Investigating the nutritional composition of cultured Asian seabass Lates calcarifer in Bangladesh's Khulna-Satkhira region: A focus on fatty acids and amino acids

Investigating the nutritional composition of cultured Asian seabass Lates calcarifer in Bangladesh's Khulna-Satkhira region: A focus on fatty acids and amino acids | Research Square window.SnipcartSettings = { analytics: { enabled: false } }; (function() { var accessVector = localStorage.getItem('access_vector') || ''; window.dataLayer = window.dataLayer || []; if (accessVector) { window.dataLayer.push({ user: { profile: { profileInfo: { snid: accessVector } } } }); } })(); (function(w,d,s,l,i){w[l]=w[l]||[];w[l].push({'gtm.start':new Date().getTime(),event:'gtm.js'});var f=d.getElementsByTagName(s)[0],j=d.createElement(s),dl=l!='dataLayer'?'&l='+l:'';j.async=true;j.src='https://www.googletagmanager.com/gtm.js?id='+i+dl;f.parentNode.insertBefore(j,f);})(window,document,'script','dataLayer','GTM-K279D39R'); Browse Preprints In Review Journals COVID-19 Preprints AJE Video Bytes Research Tools Research Promotion AJE Professional Editing AJE Rubriq About Preprint Platform In Review Editorial Policies Our Team Advisory Board Help Center Sign In Submit a Preprint Cite Share Download PDF Research Article Investigating the nutritional composition of cultured Asian seabass Lates calcarifer in Bangladesh's Khulna-Satkhira region: A focus on fatty acids and amino acids Roni Sikder, Mrs. Wahida Haque, Saikat Das, Md. Akeruzzaman Shaon, and 2 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-5750226/v1 This work is licensed under a CC BY 4.0 License Status: Posted Version 1 posted You are reading this latest preprint version Abstract Lates calcarifer , known as Asian seabass, is a brackish-water teleost species within the order Carangiformes , native to the Indo-Pacific region. In Bangladesh, this species is found in natural habitats and cultivated on a small scale in the southern coastal region, particularly in the Khulna-Satkhira area. This study analyzed the nutritional composition of cultured Asian seabass in two different size categories, 512.06 ± 14.5 g and 1003.5 ± 36.64 g, collected from this region. Our findings revealed significant differences in the proximate compositions of the two size groups, except ash content (independent t-test, p < 0.05). The smaller size group had a notably higher moisture content (74.96 ± 0.04%), whereas the larger fish exhibited significantly greater protein and fat contents (20.48 ± 0.1% and 5.34 ± 0.18%, respectively). Fatty acid analysis showed distinct differences as well; the larger size group contained higher concentrations of saturated fatty acids (SFAs) and monounsaturated fatty acids (MUFAs), while the smaller fish was richer in polyunsaturated fatty acids (PUFAs). Among the SFAs and omega-3 PUFAs, undecanoic acid and α-linolenic acid were the most prevalent in both groups. However, MUFA profiles differed: palmitoleic acid was dominant in the smaller size group, while oleic acid was more abundant in the larger size group. All essential amino acids (EAAs), except tryptophan, were detected in both groups, with lysine being the most prominent, measuring 5.53±0.44 g/100 g dry weight in smaller size group and 3.91±0.32 g/100 g dry weight in larger size group. Notably, the smaller size group demonstrated a significantly higher overall concentration of EAAs. Overall, this study highlights the nutritional richness of cultured Asian seabass, particularly in terms of fatty acid and amino acid profiles. The superior concentrations of essential amino acids, a higher EPA/DHA ratio, and a more favorable n-3/n-6 ratio in the smaller fish suggest it as a nutritionally superior and well-balanced fish option. Aquaculture and Mariculture Asian seabass Bangladesh aquaculture Cultured fish Nutritional composition Fatty acid profile Amino acid profile Proximate analysis Essential amino acids Figures Figure 1 Figure 2 Figure 3 Figure 4 1. Introduction The Asian seabass ( Lates calcarifer ), also known as barramundi or giant perch, is a highly sought-after catadromous species within the order Carangiformes, known for its culinary appeal and high demand in global markets (Haque et al., 2019 ). This species is native to the Indo-Pacific region, thriving in tropical and subtropical waters, and is widely recognized for its rich flavor and nutritional profile. In response to growing demand, commercial aquaculture of Asian seabass has expanded significantly beyond natural production, with countries such as the Philippines, Vietnam, Malaysia, India, Australia, and Saudi Arabia emerging as key producers (Haque et al., 2019 ). According to the Food and Agriculture Organization (FAO), global production of Asian seabass reached 299,810 metric tons in 2022 (FAO, 2023 ), and its market value is projected to surpass USD 1,012 million by 2024, underscoring its economic importance and promising future (FMI, 2023 ). In Bangladesh, Asian seabass has garnered considerable interest in the southern coastal regions where local markets strongly prefer this species. However, the natural production of Asian seabass in Bangladesh has declined, prompting a shift towards small-scale aquaculture to meet consumer demand. Traditionally dominated by the shrimp farming industry, the southern region has encountered significant setbacks in recent years due to disease outbreaks that have caused severe drops in shrimp production (Heal et al., 2021 ; Islam et al., 2024 ; The Financial Express, 2023 ). Given these challenges, Asian seabass has emerged as a viable aquaculture alternative offering more profit and greater stability. Furthermore, marine species are also regarded as nutritionally superior to shrimp, offering enhanced health benefits due to its well-balanced amino acids and fatty acids, particularly omega-3 fatty acids, which are known to support cardiovascular and neurological health, improve insulin sensitivity, and enhance skin health (Calder, 2015 ; De Carvalho & Caramujo, 2018 ; G. Li et al., 2011 ). Asian seabass, as a marine species, boasts a favorable nutritional composition, making it a valuable source of protein and essential nutrients (Alasalvar et al., 2002 ; Baki et al., 2015 ; Özyurt & Polat, 2006 ). However, the nutrient composition of fish is subject to substantial variability across species and even among individuals within the same species. Several factors, including size, seasonal variations, geographical location, habitat, age, sex, and whether the fish was wild-caught or farm-raised, all contribute to these nutritional differences (Alasalvar et al., 2002 ; Fuentes et al., 2010 ; Grigorakis et al., 2002a ; Krajnović-Ozretic et al., 1994 ).While earlier research has examined the nutritional profiles of wild and cultured Asian seabass concerning these variables, few studies have focused on how nutritional composition may differ across various seasons of cultured Asian seabass (Alasalvar et al., 2002 ; Baki et al., 2015 ; Fuentes et al., 2010 ; Krajnović-Ozretic et al., 1994 ). In Bangladesh, research by (Kamruzzaman, Hossain, Jewel, Khanom, Mustary, & Khatum, 2015 ; Pervin et al., 2012 ) have provided insights into the proximate composition of wild Asian seabass across various life stages, contributing to a better understanding of nutritional variations over the fish's developmental cycle. Nevertheless, a gap exists in the literature regarding the nutritional analysis of cultured Asian seabass, particularly across different size categories commonly encountered in local markets of Bangladesh. The absence of data on size-specific nutrient profiles for cultured Asian seabass underscores the need for comprehensive analysis to assist consumers in making informed dietary choices based on nutrient content. This study aims to address this gap by analyzing the nutritional composition of cultured Asian seabass from two distinct size categories, namely 512.06 ± 14.5 g and 1003.5 ± 36.64 g. These size ranges were selected based on their general market availability, ensuring that the findings are relevant and accessible to consumers. The analysis focuses on proximate nutrient composition, amino acid profile, and fatty acid profile, particularly the presence of omega-3 and other beneficial fatty acids. Understanding the nutritional benefits associated with different size classes of cultured Asian seabass has the potential to inform both consumers and producers to promote the size that supports optimal health outcomes. Moreover, it may encourage the adoption of Asian seabass as a sustainable alternative to shrimp farming, thereby enhancing food security and economic resilience in the aquaculture sector. This research aims to provide an evidence-based framework for selecting nutritionally superior fish sizes within the cultured Asian seabass and guiding aquaculture practices toward producing fish sizes with optimized nutrient profiles. 2. Materials and methods 2.1 Sample collection A total number of 8 Asian seabass were collected from different polyculture farms of Khulna and Satkhira Districts. Among them, 4 fish’s weights were around 500 g, and the remaining four fish’s weights were around 1 kg, which were collected on May 15–16, 2024. After collecting, the samples were stored in an icebox with ice until further analysis. 2.2 Laboratory analysis 2.2.1 Sample preparation At the laboratory, the samples were thawed on a stainless-steel tray. After that, individual samples were dissected, eviscerated, and filleted with a clean stainless steel knife. Then, the fillets were blended into a homogeneous pulp. The prepared samples were then stored in labeled bags at -20°C until further analysis. During these processes, all equipment was properly washed before starting a new sample to avoid cross-contamination. 