0 to
7.0, and finally, collected by centrifugation. Male Wistar rats (aged 5 weeks) were fed
with one of three types of diets for 65 days: (1) Control group (AIN-93G, 7% soybean
oil, 10% sucrose and 20% casein), (2) HFHS group (7% soybean, 27% lard, 40% sucroseand 20% casein), and (3) HFHS + WSP group (7% soybean, 27% lard, 40% sucrose,
10% casein and 10% WSP). After the feeding experiment, plasma and tissues werecollected and weighed. Also, lipid metabolism markers and adipokines (adiponectin andleptin) in the plasma and liver lipid content were measured.
[Results] There was no significant difference in total intake and total calories among thethree groups during the experimental period. The HFHS group showed a significant
increase in body weight compared to the control group, indicating the induction ofobesity. On the other hand, WSP-ingested rats tended to lose weight 30 days afteringestion, and at the end of the experiment, the body weight was about 5% lower thanthat of the HFHS group. In addition, liver weight, adipose tissue weight, and plasma
triglycerides tended to decrease in the WSP-fed group, while there was no difference
between the groups in liver cholesterol, triglycerides, and free fatty acid concentrations.
Interestingly, WSP intake improved the ratios of leptin and adiponectin, suggesting thatthe ingestion of WSP is involved in their secretion and improves obesity.

Keywords: Chronic inflammation, Obesity, water-soluble protein, metabolism.

EAFTA 2023, On-site/online, May 17 th-20 th, 2023, Morioka , Japan

Reduction of fish roe allergy riskin triploid female rainbow trout (Oncorhynchus mykiss).
○Saki Watanabe 1, Yutaka Shimizu 1, Shintaro Ishida 1, Yukiho Sasaoka 2, Takashi Todo 1,
Naoshi Hiramatsu 1, Takafumi Fujimoto 1, Gahyun Joe 1, Hiroki Saeki* 1
1, Laboratory of Marine Food Science and Technology, Faculty of Fisheries Sciences,
Hokkaido University, Minato 3, Hakodate, Hokkaido 041-8611, Japan2, Hokkaido Research Organization, Fisheries Research Department Central Fisheries
Research Institute, Hamanaka 238, Yoichi, Hokkaido 046-8555, Japan*Corresponding author: saeki@fish.hokudai.ac.jp【Background and Purpose】
Fish roe is one of the most allergenic seafood, with high incidence of food allergy inJapan. Salmon roe is the primary causative agent of fish roe allergy and its labeling is
recommended by the Japanese Food Labeling Act. The major allergen of fish roe is β'component (β'-c), one of the yolk proteins. Vitellogenin (Vg), a precursor of the yolk
proteins, also exhibits allergenicity similar to that of β'-c. Vg is synthesized in the liver ofsexually mature female fish and is transferred to oocytes via the bloodstream. As a result,if such blood remains in the muscle, fish-roe-allergic patients are at risk of developing anallergic reaction to fish meat. Therefore, the purpose of this study is to identify types of
salmonid fish that pose a lower risk of fish roe allergy in their meat. This study focusedon triploid female rainbow trout (Oncorhynchus mykiss), which are commonly available
on the market as sterile fish, and investigated their potential to cause fish roe allergy.
【Methods】
Twenty-two triploid female rainbow trout (TFRT 1-22) samples were sampled from two
trout farms in Hokkaido and analyzed in this study. Muscle, gonad, liver and kidney ofeach fish were investigated as follows: TRFT 21 and 22 were exclided in (3).
(1)Vg concentration was measured by Sandwich ELISA using anti-rainbow trout β'-crabbit IgG (anti-RTB), where the Vg concentration exceeds 0.25 µg/g indicates apotential risk of causing fish roe allergy based on Japanese Food Labeling Act.
(2)Cleavage state of Vg was analyzed using Immunobloting (IB) with anti-RTB.
(3)Development stage of oocytes was assessed by HE staining of gonad. (4)Polyploidy
was measured by flow cytometry using liver as a sample.
【Results】(1)In muscle, none of samples indicated a potential risk of causing fish roe allergy, but
four out of twenty-two samples (TFRT 10, 13, 21 and 22) showed a potential risk inorgans other than muscle. (2)Vg or its fragment were detected in 3 samples (TFRT 13,
21 and 22). (3)Gonads of 3 samples (TFRT 9, 10 and 13) showed developed oocytes atoil drop stage or yolk globule stage, which could also be observed visually. (4)Twosamples (TFRT 13 and 21) were dipoid, while all other samples were triploid.
【Conclusion】
Fish meat of triploid female rainbow trout can be eaten with a low risk of causing fishroe allergy, even though some samples may contain Vg due to triploidization failure orother reasons. However, the risk can be further minimized by visually inspecting the
gonad development and eliminating samples with developed gonads.
Keywords: fish roe allergy, β'-component, vitellogenin, triploid, rainbow trout

