Innovative Studies in Agriculture, Forestry and Aquaculture – 2025
Full text
INNOVATIVE STUDIES IN AGRICULTURE, FORESTRY AND AQUACULTURE – 2025 Editor Kenan KÖPRÜCÜ Lyon 2025
INNOVATIVE STUDIES IN AGRICULTURE, FORESTRY AND AQUACULTURE – 2025 Editor Kenan KÖPRÜCÜ Lyon 2025
Innovative Studies in Agriculture, Forestry and Aquaculture – 2025 Editor • Prof. Dr. Kenan KÖPRÜCÜ • Orcid: 0000-0002-5697-5224 Cover Design • Motion Graphics Book Layout • Motion Graphics First Published • December 2025, Lyon e-ISBN: 978-2-38236-974-6 DOI: 10.5281/zenodo.18039186 copyright © 2025 by Livre de Lyon All rights reserved. No part of this publication may be reproduced, stored in a retrieval system, or transmitted in any form or by any means, electronic, mechanical, photocopying, recording, or otherwise, without prior written permission from the Publisher. The author or authors of the relevant section are responsible for any copyright infringement that may occur due to the images and graphics used in the book. The editor or publisher does not assume responsibility in this regard. Publisher • Livre de Lyon Address • 37 rue marietton, 69009, Lyon France website • http://www.livredelyon.com e-mail • [email protected]
I PREFACE Sustainability, food security, and biotechnological innovations constitute the core focus areas of contemporary agricultural and food sciences. Rapid population growth, the limited availability of natural resources, and the impacts of climate change necessitate the adoption of smarter, more environmentally friendly, and more efficient approaches at every stage, from production to processing, packaging, and consumption. This scientific book aims to address these needs through an interdisciplinary perspective. The chapters presented in this volume cover a broad spectrum of topics, ranging from smart and sustainable seafood packaging systems aligned with the Industry 4.0 approach, to the enhancement of oxidative stability in aquatic products through the use of natural biopolymers and plant extracts; from the controlled and soilless cultivation of medicinal and aromatic plants for the production of high value-added bioactive compounds, to sustainable extraction methods and industrial applications of phytochemicals derived from these plants. In addition, the cultivation of silage corn, a key component of sustainable livestock production, is examined in detail with respect to its agroecological requirements. The primary objective of this book is to bring together scientific knowledge that prioritizes the conservation of natural resources, is supported by innovative technologies, and is transferable to industrial applications. In this respect, the book aims to serve as a reliable and up-to-date reference for academics, graduate students, researchers, and industry stakeholders. We sincerely thank all contributing authors for their valuable scientific contributions and dedicated efforts, and we hope that this book will help foster a sustainable future in the fields of agriculture, food science, fisheries, and biotechnology. Prof. Dr. Kenan Köprücü (Editor)
III CONTENTS PREFACE I CHAPTER I. SMART AND SUSTAINABLE PACKAGING FOR SEAFOOD: A PATHWAY TO INDUSTRY 4.0 1 EmrahAĞ&ÖzlemEMİRÇOBAN CHAPTER II. ENHANCING OXIDATIVE STABILITY OF RAINBOW TROUT FILLETS USING NATURAL COATINGS: CHIA SEED MUCILAGE AND HELICHRYSUM ARENARIUM EXTRACT 21 ÖzlemEMİRÇOBAN CHAPTER III. CONTROLLED AND SOILLESS CULTIVATION OF MEDICINAL AND AROMATIC PLANTS FOR BIOACTIVE COMPOUND PRODUCTION 33 İmgeİhsaneÖZCAN CHAPTER IV. MEDICINAL PLANT PHYTOCHEMICALS: SUSTAINABLE EXTRACTION, OPTIMIZATION AND INDUSTRIAL APPLICATIONS 53 İmgeİhsaneÖZCAN CHAPTER V. THE IMPORTANCE OF SILAGE CORN (Zea mays L.) PLANT IN SUSTAINABLE FORAGE PRODUCTION AND ITS CULTIVATION IN ACCORDANCE WITH ITS AGROECOLOGICAL REQUIREMENTS 81 ŞerifeAKKEÇECİ
SMART AND SUSTAINABLE PACKAGING FOR SEAFOOD: A PATHWAY . . . 7 significant advantages in terms of speed, hygiene, efficiency, and standardization in food packaging. Robots integrated with advanced technologies such as smart packaging systems perform tasks such as precise handling, sorting, filling, and labeling of food products with high accuracy (Kumari et al., 2020). Robotic automation reduces errors caused by human factors on production lines and directly supports food safety. Robotic automation enables packaging processes in the food industry to be conducted more rapidly, hygienically, economically, and securely. These systems, which operate in harmony with the digital infrastructure introduced by Industry 4.0, also integrate seamlessly with smart packaging technologies to create a holistic production environment. As robotic technologies continue to evolve, packaging processes will require less human intervention, and production lines will become more flexible and sustainable. 2.3.1.TheRoleofRoboticSystemsinPackagingProcesses In conventional packaging lines, manual operations are time-consuming, costly, and pose risks of contamination. To eliminate these issues, robotic arms, conveyor systems, and visual recognition technologies are increasingly employed. Robots are capable of performing thousands of packaging operations continuously with consistent quality, particularly in high-volume production environments. For example, robotic systems equipped with cameras and sensors can identify irregularly shaped products such as fruits and vegetables and automatically place them into appropriate packaging (Rolle et al., 2022) 2.3.2.RoboticAutomationinTermsofHygieneandFoodSafety Maintaining hygiene on food packaging lines is of paramount importance. Human contact increases the risk of cross-contamination, whereas robotic systems significantly minimize this risk. Industrial robots, with their stainless steel surfaces, easily sterilizable structures, and automated cleaning capabilities, meet hygienic production standards. Moreover, robots can operate in conjunction with environmental sensors, allowing automatic adjustments based on ambient conditions (Rodrigues et al., 2021). This feature is particularly valuable for preserving the quality of temperatureand humidity-sensitive food products. 2.3.3.FlexibilityandEfficiency Robotic systems offer greater flexibility compared to traditional fixed production lines. They can easily handle packaging of various sizes and types
8 INNOVATIVE STUDIES IN AGRICULTURE, FORESTRY AND AQUACULTURE – 2025 and quickly adapt to changes during production. In today’s market, where customized packaging demands are increasing, flexible robotic systems provide brands with a competitive edge. Furthermore, when integrated with big data and artificial intelligence, these systems can acquire learning capabilities during production, leading to enhanced efficiency and optimized resource utilization over time (García et al., 2020). 2.3.4.Human–RobotCollaboration(Cobots)andFutureOutlook In recent years, “collaborative robots” (cobots) capable of working safely alongside humans have been introduced into packaging operations. These robots share the same workspace with human operators, taking over repetitive and physically demanding tasks while leaving supervision and decision-making responsibilities to human workers. This approach promotes efficient use of labor and provides ergonomic working environments (Yılmaz & Altan, 2021). 2.4.SustainabilityandEco-FriendlyPackaging The growing global environmental challenges—particularly the detrimental impact of plastic waste on ecosystems—have brought the concept of sustainability to the forefront of the food packaging industry. In addition to extending shelf life and providing consumer information, packaging plays a direct role in shaping the environmental footprint of the production– consumption cycle. In this context, eco-friendly packaging design encompasses not only the use of recyclable materials but also practices that minimize ecological harm throughout production, transportation, and disposal processes (Marsh & Bugusu, 2007). 2.4.1.CorePrinciplesofSustainablePackaging The primary goal of sustainable packaging is to preserve product quality and safety while minimizing environmental impact. Accordingly, the use of biodegradable, recyclable, or reusable materials is of critical importance. Other key sustainability criteria include low energy consumption during production, prevention of material waste, and reduction of carbon emissions. Today, many manufacturers incorporate FSC (Forest Stewardship Council)-certified cartons, PLA (polylactic acid)-based bioplastics, and starch-based composite materials into their packaging processes (Siracusa et al., 2008). Packaging waste currently constitutes one of the largest categories of solid waste globally. Even though packaging materials may retain