2.2.2 Proximate composition analysis The moisture, protein, fat, and ash contents were analyzed using AOAC methods (AOAC, 2000 ). Each experiment was conducted in triplicate and values are expressed as mean ± SEM. (Machine) 2.2.3 Extraction and determination of fatty acids Fat extraction was carried out using the Soxhlet method (López-Bascón & De Castro, 2020 ) with petroleum ether as the solvent. The extracted fat was first trans-esterified using sodium methoxide and converted into Fatty Acid Methyl Esters (FAME). Then fatty acids were determined by Gas Chromatography-Flame Ionized Detector (GC-FID) using nitrogen gas. 2.2.4 Extraction and determination of amino acids First, 0.2 g of dry sample was homogenized with 25 ml 7 N HCL and then hydrolyzed with a hydrolyzer at 110⁰C for 24 hours. The hydrolyzed samples were neutralized with 7.5 N NaOH and the solution was prepared up to 250 ml in a volumetric flask with sample dilution buffer (pH 3.4). Then the solution is micro-filtered to remove any foreign particles if present. 100 µL of the solution was taken in a vial and 900 µL sample dilution buffer (pH 3.4) was added to make the volume 1 ml (10 times dilution). Then the amino acid was determined by high-performance liquid chromatography (HPLC) (Bidlingmeyer, 1993 ) with pre-column derivatization. 2.5 Data analysis and visualization Data analysis was conducted using R (v4.4.1.) An independent t-test was conducted to compare the means of proximate composition (e.g., moisture, ash, fat, and protein content) between the two groups. The distribution of fatty acid and amino acid concentrations across the two groups was visualized using violin plots created with the ggplot2 package, which combines kernel density estimation with a boxplot for enhanced interpretation of data distribution. 3. Results and discussions 3.1 Proximate composition The proximate compositions of the two size groups of cultured Asian seabass showed significant differences except for the ash content. Moisture content is crucial as it inversely correlates with fat content, with higher moisture levels indicating lower fat content (Dempson et al., 2004 ). Previous studies have reported that larger fish tend to store higher amounts of fat(Ahlgren et al., 1994 ; Burton & Burton, 2017 ) and this study also supports this observation, as the larger fish demonstrated greater fat accumulation, while the smaller size group exhibited higher moisture content. Additionally, protein content was higher in the larger fish group, which is noteworthy since smaller fish typically exhibit higher protein levels due to their rapid growth and greater reliance on dietary protein (Burton & Burton, 2017 ). The higher protein levels observed in larger Asian seabass can be attributed to their dietary shift, as they move from an omnivorous diet during early developmental stages to a predominantly carnivorous diet as they grow (Davis, 1985 ; Syeda & Hossain, 2012 ). Although the ash content did not differ significantly between the groups, it was slightly higher in the smaller-size group. This could be attributed to the dietary habits of smaller fish, which feed across both lower and higher trophic levels, resulting in higher ash retention. In contrast, larger fish, primarily feed at higher trophic levels, exhibited lower ash content(Aas et al., 2020 ; Hairston, 1993 ; Halver & Hardy, 2003 ). Table 1 Proximate composition of Asian seabass. Values are expressed as mean ± SEM. Values with different superscripts within the row are significantly different at p < 0.05. Proximate ingredient (%) This Study Previous studies on wild-caught Asian seabass reported by Kamruzzaman et al., ( 2015 ) & Pervin et al., ( 2012 ) Smaller size Fish (512.06 ± 14.5 g) Larger size Fish (1003.5 ± 6.64 g) Moisture 74.96 ± 0.04 a 70.75 ± 0.17 b 69% (adult male) to 74% (juvenile) Ash 2.99 ± 0.46 a 2.43 ± 0.15 a 3.25 ± 0.14% (juvenile) to 5.30 ± 0.52% (adult female) Fat 2.33 ± 0.14 a 5.34 ± 0.18 b 3.25 ± 0.14% (juvenile) to 5.30 ± 0.52% (adult female) Protein 18.72 ± 0.33 a 20.48 ± 0.1 b 17.70 ± 0.23% (spent female); 20.45 ± 0.38% (juvenile) & 22.94 ± 0.255% (adult male) When compared to the proximate composition of wild-caught Asian seabass reported by Kamruzzaman et al., ( 2015 ) & Pervin et al., ( 2012 ) (Table 1 ), the cultured fish exhibited almost a similar range of moisture, fat, and protein content except for a slightly lower fat content in the smaller one. However, the ash content was notably lower in the cultured fish. Nutrient retention in fish is governed by a complex interplay of factors including diet composition, availability of food resources, and environmental conditions (Aas et al., 2020 ; Hairston, 1993 ; Halver & Hardy, 2003 ). The reduced ash content in cultured Asian seabass may be attributed to poor feed quality, insufficient feed supply, or sub-optimal rearing conditions. Despite these challenges, the fat content remained within a normal range. This phenomenon could be explained by the catadromous nature of Asian seabass, which migrate between freshwater and marine environments and are exposed to varying environmental conditions. Such migration may lead to the development of metabolic pathways that enhance fat storage, which serves as a crucial energy reserve during periods of food scarcity or environmental stress. 3.2 Fatty acid composition The profiles of saturated fatty acids (SFA), monounsaturated fatty acids (MUFA), and polyunsaturated fatty acids (PUFA) in the flesh lipids of cultured Asian seabass of our study are presented in Figs. 1 , 2 & 3 . The results indicate that the larger size group exhibited higher total concentrations of SFA and MUFA, whereas the smaller size group demonstrated higher PUFA concentrations. These findings align with those reported by De et al. (2019) in different size groups of Tenualosa ilisha . The observed differences in fatty acid profiles between the two groups can be primarily attributed to variations in feeding behavior, which are influenced by factors such as feeding habits(Pillay & Rao, 1963 ) and dietary composition (Arzel et al., 1994 ; Chen et al., 1995 ; Grigorakis et al., 2002b ; Pirini et al., 2000 ). Therefore, the differences in fatty acid profiles between the two groups can be attributed to their feeding behavior. In the early stages, Asian seabass are omnivorous, consuming a mix of plant and animal sources. As they mature, however, they gradually shift to a predominantly carnivorous diet (Corre, 1994 ; Syeda & Hossain, 2012 ). Previous studies on seabass and related species have identified C16:0 (palmitic) and C18:0 (stearic) acids as dominant SFAs (Alasalvar et al., 2002 ; Chanmugam et al., 1986 ; Chen et al., 1995 ; Syama Dayal et al., 2019 ). In contrast, in this study, the C11:0 (undecanoic) was observed as the most dominant SFA, contributing approximately 40% to the total SFA in both groups. C11:0 is primarily derived from plant-based sources, suggesting the significant presence of plant-origin feed components in the diets of cultured seabass. Other dominating SFAs were C16:0 (palmitic), C18:0 (stearic), C14:0 (mysteric), and C10:0 (capric) acids respectively in both groups. Between the two groups, C11:0 (undecanoic), C16:0 (palmitic), C14:0 (mysteric), C10:0 (capric), C24:0 (lignoceric), C13:0 (tridecanoic), and C22:0 (behenic) acids were significantly higher in the larger size group whereas C18:0 (stearic) and C15:0 (pentadecanoic) acids were significantly higher in the smaller ones. The two groups also differed in their MUFA concentration. Within the groups, C18:1 (oleic) acid was the dominant MUFA in larger ones which has also been reported by previous studies (CITATION). On the contrary, C16:1 (palmitoleic) acid was dominant in smaller-size fishes. Differences were also observed in the groupwise distribution of MUFAs. The concentrations of C18:1 (oleic), C16:1 (palmitoleic), and C20:1 (eicosenoic) acids were significantly higher in the larger size group which is similar to the previous studies. The smaller size group, on the other hand, had a higher concentration of C17:1 (heptadecenoic) acid. In terms of polyunsaturated fatty acid (PUFA), the study identified 10 PUFAs in the smaller size group and 8 in the larger size group. Among the n-3 PUFAs, C18:3 (α-linolenic acid) was the dominant, along with significant amounts of C20:5 (Eicosapentaenoic acid, EPA) and C22:6 (Docosahexaenoic acid, DHA) in both groups. Previous studies on seabass (Alasalvar et al., 2002 ; Özyurt & Polat, 2006 ; Xu et al., 2010 ) and other similar fishes (Chen et al., 1995 ; Grigorakis et al., 2002b ; Syama Dayal et al., 2019 ) have identified EPA and DHA as the primary n-3 PUFA components. As C18:3 is predominantly sourced from plant-based sources (Mount Sinai, 2015 ), the high assimilation of C18:3 indicates a diet rich in plant-origin components. However, between the two groups, C18:3 (α-linolenic) and C20:5 (EPA) concentrations were more abundant in the smaller size group while C22:6 (DHA) concentrations were higher in the larger ones. In terms of n-6 PUFA, C18:2 (linoleic) acid was dominant in the larger size group which is similar to other research findings (Alasalvar et al., 2002 ; Syama Dayal et al., 2019 ) but C20:3 (gamma-linolenic) acid was dominant in the smaller size ones. Moreover, it is worth noting that the fatty acid profiles in both groups showed a low proportion of elongated chain fatty acids. This observation could be linked to a reduced capacity for fatty acid chain elongation, as reported by Owen et al., ( 1975 ). Among the major dominating fatty acids, C11:0 (undecanoic) exhibits antifungal and antimicrobial properties, which make it valuable in treating skin infections and promoting skin health (Ammendola et al., 2009 ). In contrast, C16:0 (palmitic) acid supports cellular membrane integrity; however, excessive intake is associated with elevated LDL cholesterol levels and inflammation (Carta et al., 2017 ; Zong et al., 2016 ); Similarly, C14:0 (myristic) acid raises both HDL (good) and LDL (bad) cholesterol levels, thus necessitating a moderate intake to maintain cardiovascular health (Calder, 2015 ; Zong et al., 2016 )​. On the other hand, C18:0 (stearic) acid has a neutral effect on cholesterol concentrations and does not significantly elevate cardiovascular risk, rendering it a safer choice among saturated fats (Zong et al., 2016 ); C15:0 (pentadecanoic) acid, meanwhile, possesses anti-inflammatory properties and supports metabolic health, potentially lowering the risk of diabetes and cardiovascular diseases (Calder, 2015 ; Zong et al., 2016 ); Furthermore, C16:1 (palmitoleic) acid is recognized for its metabolic health benefits, enhancing insulin sensitivity, and mitigating the risk of type 2 diabetes (Frigolet & Gutiérrez-Aguilar, 2017 ); In addition, C18:1 (oleic) acid supports immune function, lowers blood pressure, and reduces LDL cholesterol levels (Pravst, 2014 ); both C16:1 (palmitoleic) and C18:1 (oleic) acids possess anti-inflammatory properties that promote cardiovascular health (Frigolet & Gutiérrez-Aguilar, 2017 ; Pravst, 2014 ); C14:1 (myristoleic) acid has anti-cancer properties, is associated with benefits for bone health, prevents non-alcoholic fatty liver disease (Y.