EAFTA 2023, On-site/online, May 17th-20th, 2023, Morioka, JapanThe enhancement effects of roselle anthocyanin-based film on shrimp
texture by water distribution controlling and protein structure maintenanceJiayin Huang1,2, Yaqin Hu2*
1, College of Biosystems Engineering and Food Science, Zhejiang University, Hangzhou,
3100582, College of Food Science and Technology, Hainan Tropical Ocean University, Sanya,
572022*Corresponding author: 1004346262@qq.com
The perishable properties of food products and emerging eco-friendly requirements have
driven the development of biodegradable active packaging. Several studies have confirmed
the excellent preservation behaviors of active films by determination of primarybiochemical parameters, such as pH, TVB-N and TVC. However, investigating some otherinner quality traits are of vital importance to understand its preservative mechanism.
In

this

study,

theenhancement

effectsofpolyvinyl

alcohol/hydroxypropyl

methylcellulose/roselle anthocyanin-based (PHR) film on textural profiles of Penaeus
vannamei were studied. Results demonstrated that shrimp packaged by PHR film exhibited
better texture with higher hardness, springiness, chewiness and resilience values. The PHR
films exerted positive influences on water migration by reducing drip loss and promoting
conversion of water state. Meanwhile, the PHR film delayed protein denaturation throughincreasing myofibrillar protein and sulfhydryl group contents, decreasing trichloroacetic
acid-soluble peptide contents and maintaining protein secondary structure. Furthermore, thecorrelation analysis revealed the close relationship among texture, water and protein
profiles, indicating the better texture characteristics were resulted from the positive
influences of PHR film exerted on the water state and protein structure of shrimp.
Overall, out study proved that the PHR film was a novel potential active packaging forshrimp texture modification.
Keywords: Active film, Textural profiles, Water migration, Protein conformation,
Preservation mechanism

EAFTA 2023, On-site/online, May 17th-20th, 2023, Morioka, Japan

Involvement of MMP-9 in collagen degradation of sea bass (Lateolabrax
japonicus): cloning, expression and characterization○Ru-Qing Yang1, Yu-Lei Chen1,2, Le-Chang Sun1,2, Wei Ou1,2, Hai-Yan Liu1,2,
Ling-Jing Zhang1,2, Min-Jie Cao1,2*
1, College of Ocean Food and Biological Engineering, Jimei University, Xiamen, 361021,
China
2, Collaborative Innovation Center of Marine Food Deep Processing, Dalian Polytechnic
University, Dalian, 116034, China*Corresponding author: mjcao@jmu.edu.cn
Disintegration of intramuscular connective tissue is responsible for post-mortem
tenderization of fish muscle during chilled storage. Matrix metalloproteinase-9 (MMP-9)
was reported to be involved in this process, while the mechanism has not been identified.
In the present study, purified type I and V collagens from the connective tissue of fishmuscle were firstly prepared. These two kinds of collagens comprise three polypeptide
chains (α), forming a typical triple helical domain as determined by circular dichroism.
The complete coding region of MMP-9 containing an ORF of 2070 bp encoding 689
amino acid residues was then cloned. The recombinant MMP-9 catalytic domain(rcMMP-9) was expressed in E. coli and exhibited high hydrolyzing activity towardgelatin. rcMMP-9 degradation to type V collagen was more effective than that to type I
collagen at 4 °C. The enzymatic activity of rcMMP-9 was highly pH-dependent, and its
enzymatic activity under neutral and basic conditions were higher than that under acidic
conditions. Metal ion Ca2+ was necessary for the maintenance of rcMMP-9 activity,
whereas Zn2+ inhibited its activity. Our present study indicated that MMP-9 is
responsible for the disintegration of intramuscular connective tissues by cleaving type V
collagen during post-mortem tenderization of fish muscle.
Keywords: Lateolabrax japonicus, collagen, matrix metalloproteinase-9, enzymatic
activity, softening

