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مقاله علمی – پژوهشی:‌ تعیین باقیمانده دیازینون در بافت عضله ماهی کپور معمولی (Cyprinus carpio) پرورشی در مجتمع پرورش ماهیان گرمابی شهداء قصرشیرین در فصل صید (استان کرمانشاه) Determination of diazinon residues in the muscle tissue of common carp (Cyprinus carpio) reared in the Shohada Qasr-e Shirin warm-water fish farming complex during the harvest season (Kermanshah Province) 1 1 دیازینون یکی از آفت‌کش‌های ارگانوفسفره است که به ‌طور گسترده‌ای در فعالیت‌های کشاورزی استفاده می‌شود. ورود این سم به آبهای سطحی و تجمع  آن در بافت‌های ماهی، می‌تواند سلامت مصرف کنندگان را از طریق زنجیره ی غذایی تحت تاثیر قرار دهد. در تحقیق حاضر، میزان دیازینون در عضلات ماهی کپور معمولی پرورشی (Cyprinus carpio) در مجتمع پرورشی ماهیان گرمابی شهرستان قصرشیرین استان کرمانشاه، در فصل صید، مورد بررسی قرار گرفت. بدین‌منظور، مزارع مجتمع بر اساس فاصله از کانال اصلی آب‌رسانی منشعب از رودخانه‌ الوند به سه ناحیه ابتدایی، انتهایی و میانی تقسیم گردید. در هر ناحیه دو مزرعه نزدیک به‌هم و با ظرفیت تقریباً برابر انتخاب شد. از آب کانال آب‌رسانی هر مزرعه و بافت عضله‌ ماهیان مزارع هر ناحیه نمونه‌برداری شد. میزان دیازینون با استفاده از روش کروماتوگرافی گازی- طیف سنجی جرمی (GC-Mass) اندازه‌گیری شد. نتایج نشان داد که میزان دیازینون در آب کانال آب‌رسانی مزارع، از ناحیه 1 به ناحیه 3 کاهش می‌یابد. میانگین میزان دیازینون در نواحی 1، 2 و 3 به‌ترتیب 71/29، 67/26 و 53/24 میکروگرم در لیتر (ppb) اندازه‌گیری گردید. تفاوت معنی‌داری بین میزان دیازینون در آب کانال آب‌رسانی مزارع ناحیه 1 با کانال‌های آبرسانی ناحیه 2 و 3 مشاهده شد (05/0 >p). بر اساس نتایج حاصله، میزان دیازینون تجمع‌یافته در عضلات ماهی بستگی مستقیم به میزان دیازینون در آب داشت (79/0=R). بیشترین میزان سم در ماهیان مزارع ابتدایی 22/9 و کمترین میزان سم در ماهیان مزارع انتهایی 41/6 و در مزارع میانی، این مقدار 55/7 میکروگرم در کیلوگرم (ppb) بود. بنابراین، می‌توان نتیجه‌گیری نمود که میزان تجمع دیازینون در عضلات ماهی در زمان عرضه به بازار، در محدوده‌‌ مجاز توصیه شده از سازمان بهداشت جهانی و سازمان ایمنی غذایی اروپا، 01/0 میلی‌‌گرم در کیلوگرم برای مصرف انسانی است. 2 Introduction Diazinon is widely used non-systemic insecticide (C12H21N2O3PS) that exert its toxic effects through inhibition acetylcholinesterase, leading to accumulation of acetylcholine at synaptic and neuromuscular junctions and subsequent disruption of nerve impulse transition. Due to its water solubility and extensive use of the pesticide diazinon, residues of this pesticide enter surface waters through runoff. Accumulation of this poison in fish tissues can have negative effects on human health. The Shohada Qasr-e-Shirin Complex in Kermanshah province is considered one of the largest warm-water fish farming facilities in Iran. During the early years of fish farmers' activity in this complex, silver carp and bighead carp accounted for the largest percentage of fish produced in the farms. However, with the boom in fish exports to Iraq, during recent years, common carp (Cyprinus carpio) has become the almost dominant species in this complex due to increased export demand to Iraq. The water supply source for this complex is the Alvand River, which also supplies the water needed by many gardens and agricultural lands in the cities of Dalahu (Rijab region), Sarpol-e-Zahab, and Qasr-e-Shirin. Given the heavy use of various pesticides farmers and following irrigation and rainfall, there is a possibility that these pesticides enter the water of the Alvand River. This study aimed to assess diazinon residue levels in the muscle tissue of farmed common carp (Cyprinus carpio) from a warm-water fish farming complex, in Qasr-e Shirin city, Kermanshah Province, during the harvest season. Methodology For this purpose, Initially, to confirm the presence of diazinon in the Alvand River during the summer season, when the highest levels of this toxin were expected, three water samples were collected from the river at the junction with the main water supply canal of the aquaculture complex. The samples were transport to the toxicology laboratory under cool conditions. Subsequently, prior to the onset of the fishing season, water samples were taken from the water of the secondary canals supplying water to the farms in the designated areas. The fish farming complex was divided into three areas based on their distance from the main irrigation canal, branching off the Alvand River. In each area, two farms close to each other and with approximately equal capacity were selected. Water samples were obtained from the irrigation canal of each selected farm and muscle tissue samples were collected from fish reared in the farms. Diazinon concentrations in all water and tissue samples were determined using gas chromatography-mass spectrometry (GC-Mass). Results The results indicated that the amount of diazinon in the water of the irrigation canal supplying the farms decreased progressively from area 1 to area 3. The amount of diazinon in areas 1, 2 and 3 were29.71±0.70, 26.67±0.63 and 24.53±0.49 micrograms per liter (ppb) (Mean±SE), respectively. A significant difference was observed between the amount of diazinon in the irrigation water canal of area 1 and those of areas 2 and 3 (p<0.05). According to the results, accumulation of diazinon in fish muscles was positively correlated with its concentration in water (R=0.79). There was no significant difference in the amount of diazinon accumulated in the muscle tissue of fish from two selected farms within each area. However, a significant difference in diazinon accumulation in t fish muscle was observed -among the three studied areas (p<0.05) (Table 1). The highest concentration of diazinon in fish muscle was recorded in fish from farms located in area 1 (9.22±0.21), while the lowest amount of toxin was observed in fish from farms in area 3 (6.41±0.14), and in the intermediate farms, the concentration of diazinon was 7.55± 0.17 micrograms per kilogram (Mean ±SE). The results showed that the amount of toxin in the water was lower than the maximum residue limit (MR) recommended by the United States Environmental Protection Agency (EPA) (0.17 mg/L). The levels of diazinon detected in fish muscle tissues were lower than the maximum residue limit for diazinon in fish flesh (0.01 mg/kg) established by the World Health Organization (WHO) and the European Food Safety Authority (EFSA) for human consumption. Discussion and conclusion In this study, the mean concentration of diazinon in the water supply canal feeding the farms located in the Shohada Qasr-e-Shirin warm water fish farming complex in areas 1, 2, and 3 was measured as 29.71, 26.67, and 24.53 micrograms per liter (ppb), respectively, and in the main water supply canal at the branching point of the Alvand River was measured at 30.42 (µg/L. All measured value were less than the maximum permissible level announced by the United States Environmental Protection Agency (EPA) (0.17 mg/L) for surface waters. The results showed that the average amount of diazinon in fish from area 1 farms, which receive water from the upstream section of the main canal, was higher than that observed in fish from other areas. Notably, t this amount was very close to the maximum permissible level recommended for human consumption. A decreasing trend in diazinon accumulation in fish muscle was observed from area 1 to area 3. Considering the distance between the studied areas, the significant decrease in the amount of diazinon could be due to the decrease in the amount of toxin in the water entering the farms caused by the metabolism of the toxin under the influence of hydrolysis, photolysis, and the effect of bacteria in the water, as well as absorption by biological factors (algae, zooplankton and phytoplankton, benthos, aquatic plants, native fish) as well as absorption of the pesticide onto sediments. It can be concluded that, the level of diazinon in the fish muscle at the time market distribution is within the permissible limit (0.01 mg/kg) recommended by the World Health Organization (WHO) and the European Food Safety Authority (EFSA) for human consumption. Considering that the diazinon residue detected in fish harvested from farms in Area 1, whose water supply originates from the upstream section of the main water conveyance canal is very close to the recommended permissible limit, frequent and continuous monitoring of diazinon levels is warranted. Accordingly, systematic measurement of diazinon concentrations in the water of the Alvand River and in farmed fish from the Shohada Qasr-e Shirin warm-water fish farming complex throughout the harvesting season should be conducted by the relevant authorities and responsible organizations. Conflict of interest Conflict of interest does not exist. Acknowledgment This article is a result of (part of) the thesis entitled: Determination of diazinon toxin levels in muscle tissue of common carp (Cyprinus carpio) in the Shohada Qasr-e Shirin Complex, Kermanshah Province, during the harvesting season, in the Master's degree program in the year 2025, which was carried out with the support of the University of Kurdistan. 