-G. Kim et al., 2021 ; Kwon et al., 2015 ; S. Z. Li et al., 2023 ; Quan et al., 2020 ). Additionally, C18:3 (α-Linolenic) acid, along with other n-3 series fatty acids like EPA and DHA support cardiovascular health, reduce inflammation, aid cognitive function, and provide antioxidant benefits. The increase in α-Linolenic acid intake, ranging from 0.58 g to 2.51 g, has demonstrated improvements in cardiovascular health (Gebauer et al., 2006 ). A 2:1 EPA/DHA ratio is recommended for optimal cardiovascular outcomes, alongside a daily intake of approximately 500 mg of EPA and DHA (Gebauer et al., 2006 ), which can be obtained by consuming around 500 g of smaller fish and 600 g of larger fish. Our findings further highlight that the smaller-size group exhibited a favorable n-3/n-6 ratio of 1.37, whereas the larger-size group showed a ratio of 0.91. Both ratios meet the recommended threshold of 0.2 or higher, confirming their adequacy for maintaining optimal cardiovascular health (Syama Dayal et al., 2019 ). 3.3 Amino acid composition The amino acid profile of the muscle tissue in cultured Asian seabass is presented in Fig. 4 . This analysis identified a total of 17 amino acids, including all essential amino acids (EAA) except tryptophan. Among the EAAs, lysine was the most abundant, followed by leucine and arginine in both size groups. Interestingly, the concentrations of all EAAs were significantly higher in the smaller-size group, with the exceptions of methionine and phenylalanine, which, although not statistically significant, exhibited higher numerical values in the smaller group. Regarding non-essential amino acids (NAA) and glutamic acid emerged as the most abundant, followed by aspartic acid and alanine in both groups. Significant differences were observed in the concentrations of all NAAs between the groups, except for serine. Specifically, glutamic acid, aspartic acid, alanine, and tyrosine were significantly elevated in the smaller-size group, while glycine and proline were substantially more abundant in the larger-size group. These findings align with previous studies on seabass (Özyurt & Polat, 2006 ) and related species (Iwasaki & Harada, 1985 ; Mohanty et al., 2014 ). Özyurt & Polat, ( 2006 ) reported a total of 16 amino acids in seabass muscle, with similar dominant amino acids. They also noted that amino acid composition varies with the spawning period and feeding behavior of the fish. Similarly, Mohanty et al., ( 2014 ) highlighted that factors such as capture time and geographical location influence the quantity and types of amino acids in fish muscle. Amino acids (AAs) are recognized as essential building blocks of tissue proteins and play critical roles in synthesizing low molecular weight compounds, including glutathione, creatine, thyroid hormones, melanin, and melatonin, which are vital for various physiological processes (Blachier et al., 2011 ; J. Kim et al., 2012 ; Kong et al., 2012 ; Wu et al., 2009 ). Besides, the elevated levels of essential amino acids (EAA) in muscle are indicative of superior muscle protein quality (Young & Pellett, 1984 ). In the present study, the total EAA/total NAA ratios were 0.81 and 0.71 in the smaller and larger size groups, respectively. According toFAO/WHO, ( 1991 ) recommendations, a ratio of 0.6 or higher is considered a benchmark for high-quality protein, confirming that both groups exhibit high protein quality. Additionally, beyond essential and non-essential amino acids, a distinct group known as functional amino acids has gained recognition for their roles in regulating physiological and metabolic processes critical for health, growth, reproduction, and disease resistance (G. Li et al., 2011 ; Rezaei et al., 2013 ; Wang et al., 2013 ; Wu, 2009 , 2013 ). Functional amino acids include arginine, aspartate, cysteine, glutamine, glycine, histidine, leucine, methionine, proline, threonine, tryptophan, and tyrosine (Mohanty et al., 2014 ; Wu, 2009 , 2013 )Notably, this study found that Asian seabass muscle tissue contained all functional amino acids except tryptophan. 4. Conclusions The results of this study demonstrate that cultured Asian seabass is a highly nutritious fish, offering high levels of crude protein, fat, and ash content; an excellent source of high-quality protein, providing a well-balanced composition of essential amino acids, along with substantial amounts of α-linolenic acid, EPA, and DHA, making it a valuable dietary choice. These attributes not only benefit consumers but also support the aquaculture industry. Notably, cultured Asian seabass exhibited nutritional profiles comparable to, and in some instances surpassing, those of wild-caught seabass in terms of amino acid and fatty acid content. Significant differences in nutritional composition were observed between the two size groups, offering essential insights into their potential health benefits. The smaller size group demonstrated superior concentrations of essential amino acids, a higher EPA to DHA ratio, and a more favorable n-3 to n-6 ratio compared to the larger group. Based on these findings, it can be concluded that the smaller size group of cultured Asian seabass possesses higher nutritional value than the larger size group, making it a particularly beneficial option for dietary purposes. Thus, the results of this study will provide a framework for selecting the nutritionally superior fish, which may ultimately help in redesigning market demand and consumer preference. Declarations Frozen cultured Asian seabass were donated for use in this study by local farms during their normal harvests. Competing Interests We declare that the concerned authors and co-authors carried out the work, and all the authors have read the draft manuscript. We would also like to declare that there is no potential conflict of interest among the authors. Funding statement: The corresponding author, Roni Sikder (RS), acknowledges that the Ministry of Science and Technology, Bangladesh, partially funded this work by awarding the National Science of Technology Fellowship. The funders had no role in study design, data collection and analysis, publication decisions, or manuscript preparation. Acknowledgment The authors would like to thank the Institute of Technology Transfer and Innovation (ITTI), BCSIR for providing this study's necessary resources and facilities. Additionally, we appreciate the valuable contributions and insightful discussions from our colleagues and mentors. Data availability statement All data generated or analyzed during this study are included in this manuscript. References Aas, T. S., Sixten, H. J., Hillestad, M., Ytrestøyl, T., Sveier, H., & Åsgård, T. (2020). Feed intake, nutrient digestibility and nutrient retention in Atlantic salmon (Salmo salar L.) fed diets with different physical pellet quality. Journal of Fisheries , 8 (2), 768–776. https://doi.org/10.17017/j.fish.133 Ahlgren, G., Blomqvist, P., Boberg, M., & Gustafsson, I. (1994). Fatty acid content of the dorsal muscle—an indicator of fat quality in freshwater fish. 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Nobiletin ameliorates nonalcoholic fatty liver disease by regulating gut microbiota and myristoleic acid metabolism. Journal of Agricultural and Food Chemistry , 71 (19), 7312–7323. López-Bascón, M. A., & De Castro, M. D. L. (2020). Soxhlet extraction. In Liquid-phase extraction (pp. 327–354). Elsevier. Mohanty, B., Mahanty, A., Ganguly, S., Sankar, T. V, Chakraborty, K., Rangasamy, A., Paul, B., Sarma, D., Mathew, S., & Asha, K. K. (2014). Amino acid compositions of 27 food fishes and their importance in clinical nutrition. Journal of Amino Acids , 2014 (1), 269797. Mount Sinai. (2015). Alpha-linolenic acid. Mount Sinai. https://www.mountsinai.org/health-library/supplement/alpha-linolenic-acid. Mount Sinai . Owen, J. M., Adron, J. W., Middleton, C., & Cowey, C. B. (1975). Elongation and desaturation of dietary fatty acids in turbotScophthalmus maximus L., and rainbow trout, Salmo gairdnerii rich. Lipids , 10 (9), 528–531. Özyurt, G., & Polat, A. (2006). Amino acid and fatty acid composition of wild sea bass (Dicentrarchus labrax): a seasonal differentiation. European Food Research and Technology , 222 , 316–320. Pervin, T., Yeasmin, S., Islam, R., Rahman, A., & Sattar, A. (2012). Studies on nutritional composition and characterization of lipids of Lates calcarifer (Bhetki). In Bangladesh J. Sci. Ind. Res (Vol. 47, Issue 4). www.banglajol.info Pillay, S. R., & Rao, K. V. (1963). Observations on the biology and fishery of the Hilsa, Hilsa ilisha (Wilton) of river Godavari. IPFC Proceedings 10th Session Tech. Pap , 6 , 37–61. Pirini, M., Gatta, P. P., Testi, S., Trigari, G., & Monetti, P. G. (2000). Effect of refrigerated storage on muscle lipid quality of sea bass (Dicentrarchus labrax) fed on diets containing different levels of vitamin E. Food Chemistry , 68 (3), 289–293. Pravst, I. (2014). Oleic acid and its potential health effects . https://www.researchgate.net/publication/264503445 Quan, L. H., Zhang, C., Dong, M., Dong, M., Jiang, J., Xu, H., Yan, C., Liu, X., Zhou, H., Zhou, H., Zhang, H., Zhang, H., Chen, L., Chen, L., Zhong, F. L., Luo, Z. B., Lam, S. M., Shui, G., Li, D., … Jin, W. (2020). Myristoleic acid produced by enterococci reduces obesity through brown adipose tissue activation. Gut , 69 (7), 1239–1247. https://doi.org/10.1136/gutjnl-2019-319114 Rezaei, R., Wang, W., Wu, Z., Dai, Z., Wang, J., & Wu, G. (2013). Biochemical and physiological bases for utilization of dietary amino acids by young pigs. Journal of Animal Science and Biotechnology , 4 , 1–12. Syama Dayal, J., Ambasankar, K., Jannathulla, R., Kumuraguruvasagam, K. P., & Kailasam, M. (2019). Nutrient and fatty acid composition of cultured and wild caught gold-spot mullet Liza parsia (Hamilton, 1822). Indian Journal of Fisheries , 66 (2), 62–70. https://doi.org/10.21077/ijf.2019.66.2.84993-09 Syeda, M.