SMART AND SUSTAINABLE PACKAGING FOR SEAFOOD: A PATHWAY . . . 9 functionality after the product is consumed, they are often discarded immediately. To prevent this and establish an environmentally responsible production–consumption cycle, approaches such as green packaging, sustainable packaging, eco-friendly solutions, and zero-waste initiatives have been developed. Within this framework, environmentally compatible methods such as recycling, reuse (durable packaging), and biodegradability have gained prominence (Ersan, 2021). To clarify the concept of upcycling, it is necessary to examine its similarities and differences with recycling and reuse. Recycling refers to the process of converting post-consumer waste into raw materials or by-products through various treatments, thereby reintegrating them into production cycles (Çimen & Yılmaz, 2012). In contrast, upcycling involves creatively repurposing an object—partially or entirely retaining its original form—for a new use. For example, washing and reusing a glass bottle for the same purpose constitutes reuse, whereas transforming it into a decorative item falls under upcycling. The most significant advantage of upcycling is its considerably lower environmental impact compared to recycling. Recycling processes typically require substantial energy and resource input to convert waste into raw materials. Upcycling, on the other hand, often requires no additional energy or new materials, making it a more efficient and eco-friendly approach. When applied to packaging materials, upcycling not only conserves landfill space but also prevents the release of harmful gases and waste into the environment. 2.4.2.Bio-BasedandBiodegradablePackagingMaterials Bio-based packaging is produced using materials derived from renewable resources. Corn starch, sugarcane, cellulose, and algae are among the primary sources in this domain. Bioplastics such as PLA (polylactic acid) and PHA (polyhydroxyalkanoates), which are obtained from these materials, decompose naturally over time without harming the environment. These packaging solutions offer both protective and eco-friendly benefits, particularly for products with short shelf lives such as seafood, fruits and vegetables, and baked goods (Sharma et al., 2020). Conventional plastics widely used in food packaging are predominantly derived from petroleum-based raw materials. However, their persistence in nature and contribution to environmental pollution pose significant challenges. To mitigate this issue, various studies have focused on enhancing the biodegradability of plastics to facilitate their decomposition in natural environments. Notably, certain bacteria and yeast strains have been
10 INNOVATIVE STUDIES IN AGRICULTURE, FORESTRY AND AQUACULTURE – 2025 observed to break down plastics over time—a process referred to as “biological degradation” (Kılınç, Tomar & Çağlar, 2017). In addition to biodegradability, research is also underway to develop next-generation packaging materials with antimicrobial properties and controlled permeability to water vapor and oxygen (Orhan & Büyükgüngör, 2000). Today, the packaging industry faces multiple challenges, including the depletion of fossil resources, limited access to these materials, and their long-term persistence in the environment. As a result, manufacturers are increasingly turning to eco-friendly materials derived from renewable sources. The development of recyclable and biologically degradable packaging offers both economic and ecological benefits (Sun et al., 2017). Studies in this field highlight the prominence of natural polymers such as cellulose, starch, and proteins in the production of biodegradable packaging (Çelebi & Dehmen, 2013). Biodegradable Packaging and Films: Due to their technological advantages, plastic materials are widely used in both industrial and everyday applications. However, growing environmental concerns have led to increased interest in biodegradable packaging materials for food applications. These materials are considered environmentally responsible alternatives to conventional plastics (Kılınç, Tomar & Çağlar, 2017). Biodegradable polymers can be broken down in nature through the action of microorganisms such as bacteria, fungi, and algae. These polymers degrade via enzymes produced by microorganisms, transforming into natural components such as carbon dioxide, methane, water, biomass, and humus (Gross & Kalra, 2002). As a result, no harmful residues are left behind, and the issue of waste is significantly reduced. The use of biodegradable packaging materials is rapidly expanding, driven by growing environmental awareness and the desire to reduce reliance on petroleum-based plastics. Technological advancements have further contributed to the increased adoption of these materials. A significant portion of bioplastics is derived from natural sources such as starch, cellulose, and proteins, which broadens their applicability across various sectors. However, the primary limitations preventing bioplastics from fully competing with conventional plastics are their high production costs and limited manufacturing capacity. It is anticipated that rising crude oil prices will enhance the importance of environmentally friendly plastics produced from renewable resources, potentially leading to their replacement of traditional plastics. Biodegradable Polymers: Biopolymers are environmentally friendly materials obtained from naturally occurring biomass sources and are capable
SMART AND SUSTAINABLE PACKAGING FOR SEAFOOD: A PATHWAY . . . 11 of being degraded by microorganisms. For this reason, they are also referred to as “green polymers” (Yoruç & Uğraşkan, 2017). Biodegradable polymers are classified into three generations based on their production methods: First-generation polymers are produced by blending low-density polyethylene with 5–20% starch and various additives (e.g., pro-oxidants). However, due to their prolonged decomposition time in nature (3–5 years), they are not considered fully biodegradable. Second-generation polymers contain starch and polyethylene along with additives such as vinyl acetate and polyvinyl alcohol. Although starch itself degrades within approximately 40 days, complete decomposition of the film may take 2–3 years. Third-generation biodegradable polymers are entirely derived from natural sources or synthesized from biologically produced monomers. These materials consist either of natural polymers extracted directly from biomass or of compounds chemically synthesized from biological monomers (Erol, 2012). Synthetic polymers are composed of substances not naturally found in the environment, and microorganisms lack the enzymes necessary to break them down, resulting in significantly prolonged degradation times. In contrast, the molecular bonds in biodegradable polymers are naturally occurring, making them more susceptible to enzymatic breakdown by microorganisms and thus less harmful to the environment (Bahçegül, 2011). 2.4.3.FoodSafetyandConsumerPreferences Food safety refers to the measures taken throughout the production-toconsumption process to ensure that consumers have access to healthy and reliable food products. Consumer attitudes toward food safety directly influence purchasing decisions. In recent years, the rise in foodborne illnesses and negative media coverage has intensified the demand for trustworthy food options. Eco-friendly packaging also plays a significant role in fostering consumer trust. Consumers are increasingly attentive not only to product quality but also to the environmental impact of production and packaging processes. Particularly in European and North American markets, products with “ecolabeled” packaging are more highly preferred. This trend has prompted brands to strengthen their sustainability policies and reshape their marketing strategies accordingly (Magnier & Schoormans, 2015). A study by Bekir (2013) revealed