1 16 2025/08/27 1404/6/5 2026/03/1 1404/12/10 امین امیری Amin Amiri گروه شیلات، دانشکده منابع طبیعی، دانشگاه کردستان، سنندج، ایران Iran amiriamin921@gmail.com فرزاد غیاثی Farzad Ghiasi گروه شیلات، دانشکده منابع طبیعی، دانشگاه کردستان، سنندج، ایران Iran f.ghiasi@uok.ac.ir Diazinon common carp muscle Shohada Qasr Shirin Fish Farming Complex Cyprinus carpio دیازینون عضله کپور معمولی مجتمع پرورش ماهی شهدای قصر شیرین Cyprinus carpio Akan, J., Mohammed, Z., Jafiya, L. & Audu, S., 2013. Organophosphorus pesticide residues in different tissues of fish samples from Alau Dam, Borno State, Nigeria. 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مقاله مروری:‌ کاربرد ترکیبات زیستی دریایی برای پوشش‌دهی نانولیپوزوم‌ها با هدف بهبود خصوصیات فیزیکوشیمیایی و افزایش پایداری: مقاله مروری Application of marine biopolymers for coating nanoliposomes to improve physicochemical properties and enhance stability: A review 1 1 نانولیپوزوم‌ها به عنوان سیستم‌های حامل با توانایی بالا در محصورسازی، انتقال و رهایش کنترل‌شده ترکیبات فعال، جایگاه ویژه‌ای در زمینه‌ دارویی، غذایی و آرایشی-بهداشتی پیدا کرده‌اند. با این‌حال، حساسیت ساختاری این نانوساختارها در برابر عوامل محیطی سبب کاهش پایداری فیزیکی و شیمیایی و محدودیت در استفاده گسترده آنها می‌شود. در سال‌های اخیر، استفاده از ترکیبات زیستی دریایی به عنوان پوشش‌دهنده‌های محافظ، یک راهکار نوآورانه برای غلبه بر این چالش‌ها مطرح شده است. ترکیبات زیستی دریایی نظیر کیتوزان، آلژینات، کلاژن، ژلاتین و فوکوئیدان به دلیل ساختارهای بیولوژیک پیچیده، زیست‌سازگاری بالا و خواص عملکردی متنوع از جمله فعالیت آنتی‌اکسیدانی، ضد میکروبی و توانایی تشکیل شبکه‌های محافظت‌کننده، در پوشش‌دهی نانولیپوزوم‌ها نقش کلیدی دارند. این ترکیبات با ایجاد لایه‌های محافظ و مقاوم روی سطح نانولیپوزوم‌ها، تخریب ناشی از عوامل محیطی را به حداقل می‌رسانند و موجب افزایش پایداری ساختاری و عملکردی می‌شوند. علاوه‌براین، بهره‌گیری از پوشش‌های زیستی چندلایه می‌تواند به بهبود ویژگی‌های آزادسازی دارو از نانولیپوزوم‌ها و فراهم‌سازی رهایش زمان‌بندی‌شده و هدفمند ترکیبات فعال کمک شایانی نماید. در داروسازی، نانولیپوزوم‌های پوشش‌دهی‌شده با ترکیبات زیستی دریایی، امکان انتقال داروهای حساس را با حفظ اثربخشی و افزایش دسترسی زیستی فراهم می‌آورند. در صنایع غذایی استفاده از این نانوساختارها، پایداری و ارزش غذایی ترکیبات را در شرایط سخت فرآوری و نگهداری تضمین می‌کند. در حوزه آرایشی-بهداشتی این فناوری منجر به طراحی محصولات پیشرفته با ماندگاری بیشتر و اثرات تقویتی بر پوست و مو می‌شود. در مطالعه مروری حاضر، ترکیبات زیستی دریایی مورداستفاده در پوشش‌دهی نانولیپوزوم‌ها با تاکید بر مشخصات ساختاری، خواص عملکردی و مکانیسم‌های بهبود پایداری و رویکردهای نوین در طراحی پوشش‌های زیستی چندلایه به طور جامع بررسی شده است. 2 Introduction Nanoliposomes, recognized as one of the most advanced delivery systems for drugs and bioactive compounds, have gained significant importance across biomedical, pharmaceutical, food, and cosmetic industries. These nanosystems demonstrate remarkable efficiency due to their high encapsulation capacity, controlled release, enhanced bioavailability, and improved stability. However, the intrinsic instability of nanoliposomes under environmental factors such as oxidation, hydrolytic degradation, light exposure, temperature fluctuations, and pH variations limits their efficacy and practical applications. To overcome these challenges, coating nanoliposomes with marine biopolymers has emerged as an innovative strategy to enhance their physicochemical stability, regulate the release profile, and improve biological performance (Senadheera et al., 2023). Marine biopolymers, including chitosan, alginate, collagen, gelatin, and fucoidan, are ideal candidates for nanoliposome coating due to their high biocompatibility, antioxidant and antimicrobial properties, ability to form stable networks, and protective layer-forming capabilities. These biopolymers create protective layers that reduce structural degradation, modulate the release rate of bioactive compounds, and extend their half-life) Gómez-Guillén and Montero., 2021; Pasarin et al., 2023). Furthermore, electrostatic interactions and physicochemical bonding between biopolymers and nanoliposomes reinforce structural integrity and enhance resistance to adverse environmental conditions. This review comprehensively analyzes recent studies on the role of marine biopolymers in optimizing the structure and functionality of nanoliposomes, focusing on their impact in improving stability, controlling release, and enhancing the biological efficacy of encapsulated compounds (Tan et al., 2021). Methodology This study follows a systematic review approach, utilizing scientific articles from reputable databases such as PubMed, Scopus, and ScienceDirect. The selected articles were screened based on their direct relevance to nanoliposome coating with marine biopolymers, their effects on structural and biological stability, physicochemical properties, bioavailability, and mechanisms involved in enhancing longevity and biological performance. The extracted papers were analyzed with a focus on the functional properties of biopolymers, their protective mechanisms, and the impact of multilayer coatings. Results Extensive studies have demonstrated that coating nanoliposomes with marine biopolymers has a significant impact on enhancing the physical, chemical, and biological stability of these systems. Among these biopolymers, chitosan, as a cationic polysaccharide, interacts strongly with the nanoliposomal membrane through electrostatic interactions, leading to increased structural integrity, reduced lipid oxidation rates, and improved retention of bioactive compounds. The mechanism underlying this effect is attributed to the direct interaction of chitosan with membrane phospholipids and the formation of a stable protective layer that not only reduces oxygen permeability but also prevents lipid oxidation, thereby preserving the bioavailability of the encapsulated compounds over an extended period. Furthermore, chitosan coating decreases membrane permeability and enhances the controlled release of active compounds in biological environments )Kumar et al., 2020; Kamali et al., 2024). Alginate, as an anionic polysaccharide, has a high capacity for forming strong gel structures in the presence of calcium ions, thereby creating robust protective layers around nanoliposomes and preventing the leakage of bioactive compounds. This property is particularly crucial for protecting sensitive compounds from unstable environmental conditions, such as pH fluctuations and the presence of digestive enzymes. Recent studies have shown that alginate coatings, particularly in pharmaceutical formulations, enhance the bioavailability of active compounds and improve their absorption in biological environments. Additionally, alginate forms stable polymeric networks that enhance the mechanical stability of nanoliposomes and prevent structural changes during storage