-A.-N., & Hossain, Md. D. (2012). Seasonal Variation of Food composition and Feeding Activity of Small Adult Barramundi (Lates calcarifer, Bloch) in the South west Coastal Water near Khulna, Bangladesh . The Financial Express. (2023, September 6). Bangladesh reports drastic decline in shrimp production. The Financial Express. https://thefinancialexpress.com.bd/trade/bangladesh-reports-drastic-decline-in-shrimp-production . Wang, W., Wu, Z., Dai, Z., Yang, Y., Wang, J., & Wu, G. (2013). Glycine metabolism in animals and humans: implications for nutrition and health. Amino Acids , 45 , 463–477. Wu, G. (2009). Amino acids: Metabolism, functions, and nutrition. In Amino Acids (Vol. 37, Issue 1, pp. 1–17). https://doi.org/10.1007/s00726-009-0269-0 Wu, G. (2013). Functional amino acids in nutrition and health. In Amino acids (Vol. 45, pp. 407–411). Springer. Wu, G., Bazer, F. W., Davis, T. A., Kim, S. W., Li, P., Marc Rhoads, J., Carey Satterfield, M., Smith, S. B., Spencer, T. E., & Yin, Y. (2009). Arginine metabolism and nutrition in growth, health and disease. Amino Acids , 37 , 153–168. Xu, J., Yan, B., Teng, Y., Lou, G., & Lu, Z. (2010). Analysis of nutrient composition and fatty acid profiles of Japanese sea bass Lateolabrax japonicus (Cuvier) reared in seawater and freshwater. Journal of Food Composition and Analysis , 23 (5), 401–405. https://doi.org/10.1016/j.jfca.2010.01.010 Young, V. R., & Pellett, P. L. (1984). Amino acid composition in relation to protein nutritional quality of meat and poultry products. The American Journal of Clinical Nutrition , 40 (3), 737–742. Zong, G., Li, Y., Wanders, A. J., Alssema, M., Zock, P. L., Willett, W. C., Hu, F. B., & Sun, Q. (2016). Intake of individual saturated fatty acids and risk of coronary heart disease in US men and women: Two prospective longitudinal cohort studies. BMJ (Online) , 355 . https://doi.org/10.1136/bmj.i5796 Additional Declarations The authors declare no competing interests. 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Also discoverable on Platform About Our Team In Review Editorial Policies Advisory Board Help Center Resources Author Services Accessibility API Access RSS feed Manage Cookie Preferences © Research Square 2026 | ISSN 2693-5015 (online) Privacy Policy Terms of Service Do Not Sell My Personal Information {"props":{"pageProps":{"initialData":{"identity":"rs-5750226","acceptedTermsAndConditions":true,"allowDirectSubmit":true,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":396662268,"identity":"32c581ac-23f5-4431-a015-18296013c87c","order_by":0,"name":"Roni Sikder","email":"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAAA+UlEQVRIie3PsYrCMBjA8YRAXVpcheo7BDr4MC4RwcnA3dbBk7joUnWtIN4r9JaulxCoS7h7gFvuEe6WI3AOfnUUausmmP8Q+OD7QT6EXK47TSLKsEDk8G9jGAkRTYnHQmRKgusJVBKfhnhRDjWkv1wpZZ/GvU0oafS8fxm0l0BsnFeSrvlg2qeTaLtmbLTNDzzVWODEfFWSTmdCNaLxMDNIyiAvuABC8OI6URbIu8FzEewK/tqESPjYMPMJIYGY8qyW+IbCLeMoNZ6H00LyNyDq6i2tJPq1x1Fvk7T/0M90xvefWn3buJpcps+vbLwPzW5ZdrlcrgfpBLRPXjiLPhIuAAAAAElFTkSuQmCC","orcid":"https://orcid.org/0009-0008-9890-5137","institution":"Department of Fisheries, University of Dhaka; Dhaka-1000, Bangladesh","correspondingAuthor":true,"prefix":"","firstName":"Roni","middleName":"","lastName":"Sikder","suffix":""},{"id":396662269,"identity":"27e73dfe-0d82-402d-985f-68e0dfdd4b35","order_by":1,"name":"Mrs. Wahida Haque","email":"","orcid":"","institution":"Department of Fisheries, University of Dhaka; Dhaka-1000, Bangladesh","correspondingAuthor":false,"prefix":"Mrs.","firstName":"Wahida","middleName":"","lastName":"Haque","suffix":""},{"id":396662270,"identity":"59da2e46-eba9-4de9-8d18-2816b3626022","order_by":2,"name":"Saikat Das","email":"","orcid":"https://orcid.org/0009-0008-3178-342X","institution":"Department of Fisheries, University of Dhaka; Dhaka-1000, Bangladesh","correspondingAuthor":false,"prefix":"","firstName":"Saikat","middleName":"","lastName":"Das","suffix":""},{"id":396662271,"identity":"cff491f4-251b-43d0-9cf6-0850f52156e9","order_by":3,"name":"Md. Akeruzzaman Shaon","email":"","orcid":"https://orcid.org/0009-0007-3472-2916","institution":"Institute of Technology Transfer and Innovation (ITTI), Bangladesh Council of Scientific and Industrial Research (BCSIR); Dhaka-1205, Bangladesh. Laboratory of Nutrition and Health Research, Department of Biochemistry and Molecular Biology, University of Dhaka; Dhaka-1000, Bangladesh","correspondingAuthor":false,"prefix":"","firstName":"Md.","middleName":"Akeruzzaman","lastName":"Shaon","suffix":""},{"id":396670581,"identity":"ee386b0e-10b9-4e6e-9c40-9517d2dbf0fe","order_by":4,"name":"Md. Zafrul Islam","email":"","orcid":"https://orcid.org/0009-0005-9693-5090","institution":"Department of Fisheries, University of Dhaka; Dhaka-1000, Bangladesh","correspondingAuthor":false,"prefix":"","firstName":"Md.","middleName":"Zafrul","lastName":"Islam","suffix":""},{"id":396670582,"identity":"f9aebd58-27ca-49d5-ac01-362fd41aef25","order_by":5,"name":"Md. Rakibul Hasan","email":"","orcid":"https://orcid.org/0000-0002-2980-8690","institution":"Institute of Technology Transfer and Innovation (ITTI), Bangladesh Council of Scientific and Industrial Research (BCSIR); Dhaka-1205, Bangladesh","correspondingAuthor":false,"prefix":"","firstName":"Md.","middleName":"Rakibul","lastName":"Hasan","suffix":""}],"badges":[],"createdAt":"2025-01-02 08:43:35","currentVersionCode":1,"declarations":{"humanSubjects":false,"vertebrateSubjects":true,"conflictsOfInterestStatement":false,"humanSubjectEthicalGuidelines":false,"humanSubjectConsent":false,"humanSubjectClinicalTrial":false,"humanSubjectCaseReport":false,"vertebrateSubjectEthicalGuidelines":true},"doi":"10.21203/rs.3.rs-5750226/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-5750226/v1","draftVersion":[],"editorialEvents":[],"editorialNote":"","failedWorkflow":false,"files":[{"id":73729090,"identity":"0df4eacd-fada-4ce9-b35c-7647438da4e4","added_by":"auto","created_at":"2025-01-14 04:55:39","extension":"png","order_by":1,"title":"Figure 1","display":"","copyAsset":false,"role":"figure","size":186052,"visible":true,"origin":"","legend":"\u003cp\u003eConcentrations of saturated fatty acids (mg/100 g dry weight). Violin plot is representing data by highlighting differences between the two groups. Statistical comparisons are indicated where relevant.\u003c/p\u003e","description":"","filename":"1.png","url":"https://assets-eu.researchsquare.com/files/rs-5750226/v1/75b010b928482b15eba2f63b.png"},{"id":73729089,"identity":"778968c0-0d0d-4a98-a3ed-0ed4a324702e","added_by":"auto","created_at":"2025-01-14 04:55:39","extension":"png","order_by":2,"title":"Figure 2","display":"","copyAsset":false,"role":"figure","size":119600,"visible":true,"origin":"","legend":"\u003cp\u003eConcentrations of monounsaturated fatty acids (mg/100 g dry weight). Violin plot is representing data by highlighting differences between the two groups. Statistical comparisons are indicated where relevant.\u003c/p\u003e","description":"","filename":"2.png","url":"https://assets-eu.researchsquare.com/files/rs-5750226/v1/815210bec6f19faf37d6846a.png"},{"id":73729094,"identity":"37ea4bc2-571c-46aa-8659-a3843325565e","added_by":"auto","created_at":"2025-01-14 04:55:39","extension":"png","order_by":3,"title":"Figure 3","display":"","copyAsset":false,"role":"figure","size":166260,"visible":true,"origin":"","legend":"\u003cp\u003eConcentrations of polyunsaturated fatty acids (PUFAs) across the two groups, categorized into n-3 PUFAs (α-linolenic acid, eicosatrienoic acid, eicosapentaenoic acid, docosahexaenoic acid), n-6 PUFAs (linoleic acid, γ-linolenic acid, dihomo-γ-linolenic acid), and other PUFAs (trans-linolelaidic acid, eicosadienoic acid, docosadienoic acid). Violin plot is representing data by highlighting differences between the two groups. Statistical comparisons are indicated where relevant\u003c/p\u003e","description":"","filename":"3.png","url":"https://assets-eu.researchsquare.com/files/rs-5750226/v1/a643ca9390bff00e20231a79.png"},{"id":73729102,"identity":"02ef00cc-dc3c-4114-a669-d543bd54efc2","added_by":"auto","created_at":"2025-01-14 04:55:39","extension":"png","order_by":4,"title":"Figure 4","display":"","copyAsset":false,"role":"figure","size":194597,"visible":true,"origin":"","legend":"\u003cp\u003eConcentrations of essential and non-essential amino acids in larger-sized and smaller-sized fish, expressed in g/100g dry weight. Violin plot is illustrating data by highlighting differences between the two groups where essential amino acids are shown on the left and non-essential amino acids on the right.