12 INNOVATIVE STUDIES IN AGRICULTURE, FORESTRY AND AQUACULTURE – 2025 that consumers are most concerned about genetically modified organisms (GMOs), artificial colorants, hormone and antibiotic residues, and pesticides. These concerns have led to a decrease in the frequency of consumption of such products. Additionally, consumers tend to perceive products certified by the Ministry of Food, Agriculture and Livestock as more trustworthy. Interest in organic products has also grown in parallel with food safety concerns. In a study conducted in Kahramanmaraş, Kekeç and Seçer (2021) found that health concerns were the primary reason consumers preferred organic products. However, high prices and lack of information remain major barriers to consumption. In conclusion, consumer attitudes toward food safety shape purchasing behavior and constitute a key marketing strategy for food producers and retailers. Raising consumer awareness and offering reliable products are essential not only for protecting public health but also for enhancing consumer satisfaction. 2.4.4.LegalRegulationsandInternationalPractices The use of sustainable packaging is gaining increasing importance in terms of reducing environmental impact and promoting efficient resource utilization. Accordingly, numerous countries and international organizations have introduced legal frameworks and practical measures aimed at minimizing packaging waste and encouraging recycling. The European Union’s “Single-Use Plastics Ban” and Türkiye’s “Zero Waste Regulation” represent significant steps toward reducing packaging-related environmental burdens. Under these policies, packaging solutions that enhance recycling rates and minimize ecological harm are actively supported through government incentives (Republic of Türkiye Ministry of Environment, Urbanization and Climate Change, 2023). The European Union has taken a leading role in packaging waste management. The Packaging and Packaging Waste Regulation No. 2025/40, which came into force in 2025, introduced key provisions to reduce packaging waste and improve the recyclability and reusability of existing packaging materials. Under this regulation, all packaging placed on the EU market must be recyclable by 2030. Additionally, minimum recycled content requirements have been imposed for plastic packaging, and certain types of single-use plastic packaging have been banned. These measures not only promote environmental sustainability but also introduce labeling and consumer information obligations for producers and retailers. As a result, the environmental impact of packaging throughout its life cycle is reduced, and consumers are encouraged to make
SMART AND SUSTAINABLE PACKAGING FOR SEAFOOD: A PATHWAY . . . 13 informed choices. Globally, many countries have implemented legal regulations and practices to reduce packaging waste and promote the use of sustainable packaging. For example, countries such as Canada and Australia have established recycling targets for packaging producers and adopted extended producer responsibility frameworks. In addition, the use of single-use plastic packaging has been banned or restricted in several jurisdictions. These international practices play a crucial role in promoting sustainable packaging and reducing environmental impact. They also contribute to aligning international trade with sustainability criteria and encouraging the adoption of eco-friendly packaging solutions. 2.4.5.ApplicationExamplesinSeafoodProducts Developing environmentally friendly packaging solutions for highly perishable foods such as seafood is critical for ensuring quality control and promoting sustainability. The integration of biodegradable packaging materials with Modified Atmosphere Packaging (MAP) technology extends shelf life while reducing plastic usage. Additionally, initiatives have been launched in the fisheries sector to recycle discarded fishing nets and repurpose them as packaging materials (Lundberg et al., 2022). Further details on this topic are provided in the following section. 3. Food Packaging 4.0 in Seafood Products Seafood products are highly perishable due to their elevated water and protein content, making appropriate packaging systems essential for maintaining freshness. While conventional packaging methods can delay spoilage to some extent, they fall short of meeting the technological demands of modern food systems. With the advent of Industry 4.0, “Food Packaging 4.0” systems have revolutionized the packaging sector through innovations such as smart sensors, the Internet of Things (IoT), and cloud-based technologies (Erdem et al., 2022). These advancements transform packaging from a purely physical barrier into a digital tool that enables traceability throughout the supply chain— from production to consumption. Maintaining the freshness of seafood requires precise monitoring of temperature, humidity, and oxygen levels. IoT-enabled smart packaging provides real-time data, alerting stakeholders to temperature fluctuations and potential spoilage. Sensors embedded in the packaging transmit this data to cloud-based platforms, which can be accessed
14 INNOVATIVE STUDIES IN AGRICULTURE, FORESTRY AND AQUACULTURE – 2025 by producers, distributors, or consumers as needed (Kasım & Kasım, 2019; Erdem et al., 2022). Moreover, the integration of biosensors into seafood packaging allows for early detection of microbial spoilage. This technology plays a vital role in enhancing food safety, particularly in marine products. Active packaging systems incorporating natural substances also offer eco-friendly alternatives while extending shelf life. Films made from biopolymers such as chitosan and alginate possess natural antibacterial properties that inhibit microbial growth (Kılınç et al., 2017). Cold chain monitoring is indispensable for preserving seafood quality. Technologies such as Radio Frequency Identification (RFID) and Near Field Communication (NFC) enable real-time tracking of temperature and humidity within the packaging. These data points ensure traceability and quality control throughout the supply chain. Kocatepe and Turan (2011) emphasized that such technologies enhance quality assurance and trust in export markets. Globally, these systems are increasingly recognized as standard practices in seafood exportation (Douaki et al., 2024). Smart packaging systems also feature freshness/spoilage indicators that change color, allowing consumers to instantly assess product quality at the point of purchase. Research by Erdem et al. (2022) demonstrated that these indicators significantly extend the shelf life of seafood products. Additionally, active packaging solutions such as oxygenabsorbing labels help maintain freshness by reducing oxygen levels within the package (Olum, 2024). 3.1.Vacuum,MAP,andActivePackagingMethods One of the most widely used methods to enhance the durability of seafood products is vacuum packaging. In this technique, air is completely removed from the package, thereby reducing the impact of oxygen on microbial activity and slowing down spoilage. Another method, modified atmosphere packaging (MAP), involves introducing specific gas mixtures—such as carbon dioxide, nitrogen, and a small amount of oxygen—into the packaging environment. This modified atmosphere helps preserve the freshness of the product for a longer period. The sustainability of packaging used in seafood products is crucial not only for reducing environmental impact but also for maintaining product freshness. Bio-based materials and smart packaging technologies have been successfully applied in seafood packaging, resulting in extended shelf life and environmentally friendly solutions. Active packaging goes beyond simply protecting the product from external conditions; it interacts directly with
SMART AND SUSTAINABLE PACKAGING FOR SEAFOOD: A PATHWAY . . . 15 the product to influence microbial activity. Research in this area has focused particularly on the use of natural substances with antimicrobial properties. Biofilm packaging developed with agents such as chitosan and essential oil has been shown to slow microbial growth on seafood products and enhance consumer safety (Emir Çoban & Çoban, 2019, Olum, 2024). 3.2.UseofIoTandBlockchaininSmartPackagingSystems In Food Packaging 4.0 systems, packaging has evolved into “smart” structures through the integration of IoT and cloud-based software. Sensors embedded within the packaging continuously monitor temperature, humidity, and gas levels, transmitting this data to centralized systems. These real-time insights enable full traceability across the supply chain. In recent years, the use of blockchain technology in packaging processes has also gained momentum. Blockchain enhances data security by recording the entire journey of a product—from production to consumption—in an immutable and transparent manner. This level of traceability is particularly valuable for high-value food items such as seafood, significantly boosting consumer trust (Douaki et al., 2024).To ensure transparency, safety, and sustainability in the seafood supply chain, IoT and blockchain technologies are increasingly being integrated. These technologies allow for end-to-end traceability, from ocean to table, and play a crucial role in combating illegal, unreported, and unregulated (IUU) fishing practices (Blaha & Katafono, 2020). IoT devices collect real-time data on critical parameters such as temperature, humidity, and location throughout the seafood supply chain. For instance, maintaining optimal temperature during the transportation of fresh fish is vital for preserving product quality. IoT sensors transmit this data to centralized platforms, enabling continuous monitoring of product conditions throughout the supply chain (Gong et al., 2020). Blockchain technology ensures data security and transparency by recording every step of the supply chain in an immutable manner. As a result, information such as when, where, and by whom a fish was caught becomes verifiable by all stakeholders. For instance, the OpenSC platform utilizes blockchain to confirm that seafood products originate from sustainable sources (Simon, 2020). In Australia, blockchain-based traceability platforms enable the tracking of individual fish or fish containers through physical labels. These labels include data such as harvest time, product weight, and temperature, which consumers can access directly (Bumblauskas et al., 2020). In Malaysia, a traceability system developed for the seafood supply chain uses blockchain technology to