and biological processes (Abka-Khajouei et al., 2022). In addition to polysaccharides, structural proteins such as collagen and gelatin have also been utilized as effective nanoliposome coatings. These biopolymers, due to their strong ability to form stable polymeric networks, reinforce the mechanical integrity of nanoliposomal systems and enhance their physical stability in biological environments. Collagen and gelatin coatings improve the structural stability of nanoliposomes by forming strong intermolecular bonds, preventing undesirable changes over time. Moreover, the use of these biopolymers in nanoliposome coating improves biocompatibility and reduces toxicity, which is particularly significant for pharmaceutical and biomedical applications (Chotphruethipong et al., 2021; Naseriyeh et al., 2024). Fucoidan, a sulfated marine polysaccharide, possesses unique properties such as antioxidant, anti-inflammatory, and antimicrobial activities. Studies have demonstrated that coating nanoliposomes with fucoidan not only enhances their stability but also strengthens the biological effects of active compounds, thereby improving the efficacy of targeted drug delivery systems. This effect is attributed to fucoidan's ability to enhance cellular interactions and increase nanoliposome penetration into target tissues, which could play a crucial role in advanced pharmaceutical therapies (Rostami et al., 2018; Obiedallah et al., 2024). One of the most notable findings in this field is the synergistic effect of multilayered coatings composed of different biopolymers. Research has shown that combining chitosan and alginate in nanoliposome coatings not only enhances mechanical strength but also creates a dual-stage release system, where active compounds are gradually released under different conditions. This characteristic, particularly valuable in pharmaceutical and food applications, enhances the efficiency and effectiveness of these systems. Furthermore, coating nanoliposomes with marine biopolymers extends the stability of active compounds in biological environments, enhances cellular uptake, and prevents enzymatic degradation (Meng et al., 2024). Overall, marine biopolymers, as nanoliposome coatings, not only improve physicochemical stability but also enable precise control over compound release, thereby playing a crucial role in optimizing biological and industrial applications. Discussion and conclusion The findings of this study highlight that marine biopolymer-based coatings serve as an effective strategy for improving the biological, chemical, and physical properties of nanoliposomes. Multilayer bio-based coatings not only prevent structural degradation but also enhance membrane integrity and optimize the release kinetics of active compounds through electrostatic interactions, covalent bonding, and polymeric network formation. Coating nanoliposomes with marine biopolymers has demonstrated significant potential in regulating the gradual release of bioactive compounds and broadening their applications. In drug delivery systems, this technology can enhance the efficiency of hydrophilic and lipophilic drug transport while improving their bioavailability) Gómez-Guillén and Montero, 2021; Pasarin et al., 2023; Meng et al., 2024). In the food industry, coated nanoliposomes help preserve sensitive bioactive compounds such as vitamins, carotenoids, and polyphenols, thereby extending their shelf life. Additionally, in cosmetic formulations, these nanocarriers improve skin absorption and prolong the effectiveness of active ingredients (Ajeeshkumar et al., 2021). Comparative analyses indicate that integrating different biopolymers into multilayer coatings enhances mechanical and chemical stability under various environmental conditions. This advantage underscores the importance of this technology in dose control and therapeutic efficacy optimization. Furthermore, advancements in encapsulation methods particularly the development of novel bio-based materials and optimization of production processes can further enhance the performance of coated nanoliposomes in industrial applications (Meng et al., 2024; Gan et al., 2024) Overall, nanoliposome coating with marine biopolymers is an emerging technology that facilitates the development of controlled-release delivery systems and the protection of sensitive compounds. Future research should focus on structural modifications of biopolymers, optimization of formulation conditions, and molecular-level investigations of interactions between bio-based coatings and nanoliposomes. Such studies can deepen our understanding of the stabilization and controlled-release mechanisms of bioactive compounds and accelerate the commercialization of this technology. Conflict of interest The authors declare no conflict of interest in this study. Acknowledgment  This review study was conducted based on an extensive search and analysis of reliable scientific resources. The authors would like to express their gratitude to all researchers and authors whose studies formed the basis of this research. 17 40 2025/08/272025/02/18 1403/11/30 2026/03/12026/03/1 1404/12/10 زینب رستمی Zeynab Rostami گروه شیلات، دانشکده علوم دامی و شیلات، دانشگاه علوم کشاورزی و منابع طبیعی ساری، ساری، ایران Iran fazele.K.ROSTAMI@GMAIL.COM مینا اسمعیلی خاریکی Mina Esmaeili گروه شیلات، دانشکده علوم دامی و شیلات، دانشگاه علوم کشاورزی و منابع طبیعی ساری، ساری، ایران Iran mina.smaily@gmail.com Marine Biopolymers biocompatibility nanoliposomes liposome coating controlled-release. ترکیبات زیستی دریایی زیست‌سازگاری نانولیپوزوم‌ پوشش‌دهی لیپوزوم‌ها رهایش کنترل‌شده Abka-Khajouei, R., Tounsi, L., Shahabi, N., Patel, A. K., Abdelkafi, S., & Michaud, P., 2022. Structures, properties and applications of alginates. Marine drugs, 20(6): 364. 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مقاله علمی – پژوهشی:‌ شناسایی و اولویت‌بندی بازارهای صادراتی میگوی ایران Identification and prioritization of export markets for Iranian shrimp 1 1 محصولات دریایی به‌ویژه میگو، از منابع مهم تأمین پروتئین و مواد مغذی در جهان به‌شمار می‌روند. افزایش تقاضای جهانی برای میگو و وابستگی برخی کشورها به واردات، فرصتی ارزشمند برای کشورهای تولیدکننده فراهم کرده است تا با برنامه‌ریزی هدفمند، سودآوری خود را افزایش دهند. در پژوهش حاضر، با تمرکز بر صادرات میگو از ایران و با بهره‌گیری از روش‌های تصمیم‌گیری چندمعیاره، بازارهای هدف شناسایی و رتبه‌بندی شدند. برای این منظور، شاخص‌هایی همچون ضریب جینی، فاصله جغرافیایی، درآمد سرانه، جمعیت، واردات میگوی هر کشور، نرخ رشد واردات میگوی هر کشور، تورم کشورها، ثبات سیاسی کشورها و میزان کل صادرات کشور مبدأ، مورد بررسی قرار گرفتند و با استفاده از روش بهترین-بدترین وزن‌دهی شدند. سپس از روش واسپاس جهت اولویت‌بندی کشورها استفاده شد. با ارزیابی ۱۹۴ کشور به عنوان بازارهای بالقوه صادراتی میگوی ایران، ۲۰ کشور به عنوان بازارهای اولویت‌دار شناسایی شدند که در میان آنها، چین و امارات متحده عربی به عنوان جذاب‌ترین مقاصد صادراتی تعیین گردیدند. بر اساس این نتایج، پیشنهاد می‌شود سیاست‌گذاران و فعالان صنعت آبزی‌پروری، با تمرکز بر توسعه روابط تجاری و کاهش موانع صادراتی در بازارهای اولویت‌دار مشخص شده به‌ویژه چین و امارات، اقدام کنند تا ظرفیت صادرات میگوی ایران افزایش یابد و سودآوری این بخش ارتقاء یابد. 2 Introduction Shrimp is among the most valuable aquatic commodities worldwide due to its high nutritional value and economic significance. In recent decades, shrimp aquaculture has become a major driver of employment, income generation, and export growth in coastal nations (Ray et al., 2021). The global shrimp market continues to expand, supported by population growth, urbanization, and lifestyle changes that have increased demand for high-protein and low-calorie foods (N’Souvi et al., 2024). Shrimp is rich in essential nutrients, including iodine, zinc, selenium, phosphorus, and vitamin B12, which support immune function, wound healing, and overall health (USDA, 2019; Berkheiser, 2023; Sass, 2024). Among farmed species, the Pacific white shrimp (Penaeus vannamei) dominates