EAFTA 2023, On-site/online, May 17th-20th, 2023, Morioka, Japan

Expression of bioactive polyphenol oxidase of Litopenaeus vannamei and its
characterization○Qian Hong1, Yu-Lei Chen1, Duanquan Lin1, Ling-Jing Zhang1, Kai-Yuan Cao2, Yi-Ming
Liu1, Ling Weng1, Le-Chang Sun1, Min-Jie Cao1*
1, College of Ocean Food and Biological Engineering, Jimei University, Xiamen, China,
361021
2, Department of Biological Science, National University of Singapore, Singapore, 117558*Corresponding author: mjcao@jmu.edu.cn
Polyphenol oxidase (PPO) is an enzyme that plays a critical role in decrement of shrimp
quality. Purification of native PPO is challenging because of its low content and unstability.
To obtain active PPO and elucidate its enzymatic properties, PPO from Litopenaeus vannamei(Lv-PPO) was cloned, expressed by constructing an Escherichia coli prokaryotic expression
system and then purified by affinity column chromatography. The Lv-PPO gene was 2076 bp
in length and encoded 691 amino acid residues with deduced molecular weight of 78.8 kDa.
The recombinant Lv-PPO (rLv-PPO) with a molecular weight of ~85.0 kDa was partiallydissolved while mainly in inclusion body, and its sequence was verified by LC-MS/MS. rLvPPO was biologically active with an optimal temperature of 40℃ and an optimal pH of 6.0.
Its denaturation temperature was 54.2 ± 0.7℃ as measured by circular dichroism. Metal ionssuch as Cu2+ and Zn2+ altered the activity of rLv-PPO by influencing its secondary and tertiary
structures. rLv-PPO showed catalytic activity towards L-Dopa and catechol. Its enzymaticcharacteristics were studied by kinetic analysis. A specific polyclonal antibody against rLvPPO was prepared. Western blot analysis revealed that PPO levels were highest in
hemolymph, followed by telson, carapace, and eyestalk. A trivial amount of PPO was
identified in muscle while it could not be detected in hepatopancreas and midgut. Our present
study expressed a biological active rLv-PPO for the first time. A specific polyclonal antibody
against rLv-PPO was prepared for the prospect of assisting future studies on the enzymatic
properties of PPO, and exploring its action mechanism in shrimp melanosis.
Keywords ： polyphenol oxidase, Litopenaeus vannamei, melanosis, protein expression,
enzymatic property, polyclonal antibody

EAFTA 2023, On-site/online, May 17th-20th, 2023, Morioka, Japan

Properties of Pacific white shrimp (Litopenaeus vannamei) collagen and its
degradation by endogenous proteinases during cold storage○Xu-Chen Xiao1#, Duanquan Lin1#, Kai-Yuan Cao2, Le-Chang Sun1, Yu-Lei Chen1, Ling
Weng1, Ling-Jing Zhang1, Min-Jie Cao1*
1, College of Ocean Food and Biological Engineering, Jimei University, Xiamen, China,
361021
2, Department of Biological Science, National University of Singapore, Singapore, 117558#
These two authors contributed equally to this manuscript.
*Corresponding author: mjcao@jmu.edu.cn

Many factors are responsible for the diminished quality of shrimp during cold storage,
while the role of collagen has rarely been studied. This study therefore investigated therelationship between collagen degradation and changes of textural properties of Pacificwhite shrimp, and its hydrolysis by endogenous proteinases. The textural properties ofshrimp decreased gradually along with disruption of shrimp muscle tissues, and thechewiness property of shrimp muscle showed a linear relationship with collagen contents
in muscle during 6-day-storage at 4ºC. Pepsin-solubilized collagen in shrimp muscle
consisted of one α1 chain and two α2 chains, revealing a typical tripeptide sequence (i.e.,
Gly-X-Y) in their molecules. In addition, collagen could be hydrolyzed by crudeendogenous proteinases extracted from shrimp hepatopancreas, and serine proteinase
plays a critical role in the process. These findings strongly suggested that the quality
reduction of shrimp during cold storage is closely associated with collagen degradation.
Keywords: collagen degradation; endogenous proteinase; shrimp preservation; cold
storage; quality degradation.