16 INNOVATIVE STUDIES IN AGRICULTURE, FORESTRY AND AQUACULTURE – 2025 ensure data integrity and transparency. This system allows all stakeholders in the supply chain to input data and securely share it (Simon, 2020). The integration of blockchain and IoT technologies provides consumers with reliable information about the origin and processing of seafood products, thereby promoting sustainable and ethical fishing practices. This level of transparency enhances consumer trust and contributes to reducing environmental impact (Vasconcelos et al., 2020). 3.3.PackagingwithNaturalMaterialsandEnvironmentalImpacts With the rise of Industry 4.0, innovation has extended beyond digitalization to include environmentally friendly production and sustainability-focused advancements. As awareness grows regarding the environmental harm caused by plastic use in food packaging, interest in biodegradable packaging made from natural materials has significantly increased. These next-generation packaging systems are not only effective in ensuring product safety but also decompose harmlessly into the environment after use. Materials derived from biological sources—such as chitosan, starch, cellulose, alginate, and gelatin—stand out as renewable and easily biodegradable alternatives to plastics. For example, chitosan extracted from shellfish exhibits antimicrobial properties and can be used in food packaging to inhibit microbial growth. Similarly, starch-based films reduce oxygen permeability, thereby extending shelf life, and their natural composition allows for easy degradation in nature (Kılınç, Tomar & Çağlar, 2017). Starch-based packaging is typically produced using starch from agricultural sources such as corn, potatoes, and wheat. These materials are readily biodegradable and can break down without harming the environment (Ghadge, 2020). Among plant-based biodegradable packaging materials, cellulose and its derivatives are widely preferred polymers in food packaging due to their high mechanical strength and biodegradability (Çelebi, 2018). Packaging made from these natural materials not only offers environmental benefits but also provides functional advantages such as preserving food freshness, extending shelf life, and protecting consumer health. Moreover, the use of renewable resources in their production supports sustainable agriculture and waste management practices. In conclusion, packaging systems developed from natural materials play a vital role in reducing environmental impact and achieving sustainability goals. Continued research and technological advancements in this field are
ENHANCING OXIDATIVE STABILITY OF RAINBOW TROUT FILLETS . . . 23 All groups were packaged in polystyrene trays and stored at +4 °C. Rainbow trout weighing 250–300 g were used in the study. Analyses were performed in duplicate with three replicates. Sampling was conducted every three days until the end of the storage period. Proximate composition analyses were performed only on the production day. 2.2.Methods 2.2.1.PreparationofChiaSeedMucilage Chia seed mucilage was obtained by hydration. Seeds were dispersed in distilled water at a ratio of 1:20 and stirred using a magnetic stirrer with heating for 2 h at 25 °C. Glycerol (1%) was added as a plasticizer, followed by an additional 15 min of stirring at 25 °C. Insoluble particles were removed by centrifugation, and residual seeds were eliminated by filtration. 2.2.2.IncorporationofHelichrysumarenariumExtract Two concentrations of Helichrysum arenarium extract were prepared. The extract was solubilized in Tween 20 (0.1%, w/w) and homogenized using a digital homogenizer at 20,000 rpm for 1 min, then incorporated into the prepared chia seed mucilage (Muñoz et al., 2012). 2.2.3.CoatingofFillets Fillets were immersed for 15 s in the three coating solutions, then dried in a cabinet dryer at 10 °C for 1 h. The coated samples were packaged in polystyrene trays and stored at +4 °C. 2.2.4.AnalyticalProcedures The proximate composition of rainbow trout fillets was determined using standard analytical procedures. Moisture content was measured by oven-drying according to AOAC 950.46, and dry matter was calculated by subtracting the moisture percentage from 100 (AOAC, 2002a). Crude protein was quantified using the micro-Kjeldahl method (AOAC 928.08) (AOAC, 2002b). Crude fat was determined by Soxhlet extraction following AOAC 960.39 (AOAC, 2002c). Crude ash was measured by incineration according to AOAC 920.153 (AOAC, 2002d). Carbohydrate content was not directly analyzed but calculated by difference, subtracting protein, fat, and ash from the dry matter (Akkılıç & Sürmen, 1979).
24 INNOVATIVE STUDIES IN AGRICULTURE, FORESTRY AND AQUACULTURE – 2025 pH Measurement: Ten grams of sample homogenized with 100 ml distilled water for 1 min; pH measured using a pH meter (AOAC, 2002e). Thiobarbituric Acid (TBA) Value: Determined according to Tarladgis et al. (1960). Malondialdehyde formed during lipid oxidation reacts with 2-thiobarbituric acid in acetic acid medium, producing a red color measured at 538 nm. Absorbance values were multiplied by 7.8 to calculate MDA concentration. Peroxide Value: Determined using the Wheeler method modified by Hadorn et al. (Varlık et al., 1993). After extraction, peroxide value was expressed as millimoles of oxygen per kilogram of fat, calculated using the following formula: PV (mmolkg-1 numune)=(a-b) F100/E where: a = volume (ml) of 0.01 N sodium thiosulfate used in the sample titration, b = volume (ml) of 0.01 N sodium thiosulfate used in the blank titration, F = correction factor of 0.01 N sodium, thiosulfate solution, E = fat content (g) in 20 ml solution 2.2.5.StatisticalAnalysis All data were analyzed using IBM SPSS® 22 (SPSS Inc., Chicago, IL, USA). Differences among groups and storage days were evaluated by analysis of variance (ANOVA). Statistical significance was determined according to Özdamar (2001). 3. Results and Discussion The proximate composition obtained in this study aligns closely with previous reports. Emir Çoban & Jamshidi (2023) reported rainbow trout fillets with moisture around 77% and protein near 18%, values comparable to our findings. Öz & Dikel (2024) highlighted seasonal variability in fat content, ranging from 2% to 8%, with lower values in leaner farmed fillets, consistent with the 2% fat observed here. Kavusan et al. (2023) emphasized that proximate composition remains largely unaffected by edible coatings, but coatings can indirectly preserve functional attributes such as water-holding capacity and oxidative stability by reducing drip loss and oxygen ingress.
ENHANCING OXIDATIVE STABILITY OF RAINBOW TROUT FILLETS . . . 25 Figure 1. Proximate composition of rainbow trout fillets. Thiobarbituric acid reactive substances (TBARS) are widely used as indicators of secondary lipid oxidation in fish muscle. In the literature, the sensory acceptability threshold for TBARS in fish fillets is generally reported around 2 mg MDA/kg, above which rancid odours and flavours become perceptible to consumers (Emir Çoban & Çoban, 2019; Kurek et al., 2024). Values between 2–3 mg MDA/kg are considered borderline, while levels exceeding 3–5 mg MDA/kg indicate pronounced rancidity and unacceptable quality (Soni et al., 2025). These thresholds provide a practical framework for evaluating oxidative stability during storage. In the present study, TBARS values increased progressively in all groups during 15 days of refrigerated storage. The control group (C) exceeded the acceptability limit by day 9 and reached the highest levels by day 15, confirming rapid progression of secondary oxidation in uncoated fillets (Fig 2a). Chia seed mucilage (CSM) coatings delayed TBARS accumulation, with values approaching the limit only in later storage days. More pronounced suppression was observed in groups enriched with Helichrysum arenarium extract.