\u003c/p\u003e","description":"","filename":"4.png","url":"https://assets-eu.researchsquare.com/files/rs-5750226/v1/bc9011e4e3ce17d07ac82864.png"},{"id":73730489,"identity":"c1d3ea84-bf1a-4767-a762-58be499d820d","added_by":"auto","created_at":"2025-01-14 05:27:44","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":1333711,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-5750226/v1/51b75b5c-fa9d-4c23-a0bb-47b03a5c2917.pdf"}],"financialInterests":"The authors declare no competing interests.","formattedTitle":"\u003cp\u003e\u003cstrong\u003eInvestigating the nutritional composition of cultured Asian seabass \u003c/strong\u003e\u003cem\u003e\u003cstrong\u003eLates calcarifer\u003c/strong\u003e\u003c/em\u003e\u003cstrong\u003e in Bangladesh's Khulna-Satkhira region: A focus on fatty acids and amino acids\u003c/strong\u003e\u003c/p\u003e","fulltext":[{"header":"1. Introduction","content":"\u003cp\u003eThe Asian seabass (\u003cem\u003eLates calcarifer\u003c/em\u003e), also known as barramundi or giant perch, is a highly sought-after catadromous species within the order Carangiformes, known for its culinary appeal and high demand in global markets (Haque et al., \u003cspan citationid=\"CR29\" class=\"CitationRef\"\u003e2019\u003c/span\u003e). This species is native to the Indo-Pacific region, thriving in tropical and subtropical waters, and is widely recognized for its rich flavor and nutritional profile. In response to growing demand, commercial aquaculture of Asian seabass has expanded significantly beyond natural production, with countries such as the Philippines, Vietnam, Malaysia, India, Australia, and Saudi Arabia emerging as key producers (Haque et al., \u003cspan citationid=\"CR29\" class=\"CitationRef\"\u003e2019\u003c/span\u003e). According to the Food and Agriculture Organization (FAO), global production of Asian seabass reached 299,810 metric tons in 2022 (FAO, \u003cspan citationid=\"CR19\" class=\"CitationRef\"\u003e2023\u003c/span\u003e), and its market value is projected to surpass USD 1,012\u0026nbsp;million by 2024, underscoring its economic importance and promising future (FMI, \u003cspan citationid=\"CR21\" class=\"CitationRef\"\u003e2023\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eIn Bangladesh, Asian seabass has garnered considerable interest in the southern coastal regions where local markets strongly prefer this species. However, the natural production of Asian seabass in Bangladesh has declined, prompting a shift towards small-scale aquaculture to meet consumer demand. Traditionally dominated by the shrimp farming industry, the southern region has encountered significant setbacks in recent years due to disease outbreaks that have caused severe drops in shrimp production (Heal et al., \u003cspan citationid=\"CR30\" class=\"CitationRef\"\u003e2021\u003c/span\u003e; Islam et al., \u003cspan citationid=\"CR31\" class=\"CitationRef\"\u003e2024\u003c/span\u003e; The Financial Express, \u003cspan citationid=\"CR54\" class=\"CitationRef\"\u003e2023\u003c/span\u003e). Given these challenges, Asian seabass has emerged as a viable aquaculture alternative offering more profit and greater stability. Furthermore, marine species are also regarded as nutritionally superior to shrimp, offering enhanced health benefits due to its well-balanced amino acids and fatty acids, particularly omega-3 fatty acids, which are known to support cardiovascular and neurological health, improve insulin sensitivity, and enhance skin health (Calder, \u003cspan citationid=\"CR11\" class=\"CitationRef\"\u003e2015\u003c/span\u003e; De Carvalho \u0026amp; Caramujo, \u003cspan citationid=\"CR17\" class=\"CitationRef\"\u003e2018\u003c/span\u003e; G. Li et al., \u003cspan citationid=\"CR39\" class=\"CitationRef\"\u003e2011\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eAsian seabass, as a marine species, boasts a favorable nutritional composition, making it a valuable source of protein and essential nutrients (Alasalvar et al., \u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e2002\u003c/span\u003e; Baki et al., \u003cspan citationid=\"CR7\" class=\"CitationRef\"\u003e2015\u003c/span\u003e; \u0026Ouml;zyurt \u0026amp; Polat, \u003cspan citationid=\"CR45\" class=\"CitationRef\"\u003e2006\u003c/span\u003e). However, the nutrient composition of fish is subject to substantial variability across species and even among individuals within the same species. Several factors, including size, seasonal variations, geographical location, habitat, age, sex, and whether the fish was wild-caught or farm-raised, all contribute to these nutritional differences (Alasalvar et al., \u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e2002\u003c/span\u003e; Fuentes et al., \u003cspan citationid=\"CR23\" class=\"CitationRef\"\u003e2010\u003c/span\u003e; Grigorakis et al., \u003cspan citationid=\"CR25\" class=\"CitationRef\"\u003e2002a\u003c/span\u003e; Krajnović-Ozretic et al., \u003cspan citationid=\"CR37\" class=\"CitationRef\"\u003e1994\u003c/span\u003e).While earlier research has examined the nutritional profiles of wild and cultured Asian seabass concerning these variables, few studies have focused on how nutritional composition may differ across various seasons of cultured Asian seabass (Alasalvar et al., \u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e2002\u003c/span\u003e; Baki et al., \u003cspan citationid=\"CR7\" class=\"CitationRef\"\u003e2015\u003c/span\u003e; Fuentes et al., \u003cspan citationid=\"CR23\" class=\"CitationRef\"\u003e2010\u003c/span\u003e; Krajnović-Ozretic et al., \u003cspan citationid=\"CR37\" class=\"CitationRef\"\u003e1994\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eIn Bangladesh, research by (Kamruzzaman, Hossain, Jewel, Khanom, Mustary, \u0026amp; Khatum, \u003cspan citationid=\"CR33\" class=\"CitationRef\"\u003e2015\u003c/span\u003e; Pervin et al., \u003cspan citationid=\"CR46\" class=\"CitationRef\"\u003e2012\u003c/span\u003e) have provided insights into the proximate composition of wild Asian seabass across various life stages, contributing to a better understanding of nutritional variations over the fish's developmental cycle. Nevertheless, a gap exists in the literature regarding the nutritional analysis of cultured Asian seabass, particularly across different size categories commonly encountered in local markets of Bangladesh. The absence of data on size-specific nutrient profiles for cultured Asian seabass underscores the need for comprehensive analysis to assist consumers in making informed dietary choices based on nutrient content.\u003c/p\u003e \u003cp\u003eThis study aims to address this gap by analyzing the nutritional composition of cultured Asian seabass from two distinct size categories, namely 512.06\u0026thinsp;\u0026plusmn;\u0026thinsp;14.5 g and 1003.5\u0026thinsp;\u0026plusmn;\u0026thinsp;36.64 g. These size ranges were selected based on their general market availability, ensuring that the findings are relevant and accessible to consumers. The analysis focuses on proximate nutrient composition, amino acid profile, and fatty acid profile, particularly the presence of omega-3 and other beneficial fatty acids. Understanding the nutritional benefits associated with different size classes of cultured Asian seabass has the potential to inform both consumers and producers to promote the size that supports optimal health outcomes. Moreover, it may encourage the adoption of Asian seabass as a sustainable alternative to shrimp farming, thereby enhancing food security and economic resilience in the aquaculture sector.\u003c/p\u003e \u003cp\u003eThis research aims to provide an evidence-based framework for selecting nutritionally superior fish sizes within the cultured Asian seabass and guiding aquaculture practices toward producing fish sizes with optimized nutrient profiles.\u003c/p\u003e"},{"header":"2. Materials and methods","content":"\u003cdiv id=\"Sec3\" class=\"Section2\"\u003e \u003ch2\u003e2.1 Sample collection\u003c/h2\u003e \u003cp\u003eA total number of 8 Asian seabass were collected from different polyculture farms of Khulna and Satkhira Districts. Among them, 4 fish\u0026rsquo;s weights were around 500 g, and the remaining four fish\u0026rsquo;s weights were around 1 kg, which were collected on May 15\u0026ndash;16, 2024. After collecting, the samples were stored in an icebox with ice until further analysis.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec4\" class=\"Section2\"\u003e \u003ch2\u003e2.2 Laboratory analysis\u003c/h2\u003e \u003cdiv id=\"Sec5\" class=\"Section3\"\u003e \u003ch2\u003e2.2.1 Sample preparation\u003c/h2\u003e \u003cp\u003eAt the laboratory, the samples were thawed on a stainless-steel tray. After that, individual samples were dissected, eviscerated, and filleted with a clean stainless steel knife. Then, the fillets were blended into a homogeneous pulp. The prepared samples were then stored in labeled bags at -20\u0026deg;C until further analysis. During these processes, all equipment was properly washed before starting a new sample to avoid cross-contamination.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec6\" class=\"Section3\"\u003e \u003ch2\u003e2.2.2 Proximate composition analysis\u003c/h2\u003e \u003cp\u003eThe moisture, protein, fat, and ash contents were analyzed using AOAC methods (AOAC, \u003cspan citationid=\"CR5\" class=\"CitationRef\"\u003e2000\u003c/span\u003e). Each experiment was conducted in triplicate and values are expressed as mean\u0026thinsp;\u0026plusmn;\u0026thinsp;SEM. (Machine)\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec7\" class=\"Section3\"\u003e \u003ch2\u003e2.2.3 Extraction and determination of fatty acids\u003c/h2\u003e \u003cp\u003eFat extraction was carried out using the Soxhlet method (L\u0026oacute;pez-Basc\u0026oacute;n \u0026amp; De Castro, \u003cspan citationid=\"CR41\" class=\"CitationRef\"\u003e2020\u003c/span\u003e) with petroleum ether as the solvent. The extracted fat was first trans-esterified using sodium methoxide and converted into Fatty Acid Methyl Esters (FAME). Then fatty acids were determined by Gas Chromatography-Flame Ionized Detector (GC-FID) using nitrogen gas.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec8\" class=\"Section3\"\u003e \u003ch2\u003e2.2.4 Extraction and determination of amino acids\u003c/h2\u003e \u003cp\u003eFirst, 0.2 g of dry sample was homogenized with 25 ml 7 N HCL and then hydrolyzed with a hydrolyzer at 110⁰C for 24 hours. The hydrolyzed samples were neutralized with 7.5 N NaOH and the solution was prepared up to 250 ml in a volumetric flask with sample dilution buffer (pH 3.4). Then the solution is micro-filtered to remove any foreign particles if present. 100 \u0026micro;L of the solution was taken in a vial and 900 \u0026micro;L sample dilution buffer (pH 3.4) was added to make the volume 1 ml (10 times dilution). Then the amino acid was determined by high-performance liquid chromatography (HPLC) (Bidlingmeyer, \u003cspan citationid=\"CR8\" class=\"CitationRef\"\u003e1993\u003c/span\u003e) with pre-column derivatization.\u003c/p\u003e \u003c/div\u003e \u003c/div\u003e \u003cdiv id=\"Sec9\" class=\"Section2\"\u003e \u003ch2\u003e2.5 Data analysis and visualization\u003c/h2\u003e \u003cp\u003eData analysis was conducted using R (v4.4.1.) An independent t-test was conducted to compare the means of proximate composition (e.g., moisture, ash, fat, and protein content) between the two groups. The distribution of fatty acid and amino acid concentrations across the two groups was visualized using violin plots created with the ggplot2 package, which combines kernel density estimation with a boxplot for enhanced interpretation of data distribution.