global aquaculture production, accounting for about 90% of shrimp farms worldwide (Dugassa and Gaetan, 2018). Its global production reached 5.8 million tons in 2020 (Asmild et al., 2024). Leading producers, including Ecuador, China, India, Vietnam, and Indonesia, collectively supply about 74% of global shrimp output (FAO, 2023; Jory, 2023; Mandal and Singh, 2025). Iran, with suitable climatic conditions and access to southern and northern coastal waters, has emerged as a significant shrimp producer (Mirzaei et al., 2021). In 2023, the country’s total aquaculture production reached 639,936 tons, while total marine catch was 778,278 tons (IFO, 2023). Major shrimp-producing provinces include Bushehr, Hormozgan, and Golestan. These figures highlight Iran’s strong potential for expanding shrimp exports. However, realizing this potential depends on accurately identifying and prioritizing export markets. Incorrect market targeting may reduce profitability and market share (Mora, 2023). Although several studies have analyzed seafood exports using multicriteria decision-making (MCDM) approaches such as AHP, TOPSIS, and fuzzy extensions (Çelik & Akmermer, 2021; Adeli, 2022; Majidian et al., 2025), few have focused on shrimp exports. The present study addresses this gap by employing a hybrid framework integrating the Best–Worst Method (BWM) and Weighted Aggregated Sum Product Assessment (WASPAS) to evaluate and rank 194 countries based on nine key criteria, offering a robust model for strategic export market selection. Methodology This applied study employed a descriptive, analytical, and quantitative design aimed at identifying and prioritizing export markets for Iranian shrimp. Initial criteria were extracted through a comprehensive literature review covering aquaculture studies and multicriteria decision-making frameworks. To ensure the validity and adequacy of the selected criteria, a modified Delphi method was conducted in two rounds with 12 experts from the aquaculture and export sectors. The Kendall coefficient of 0.937 indicated a strong consensus among participants. Consequently, nine indicators with the highest agreement were retained: Gini coefficient, geographical distance, population, income per capita, shrimp import volume, shrimp import growth rate, inflation rate, political stability, and total exports. The relative weights of these indicators were calculated using the Best–Worst Method (BWM) as proposed by Rezaei (2015; 2016). Subsequently, the WASPAS method (Zavadskas et al., 2012; Chakraborty et al., 2015) was applied to rank 194 countries. Raw criteria data were normalized using a fuzzy scaling method, and both Weighted Sum Model (WSM) and Weighted Product Model (WPM) scores were computed. Final WASPAS scores were derived by linearly combining WSM and WPM results with λ = 0.5, providing a comprehensive prioritization of potential shrimp export destinations. Results Data from 194 shrimp-importing countries were analyzed for the years 2017–2023, with average values calculated for each of the nine selected criteria. According to expert evaluations and the Delphi method, total exports of the origin country emerged as the most critical criterion (weight = 0.3248), emphasizing the role of existing trade relationships in market selection. Shrimp import volume was the second most significant factor (weight = 0.2046), reflecting the importance of market size and demand dynamics. In contrast, the Gini coefficient received the lowest weight (0.0267), indicating minimal influence on market prioritization. The weighted criteria were subsequently used in the WASPAS method. Fuzzy normalization was applied to raw data, and WSM and WPM scores were computed for each country. Final WASPAS scores were obtained by linearly combining these two components. Based on these scores, the countries were ranked in descending order to identify the top 20 destinations for Iranian shrimp exports. China was ranked first (0.7044), establishing it as the most attractive market, followed by the United Arab Emirates (0.2819), the United States (0.2449), Iraq (0.2043), and India (0.2003). Other high-ranking countries included Turkey, South Korea, Japan, Germany, and Spain. The presence of both Asian and European countries among the top twenty highlights the geographical and structural diversity of promising markets. These findings suggest that, alongside traditional markets such as China and the UAE, emerging markets like Japan and Germany present opportunities for diversification, enhancing export stability and reducing dependency on a limited number of destinations. The integrated BWM-WASPAS framework demonstrated reliability in evaluating countries based on multiple economic, demographic, and political indicators, providing a robust basis for strategic export decisions. Conclusion and discussion The integration of BWM and WASPAS methods identified China as the most attractive export destination for Iranian shrimp, driven by its large population and sustained shrimp demand (average annual import growth rate = 0.385). The UAE ranked second, reflecting its geographic proximity and established trade relations with Iran. Historical trade data confirm that China has been Iran’s primary shrimp export market from 2017 to 2021, except 2020 when the UAE temporarily led (Saeedi et al., 2025). Recent Iranian customs and fisheries statistics indicate that shrimp exports have predominantly targeted China, the UAE, and Russia (IRICA & IFO, 2024). Russia, despite being a major current importer, ranked twelfth in this analysis, illustrating the distinction between descriptive past statistics and the forward-looking assessment provided by a multi-criteria analytical approach. Factors such as high inflation, long distances, and political instability negatively affected its ranking. Conversely, the United States, Iraq, and India were identified among the top five markets due to favorable population size and structural advantages. European and East Asian countries, including Germany, Japan, and South Korea, also demonstrate substantial potential for future expansion (EUMOFA, 2023; FAO, 2024; OECD-FAO, 2023; ITC, 2023). These findings highlight that combining traditional markets, such as China and the UAE, with emerging destinations in Europe and East Asia—including Japan, South Korea, Germany, and Spain—can enhance export stability, competitiveness, and risk diversification. Total exports of the origin country, reflecting the depth of trade, were the most influential criterion. Meanwhile, import volume and growth also affected rankings, indicating that both demand and growth trends significantly influence export success. This approach enables policymakers to select an optimal blend of regional and non-regional markets, reducing dependence on limited destinations. Overall, it provides a data-driven basis for strategies in the Iranian shrimp industry, supporting diversification and strengthening global competitiveness. Conflict of interest The authors declare no conflict of interest related to this research or the publication of this manuscript. Acknowledgment The authors gratefully acknowledge the valuable comments and suggestions that improved the quality of this research. 41 54 2025/08/272025/02/182025/06/30 1404/4/9 2026/03/12026/03/12026/03/1 1404/12/10 نجمه مرزوقی Najmeh Marzooghi دانشکده مهندسی صنایع، دانشگاه صنعتی شیراز، شیراز، ایران Iran najmeh.marzoogi@gmail.com جواد زارعی Javad Zarei دانشکده مهندسی صنایع، دانشگاه صنعتی شیراز، شیراز، ایران Iran j.zarei@sutech.ac.ir Iranian Shrimp Export Markets Multi-Criteria Decision Making Best–Worst Method WASPAS Method. میگوی ایران بازارهای صادراتی تصمیم‌گیری چندمعیاره روش بهترین-بدترین روش واسپاس Adeli, O., 2022. Prioritization of Iranian shrimp export target markets. Journal of Animal Environment, 14(2):317–324. DOI:10.22034/AEJ.2021.278494.2484##Akmermer, B. and Çelik, P., 2021. Contribution of fishery and aquaculture products to Turkish foreign trade: An evaluation by a hybrid multi-criteria decision making method. 