26 INNOVATIVE STUDIES IN AGRICULTURE, FORESTRY AND AQUACULTURE – 2025 a b c Figüre 2. Changes in peroxide values (TBA,TVB-N, PV) of rainbow trout fillets under different coating treatments during 15 days of refrigerated storage at +4 °C.
ENHANCING OXIDATIVE STABILITY OF RAINBOW TROUT FILLETS . . . 27 The CSM+0.5%HA group maintained TBARS values near or below the threshold until mid-storage, while the CSM+1%HA group consistently remained below the critical limit throughout the storage period. These results demonstrate that chia–Helichrysum coatings effectively extend the oxidative stability and sensory acceptability of rainbow trout fillets under +4 °C storage conditions. The observed TBARS suppression is consistent with previous reports on edible coatings in fish preservation. Kurek et al. (2024) highlighted that plant-based films reduce oxygen permeability and moisture loss, thereby limiting aldehyde formation. Similarly, Soni et al. (2025) emphasized the dosedependent antioxidant effects of sage and rosemary extracts in fish and meat systems, which parallels the concentration-dependent efficacy observed in the present study. Kavusan et al. (2023) demonstrated that chia mucilage infused with sage extract significantly reduced TBARS in turkey fillets during chilled storage. Although the species and antioxidant source differ, the mechanism— phenolic radical scavenging and metal chelation—is comparable to the action of Helichrysum arenarium. The strong antioxidant capacity of Helichrysum extracts, confirmed by Uğur & Güzel (2025) through LC-MS/MS analysis, supports the sustained TBARS suppression observed in the 1% extract group. Furthermore, El Alami El Hassani et al. (2024) reviewed natural preservatives in fish systems and concluded that phenolic-rich coatings delay secondary oxidation and maintain sensory quality. The present findings extend this evidence by showing that chia–Helichrysum coatings maintain TBARS values below the acceptability threshold even under +4 °C storage, which is more challenging than the 2 °C conditions commonly used in similar studies. Peroxide value (PV) results in rainbow trout fillets clearly demonstrate oxidative progress during 15 days of cold storage (Fig 2b). The control group (C) exhibited the highest PV values, exceeding the spoilage threshold on day 9. Chia seed mucilage (CSM) coatings reduced PV accumulation, while the groups enriched with helichrysum extract exhibited antioxidant activity depending on the concentration used. In particular, the CSM+1%HA group maintained PV values below the critical limit throughout the entire period, demonstrating the highest oxidative stability. These findings are consistent with recent studies emphasizing the dual effects of edible coatings: chia mucilage creates a physical barrier that limits oxygen transmission, while Helichrysum arenarium extract delays lipid peroxidation by providing phenolic compounds that can capture peroxyl radicals (Emir Çoban & Jamshidi, 2023; Eren et al., 2023). Additionally, Kurek et al. (2024) demonstrated the barrier effect of edible coatings, which limits oxygen transmission and delays oxidation. Similarly, Çoban & Emir Çoban (2020)
28 INNOVATIVE STUDIES IN AGRICULTURE, FORESTRY AND AQUACULTURE – 2025 demonstrated that a combination of chia mucilage and propolis extract reduced PV and extended shelf life in sea bass fillets. The current study demonstrates that Helichrysum arenarium, a phenolic-rich plant extract, can be successfully integrated into chia-based coatings. Recent studies emphasize the importance of plant-derived antioxidants. Pandiyan et al. (2025) reported that phenolic compounds in seafood delay PV accumulation and increase storage stability. Furthermore, Uğur & Güzel (2025) confirmed the high phenolic content and strong radical scavenging activity of helichrysum extract using LC-MS/MS analyses, supporting the fact that the 1% extract group maintained low PV even after 15 days. In addition to TBA and TVB-N analyses, pH changes in rainbow trout fillets were monitored during 15 days of cold storage under different coating treatments, and the results are presented in Figure 2c. The initial pH in all groups was approximately 6.4–6.5, which is consistent with the range reported in the literature for fresh fish muscle (Emir Çoban Jamshidi,). As storage progressed, a significant increase in pH was observed, especially in the uncoated control group (C), reaching above 7.0 by day 15. In contrast, the pH increase remained significantly lower in the groups coated with chia seed mucilage (CSM) and enriched with helichrysum arenarium extract (CSM+0.5%HA and CSM+1%HA); the CSM+1%HA group, in particular, maintained the most stable pH profile throughout the entire storage period. These findings are consistent with the literature reporting that pH increases in fish muscle during cold storage are associated with microbial activity and the accumulation of alkaline metabolites such as ammonia and trimethylamine (Kurek et al., 2024). The rapid pH increase in the control group reflects the typical spoilage process in fish with high moisture and protein content, such as rainbow trout. In contrast, edible coatings can slow pH changes by delaying microbial growth and enzymatic degradation. Hassan et al. (2025) reported that vacuum packaging of carp fillets caused a slower pH increase than atmospheric packaging, highlighting the role of oxygen limitation in controlling spoilage. The pH stabilization effect of chia mucilage coatings is attributed to their semi-permeable barrier properties, which reduce oxygen transmission and moisture loss. The addition of helichrysum extract further enhanced this effect due to the antimicrobial and antioxidant capacity of phenolic compounds. The dose-dependent effect observed in this study was similar to that observed in El Hassani et al. (2024) reported that the effectiveness of natural preservatives depends on concentration.
ENHANCING OXIDATIVE STABILITY OF RAINBOW TROUT FILLETS . . . 29 Statistical analyses revealed significant differences (p<0.05) between groups and days, with the CSM+1%HA group clearly distinguishing itself from the control group starting on day 6. These results indicate that chia-gold herb coatings not only suppress oxidative markers (PV, TBARS) but also control pH changes, contributing to extending shelf life and maintaining quality in rainbow trout fillets. Conclusion This study demonstrated that edible coatings prepared with chia seed mucilage and Helichrysum arenarium extract effectively delayed oxidative deterioration and pH changes in rainbow trout fillets during refrigerated storage. In particular, coatings containing 1% extract maintained PV and TBARS values below critical limits and stabilized pH, thereby extending shelf life and preserving quality. These findings highlight the chia–Helichrysum combination as efficient preservation strategy for seafood products. Acknowledgment This study is grateful to the Firat University Scientific Research Projects Coordination Unit (FÜBAP) for the financial support provided under project number SÜF.24.15. References Akkılıç, M., & Sürmen, S. (1979). Yem Maddeleri ve Hayvan Besleme. Ankara Üniversitesi Veteriner Fakültesi Yayınları No: 357. Van Soest A.Ü. Basımevi, Ankara. AOAC. (2002a). Moisture content (Method 950.46). In Official Methods of Analysis (17th ed.). Association of Official Analytical Chemists, Gaithersburg, Maryland. AOAC. (2002b). Crude protein (Method 928.08). In Official Methods of Analysis (17th ed.). Association of Official Analytical Chemists, Gaithersburg, Maryland. AOAC. (2002c). Fat content in meat (Method 960.39). In Official Methods of Analysis (17th ed.). Association of Official Analytical Chemists, Gaithersburg, Maryland. AOAC. (2002d). Ash content in meat (Method 920.153). In Official Methods of Analysis (17th ed.). Association of Official Analytical Chemists, Gaithersburg, Maryland.