\u003c/p\u003e \u003c/div\u003e"},{"header":"3. Results and discussions","content":"\u003cdiv id=\"Sec11\" class=\"Section2\"\u003e \u003ch2\u003e3.1 Proximate composition\u003c/h2\u003e \u003cp\u003eThe proximate compositions of the two size groups of cultured Asian seabass showed significant differences except for the ash content. Moisture content is crucial as it inversely correlates with fat content, with higher moisture levels indicating lower fat content (Dempson et al., \u003cspan citationid=\"CR18\" class=\"CitationRef\"\u003e2004\u003c/span\u003e). Previous studies have reported that larger fish tend to store higher amounts of fat(Ahlgren et al., \u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e1994\u003c/span\u003e; Burton \u0026amp; Burton, \u003cspan citationid=\"CR10\" class=\"CitationRef\"\u003e2017\u003c/span\u003e) and this study also supports this observation, as the larger fish demonstrated greater fat accumulation, while the smaller size group exhibited higher moisture content. Additionally, protein content was higher in the larger fish group, which is noteworthy since smaller fish typically exhibit higher protein levels due to their rapid growth and greater reliance on dietary protein (Burton \u0026amp; Burton, \u003cspan citationid=\"CR10\" class=\"CitationRef\"\u003e2017\u003c/span\u003e). The higher protein levels observed in larger Asian seabass can be attributed to their dietary shift, as they move from an omnivorous diet during early developmental stages to a predominantly carnivorous diet as they grow (Davis, \u003cspan citationid=\"CR16\" class=\"CitationRef\"\u003e1985\u003c/span\u003e; Syeda \u0026amp; Hossain, \u003cspan citationid=\"CR53\" class=\"CitationRef\"\u003e2012\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eAlthough the ash content did not differ significantly between the groups, it was slightly higher in the smaller-size group. This could be attributed to the dietary habits of smaller fish, which feed across both lower and higher trophic levels, resulting in higher ash retention. In contrast, larger fish, primarily feed at higher trophic levels, exhibited lower ash content(Aas et al., \u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e2020\u003c/span\u003e; Hairston, \u003cspan citationid=\"CR27\" class=\"CitationRef\"\u003e1993\u003c/span\u003e; Halver \u0026amp; Hardy, \u003cspan citationid=\"CR28\" class=\"CitationRef\"\u003e2003\u003c/span\u003e).\u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab1\" border=\"1\"\u003e \u003ccaption language=\"En\"\u003e \u003cdiv class=\"CaptionNumber\"\u003eTable 1\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cp\u003eProximate composition of Asian seabass. Values are expressed as mean\u0026thinsp;\u0026plusmn;\u0026thinsp;SEM. Values with different superscripts within the row are significantly different at p\u0026thinsp;\u0026lt;\u0026thinsp;0.05.\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"4\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003eProximate ingredient\u003c/p\u003e \u003cp\u003e(%)\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colspan=\"2\" nameend=\"c3\" namest=\"c2\"\u003e \u003cp\u003eThis Study\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c4\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003ePrevious studies on wild-caught Asian seabass reported by Kamruzzaman et al., (\u003cspan citationid=\"CR33\" class=\"CitationRef\"\u003e2015\u003c/span\u003e) \u0026amp; Pervin et al., (\u003cspan citationid=\"CR46\" class=\"CitationRef\"\u003e2012\u003c/span\u003e)\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c2\"\u003e \u003cp\u003eSmaller size Fish\u003c/p\u003e \u003cp\u003e(512.06\u0026thinsp;\u0026plusmn;\u0026thinsp;14.5 g)\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c3\"\u003e \u003cp\u003eLarger size Fish\u003c/p\u003e \u003cp\u003e(1003.5\u0026thinsp;\u0026plusmn;\u0026thinsp;6.64 g)\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eMoisture\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e74.96\u0026thinsp;\u0026plusmn;\u0026thinsp;0.04\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e70.75\u0026thinsp;\u0026plusmn;\u0026thinsp;0.17\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e69% (adult male) to 74% (juvenile)\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eAsh\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e2.99\u0026thinsp;\u0026plusmn;\u0026thinsp;0.46\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e2.43\u0026thinsp;\u0026plusmn;\u0026thinsp;0.15\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e3.25\u0026thinsp;\u0026plusmn;\u0026thinsp;0.14% (juvenile) to 5.30\u0026thinsp;\u0026plusmn;\u0026thinsp;0.52% (adult female)\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eFat\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e2.33\u0026thinsp;\u0026plusmn;\u0026thinsp;0.14\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e5.34\u0026thinsp;\u0026plusmn;\u0026thinsp;0.18\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e3.25\u0026thinsp;\u0026plusmn;\u0026thinsp;0.14% (juvenile) to 5.30\u0026thinsp;\u0026plusmn;\u0026thinsp;0.52% (adult female)\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eProtein\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e18.72\u0026thinsp;\u0026plusmn;\u0026thinsp;0.33\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e20.48\u0026thinsp;\u0026plusmn;\u0026thinsp;0.1\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e17.70\u0026thinsp;\u0026plusmn;\u0026thinsp;0.23% (spent female); 20.45\u0026thinsp;\u0026plusmn;\u0026thinsp;0.38% (juvenile) \u0026amp; 22.94\u0026thinsp;\u0026plusmn;\u0026thinsp;0.255% (adult male)\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003cp\u003eWhen compared to the proximate composition of wild-caught Asian seabass reported by Kamruzzaman et al., (\u003cspan citationid=\"CR33\" class=\"CitationRef\"\u003e2015\u003c/span\u003e) \u0026amp; Pervin et al., (\u003cspan citationid=\"CR46\" class=\"CitationRef\"\u003e2012\u003c/span\u003e) (Table\u0026nbsp;\u003cspan refid=\"Tab1\" class=\"InternalRef\"\u003e1\u003c/span\u003e), the cultured fish exhibited almost a similar range of moisture, fat, and protein content except for a slightly lower fat content in the smaller one. However, the ash content was notably lower in the cultured fish. Nutrient retention in fish is governed by a complex interplay of factors including diet composition, availability of food resources, and environmental conditions (Aas et al., \u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e2020\u003c/span\u003e; Hairston, \u003cspan citationid=\"CR27\" class=\"CitationRef\"\u003e1993\u003c/span\u003e; Halver \u0026amp; Hardy, \u003cspan citationid=\"CR28\" class=\"CitationRef\"\u003e2003\u003c/span\u003e). The reduced ash content in cultured Asian seabass may be attributed to poor feed quality, insufficient feed supply, or sub-optimal rearing conditions. Despite these challenges, the fat content remained within a normal range. This phenomenon could be explained by the catadromous nature of Asian seabass, which migrate between freshwater and marine environments and are exposed to varying environmental conditions. Such migration may lead to the development of metabolic pathways that enhance fat storage, which serves as a crucial energy reserve during periods of food scarcity or environmental stress.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec12\" class=\"Section2\"\u003e \u003ch2\u003e3.2 Fatty acid composition\u003c/h2\u003e \u003cp\u003eThe profiles of saturated fatty acids (SFA), monounsaturated fatty acids (MUFA), and polyunsaturated fatty acids (PUFA) in the flesh lipids of cultured Asian seabass of our study are presented in Figs.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003e,\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e2\u003c/span\u003e \u0026amp; \u003cspan refid=\"Fig3\" class=\"InternalRef\"\u003e3\u003c/span\u003e. The results indicate that the larger size group exhibited higher total concentrations of SFA and MUFA, whereas the smaller size group demonstrated higher PUFA concentrations. These findings align with those reported by De et al. (2019) in different size groups of \u003cem\u003eTenualosa ilisha\u003c/em\u003e. The observed differences in fatty acid profiles between the two groups can be primarily attributed to variations in feeding behavior, which are influenced by factors such as feeding habits(Pillay \u0026amp; Rao, \u003cspan citationid=\"CR47\" class=\"CitationRef\"\u003e1963\u003c/span\u003e) and dietary composition (Arzel et al., \u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e1994\u003c/span\u003e; Chen et al., \u003cspan citationid=\"CR14\" class=\"CitationRef\"\u003e1995\u003c/span\u003e; Grigorakis et al., \u003cspan citationid=\"CR26\" class=\"CitationRef\"\u003e2002b\u003c/span\u003e; Pirini et al., \u003cspan citationid=\"CR48\" class=\"CitationRef\"\u003e2000\u003c/span\u003e). Therefore, the differences in fatty acid profiles between the two groups can be attributed to their feeding behavior. In the early stages, Asian seabass are omnivorous, consuming a mix of plant and animal sources. As they mature, however, they gradually shift to a predominantly carnivorous diet (Corre, \u003cspan citationid=\"CR15\" class=\"CitationRef\"\u003e1994\u003c/span\u003e; Syeda \u0026amp; Hossain, \u003cspan citationid=\"CR53\" class=\"CitationRef\"\u003e2012\u003c/span\u003e).