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مقاله علمی – پژوهشی:‌ تأثیر عصاره هیدروالکلی ریزپوشانی‌شده برگ گیاه آقطی سفید (Sambucus ebulus) بر شاخص‌های خونی، بیوشیمیایی، ایمنی و مقاومت قزل‌آلای رنگین‌کمان (Oncorhynchus mykiss) در برابر باکتری Yersinia ruckeri Effect of microencapsulated hydroalcoholic extract of dwarf elder (Sambucus ebulus) leaf on hematological, biochemical, immunological parameters and disease resistance of rainbow trout (Oncorhynchus mykiss) against Yersinia ruckeri 1 1 با توجه به نقش کلیدی قزل‌آلا در صنعت آبزی‌پروری ایران و کاهش اثربخشی آنتی‌بیوتیک‌ها در کنترل بیماری‌هایی مانند Yersiniosis، استفاده از گیاهان دارویی به ‌عنوان راهکاری نوین برای تقویت ایمنی و افزایش مقاومت ماهیان، ضروری به‌نظر می‌رسد. هدف از مطالعه حاضر، ارزیابی اثرات عصاره هیدروالکلی کپسوله شده گیاه آقطی بر شاخص‌‌های ایمنی، خون، بیوشیمیایی و مقاومت در برابر باکتری Yersinia ruckeri بوده است. بدین‌منظور، عصاره هیدروالکلی برگ گیاه آقطی به ‌صورت ریزپوشانی‌شده با ژلاتین در غلظت‌های 0 (شاهد)، 1/0، 25/0 و 5/0 درصد به جیره غذایی ماهیان قزل‌آلای رنگین‌کمان (میانگین وزنی 55/2±11/59 گرم) افزوده شده و ماهیان به مدت 8 هفته تغذیه شدند. در پایان هفته‌های 4 و 8 آزمایش، از 8 عدد برای سنجش شاخص‌های خونی، بیوشیمیایی و ایمنی خون‌گیری انجام گرفت. در پایان هفته 8 نیز به 30 عدد ماهی از هر تیمار، 1/0 میلی‌لیتر از باکتری Y. ruckeri به ‌صورت داخل ‌صفاقی تزریق شد. نتایج این بررسی نشان داد که تعداد گلبول‌های قرمز، هماتوکریت، هموگلوبین، حجم متوسط گلبولی، میزان هموگلوبین گلبولی و غلظت متوسط هموگلوبین گلبولی ، آلبومین، آسپارتات آمینوترانسفراز تفاوت معنی داری در پایان هفته‌های 4 و 8 آزمایش بین شاهد و تیمارها نشان ندادند. تعداد گلبول‌های سفید، کلسترول، تری‌‌گلیسرید، پروتئین تام سرم، آلانین آمینوترانسفراز، IgM تام سرم، لیزوزیم، انفجار تنفسی، در گروه‌‌های تیمار در مقایسه با شاهد بهبود معنی‌‌داری داشتند. در مواجهه با باکتری، درصد بازماندگی در تیمارهای 0 (شاهد)، 1/0، 25/0 و 5/0 درصد به‌ترتیب 3/23، 7/56، 7/66 و 3/73 درصد بودند. نتایج این بررسی نشان داد که عصاره گیاه آقطی از توانائی بسیار خوبی در بهبود شاخص‌های بیوشیمیائی، ایمنی و مقاومت ماهیان قزل آلا دارد در برابر باکتری بیماریزا برخوردار بوده و بهترین دوز مصرفی 25/0 درصد است. 2 Introduction Rainbow trout (Oncorhynchus mykiss) is one of the most widely farmed fish species in Iran and globally, valued for its economic and nutritional significance. However, intensive aquaculture practices often lead to physiological stress, liver dysfunction, and increased susceptibility to bacterial infections such as yersiniosis, caused by Yersinia ruckeri. The emergence of biotype 2 strains with reduced sensitivity to conventional vaccines has raised concerns about vaccine failure and disease outbreaks in trout farms (Austin et al., 2003; Kumar et al., 2017). In recent years, medicinal plants have gained attention as natural immunostimulants and hepatoprotective agents in aquaculture. Elderberry (Sambucus ebulus) is one such plant, known for its flavonoid compounds with antimicrobial, antioxidant, anti-inflammatory, and immune-boosting properties (Ivanova et al., 2014; Ngugi et al., 2015). Previous studies have demonstrated the positive effects of dietary elderberry leaf supplementation on blood parameters, serum biochemistry, immune response, and disease resistance in rainbow trout (Ghiasi et al., 2023). Despite promising findings, previous research suggests that this plant has strong potential to be introduced as a dietary immunostimulant and growth enhancer in aquaculture. The aim of this study was to evaluate the effects of encapsulated elderberry leaf extract as a dietary supplement to promote growth and immunity. As a complementary strategy alongside conventional vaccines, it may play a significant role in reducing mortality and enhancing trout production in Iran. Methodology  To prepare the herbal extract, S. ebulus leaves were collected from Sefrabad (Sari, Iran) and authenticated by the Semnan Agricultural Research Center. After shade-drying, the leaves were extracted using a modified Soxhlet method with 70% ethanol (Păvăloiu et al., 2020). The concentrated extract was freeze-dried and microencapsulated with gelatin through high-speed homogenization. Encapsulation efficiency and extract yield were assessed spectrophotometrically (Shu et al., 2006). Experimental diets were formulated weekly by spraying the dissolved microcapsules onto commercial trout feed (GF1, Beyza Co., Iran), followed by air-drying and cold storage. A total of 280 rainbow trout (O. mykiss, 59.11 ± 2.55 g) were randomly assigned to 12 fiberglass tanks. After a 2-week acclimation, fish were fed for 8 weeks with diets containing 0, 0.1, 0.25, and 0.5% extract, adjusted to 2–2.5% of body weight daily. At weeks 4 and 8, blood samples were collected from anesthetized fish to evaluate hematological and immunological responses. Standard methods were used to measure RBC, WBC, Hct, Hb, MCV, MCH, and MCHC (Blaxhall and Daisley, 1973; Seiverd, 1964), while differential leukocyte counts were performed on Giemsa-stained smears (Lee et al., 1998). Respiratory burst activity was assessed using luminol-based chemiluminescence (Binaii et al., 2022). Serum levels of TP, Alb, Chol, Tri, ALT, AST, total IgM, and lysozyme activity were measured using commercial kits and turbidimetric assays (Ellis, 1990; Binaii et al., 2014). At the end of the trial, fish were challenged with Y. ruckeri strain MT968739 via intraperitoneal injection (Ghiasi et al., 2023). Mortality was monitored for 14 days, and survival rates were calculated. All data were statistically analyzed using one-way ANOVA followed by Duncan’s multiple range tests at a 95% confidence level (Zar, 1994). Results Microencapsulation showed strong formulation performance, yielding 92% with an encapsulation efficiency of 78%. Hematological profiles at weeks 4 and 8 revealed no significant differences in RBC count, hemoglobin, hematocrit, MCV, MCH, or MCHC between treated and control groups, though treated fish tended to display slightly higher values. In the 0.25% group, WBC count and neutrophil percentage increased, accompanied by a reduction in lymphocytes, while other treatments followed similar but non-significant trends. Serum biochemistry remained stable for AST and albumin across all groups, yet treated fish generally exhibited improved profiles. Cholesterol declined slightly at week 4 and significantly by week 8 in all treatments. Triglycerides consistently decreased, while total serum protein rose in the 0.25% and 0.5% groups. ALT levels were lower in all treatments, suggesting enhanced liver function. Immunological responses were strengthened, with IgM levels significantly elevated in all treated groups at both time points. Lysozyme activity increased in the 0.25% group at week 4 and further in the 0.25% and 0.5% groups by week 8, while the 0.1% group showed a non-significant rise. Respiratory burst activity was consistently higher, with the 0.25% group showing the strongest response at week 4 and sustained elevations across treatments by week 8. Upon challenge with Y. ruckeri, survival improved markedly in all supplemented groups. Fish receiving 0.1%, 0.25%, and 0.5% supplementation achieved survival rates of 56.67%, 73.33%, and 66.67%, respectively, compared to only 23.33% in controls. Discussion and conclusion  This study shows that dietary supplementation with microencapsulated herbal extract can enhance both physiological and immunological status in rainbow trout. While hematological indices did not differ significantly, treated fish generally displayed improved profiles. Biochemical changes, including reduced ALT and triglycerides together with higher total protein, point to better liver function and metabolic stability. Immunological outcomes such as elevated IgM, lysozyme activity, and respiratory burst highlight strengthened innate immunity, especially in the 0.25% and 0.5% groups. Most importantly, survival after Y.ruckeri challenge was markedly higher in all supplemented groups, confirming the protective effect of the extract. Overall, the formulation proved effective in supporting fish health and disease resistance, with the 0.25% dose delivering the most consistent benefits Conflict of interest The authors declare that they have no conflict of interest. Acknowledgment This article is derived from a dedicated research project entitled “Evaluation of the effects of free and microencapsulated hydroalcoholic extract of elderberry (S. ebulus) leaves on growth, immunity, and disease resistance of  rainbow trout against Y. ruckeri”, approved under project number 001292-101-12-76-2. We sincerely thank all colleagues at the Iranian Fisheries Science Research Institute and the Caspian Sea Ecology Research Center for their valuable scientific and laboratory support throughout this study. 