30 INNOVATIVE STUDIES IN AGRICULTURE, FORESTRY AND AQUACULTURE – 2025 AOAC. (2002e). pH (Method 981.12). In Official Methods of Analysis (17th ed.). Association of Official Analytical Chemists, Gaithersburg, Maryland. Çoban, M. Z., & Emir Çoban, Ö. (2020). Potency and use of chia mucilage coating containing propolis liquid extract for improving shelf-life of sea bass fillets. Acta Sci. Pol. Technol. Aliment., 19(3), 255–260. https://doi. org/10.17306/J.AFS.2020.0843 El Alami El Hassani, N., Baraket, A., & Alem, C. (2024). Recent advances in natural food preservatives: a sustainable solution for food safety and shelf life extension. Journal of Food Measurement and Characterization, 19(1), 293–315. https://doi.org/10.1007/s11694-024-02969-x Emir Çoban, Ö., & Jamshidi, A. (2023). Development of bionanocomposite film based on chia seed mucilage incorporated with ZnO nanoparticles for preserving rainbow trout fillets. Journal of Food Measurement and Characterization, 18(4), 1000–1011. https://doi.org/10.1007/s11694-02302263-2 Emir Çoban, Ö. ve Çoban, M.Z., (2019). Meyan Kökü Ekstraktı İle Zenginleştirilmiş Kitosan Kaplamanın Alabalık (Oncorhynchus mykiss) Filetolarının Kalitesi Üzerine Etkisi, Ecological Life Sciences (NWSAELS), 14(4):83-92 Eren, A., İnci, Ş., Saleh, K. K., Kırbağ, S., & Güven, K. (2023). Helichrysum arenarium subsp. aucheri ekstraktlarının antimikrobiyal ve antioksidan aktiviteleri. Turkish Journal of Science and Technology, 18(2), 345–351. Kurek, M., Pišonić, P., Ščetar, M., Janči, T., Čanak, I., Vidaček Filipec, S., Benbettaieb, N., Debeaufort, F., & Galić, K. (2024). Edible coatings for fish preservation: Literature data on storage temperature, product requirements, antioxidant activity, and coating performance. Antioxidants, 13(11), 1417. Muñoz, L. A., Cobos, A., Díaz, O., & Aguilera, J. M. (2012). Chia seeds: Microstructure, mucilage extraction and hydration. Journal of Food Engineering, 108(1), 216–224. https://doi.org/10.1016/j.jfoodeng.2011.06.037 OUP (Oxford University Press). (2024). Biopolymer-based edible films and coatings: Toward eco-friendly and sustainable food preservation. International Journal of Food Science & Technology, 60(2), vvaf213. https://doi.org/10.1093/ ijfst/vvaf213 Özdamar, K. (2001). SPSS ile Biyoistatistik (4th ed.). Kaan Kitabevi Yayın No: 3, Eskişehir. Pandiyan, P., et al. (2025). Plant-derived antioxidants in seafood preservation: A review. International Journal of Research in Agronomy, 8(4), 49–56
ENHANCING OXIDATIVE STABILITY OF RAINBOW TROUT FILLETS . . . 31 Popa, G., Dănăilă-Guidea, S., Damian, E., Mărgărit, G., & GroposilăConstantinescu, D. (2023). Evaluation of bioactive compounds with antioxidant activity of Helichrysum arenarium inflorescences. Scientific Papers Series B, Horticulture, 67(2), 145–152. Sharefiabadaa, E., Kavusan, H. S., & Serdaroğlu, M. (2023). Innovative coating approach: vacuum impregnation with chia mucilage and sage infusion for turkey fillets. Meat Technology, 64(2), 67–75. https://doi.org/10.18485/ meattech.2023.64.2.67 Tarladgis, B. G., Watts, B. M., Younathan, M. T., & Dugan, L. R. Jr. (1960). A distillation method for the quantitative determination of malonaldehyde in rancid foods. Journal of the American Oil Chemists’ Society, 37(1), 44–48. https://doi.org/10.1007/BF02630824 Uğur, Y., & Güzel, A. (2025). Phenolic profile and antioxidant capacity of Helichrysum arenarium extracts: A comprehensive LC-MS/MS analysis. KSU Journal of Agriculture and Nature, 28(1), 1–8. Varlık, C., Uğur, M., Gökoğlu, N., & Gün, H. (1993). Su Ürünlerinde Kalite Kontrol İlke ve Yöntemleri. Gıda Teknolojisi Derneği Yayın No: 17, İstanbul, 174 pp
CONTROLLED AND SOILLESS CULTIVATION OF MEDICINAL . . . 39 Biological elicitors stimulate plant defense responses through microorganisms or their cell wall components. This approach not only enhances metabolite production but also contributes to strengthening plant stress tolerance. 4.5.StrategicImportanceofElicitationinControlledSystems Controlled environment agriculture and soilless cultivation systems provide an ideal infrastructure for implementing elicitation strategies in a species and target metabolite specific manner. The temporal management of light, nutrient supply, and stress factors enables the selective and reproducible enhancement of secondary metabolites (Dsouza et al., 2025). In conclusion, when carefully applied under controlled environmental conditions, elicitation strategies represent a powerful tool that enables the enhancement of secondary metabolite production in medicinal and aromatic plants without causing a substantial suppression of growth. 5. Nutrient Solution and Secondary Metabolite Optimization In hydroponic and soilless cultivation systems, the nutrient solution functions not merely as a passive component supporting plant growth but as a key regulator directing secondary metabolite biosynthesis. The ionic composition of the nutrient solution, its pH value, and particularly its electrical conductivity exert direct effects on carbon nitrogen balance and metabolic priorities within the plant (Rusu et al., 2021; Hazrati et al., 2024). 5.1.MetabolicRoleoftheHydroponicNutrientSolution The hydroponic nutrient solution enables the controlled supply of macro and microelements, thereby allowing the establishment of a balance among photosynthetic efficiency, root development, and secondary metabolite production. In particular, variations in the ratios of nitrogen, phosphorus, and potassium directly influence the biosynthesis of metabolite classes such as phenolic compounds, flavonoids, and terpenoids (Yonesi et al., 2024). 5.2.ElectricalConductivityManagementandMetaboliteResponse The electrical conductivity of the nutrient solution determines the osmotic potential of the root zone, thereby shaping ion uptake kinetics and metabolic orientation. Low to moderate electrical conductivity levels can induce mild
40 INNOVATIVE STUDIES IN AGRICULTURE, FORESTRY AND AQUACULTURE – 2025 nutrient stress, which may promote secondary metabolite accumulation within the framework of the balance between growth and defense. This effect has been clearly demonstrated in the cultivation of Glehnia littoralis, where low electrical conductivity conditions were reported to increase both biomass production and the accumulation of pharmacologically important compounds such as xanthotoxin, bergapten, and imperatorin when compared with higher electrical conductivity levels (Yeom & Oh, 2023). Similarly, in Acmella oleracea, variations in electrical conductivity levels have been reported to exert pronounced effects on essential oil content and composition (Carmo et al., 2024). 5.3.IonRatiosandtheRegulationofChemicalProfiles The relative proportions of macro and microelements in the nutrient solution directly influence the activity of biochemical pathways involved in secondary metabolite biosynthesis. In particular, nitrogen forms and potassium balance play a decisive role in regulating phenylpropanoid and terpenoid metabolism. Studies conducted on basil (Ocimum basilicum L.) have shown that linalool content reaches its maximum level under low ion concentration conditions. This finding demonstrates that nutrient management in hydroponic systems should be addressed not only in terms of absolute amounts but also on a ratio based basis (Hazrati et al., 2024). 5.4.FertilizerFormandOrganic-InorganicInteractions In the optimization of secondary metabolite production, the form of fertilizer is as important as the total nutrient supply. It has been reported that a balanced combination of organic and inorganic fertilizers enhances the accumulation of ginsenoside monomers such as Rb1, Re, and Rg1 in saponin producing medicinal plants (Lv et al., 2025). Similarly, in Sideritis cypria, phosphorus levels have been shown to interact with foliar applied iron and zinc, thereby regulating total phenolic and flavonoid content (Neofytou et al., 2025). These interactions indicate that micronutrients can function as metabolic modulators. In conclusion, optimization of the hydroponic nutrient solution in terms of electrical conductivity, ion ratios, and fertilizer form enables the species specific and target compound oriented regulation of secondary metabolite production in medicinal and aromatic plants.