EAFTA 2023, On-site/online, May 17th-20th, 2023, Morioka, Japan

Flash Boil-Shucking of Yesso Scallop (Mizuhopecten yessoensis) and Its
Effect on Biochemical, Sensorial and Ultrastructural Properties of
Adductor Muscle During Refrigeration○Yabin Niu1, Shiliang Dong1, Nobuyoshi Wada2, Huamao Wei3, Tetsuro Yamashita2,
Koichi Takaki4,5, Chunhong Yuan2,5*
1 United Graduate School of Agricultural Sciences, Iwate University, Ueda 3-18-8, Morioka,
Iwate 020-8550, Japan
2 Faculty of Agriculture, Iwate University, Ueda 3-18-8, Morioka, Iwate 020-8550, Japan
3 College of Food and Pharmaceutical Sciences, Ningbo University, Ningbo, Zhejiang
315211, China
4 Faculty of Science and Engineering, Iwate University, Ueda 4-3-5, Morioka, Iwate 0208551, Japan
5 Agri-Innovation Center, Iwate University, Ueda 3-18-8, Morioka, Iwate 020-8550, Japan.
*Corresponding author:chyuan@iwate-u.ac.jp
Scallop adductor muscle has great popularity for raw consumption in Japan. However, thetraditional method of shucking live scallops with a knife requires skill and is time-consuming.
Sushi restaurants have adopted an alternative technique known as instantaneous boiling,which offers a convenient shucking method for live scallops. However, it is not clear about
the effects of this treatment on the quality of its main edible part, adductor muscle.
The objective of this study was to examine the effects of instantaneous boiling on the qualityof raw-edible scallop adductor muscle, with a specific focus on flavor-enhancing components
and ultrastructure. It was observed that the thermal treatment employed during instantaneous
boiling contributed to the formation of flavor-enhancing nucleotides primarily in the externallayer of the adductor muscle. Adenosine monophosphate content increased with the boilingtime, which significantly increased to 1.47 ± 0.42 µmol/g in the 30 s-boiled group comparedwith 0.38 ± 0.13 µmol/g in the control group (p ˂ 0.05). The rapid adenosine monophosphate
accumulation after boiling, leads to a more rapid accumulation of inosine and hypoxanthineduring subsequent refrigeration, which could bring bitterness to aquatic products. Despite thepossibility of a decrease in total free amino acids, the proportion of free amino acids relevantto the flavor of scallop adductor muscle remained largely unaffected by instantaneous boiling.
Instantaneous boiling has been found to potentially induce a hardening effect on the externallayer of scallop adductor muscle. At the ultrastructural level, the muscle fibers demonstrate atendency to aggregate transversely and contract longitudinally, with the extent of thesechanges depending on the duration of heating. Based on sensory evaluation, the 30 s-boiled
samples were most suitable for immediate consumption, otherwise boiling for 8 seconds has
been suggested as an alternative.
Keywords: Shellfish, Shucking, Ultrastructure, Flavor components, Sensory evaluation