55 72 2025/08/272025/02/182025/06/302025/10/8 1404/7/16 2026/03/12026/03/12026/03/12026/03/1 1404/12/10 مریم قیاسی Maryam Ghiasi پژوهشکده اکولوژی دریای خزر، موسسه تحقیقات علوم شیلاتی کشور، سازمان تحقیقات، آموزش و ترویج کشاورزی، ساری، ایران Iran ghiasimaryam4@gmail.com فرزانه بهادری Farzaneh Bahadori مرکز تحقیقات و آموزش کشاورزی و منابع طبیعی استان سمنان، سازمان تحقیقات، آموزش و ترویج کشاورزی، سمنان، ایران Iran farbahadori@gmail.com راضیه عظیمی اترگله Razieh Azimi Atargoleh موسسه تحقیقات جنگل‌ها و مراتع کشور، سازمان تحقیقات، آموزش و ترویج کشاورزی، تهران، ایران Iran razimi@rifr-ac.ir محمد بینائی Mohamad Binaii پژوهشکده اکولوژی دریای خزر، موسسه تحقیقات علوم شیلاتی کشور، سازمان تحقیقات، آموزش و ترویج کشاورزی، ساری، ایران Iran bbinaii@gmail.com فرشیده حبیبی Farshideh Habibi پژوهشکده اکولوژی دریای خزر، موسسه تحقیقات علوم شیلاتی کشور، سازمان تحقیقات، آموزش و ترویج کشاورزی، ساری، ایران Iran farshidehhabibi@gmail.com محمد جواد تقوی Mohammad Javad Taghavi پژوهشکده اکولوژی دریای خزر، موسسه تحقیقات علوم شیلاتی کشور، سازمان تحقیقات، آموزش و ترویج کشاورزی، ساری، ایران Iran j_taghavi2001@yahoo.com Sambucus ebulus Oncorhynchus mykiss Yersinia ruckeri total serum protein immunity lysozyme آقطی قزل آلای رنگین کمان Yersinia ruckeri پروتئین تام سرم ایمنی لیزوزیم Abdel-Latif, H.M.R., Citarasu, T., Turgay, E., Yilmaz, E., Yousefi, M., Hosseini Shekarabi, P., Ahmadifar, E., Nowosad, J., Kucharczyk, D., and Yilmaz, S., 2025. 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مقاله علمی – پژوهشی:‌ مقایسه اثرات منابع سلنیوم آلی و معدنی بر عملکرد رشد، ترکیب لاشه و مقاومت به استرس شوری، قطع هوادهی و افزایش ناگهانی تراکم جمعیت در کپور معمولی (Cyprinus carpio Linnaeus, 1758) Comparison of the effects of organic and inorganic selenium resources on growth performance, carcass composition, and resistance to salinity stress, aeration interruption, and sudden increase in population density in common carp (Cyprinus carpio Linnaeus, 1758) 1 1 سلنیوم (Se) به عنوان یک ریزمغذی حیاتی، در فرآیندهای متابولیک، سیستم ایمنی و تولیدمثل ماهیان مؤثر است. این مطالعه به‌ منظور ارزیابی تأثیر منابع و سطوح مختلف سلنیوم بر رشد و مقاومت به استرس در ماهی کپور معمولی انجام شد. اثرات شش تیمار غذایی شامل: 1- حاوی 3/0 میلی‌‌گرم سلنیوم اویلاسلنیوم در هر کیلوگرم غذا، 2- حاوی 1 میلی‌‌گرم سلنیوم اویلاسلنیوم در هر کیلوگرم غذا، 3- حاوی 3/0 میلی‌‌گرم سلنیوم سل‌‌استار در هر کیلوگرم غذا، 4- حاوی 3/0 میلی‌‌گرم سلنیوم سلنیت سدیم در هر کیلوگرم غذا، 5- حاوی 1 میلی‌‌گرم سلنیوم سلنیت سدیم در هر کیلوگرم غذا و 6- شاهد فاقد مکمل سلنیومی بر گونه کپور معمولی (Cyprinus carpio) مورد ارزیابی قرار گرفت. آزمایش در قالب یک طرح کاملاً تصادفی با ۲ تکرار برای هر تیمار و به مدت ۵۲ روز در مخازن ۲۵۰ لیتری با تراکم ۳۱ عدد ماهی در هر مخزن (میانگین وزن اولیه: 33/0±62/1 گرم) انجام شد. تفاوت معنی‌داری در وزن نهایی، طول استاندارد و ضریب چاقی بین تیمارها مشاهده نشد (05/0<p). سلنیوم اثر معنی‌‌داری بر نرخ زنده‌‌مانی طی دوره پرورش، درصد ماده خشک، پروتئین خام و خاکستر لاشه نداشت (05/0<p). تغذیه با اویلاسلنیوم یا سلنیت سدیم به میزان 3/0 میلی‌‌گرم در هر کیلوگرم غذا سبب کاهش معنی‌‌دار درصد چربی خام لاشه گردید (05/0>p). پس از استرس شوری و قطع هوادهی، تفاوت معنی‌داری در مدت زنده‌مانی مشاهده نشد (05/0<p). بعد از استرس شوری همراه با هوادهی، تیمار سلنیت سدیم به طور معنی‌‌داری مدت زنده ماندن را افزایش داد (05/0>p). استفاده از اویلاسلنیوم بهبودی نسبی در زنده‌مانی ایجاد کرد، اما در مقایسه با شاهد اثر معنی‌داری نداشت (05/0<p) درحالی‌که استفاده از سلنیوم سل‌‌استار موجب کاهش معنی‌‌دار مدت زنده ماندن گردید (05/0>p). در مجموع، بهترین نتیجه با مصرف سلنیت سدیم در سطح بهینه 3/0 میلی‌‌گرم سلنیوم در هر کیلوگرم جیره به‌‌دست آمد. 2 Introduction Aquaculture makes a significant contribution to food production for the world population, plays a decisive role in international trade, and offers potential solutions to food security challenges (Mihaly-Karnai et al., 2025). Due to overfishing and in order to restore the stocks of common carp (Cyprinus carpio), responsible institutions are taking steps to artificially propagate this fish and release it into the wild. Unfortunately, in recent years, due to various reasons including reduced rainfall and excessive water withdrawal for agricultural purposes, it is not far-fetched that when releasing juvenile fish into the wild, they may encounter salinity stress (Roohi et al., 2017). Frequent power outages and subsequent aeration interruptions in recent years have caused oxygen deficiency stress, especially in intensive farming. With the continuous increase in the intensity of fish population density in advanced aquaculture, unfavorable conditions such as overpopulation stress, oxygen deficiency and poor nutrition have been created in the aquaculture process, and these will easily lead to oxidative stress. The damage of oxidative stress is very prominent in production and therefore it is necessary to solve the problem of stress in aquaculture animals (Li et al., 2023). Selenium (Se) improves the body's antioxidant capacity, prevents oxidative stress, enhances immunity and improves body health (Jingyuan et al., 2020; Wischhusen et al., 2020). Many studies have been conducted on the effects of selenium on various fish species, but few studies have been reported on common carp, Cyprinus carpio, (Mustafa and Omar, 2024) although common carp is one of the most important farmed fish species in the world (Adineh et al., 2018). Availa-Se is a zinc-L-selenomethionine complex and is used as a food additive to provide organic selenium in animal feed. Selstar is a type of organic selenium chelated with a mixture of amino acids for adding to the feed of various animals. Today, some researchers believe that the use of selenite in industry is questionable due to the high toxicity of inorganic selenium, its pro-oxidant quality and its bioavailability (Moustafa et al., 2024). The aim of the present study was to determine the effect of sodium selenite on the common carp species in comparison with the two commercial organic selenium compounds mentioned above. For this purpose, growth performance, carcass composition and resistance to salinity stress and oxygen deficiency in common carp were investigated. Methodology The effects of six dietary treatment containing 0.3 mg Se from Availa-Se (Treatment 1), 1 mg Se from Availa-Se (Treatment 2), 0.3 mg Se from Selstar (Treatment 3), 0.3 mg Se from sodium selenite (Treatment 4), 1 mg Se from sodium selenite (Treatment 5) per kg of diet, and a control diet without Se supplementation (Treatment 6) were investigated in common carp for 52 days. Each dietary treatment was studied in 2 tanks (replicates) and each tank contained 31 fish with an initial weight of 1.62±0.33 g (mean ± SD) in a completely randomized design. Fish were fed three times a day ad-libitum. Temperature and dissolved oxygen were measured once a day and salinity and pH were recorded once a week in all tanks. The study of the response to salinity stress has been validated by various researchers to investigate the physiological status and health of fish (Jelkic et al., 2014; Roohi et al., 2017; Talebian Nik and Alamdari, 