CONTROLLED AND SOILLESS CULTIVATION OF MEDICINAL . . . 41 6. Effects of Light Spectrum and Intensity on Secondary Metabolite Profiles Light functions not only as an energy source supporting photosynthesis in plants but also as a key environmental signal regulating secondary metabolite biosynthesis. In controlled environment agriculture and soilless cultivation systems, the precise adjustment of light quality, intensity, and photoperiod enables the development of metabolite focused production strategies (Hashim et al., 2021). 6.1.MetabolicRegulatoryRoleofLight Plants perceive light signals through photoreceptors such as phytochromes, cryptochromes, and phototropins, and these signals directly influence the expression of genes involved in secondary metabolite biosynthesis. In particular, phenylpropanoid and terpenoid pathways are among the metabolic networks that are highly sensitive to light quality and intensity (Hashim et al., 2021). This indicates that light can be deliberately used as an elicitor under controlled conditions. 6.2.LEDLightSpectrumandChemicalComposition Light emitting diode technology enables targeted regulation of plant metabolism by providing narrow band and adjustable light spectra. Combinations of red and blue light have been reported to enhance the accumulation of phenolic compounds and flavonoids, while green and far red light can exert indirect metabolic effects through morphological responses (Hashim et al., 2021). Under hydroponic conditions, Lippia palmeri cultivated under full spectrum LED lighting showed an increase in carvacrol content of up to 47%, and this shift was reflected in the biological activity of the extracted essential oil (Bringas‐Burgos et al., 2023). This example clearly demonstrates a direct relationship between light spectrum and chemical profile. 6.3.LightIntensity,Photoperiod,andDailyLightIntegral Light intensity and photoperiod influence metabolite production through the Daily Light Integral, which represents the total amount of light received by a plant per day. In plant factories with artificial lighting, high Daily Light Integral strategies have been reported to enhance both yield and quality in phenolic rich plant species (Xu et al., 2021).
42 INNOVATIVE STUDIES IN AGRICULTURE, FORESTRY AND AQUACULTURE – 2025 Studies conducted on Nasturtium officinale have shown that the combination of continuous lighting and high Daily Light Integral increases total phenolic content and enhances the functional value of the product (Xu et al., 2021). These findings indicate that light management can be considered a quantitative control parameter in metabolite oriented production systems. 6.4.LightWaterInteractionsandMetaboliteDynamics The effects of light intensity on metabolite production should often be evaluated in conjunction with water availability. Particularly in medicinal plants containing alkaloids, the interaction between light intensity and water stress can play a decisive role in shaping metabolic responses. Studies conducted on Mitragyna speciosa have reported that the combination of low light intensity and moderate water stress results in the highest total alkaloid content. In addition, young leaves were found to exhibit higher alkaloid accumulation compared with older leaves (Leksungnoen et al., 2025). These results indicate that light management should be addressed in a tissue specific and developmental stage dependent manner. In summary, the controlled management of light spectrum, intensity, and photoperiod enables the targeted and reproducible optimization of secondary metabolite production in medicinal and aromatic plants. 7. Biotechnological Approaches and Cell Culture Applications The sustainable and standardized production of high value secondary metabolites has become increasingly difficult using traditional approaches based on raw material supply from natural populations. In this context, biotechnological production systems offer a strategic alternative for medicinal and aromatic plants by providing controlled, year round production that is independent of environmental variability (Wawrosch & Zotchev, 2021; Danova & Pistelli, 2022). 7.1.StrategicImportanceofInVitroProductionSystems In vitro culture systems enable the precise regulation of metabolite biosynthesis through controlled manipulation of culture media composition, hormone balance, and environmental stimuli using callus, cell suspension, organ, and root cultures. These systems offer production models that meet pharmaceutical quality requirements by ensuring genetic and chemical consistency (Niazian & Sabbatini, 2021).
CONTROLLED AND SOILLESS CULTIVATION OF MEDICINAL . . . 43 In particular, for species that are sensitive to environmental stresses or exhibit slow growth under field conditions, in vitro systems provide the opportunity to optimize the production of target metabolites independently of biomass accumulation. 7.2. Adventitious Root Cultures as Metabolite Focused Production Platforms Adventitious root cultures are among the high efficiency biotechnological platforms for plant species in which secondary metabolites predominantly accumulate in root tissues. These systems offer advantages such as rapid growth, genetic stability, and limited hormone requirements, thereby providing economically feasible production model (Khanam et al., 2022). Root cultures are particularly prominent for the production of alkaloids, naphthoquinones, and phenolic compounds, enabling these metabolites to be obtained at higher and more reproducible levels compared with field grown plants. 7.3.MicrobialElicitorsandSymbioticInteractions In biotechnological production systems, microbial elicitors are considered effective tools for enhancing secondary metabolite production by activating plant defense mechanisms. Symbiotic microorganisms such as arbuscular mycorrhizal fungi can guide metabolic responses in addition to improving nutrient uptake in plants. In Alkanna tinctoria cultivated under partially hydroponic systems, inoculation with arbuscular mycorrhizal fungi has been reported to increase the production of alkannin and shikonin derivatives, and this effect has been associated with the regulation of genes involved in biosynthetic pathways (Zhao et al., 2023). 7.4.NanotechnologicalApproachesasNextGenerationElicitors Nanotechnology has attracted increasing interest in the field of plant biotechnology. It has been reported that the application of metal and metal oxide nanoparticles at low doses can stimulate secondary metabolite production by activating plant defense metabolism (Kralova & Jampilek, 2021; Punetha et al., 2022). However, the accumulation potential of nanoparticles in plant tissues, their bioavailability, and implications for final product safety require careful
44 INNOVATIVE STUDIES IN AGRICULTURE, FORESTRY AND AQUACULTURE – 2025 evaluation. Therefore, despite its high potential, this approach is currently considered to be at an experimental and optimization stage. 7.5.InteractionBetweenGeneticBackgroundandCultivationSystems Secondary metabolite production is closely linked not only to the cultivation system but also to the genetic background of the plant. Comparative studies conducted on diploid and tetraploid lines of Bacopa monnieri have shown that metabolite production varies depending on ploidy level and cultivation conditions. It has also been reported that hydroponic systems can substantially reduce the risk of heavy metal and toxic substance contamination, even in cases where metabolite content is lower than that observed under field conditions (Inthima & Supaibulwatana, 2024). This situation demonstrates that biotechnological and controlled cultivation systems play a critical role in ensuring product safety. In conclusion, biotechnological production systems and cell culture applications offer complementary strategies that enable the standardized and sustainable production of secondary metabolites in medicinal and aromatic plants, independently of environmental uncertainties. 8. Precision Cultivation and Organ Specific Metabolite Optimization In medicinal and aromatic plants, secondary metabolites are often produced and stored in specific organs such as leaves, flowers, roots, or fruits. Therefore, metabolite focused production strategies increasingly aim to optimize the growth of the target organ rather than overall plant biomass. In this context, precision cultivation refers to an advanced approach that focuses on adjusting environmental conditions to enhance metabolite production at the organ level (Atherton & Li, 2023). 8.1.OrganSpecificLocalizationofSecondaryMetabolites The accumulation of secondary metabolites at different concentrations among plant organs is closely associated with plant defense strategies and developmental physiology. Phenolic compounds and flavonoids are predominantly concentrated in leaf and flower tissues, whereas alkaloids and certain terpenoid derivatives are often found at higher concentrations in root tissues. This organ specific distribution indicates that the biosynthesis and storage of secondary metabolites are regulated in a tissue dependent manner (Wink, 2008).