EAFTA 2023, On-site/online, May 17th-20th, 2023, Morioka, Japan

Exploring Wakame Recipe Information and Cross-Region Food culture Analysis
for seaweed marketing○Natsumi Shimoyama1, Kazuma Sugawara2,Chunhong Yuan3
1, Fishery Innovation Program, Division of Regional Innovation and Management, Graduate
School of Arts and Sciences, Iwate University, Ueda 3-18-8, Morioka, Iwate 020-8550, Japan2, Marine Chemical Resource Development, Faculty of Fisheries Sciences, Hokkaido
University, Hakodate 041-8611, Hokkaido, Japan3, Faculty of Agriculture, Iwate University, Ueda 3-18-8, Morioka, Iwate 020-8550, Japan*Corresponding author:chyuan@iwate-u.ac.jp
Iwate Prefecture is one of the leading producers of wakame seaweed in Japan, but the
production volume has been declining in recent years due to the low unit price. In order toimprove the market value of high quality wakame, there is possibility to develop a marketingstrategy for export which is rarely done at present. However, wakame is mainly consumed inJapan and South Korea, and many countries do not have a food culture. In this study, we
analyzed the existing seaweed recipes from the global recipe service, and investigated how
seaweed is eaten and consumer preference corelated to its food culture of each country for
expanding overseas markets of seaweed.
The recipe dataset (n=494,963) provided to Kaggle in 2017 by Yummly, a global online
cooking recipe service was used. The data were categorized according to, (1) regions, (2)search words, (3) frequency of use of ingredients and seasonings, and (4) types of dishes
included in the dish names for seaweed recipes. Using R, we classified? words such as
ingredient names and regions included in each variable, and judged how seaweed was cookedfrom the frequency.
As a result of extracting data containing “wakame seaweed” in the variable “ingredient”
from the dataset, 73 cases (0.015%) were obtained. From this, it turned out that seaweed
dishes are rarely posted on English-speaking recipe sites. On the other hand, there were 281
cases on “seaweed”, which include another species. (1) We analyzed the area and other searchwords from the variable "search tag". As a result, “Japanese'” was the most common area with
35 search tags, followed by “Chinese” with 3, “Korean” with 2, “Cajun”, “Irish” and“French” with 1. (2) The most frequently searched words were “vegetarian” in 28 cases,“dinner” in 26 cases, “soup” in 25 cases, “vegan” in 23 cases, and "salad'' in 21 cases. It cansee the keywords that are easy for users to search for seaweed recipes. This result shows that
there are many tags indicating that it is a Japanese-style dish. Since there are many words
such as vegetarian in the tag, it is thought that wakame recipes is targeting vegetarians. (3) As
a result of examining the frequency of use of each ingredient and seasoning from the variable“ingredients”, the food ingredients were “water (30 cases)”, “onion” (26 cases), “garlic” (16cases), “carrot” (13 cases), “green onion” (13 cases). Seasonings: “soy sauce” & “shoyu” (42cases), “salt” (28 cases), “pepper” (25 cases). It is thought that the frequency increased
because water is often used as a soup. And Few combinations with meat or fish. (4) As aresult of analyzing the type of recipe from the variable "name", there were 25 "soup" recipes,
15 of which used miso. There were 22 cases for “salad” and 7 cases for “noodle”.
Based on these results, recipes using wakame seaweed are limited on English recipe sites,
and it is thought that the development of recipes that match the taste of each country has notprogressed. There are many search tags with vegetables and recipes for vegetarians, and it isconsidered that the current wakame is for people who like vegetables. To next step, we would
like to conduct the same analysis on recipe information in formats other than English andclarify the differences in recipe information for each country.
Keywords: Wakame seaweed, Recipe, Data analysis, Food culture

EAFTA 2023, On-site/online, May 17th-20th, 2023, Morioka, Japan

Efect of " uzumaki process " on post mortem quality changes incultured red sea bream at 4 ℃○Koma Nakahata1, Keisuke Katsura2, Xin Lu3, Chunhong Yuan*1.

1.
2.

Faculty of Agriculture, Iwate University, Ueda 3-18-8, Morioka, Iwate 020-8550,
Japan
Japan Uzumaki Association, 726 Hachijima-cho, Nishi-ku, Kumamoto, 860-0057

3.

Faculty of Science and Engineering, Iwate University, Ueda 4-3-5, Morioka, Iwate 020-8551,
Japan*Corresponding author:chyuan@iwate-u.ac.jp
In Japan, freshness preservation by Spinal cord destruction is widely practiced when wild or farmed
live fish are landed. This slaughtering technique involves the destruction of both the brain and spinal
cord, but it requires a high level of skill to perform.
To address this challenge, the Uzumaki process was developed. This method involves using a tube tosuction the spinal cord out of the brain and is considered more reliable than traditional spinal cord
destruction. In our study, we analyzed the postmortem changes in ATP, pH, rigor index, andimpedance in red sea bream, a popular Japanese marine product.
Ten live cultured red sea bream were transported to lab and stored at 4°C for 72 hours. Four of the
fish underwent spinal cord destruction while the remaining six were processed using the Uzumaki
method. One half of each group was used to measure ATP and pH levels, while the other half wasused for rigor index and impedance measurements and SEM image observation. HPLC (Asahipak
GS-320HQ, 25°C, 0.6 mL/min) was used to measure ATP-related compounds, and a Fish Analyzer(Yamato seikou, DFA100) was used to measure impedance values.
The results revealed that the onset of rigor mortis was delayed by approximately 10 hours in the
Uzumaki process group compared to the spinal cord destruction group. Additionally, the decrease in
ATP content was slower in the Uzumaki process group, indicating a slower progression of
postmortem rigor. While the pH decreased to 6.3 and 6.4 in both groups, there was no obvious
difference observed. SEM images showed muscle contraction and cell membrane degradation, which
could be associated with impedance fluctuations. Based on these findings, the Uzumaki process wasdeemed more effective in preserving the freshness of the fish compared to spinal cord destruction.
However, it is necessary to further investigate the relationship between freshness retention and spinal
cord removal rate in future studies.
Keywords: red sea bream, freshness, Spinal cord destruction, bioimpedance, ATP related
compounds