\u003c/p\u003e \u003cp\u003ePrevious studies on seabass and related species have identified C16:0 (palmitic) and C18:0 (stearic) acids as dominant SFAs (Alasalvar et al., \u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e2002\u003c/span\u003e; Chanmugam et al., \u003cspan citationid=\"CR13\" class=\"CitationRef\"\u003e1986\u003c/span\u003e; Chen et al., \u003cspan citationid=\"CR14\" class=\"CitationRef\"\u003e1995\u003c/span\u003e; Syama Dayal et al., \u003cspan citationid=\"CR52\" class=\"CitationRef\"\u003e2019\u003c/span\u003e). In contrast, in this study, the C11:0 (undecanoic) was observed as the most dominant SFA, contributing approximately 40% to the total SFA in both groups. C11:0 is primarily derived from plant-based sources, suggesting the significant presence of plant-origin feed components in the diets of cultured seabass. Other dominating SFAs were C16:0 (palmitic), C18:0 (stearic), C14:0 (mysteric), and C10:0 (capric) acids respectively in both groups. Between the two groups, C11:0 (undecanoic), C16:0 (palmitic), C14:0 (mysteric), C10:0 (capric), C24:0 (lignoceric), C13:0 (tridecanoic), and C22:0 (behenic) acids were significantly higher in the larger size group whereas C18:0 (stearic) and C15:0 (pentadecanoic) acids were significantly higher in the smaller ones.\u003c/p\u003e \u003cp\u003eThe two groups also differed in their MUFA concentration. Within the groups, C18:1 (oleic) acid was the dominant MUFA in larger ones which has also been reported by previous studies (CITATION). On the contrary, C16:1 (palmitoleic) acid was dominant in smaller-size fishes. Differences were also observed in the groupwise distribution of MUFAs. The concentrations of C18:1 (oleic), C16:1 (palmitoleic), and C20:1 (eicosenoic) acids were significantly higher in the larger size group which is similar to the previous studies. The smaller size group, on the other hand, had a higher concentration of C17:1 (heptadecenoic) acid.\u003c/p\u003e \u003cp\u003eIn terms of polyunsaturated fatty acid (PUFA), the study identified 10 PUFAs in the smaller size group and 8 in the larger size group. Among the n-3 PUFAs, C18:3 (α-linolenic acid) was the dominant, along with significant amounts of C20:5 (Eicosapentaenoic acid, EPA) and C22:6 (Docosahexaenoic acid, DHA) in both groups. Previous studies on seabass (Alasalvar et al., \u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e2002\u003c/span\u003e; \u0026Ouml;zyurt \u0026amp; Polat, \u003cspan citationid=\"CR45\" class=\"CitationRef\"\u003e2006\u003c/span\u003e; Xu et al., \u003cspan citationid=\"CR59\" class=\"CitationRef\"\u003e2010\u003c/span\u003e) and other similar fishes (Chen et al., \u003cspan citationid=\"CR14\" class=\"CitationRef\"\u003e1995\u003c/span\u003e; Grigorakis et al., \u003cspan citationid=\"CR26\" class=\"CitationRef\"\u003e2002b\u003c/span\u003e; Syama Dayal et al., \u003cspan citationid=\"CR52\" class=\"CitationRef\"\u003e2019\u003c/span\u003e) have identified EPA and DHA as the primary n-3 PUFA components. As C18:3 is predominantly sourced from plant-based sources (Mount Sinai, \u003cspan citationid=\"CR43\" class=\"CitationRef\"\u003e2015\u003c/span\u003e), the high assimilation of C18:3 indicates a diet rich in plant-origin components. However, between the two groups, C18:3 (α-linolenic) and C20:5 (EPA) concentrations were more abundant in the smaller size group while C22:6 (DHA) concentrations were higher in the larger ones. In terms of n-6 PUFA, C18:2 (linoleic) acid was dominant in the larger size group which is similar to other research findings (Alasalvar et al., \u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e2002\u003c/span\u003e; Syama Dayal et al., \u003cspan citationid=\"CR52\" class=\"CitationRef\"\u003e2019\u003c/span\u003e) but C20:3 (gamma-linolenic) acid was dominant in the smaller size ones. Moreover, it is worth noting that the fatty acid profiles in both groups showed a low proportion of elongated chain fatty acids. This observation could be linked to a reduced capacity for fatty acid chain elongation, as reported by Owen et al., (\u003cspan citationid=\"CR44\" class=\"CitationRef\"\u003e1975\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eAmong the major dominating fatty acids, C11:0 (undecanoic) exhibits antifungal and antimicrobial properties, which make it valuable in treating skin infections and promoting skin health (Ammendola et al., \u003cspan citationid=\"CR4\" class=\"CitationRef\"\u003e2009\u003c/span\u003e). In contrast, C16:0 (palmitic) acid supports cellular membrane integrity; however, excessive intake is associated with elevated LDL cholesterol levels and inflammation (Carta et al., \u003cspan citationid=\"CR12\" class=\"CitationRef\"\u003e2017\u003c/span\u003e; Zong et al., \u003cspan citationid=\"CR61\" class=\"CitationRef\"\u003e2016\u003c/span\u003e); Similarly, C14:0 (myristic) acid raises both HDL (good) and LDL (bad) cholesterol levels, thus necessitating a moderate intake to maintain cardiovascular health (Calder, \u003cspan citationid=\"CR11\" class=\"CitationRef\"\u003e2015\u003c/span\u003e; Zong et al., \u003cspan citationid=\"CR61\" class=\"CitationRef\"\u003e2016\u003c/span\u003e)​. On the other hand, C18:0 (stearic) acid has a neutral effect on cholesterol concentrations and does not significantly elevate cardiovascular risk, rendering it a safer choice among saturated fats (Zong et al., \u003cspan citationid=\"CR61\" class=\"CitationRef\"\u003e2016\u003c/span\u003e); C15:0 (pentadecanoic) acid, meanwhile, possesses anti-inflammatory properties and supports metabolic health, potentially lowering the risk of diabetes and cardiovascular diseases (Calder, \u003cspan citationid=\"CR11\" class=\"CitationRef\"\u003e2015\u003c/span\u003e; Zong et al., \u003cspan citationid=\"CR61\" class=\"CitationRef\"\u003e2016\u003c/span\u003e); Furthermore, C16:1 (palmitoleic) acid is recognized for its metabolic health benefits, enhancing insulin sensitivity, and mitigating the risk of type 2 diabetes (Frigolet \u0026amp; Guti\u0026eacute;rrez-Aguilar, \u003cspan citationid=\"CR22\" class=\"CitationRef\"\u003e2017\u003c/span\u003e); In addition, C18:1 (oleic) acid supports immune function, lowers blood pressure, and reduces LDL cholesterol levels (Pravst, \u003cspan citationid=\"CR49\" class=\"CitationRef\"\u003e2014\u003c/span\u003e); both C16:1 (palmitoleic) and C18:1 (oleic) acids possess anti-inflammatory properties that promote cardiovascular health (Frigolet \u0026amp; Guti\u0026eacute;rrez-Aguilar, \u003cspan citationid=\"CR22\" class=\"CitationRef\"\u003e2017\u003c/span\u003e; Pravst, \u003cspan citationid=\"CR49\" class=\"CitationRef\"\u003e2014\u003c/span\u003e); C14:1 (myristoleic) acid has anti-cancer properties, is associated with benefits for bone health, prevents non-alcoholic fatty liver disease (Y.-G. Kim et al., \u003cspan citationid=\"CR35\" class=\"CitationRef\"\u003e2021\u003c/span\u003e; Kwon et al., \u003cspan citationid=\"CR38\" class=\"CitationRef\"\u003e2015\u003c/span\u003e; S. Z. Li et al., \u003cspan citationid=\"CR40\" class=\"CitationRef\"\u003e2023\u003c/span\u003e; Quan et al., \u003cspan citationid=\"CR50\" class=\"CitationRef\"\u003e2020\u003c/span\u003e). Additionally, C18:3 (α-Linolenic) acid, along with other n-3 series\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003cp\u003efatty acids like EPA and DHA support cardiovascular health, reduce inflammation, aid cognitive function, and provide antioxidant benefits. The increase in α-Linolenic acid intake, ranging from 0.58 g to 2.51 g, has demonstrated improvements in cardiovascular health (Gebauer et al., \u003cspan citationid=\"CR24\" class=\"CitationRef\"\u003e2006\u003c/span\u003e). A 2:1 EPA/DHA ratio is recommended for optimal cardiovascular outcomes, alongside a daily intake of approximately 500 mg of EPA and DHA (Gebauer et al., \u003cspan citationid=\"CR24\" class=\"CitationRef\"\u003e2006\u003c/span\u003e), which can be obtained by consuming around 500 g of smaller fish and 600 g of larger fish.\u003c/p\u003e \u003cp\u003eOur findings further highlight that the smaller-size group exhibited a favorable n-3/n-6 ratio of 1.37, whereas the larger-size group showed a ratio of 0.91. Both ratios meet the recommended threshold of 0.2 or higher, confirming their adequacy for maintaining optimal cardiovascular health (Syama Dayal et al., \u003cspan citationid=\"CR52\" class=\"CitationRef\"\u003e2019\u003c/span\u003e).\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec13\" class=\"Section2\"\u003e \u003ch2\u003e3.3 Amino acid composition\u003c/h2\u003e \u003cp\u003eThe amino acid profile of the muscle tissue in cultured Asian seabass is presented in Fig.\u0026nbsp;\u003cspan refid=\"Fig4\" class=\"InternalRef\"\u003e4\u003c/span\u003e. This analysis identified a total of 17 amino acids, including all essential amino acids (EAA) except tryptophan. Among the EAAs, lysine was the most abundant, followed by leucine and arginine in both size groups. Interestingly, the concentrations of all EAAs were significantly higher in the smaller-size group, with the exceptions of methionine and phenylalanine, which, although not statistically significant, exhibited higher numerical values in the smaller group. Regarding non-essential amino acids (NAA) and glutamic acid emerged as the most abundant, followed by aspartic acid and alanine in both groups. Significant differences were observed in the concentrations of all NAAs between the groups, except for serine. Specifically, glutamic acid, aspartic acid, alanine, and tyrosine were significantly elevated in the smaller-size group, while glycine and proline were substantially more abundant in the larger-size group.\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003cp\u003eThese findings align with previous studies on seabass (\u0026Ouml;zyurt \u0026amp; Polat, \u003cspan citationid=\"CR45\" class=\"CitationRef\"\u003e2006\u003c/span\u003e) and related species (Iwasaki \u0026amp; Harada, \u003cspan citationid=\"CR32\" class=\"CitationRef\"\u003e1985\u003c/span\u003e; Mohanty et al., \u003cspan citationid=\"CR42\" class=\"CitationRef\"\u003e2014\u003c/span\u003e). \u0026Ouml;zyurt \u0026amp; Polat, (\u003cspan citationid=\"CR45\" class=\"CitationRef\"\u003e2006\u003c/span\u003e) reported a total of 16 amino acids in seabass muscle, with similar dominant amino acids. They also noted that amino acid composition varies with the spawning period and feeding behavior of the fish. Similarly, Mohanty et al., (\u003cspan citationid=\"CR42\" class=\"CitationRef\"\u003e2014\u003c/span\u003e) highlighted that factors such as capture time and geographical location influence the quantity and types of amino acids in fish muscle.