2020). In the present study, in addition to studying growth performance and carcass composition (AOAC, 2000), for the first time, a combination of salinity stress, oxygen deficiency, and a sudden increase in population density was used to investigate the level of fish resistance. Two tests were used to measure the resistance of fish to death: a) salinity stress with cessation of aeration and b) salinity stress with aeration. In the first test, 10 fish from each tank were suddenly transferred to a container containing 2 liters of water with a salinity of 18 g/L and without aeration. In the second test, 10 fish from each tank were suddenly placed to a container containing 14 liters of water with a salinity of 18 g/L and with aeration. Dead fish were removed and the number of deaths per minute was recorded for until 5 hours. Results The average values of temperature, dissolved oxygen, salinity and pH were 24.6-25.5 °C, 7.4-7.6 mg/L, 2.0-2.1 g/L and 7.6-7.9, respectively. As shown in Table 5, no significant differences were observed in terms of fish weight, standard length, and condition factor between different dietary treatments (p>0.05). Table 6 shows that Se supplements had no significant effect on the survival rate during the rearing period (p>0.05). The dry matter, crude protein and ash contents of carcass did not affect (p>0.05). Feeding with 0.3 mg Availa-Se or sodium selenite per kg diet significantly reduced the crude fat content in the carcass (p<0.05). Based on Table 7, no significant differences were observed in terms of survival time after exposure to salinity stress without aeration (higher-intensity stress) (p>0.05). After salinity stress with aeration (lower-intensity stress), the use of sodium selenite significantly increased survival time (p<0.05) and although the use of Availa-Se improved this index, it had no significant effect compared to the control (p>0.05), while the use of Selstar significantly reduced survival time (p<0.05). Discussion and conclusion The insignificant difference in the survival rate of the fish indicated that the living conditions were suitable for them (Hassan and Mohammad, 2023). There are conflicting reports on the effect of selenium supplementation on the growth and carcass analysis in different fish. It depends on many factors such as water quality, feed and genetic factors (Sumana et al., 2023). Selenium is absorbed more effectively in the presence of certain nutrient compounds and they affect the bioavailability of selenium (Mustafa and Omar, 2024). Selenium affects lipid, sugar and amino acid metabolism in aquatic animals (Li et al., 2023). In the present study, increasing the amount of selenium in the diet did not affect growth parameters and some biochemical carcass indices. Accordingly, higher doses of Se may be required to observe effects on growth parameters (Durigon et al., 2019) and carcass analysis. However, according to new standards, the amount of selenium supplementation in animal feed is limited (Sumana et al., 2023). Studies have shown that varying selenium doses and stress conditions even within the same fish species, affect the severity of stress responses (Mechlaoui et al., 2019). The present study showed that although fish fed a diet without selenium supplementation had good growth and a high survival rate during the rearing period compared to other fish, the amount of selenium in the control diet was not sufficient to protect the fish against stressful conditions. The level of resistance to salinity stress in common carp was higher when sodium selenite than organic selenium with the brand name Availa-Se or Selstar were consumed. In general, the use of Selstar organic selenium is not recommended and 0.3 mg/Kg selenium from sodium selenite is optimal dose in common carp diet. Conflict of interest The authors declare that they have no conflict of interest. Acknowledgement This research was conducted with the financial support of Darya Shilan Dez Company. 73 84 2025/08/272025/02/182025/06/302025/10/82025/08/3 1404/5/12 2026/03/12026/03/12026/03/12026/03/12026/03/1 1404/12/10 حجت اله علمداری Hojjatollah Alamdari گروه شیلات، دانشکده منابع طبیعی، دانشگاه صنعتی خاتم‌‌الانبیاء‌‌ (ص) بهبهان، بهبهان، ایران Iran alamdari671@yahoo.com نرگس بحرینی پور Narges Bahreinipour گروه شیلات، دانشکده منابع طبیعی، دانشگاه صنعتی خاتم‌‌الانبیاء‌‌ (ص) بهبهان، بهبهان، ایران Iran narges.bahrainipour@gmail.com Availa-Se Fish Selstar Sodium selenite Stress استرس اویلاسلنیوم سل‌‌استار سلنیت سدیم ماهی Adineh, H., Harsij, M. and Nazer, A., 2018. Effects of Yucca schidigera extract on the growth performance, feed efficiency, body composition of Common Carp (Cyprinus carpio, Linnaeus 1758) and culture water quality. Iranian Scientific Fisheries Journal, 27(3):11-22. DOI:10.22092/ISFJ.2018.116923 (in Persian)##Antony Jesu Prabhu, P., Holen, E., Espe, M., Silva, M. 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مقاله علمی – پژوهشی:‌ تأثیر برخی شاخص‌های فیزیکی و شیمیایی بر فعالیت آنزیم‌ تریپسین امعاء و احشاء ماهی قزل‌آلای رنگین‌کمان (Oncorhynchus mykiss) Effect of some physical and chemical factors on the trypsin activity from the viscera of rainbow trout (Oncorhynchus mykiss) 1 1 در مطالعه حاضر، اثر برخی شاخص‌های فیزیکی و شیمیایی بر فعالیت آنزیم تریپسین در عصاره خام تهیه شده از ضمائم پیلوریک و روده ماهی قزل‌آلای رنگین‌کمان (Oncorhynchus mykiss) بررسی گردید. امعاء و احشاء ماهی بعد از انتقال به آزمایشگاه و جداسازی ضمائم پیلوریک و روده، در بافر 50 میلی‌مولار HCl Tris- همگن شده و در دمای 4 درجه سانتی‌گراد سانتریفیوژ گردید. سپس مایع‌رویی حاصله برای بررسی شاخص‌های فیزیکی و شیمیایی شامل دمای بهینه، پایداری دمایی، pH بهینه، پایداری pH، بازدارنده‌ها و یون‌های فلزی بر فعالیت آنزیم استفاده گردید. یافته‌ها نشان داد دما و pH بهینه فعالیت آنزیم از ضمائم پیلوریک و روده به‌ترتیب 55 و 5/8  درجه سانتی‌گراد بود و در دمای 50-10 درجه سانتی‌گراد و pH 11-5 نیز پایدار بود. بازدارنده‌های SBTI، TLCK،PMSF، پپاستاتین A و یدواستیک اسید، بازدارندگی معنی‌داری بر فعالیت آنزیم داشتند (05/0>p) درحالی‌که در حضور TPCK و EDTA مهارکنندگی بر فعالیت آنزیم مشاهده نگردید. فعالیت آنزیم تریپسین در ضمائم پیلوریک و روده در حضور یون‌های Cu2+، Ba2+،  Zn2+و Al3+ کاهش معنی‌دار و در حضور یون‌های Ca2+ وMg2+ افزایش معنی‌داری داشت (05/0>p). بر اساس نتایج به‌دست آمده، فعالیت آنزیم‌ تریپسین از ضمائم پیلوریک و روده قزل‌آلای رنگین‌کمان به طور معنی‌داری تحت تأثیر شاخص‌های فیزیکی و شیمیایی مورد مطالعه قرار داشت که می‌تواند بر فعالیت بهینه این آنزیم در فیزیولوژی گوارش مؤثر باشد. 2 Introduction Fish farming industry relies largely on the development of formulated compound diets for promoting growth (Tacon, 2020). However, the ability of fish to digest compound diets is mostly related to the existence of the digestive enzymes in different parts of the gastrointestinal tract (Yúfera and Darias, 2007; Nolasco‐Soria, 2021). In gastric fish species, protein is digested along the gastrointestinal tract by several proteases like pepsin, trypsin, and chymotrypsin. Trypsin, as an alkaline protease, has a key role in the digestion of protein, hydrolyzing them to free amino acids and small peptides for intestinal absorption; therefore, the activity of trypsin has been widely used as a valuable indicator of digestive capacity in fish (Nazdar et al., 2018). Hence, a better understanding of the properties of trypsin is necessary to generate valuable information for protein degradation in the fish digestive tract. In the present work, it was aimed to evaluate the effect of some physicochemical factors on the trypsin activity from pyloric caeca and intestine of rainbow trout (Oncorhynchus mykiss). Methodology Viscera from 10 specimens of rainbow trout with an average weight of 800 ± 31 g were obtained from a local market. Those samples were packed in polyethylene bags, placed in ice with the sample/ice ratio of approximately 1:2(w/w), and directly transported to the laboratory. Upon arrival, the pyloric caeca and intestine were removed from the rest of the