Investigation of evaluation index of Wakame by Measuring Physical
Properties and Analyzing Functional Components.
○

Tomoya Suzuki1, Kazuma Sugawara1, Lu Xin2,

Gouki Mikawa1, Chunhong Yuan1
1. Faculty of Agriculture, Iwate University
2. Faculty of Science and Engineering, Iwate University
Objective: The market has introduced a system to certify wakame of the first grade landed
in Iwate Prefecture. However, the grading of wakame is based on the findings of field
staff based on their many years of experience. There are very few examples of wakame
grading based on quantitative methods, so recently there has been a need to introduce anevaluation method that is more objective and quicker through mechanization andnumerical methods. In order to develop a grading system for wakame seaweed using deeplearning by correlating image data with the results of biochemical analysis, we conducted
physical property measurements and biochemical analysis on wakame seaweed fromseveral production areas and examined effective indices for grading evaluation.
Methods: Salted and frozen wakame from Iwate and Miyagi prefectures were used forthe analysis. 1 x 3 cm pieces of wakame were cut and used for tensile strength
measurements (rheometer, speed 300 mm/min). SPAD and L*a*b* values were measured
to determine color-tone. Lyophilized powdered wakame seaweed was used for component
analysis. The Folin-Ciocalteu method was used to determine total polyphenol
concentration. Fucoxanthin was extracted by the same method as polyphenols, and its
content was calculated by measuring the absorbance value at 449 nm.
Results: Correlation analysis of the measured indices showed a significant correlation
between SPAD and b* values, SPAD is a simple measure of chlorophyll content and Lab
is a value indicating the optical properties of the object. Since both indicators are related
to color, it was suggested that they could be correlated with image data. Significant
regional differences were observed in the correlation between optical properties andfucoxanthin concentration. In addition, there are some differences in correlations due todifferent trends in component content and physical properties depending on the region of
production, so it is necessary to form an evaluation system that takes regional
characteristics of wakame into account.
Keywords: Wakame, Seaweed, Food technology, Iwate prefecture

EAFTA 2023, On-site/online, May 17th-20th, 2023, Morioka, Japan

Effect of short-term rearing on changes in ATP related compounds andglycogen content during storage at -1℃
Sota Hayashi1
1, Faculty of Agriculture, Iwate University, Ueda 3-18-8, Morioka, Iwate 020-8550, Japan*Corresponding author: g0123041@iwate-u.ac.jp

Short-term rearing is carried out with the aim of reducing fishing stress at the time oflanding, thereby reducing the loss of vitality of the product by the fishery product.
Among bivalves, there is a report that confirms the effect of rearing scallops, but there islittle knowledge about oysters. In this study, we examined the effect of short-term
rearing on oysters (Crassostrea gigas). Rearing oysters and non-reared oysters werestored in a refrigerator at -1°C for 14 days, and it were experimented on storage days 0, 1,
3, 7, and 14. ATP-related compounds, arginine phosphate, and glycogen were extracted,
and changes in vitality over time during storage were examined. The ATP content was
clearly higher in the 3 conditions with rearing oyster than without rearing on the 0th to
1st days of storage. The A.E.C value was higher for the oysters that were reared on the
0th day of storage. Compared to the arginine phosphate content in scallops, the arginine
phosphate content was much lower, and we were unable to find a relationship withchanges in ATP content over time. Glycogen was measured in this experiment as anindirect indicator of vitality, given that ATP is produced by glycogen. No relationshipwas found with ATP-related compounds or arginine phosphate, and no relationship withvitality could be found. It was suggested that short-term rearing might reduce thedecrease in vitality of oysters on the 0th day of storage from the results of ATP-related
compounds.
Keywords: oyster, short-term rearing, ATP-related, glycogen, A.E.C. value, vitality,
syorage

EAFTA 2023, On-site/online, May 17th-20th, 2023, Morioka, Japan

Detection of ATP related compounds for judging Anisakis death killed byliquid nitrogen freezing○Yuya Ota1