\u003c/p\u003e \u003cp\u003eAmino acids (AAs) are recognized as essential building blocks of tissue proteins and play critical roles in synthesizing low molecular weight compounds, including glutathione, creatine, thyroid hormones, melanin, and melatonin, which are vital for various physiological processes (Blachier et al., \u003cspan citationid=\"CR9\" class=\"CitationRef\"\u003e2011\u003c/span\u003e; J. Kim et al., \u003cspan citationid=\"CR34\" class=\"CitationRef\"\u003e2012\u003c/span\u003e; Kong et al., \u003cspan citationid=\"CR36\" class=\"CitationRef\"\u003e2012\u003c/span\u003e; Wu et al., \u003cspan citationid=\"CR58\" class=\"CitationRef\"\u003e2009\u003c/span\u003e). Besides, the elevated levels of essential amino acids (EAA) in muscle are indicative of superior muscle protein quality (Young \u0026amp; Pellett, \u003cspan citationid=\"CR60\" class=\"CitationRef\"\u003e1984\u003c/span\u003e). In the present study, the total EAA/total NAA ratios were 0.81 and 0.71 in the smaller and larger size groups, respectively. According toFAO/WHO, (\u003cspan citationid=\"CR20\" class=\"CitationRef\"\u003e1991\u003c/span\u003e) recommendations, a ratio of 0.6 or higher is considered a benchmark for high-quality protein, confirming that both groups exhibit high protein quality.\u003c/p\u003e \u003cp\u003eAdditionally, beyond essential and non-essential amino acids, a distinct group known as functional amino acids has gained recognition for their roles in regulating physiological and metabolic processes critical for health, growth, reproduction, and disease resistance (G. Li et al., \u003cspan citationid=\"CR39\" class=\"CitationRef\"\u003e2011\u003c/span\u003e; Rezaei et al., \u003cspan citationid=\"CR51\" class=\"CitationRef\"\u003e2013\u003c/span\u003e; Wang et al., \u003cspan citationid=\"CR55\" class=\"CitationRef\"\u003e2013\u003c/span\u003e; Wu, \u003cspan citationid=\"CR56\" class=\"CitationRef\"\u003e2009\u003c/span\u003e, \u003cspan citationid=\"CR57\" class=\"CitationRef\"\u003e2013\u003c/span\u003e). Functional amino acids include arginine, aspartate, cysteine, glutamine, glycine, histidine, leucine, methionine, proline, threonine, tryptophan, and tyrosine (Mohanty et al., \u003cspan citationid=\"CR42\" class=\"CitationRef\"\u003e2014\u003c/span\u003e; Wu, \u003cspan citationid=\"CR56\" class=\"CitationRef\"\u003e2009\u003c/span\u003e, \u003cspan citationid=\"CR57\" class=\"CitationRef\"\u003e2013\u003c/span\u003e)Notably, this study found that Asian seabass muscle tissue contained all functional amino acids except tryptophan.\u003c/p\u003e \u003c/div\u003e"},{"header":"4. Conclusions","content":"\u003cp\u003eThe results of this study demonstrate that cultured Asian seabass is a highly nutritious fish, offering high levels of crude protein, fat, and ash content; an excellent source of high-quality protein, providing a well-balanced composition of essential amino acids, along with substantial amounts of α-linolenic acid, EPA, and DHA, making it a valuable dietary choice. These attributes not only benefit consumers but also support the aquaculture industry. Notably, cultured Asian seabass exhibited nutritional profiles comparable to, and in some instances surpassing, those of wild-caught seabass in terms of amino acid and fatty acid content.\u003c/p\u003e \u003cp\u003eSignificant differences in nutritional composition were observed between the two size groups, offering essential insights into their potential health benefits. The smaller size group demonstrated superior concentrations of essential amino acids, a higher EPA to DHA ratio, and a more favorable n-3 to n-6 ratio compared to the larger group. Based on these findings, it can be concluded that the smaller size group of cultured Asian seabass possesses higher nutritional value than the larger size group, making it a particularly beneficial option for dietary purposes. Thus, the results of this study will provide a framework for selecting the nutritionally superior fish, which may ultimately help in redesigning market demand and consumer preference.\u003c/p\u003e"},{"header":"Declarations","content":"\u003cp\u003eFrozen cultured Asian seabass were donated for use in this study by local farms during their normal harvests.\u003c/p\u003e\n\u003cp\u003eCompeting Interests\u003c/p\u003e\n\u003cp\u003eWe declare that the concerned authors and co-authors carried out the work, and all the authors have read the draft manuscript. We would also like to declare that there is no potential conflict of interest among the authors.\u003c/p\u003e\n\u003cp\u003eFunding statement:\u003c/p\u003e\n\u003cp\u003eThe corresponding author, Roni Sikder (RS), acknowledges that the Ministry of Science and Technology, Bangladesh, partially funded this work by awarding the National Science of Technology Fellowship. The funders had no role in study design, data collection and analysis, publication decisions, or manuscript preparation.\u003c/p\u003e\n\u003cp\u003eAcknowledgment\u003c/p\u003e\n\u003cp\u003eThe authors would like to thank the Institute of Technology Transfer and Innovation (ITTI), BCSIR for providing this study\u0026apos;s necessary resources and facilities. Additionally, we appreciate the valuable contributions and insightful discussions from our colleagues and mentors.\u003c/p\u003e\n\u003cp\u003eData availability statement\u003c/p\u003e\n\u003cp\u003eAll data generated or analyzed during this study are included in this manuscript.\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\n \u003cli\u003eAas, T. S., Sixten, H. J., Hillestad, M., Ytrest\u0026oslash;yl, T., Sveier, H., \u0026amp; \u0026Aring;sg\u0026aring;rd, T. (2020). 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B., \u0026amp; Sun, Q. (2016). Intake of individual saturated fatty acids and risk of coronary heart disease in US men and women: Two prospective longitudinal cohort studies. \u003cem\u003eBMJ (Online)\u003c/em\u003e, \u003cem\u003e355\u003c/em\u003e. https://doi.org/10.1136/bmj.i5796\u003c/li\u003e\n\u003c/ol\u003e"}],"fulltextSource":"","fullText":"","funders":[{"identity":"ab131fc8-d7a1-4384-bd2d-b1d4763f66bd","identifier":"10.13039/501100008804","name":"Ministry of Science and Technology, Government of the People’s Republic of Bangladesh","awardNumber":"NST Fellowship","order_by":0}],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":true,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":true,"hideJournal":true,"highlight":"","institution":"University of Dhaka","isAcceptedByJournal":false,"isAuthorSuppliedPdf":false,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":false,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true},"keywords":"Asian seabass, Bangladesh aquaculture, Cultured fish, Nutritional composition, Fatty acid profile, Amino acid profile, Proximate analysis, Essential amino acids","lastPublishedDoi":"10.21203/rs.3.rs-5750226/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-5750226/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003e\u003cem\u003eLates calcarifer\u003c/em\u003e, known as Asian seabass, is a brackish-water teleost species within the order \u003cem\u003eCarangiformes\u003c/em\u003e, native to the Indo-Pacific region. In Bangladesh, this species is found in natural habitats and cultivated on a small scale in the southern coastal region, particularly in the Khulna-Satkhira area. This study analyzed the nutritional composition of cultured Asian seabass in two different size categories, 512.06 ± 14.5 g and 1003.5 ± 36.64 g, collected from this region. Our findings revealed significant differences in the proximate compositions of the two size groups, except ash content (independent t-test, p \u0026lt; 0.05). The smaller size group had a notably higher moisture content (74.96 ± 0.04%), whereas the larger fish exhibited significantly greater protein and fat contents (20.48 ± 0.1% and 5.34 ± 0.18%, respectively). Fatty acid analysis showed distinct differences as well; the larger size group contained higher concentrations of saturated fatty acids (SFAs) and monounsaturated fatty acids (MUFAs), while the smaller fish was richer in polyunsaturated fatty acids (PUFAs). Among the SFAs and omega-3 PUFAs, undecanoic acid and α-linolenic acid were the most prevalent in both groups. However, MUFA profiles differed: palmitoleic acid was dominant in the smaller size group, while oleic acid was more abundant in the larger size group. All essential amino acids (EAAs), except tryptophan, were detected in both groups, with lysine being the most prominent, measuring 5.53±0.44\u003csup\u003e \u003c/sup\u003eg/100 g dry weight in smaller size group and 3.91±0.32 g/100 g dry weight in larger size group. Notably, the smaller size group demonstrated a significantly higher overall concentration of EAAs. Overall, this study highlights the nutritional richness of cultured Asian seabass, particularly in terms of fatty acid and amino acid profiles. The superior concentrations of essential amino acids, a higher EPA/DHA ratio, and a more favorable n-3/n-6 ratio in the smaller fish suggest it as a nutritionally superior and well-balanced fish option.\u003c/p\u003e","manuscriptTitle":"Investigating the nutritional composition of cultured Asian seabass Lates calcarifer in Bangladesh's Khulna-Satkhira region: A focus on fatty acids and amino acids","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2025-01-14 04:55:34","doi":"10.21203/rs.3.rs-5750226/v1","editorialEvents":[{"type":"communityComments","content":0}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true}}],"origin":"","ownerIdentity":"ea5af54a-9238-450e-ae22-73ab5acd3fd1","owner":[],"postedDate":"January 14th, 2025","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"posted","subjectAreas":[{"id":42257829,"name":"Aquaculture and Mariculture"}],"tags":[],"updatedAt":"2025-01-14T04:55:34+00:00","versionOfRecord":[],"versionCreatedAt":"2025-01-14 04:55:34","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-5750226","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-5750226","identity":"rs-5750226","version":["v1"]},"buildId":"8U1c8b4HqxoKbykW_rLl7","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}