collected viscera, washed with cold distilled water (4°C), pooled and stored at -80°C for further analysis (Zamani et al., 2023). The frozen samples were partially thawed in the refrigerator at 4°C for 2h and then cut into small pieces. Those pieces were homogenized in 50mM Tris–HCl buffer, filtered with a cheese cloth for separation of the floating fat phase and then centrifuged at 4°C (Zamani et al., 2014). The resulting supernatant from each sample was collected and finally used for assessment of the physicochemical factors including optimum temperature and thermostability (from 10 to 70 °C), optimum pH and pH stability (from 4.0 to 11.0), inhibitors (SBTI, TLCK, TPCK, PMSF, pepstatin A, iodoacetic acid, and EDTA) and metal ions (K+, Na+, Ca2+, Mg2+, Cu2+, Ba2+, Zn2+, and Al3+) on the trypsin activity according to the method described by Zamani et al. (2023). Results According to the obtained results, optimum temperature and pH of the trypsin from pyloric caeca and intestine were recorded at 55°C and 8.5, respectively. The stability of the trypsin from the both samples was well preserved at temperatures of up to 50°C and pH from 5.0 to 11.0. The enzyme activity from the both samples was significantly inhibited in presence of SBTI, TLCK, PMSF, pepstatin A and iodoacetic acid (p<0.05), while TPCK and EDTA showed no inhibitory effect on the enzyme activity. The enzyme activity from the both samples was significantly increased in the presence of Ca2+ and Mg2+ and decreased by Cu2+, Ba2+, Zn2+, and Al3+ (p<0.05). However, Na+ and K+ did not show any significant effect on the activity of both samples. Discussion and conclusion Enzymes are one of the main biological macromolecules that their maximum activity depend on an optimum temperature to make them functional. The trypsin from pyloric caeca and intestine of rainbow trout had optimum temperature of 55°C. However, an obvious decrease in the trypsin activity of both samples was observed at temperatures above 60°C, probably due to thermal inactivation of this enzyme caused by protein unfolding (Zamani et al., 2014). Similar optimum temperature (55°C) was recorded for trypsins in silver mojarra, cuttlefish, unicorn leatherjacket, and beluga and sevruga (Silva et al., 2011; Balti et al., 2012; Zamani and Benjakul, 2016; Zamani et al., 2023). The optimal temperature of trypsin is in the range of 30-60°C and the differences could be attributed to the temperature of fish habitat or experimental conditions used in assessments (Klomklao and Benjakul, 2018). The stability of trypsin from both samples was highly maintained up to 50°C. These results were in accordance with those of mandarin fish, grey triggerfish, mrigal carp, catfish, albacore tuna, common dolphinfish, and beluga and sevruga (Lu et al., 2008; Jellouli et al., 2009; Khangembam and Chakrabarti, 2015; Dos Santos et al., 2016; Klomklao and Benjakul, 2018; dos Santos et al., 2020; Zamani et al., 2023). In general, thermostability of the trypsin enzyme might vary by some factors such as fish species and experimental conditions (Kanno et al., 2010). Trypsin from both samples had a maximal activity at pH 8.5. The optimum pH (8.5) recorded for the enzyme in both samples was similar with data reported from the brownstripe red snapper, masu salmon, albacore tuna, and Asian seabass (Kanno et al., 2010; Khantaphant and Benjakul, 2010; Klomklao and Benjakul, 2018; Patil et al., 2023). The stability of trypsin from both samples was highly preserved at pH values comprised between 5.0 and 11.0. Similar results were reported for trypsins from grey triggerfish, masu salmon, zebra blenny, catfish, albacore tuna, common dolphinfish, and beluga and sevruga (Jellouli et al., 2009; Kanno et al., 2010; Ktari et al., 2012; Dos Santos et al., 2016; Klomklao and Benjakul, 2018; dos Santos et al., 2020; Zamani et al., 2023). The high ranges of pH may change the net charge and conformation of an enzyme and inhibit to bind to substrate properly, resulting in the abrupt loss of enzymatic activity (Klomklao and Benjakul, 2018). Trypsins are mainly known to be more activity within a range of pH values comprised between 7.5 and 10.5 (Simpson, 2000). The sensitivity of protease enzymes to various inhibitors is a valuable tool for their proper functional characterization (Dos Santos et al., 2020). The results of this study showed that specific inhibitors including SBTI and TLCK had a complete inhibitory effect on the enzyme activity from both samples while the other inhibitors partially inhibited the enzyme activity or had no inhibitory effect on the enzyme activity. Our findings was in agreement with data reported in the brownstripe red snapper, masu salmon, silver mojarra, zebra blenny, mrigral carp, albacore tuna, common dolphinfish, Asian seabass, and beluga and sevruga (Kanno et al., 2010; Khantaphant and Benjakul, 2010; Silva et al., 2011; Ktari et al., 2012; Khangembam and Chakrabarti, 2015; Klomklao and Benjakul, 2018; dos Santos et al., 2020; Zamani et al., 2023; Patil et al., 2023). SBTI is a single polypeptide chain that acts as a reversible competitive inhibitor of trypsin and forms a stable, enzymatically inactive complex with trypsin, resulting in reduction of the enzyme availability (Senphan et al., 2015). TLCK is an irreversible inhibitor of trypsin and trypsin-like serine protease that deactivates these enzymes through the formation of a covalent bond with histidine residue in the catalytic site of the enzyme and blocks the active center of the enzyme for binding to substrate (Eun-Sil et al., 1998). The obtained results from the metal ions showed that Ca2+ and Mg2+ increased the enzyme activity from the both samples while Cu2+, Ba2+, Zn2+, and Al3+ decreased the enzyme activity and Na+ and K+ had no effect on the enzyme activity. Our results were in agreement with data reported in mandarin fish, zebra blenny, common dolphinfish, and beluga and sevruga (Lu et al., 2008; Ktari et al., 2012; dos Santos et al., 2020; Zamani et al., 2023). Metal ions can affect enzyme-catalyzed reactions by changing the electron flow in a substrate or enzyme and play a key role in binding to the substrate, depending on the functional groups present in the active site of enzyme. Abita et al. (1969) showed that Ca2+ can bind to the N-terminal of trypsinogen and, without changing its structure, increase the affinity of the lysine-isoleucine bond for hydrolysis by trypsin. Of course, the effect of metal ions on the activity of trypsin enzyme may depend on fish species as well as dietary and environmental adaptations (Khangembam and Chakrabarti, 2015). Based on the obtained results, the trypsin enzyme activity from the pyloric caeca and intestine of rainbow trout was significantly affected by the tested physicochemical factors which can affect the optimal activity of the enzyme in digestive physiology. Conflict of interest The authors have no conflicts of interest to declare. Acknowledgment The author would like to thank the staff of the Fisheries Laboratory at Malayer University for their helpful assistance during the research process. 85 98 2025/08/272025/02/182025/06/302025/10/82025/08/32025/08/10 1404/5/19 2026/03/12026/03/12026/03/12026/03/12026/03/12026/03/1 1404/12/10 عباس زمانی Abbas Zamani گروه علوم و مهندسی شیلات، دانشکده منابع طبیعی و محیط زیست، دانشگاه ملایر، ملایر، ایران Iran a.zamani@malayeru.ac.ir Digestion Enzymatic inhibitors Pyloric caeca Rainbow trout Trypsin بازدارنده‌های آنزیمی تریپسین ضمائم پیلوریک قزل‌آلای رنگین‌کمان گوارش Abita, J.P., Delaage, M., Lazdunski, M. and Savrda, J., 1969. The Mechanism of Activation of Trypsinogen: The Role of the Four N‐Terminal Aspartyl Residues. European Journal of Biochemistry, 8(3): 314-324. DOI: 10.1111/j.1432-1033.1969.tb00530.x##Balti, R., Bougherra, F., Bougatef, A., Hayet, B.K., Nedjar-Arroume, N., Dhulster, P., Guillochon, D. and Nasri, M., 2012. 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