1Faculty of Agriculture, Iwate University, Ueda 3-18-8, Morioka, Iwate 020-8550, Japan
Anisakis is a dangerous parasite that poses a major problem when eating marine products
raw in Japan, and is counted individually in food poisoning statistics. Furthermore, with theaddition of Japanese food to the list of intangible cultural heritage, there is a possibility thatthe incidence of anisakiasis will increase as opportunities to eat raw fish increase worldwide.
For this reason, guidelines have been established in various countries. For example, in the US
FDA, Anisakis can be killed by freezing at -20°C or below for 7 days, freezing to -35°C andstoring it at that temperature for 15 hours, or - It indicates that 24 hours of storage at 20°C or
below is necessary but not suitable for large fish (e.g. fleshy fish over 6 inches). If theguidelines are followed, long-term frozen storage is required, and depending on the fish
species, the quality will drop all at once.
Therefore, if Anisakis can be killed instantaneously by freezing liquid nitrogen,denaturation due to freezing will be less and it will be possible to eat something closer to thefinal taste and texture. However, there are few studies on the death of Anisakis. In previous
research, the emphasis was placed on visual inspection to determine death, and there is also amethod that uses malachite green to stain dead or injured individuals, but the result is that the
injured part There is no certainty because it is not dyed. Therefore, this time, we compared
Anisakis from ATP-related compounds under various conditions and verified whether there
was a difference in vitality.
Anisakis was harvested from donko and squid. Freezing was frozen at -70°C for 3, 6, and15 minutes using a liquid nitrogen freezer. As a control, ATP-related compounds were also
extracted from unfrozen Anisakis.
As a result, only ATP and ADP were detected in Anisakis without freezing. In addition to
ATP and ADP, AMP was also detected in 3 and 15 min freezing. However, after freezing for 6
minutes, the percentage of ATP decreased, but AMP was not detected.
From these results, it was found that there was a difference in vitality between those that werefrozen and those that were not. However, the fact that the ratio of the 6 min individuals to theunfrozen specimens was closer to that of the other frozen specimens may be due to thedifference in the vitality of Anisakis before freezing.
Therefore, it was suggested that the difference in the vitality of Anisakis could be observedby analyzing the components of ATP-related compounds. In the future, by establishing amethod for extracting ATP-related compounds from Anisakis, it is expected that ATP-related
compounds will be used as indicators of vitality in Anisakis and as a method for determining
life and death.
Keywords: Anisakis, liquid nitrogen freezing,Effect of ultrasound in conjunction with bromelain on the tenderness of
Litopenaeus Vannamei
Xinyi Zhou1, Lanxiang Su1, Huamao Wei1, Wenge Yang1, Tao Huang1, Chunhong
Yuan2(1. College of Food and Pharmaceutical Sciences, Ningbo University, 315211; 2.
Faculty of Agriculture, Iwate University, Ueda 3-18-8, Morioka, Iwate 020-8550,
Japan)[Objective] Penaeus vanname is one of the most valuable aquaculture commodities globally
due to its high nutritional and economic value. In China, the aquaculture output of Penaeus
vanname has shown an upward trend in the past three years, reaching 1.98 million tons in
2021. In recent years, shrimp can be processed into dried, frozen, boiled, and fried shrimp
products. Among them, dried shrimps are more and more popular with consumers, because itis easy to carry, eat and store, as well as have unique flavor. However, the meat isaccompanied by physical and chemical changes during the heating process, such as protein
aggregation, severe oxidation, cross-linking, or increased disulfide (S-S) content, which
increase the firmness of the meat and decrease the susceptibility of proteins during
gastrointestinal digestion. Previous studies have found that sonication or enzymatic treatment
can improve digestibility by increasing tenderness. To our knowledge, numerous publications
are following closely in the poultry meat industry, but few have focused on the effect ofultrasound combined with enzyme treatments on aquatic products. So, this study aimed toinvestigate the combined effect of ultrasound with bromelain on the tenderization of dried
shrimp and analyze the possible mechanism of tenderization.
[Methods] The samples were immersed in a bromelain solution (concentration 10 to 30 U/gwith 2% salt, sample-to-solution ratio 1:1 (w/v)). At the same time, the packed meat wassonicated for 5 minutes (power 50-250 W, temperature 30-70 °C). During sonication, thetemperature point was maintained by adjusting the temperature of the sonicator. After
ultrasonic treatment, the meat was spread evenly on a wire mesh and dried using a hot air
dryer. Drying is carried out at 100ºC until the moisture content reaches 50%. After drying, themeat was subjected to textural analysis (TPA), shear force, and microstructural analysis.
Changes in biochemical properties of raw shrimp without drying were determined bymeasuring SDS-PAGE, MFI, and active sulfhydryl content.
[Results] A single-factor experiment was used to determine the optimal processing conditions
for the tenderization of dried shrimp meat. The result showed that the hardness and shear
force of samples were decreased sharply and that the dried shrimp become redder in color.
The optimal treatment conditions to achieve dried shrimp meat with minimum firmness andshear force were obtained: power of 100 W, temperature of 50℃, and bromelain
concentration of 20 U/g. Under these conditions, the predicted value of the MFI was highest
among the examined groups. The bromelain has the ability to destroy myofibril protein andconnective tissue into smaller peptides and amino acids. Whereas ultrasound treatment not
only facilitates the penetration of the enzyme, but ultrasound itself has the effect of disrupting
cell membranes and basic structures. The combination of ultrasound and enzymes causes
protein degradation in shrimp meat. In addition, the tissue cells exhibited severe damage, withfractures and large spacing within loose muscle fibers. From among the treatments exploredthroughout this study, the combined ultrasound and bromelain yielded the best tenderness
results, especially when an ultrasound radiation of 100 W was applied at 50 ℃ and 20 U/g
concentration of bromelain. Therefore, it can be concluded that the combined treatment withultrasonic radiation and bromelain could be a promising technique for the production of dried
shrimp meat and may have a high potential for application in the meat industry.
Keywords：Penaeus vanname，tenderness，bromelainEffect of protease treatment on the quality of Pacific oysters during refrigeration
Lanxiang Su1，Xinyi Zhou1, Huamao Wei1, Wenge Yang1, Tao Huang1, Chunhong Yuan2(1. College of Food and Pharmaceutical Sciences, Ningbo University, 315211; 2. Faculty of Agriculture, Iwate
University, Ueda 3-18-8, Morioka, Iwate 020-8550, Japan)[Objective] Pacific oysters (Crassostrea gigas) are one of the main economic fish species
in the East China Sea region and their popularity is mainly due to their flavor andfreshness. Currently, Pacific oysters are usually distributed as shucked meat and their highmoisture content tends to cause the freshness of the product to deteriorate rapidly. The
content of ATP-related compounds in oysters varied with freshness, with AMP and IMP
contributing to flavor. Consumers demand for fresher, more flavorful oyster products isincreasing daily. Therefore, the enhancement of oyster flavor during the preservation
process has become a major research priority. Many studies have shown that neutral
proteases are beneficial in increasing peak flavor nucleotide (IMP) levels in the short term.
However, the introduction of enzymatic hydrolysis techniques has accelerated theautolysis process in oysters, ultimately leading to a reduction in shelf life. Therefore, it isessential to investigate the shelf life of enzyme-treated oysters.
[Methods] Fresh Pacific oysters were bought from the aquatic market, shucked andimmersed in diluted water containing 0.5% neutral protease (w/v 1:2) for 1.5 h. The
control group was immersed directly in diluted water. After washing with sterile saline,
the oysters were packed in sealed bags containing 3.5% sterilized saline and stored at 4 °C
for 7 days. Biochemical parameters (glycogen, pH, ATP-associated compounds, TVB-N
and total viable count) were measured on days 0, 1, 3, 5 and 7 for the four parts of the
mantle, adductor muscle, gills and body trunk, respectively, as well as for the whole oyster.
[Results] AMP and IMP content in mantle, adductor muscle, gill and body trunk
increased after protease treatment. During refrigeration, all four fractions of the
enzymatic hydrolysis group exhibited higher levels of IMP. The pH of each groupdecreased with increasing refrigeration time, with the enzymatic hydrolysis (EH) group
showing a more pronounced downward trend, probably due to the accelerated autolysis
process by neutral proteases. Glycogen content decreased in the mantle and adductor
muscle but did not change significantly in the gill and body trunk. In terms of TVB-N
value, those of the control group and EH group increased from 5.85 mg/100 g and 5.37
mg/100 g to 10.19 mg/100 g and 12.74 mg/100 g, respectively. On day 5, the total viablecount reached 10 mg/100 g in the control and enzymatic hydrolysis groups. The overall
results indicated a minimum shelf life of 3 days for the muscle of Pacific oysters(Crassostrea gigas) treated under experimental conditions, with increasing levels of AMP
and IMP.
Keyword: Crassostrea gigas, neutral proteases, AMP, IMP