CONTROLLED AND SOILLESS CULTIVATION OF MEDICINAL . . . 45 Organ specific localization of metabolites is not determined solely by the genetic background of the plant but is also sensitive to environmental factors such as light conditions, nutrient availability, and stress related signals. In controlled environment agriculture and soilless cultivation systems, the precise management of these parameters enables the development of strategies aimed at increasing both the biomass and metabolic activity of the organ in which the target metabolite is concentrated (Ćavar Zeljković et al., 2022). In this context, metabolite focused production approaches go beyond traditional cultivation practices that aim to optimize the entire plant uniformly and instead require the adoption of precision cultivation strategies specifically targeted at selected plant organs. 8.2.VerticalSystemsandOrganSpecificProductionPerformance Vertical hydroponic systems enable the selective promotion of specific plant organs through optimized light distribution and precise microclimate control. The ability to adjust light spectrum and intensity across vertical layers can directly influence the biomass and chemical composition of metabolite rich organs such as leaves and flowers (Atherton & Li, 2023; Dsouza et al., 2025) In studies targeting apigenin accumulation, flowers of Matricaria chamomilla and leaves of Petroselinum crispum grown under vertical hydroponic conditions have demonstrated high production performance. In particular, the Bodegold chamomile cultivar exhibited apigenin accumulation reaching 0.70 milligrams per gram of dry tissue in flower organs, highlighting the decisive role of the interaction among genotype, organ, and cultivation system in metabolite optimization (Maynard et al., 2025). 8.3.PlantDensity,HarvestTiming,andMetaboliteDynamics Plant density and harvest timing often represent a trade off in secondary metabolite production. Under high density conditions, competition for light and nutrients can limit metabolite accumulation, whereas low density conditions may reduce yield per unit area (Atherton & Li, 2023). Studies conducted on Andrographis paniculata have shown that a moderate plant density of 30 plants per square meter combined with harvesting at 90 days maximizes andrographolide accumulation (Chutimanukul et al., 2022). These findings demonstrate that the concept of an optimal range is critical for metabolite focused production within precision cultivation strategies.
46 INNOVATIVE STUDIES IN AGRICULTURE, FORESTRY AND AQUACULTURE – 2025 8.4.IntegrationofPhenomicsandMetabolomicsApproaches The integration of phenomics and metabolomics approaches in the development of precision cultivation strategies enables high resolution characterization of the relationships between environmental conditions and metabolic responses. These methods allow the simultaneous assessment of not only morphological traits but also metabolic reprogramming processes (Dsouza et al., 2025). Studies conducted under semi hydroponic systems have shown that phenolic and terpenoid metabolism can be characterized in detail in Mentha species and Ocimum basilicum genotypes under conditions of low nitrate availability and mild salinity stress (Ćavar Zeljković et al., 2022). These approaches provide powerful analytical tools for predicting future cultivation scenarios. Overall, precision cultivation approaches enable the development of optimized production models for target compounds in medicinal and aromatic plants by jointly considering organ specific metabolite localization, cultivation system selection, plant density, and harvest timing. 9. Conclusion Traditional soil based cultivation systems are not always sufficient for the production of consistent and reliable plant raw materials rich in bioactive compounds due to environmental variability, resource limitations, and contamination risks. In contrast, controlled environment and soilless cultivation systems enable precise regulation of the root environment, nutrient management, and environmental conditions, thereby allowing the directed control of the relationship between plant growth and bioactive compound accumulation. Hydroponic and aquaponic systems contribute to improved resource use efficiency while supporting the production of plant raw materials that are biologically active and chemically more consistent. In aquaponic systems, microbial interactions provide an additional advantage by further supporting the synthesis of bioactive compounds. Elicitation strategies based on environmental manipulation emerge as effective tools for enhancing bioactive compound production when applied under controlled conditions. However, it is clear that such applications must be carefully evaluated with respect to dosage, duration, and product safety. Nutrient solution management and the optimization of light spectrum and intensity represent core technical components of bioactive compound
CONTROLLED AND SOILLESS CULTIVATION OF MEDICINAL . . . 47 focused cultivation. The integrated consideration of these parameters enables the development of species specific and target compound oriented production strategies. Biotechnological approaches and in vitro culture systems support the sustainable production of bioactive compounds by offering standardized production that is independent of environmental conditions. These systems also contribute to reducing harvesting pressure on natural populations. In conclusion, the integrated use of controlled environment systems, soilless cultivation, and biotechnological approaches provides a robust and practical roadmap for sustainable, reliable, and standardized production of bioactive compounds in medicinal and aromatic plants. References Abdalla, M. A., Li, F., Wenzel-Storjohann, A., Sulieman, S., Tasdemir, D., & Mühling, K. H. (2021). Comparative Metabolite Profile, Biological Activity and Overall Quality of Three Lettuce (Lactuca sativa L., Asteraceae) Cultivars in Response to Sulfur Nutrition. Pharmaceutics, 13(5), 713. https:// doi.org/10.3390/pharmaceutics13050713 Atherton, H. R., & Li, P. (2023). Hydroponic Cultivation of Medicinal Plants—Plant Organs and Hydroponic Systems: Techniques and Trends. Horticulturae, 9(3), 349. https://doi.org/10.3390/horticulturae9030349 Bringas‐Burgos, B. F., Martínez‐Robinson, K. G., Toledano‐Magaña, Y., García‐Ramos, J. C., Ovando‐Martínez, M., & López‐Elías, J. (2023). Antiproliferative Effect of Essential Oil Obtained from Oregano ( Lippia palmeri S. Watson) Leaves Grown in Hydroponics and LED Light. Chemistry & Biodiversity, 20(4), e202201076. https://doi.org/10.1002/cbdv.202201076 Carmo, A. P. M. D., Freitas, M. S. M., Machado, L. C., Silva, L. D. S., Petri, D. J. C., Vimercati, J. C., Matos, C. R. R., Mathias, L., Vieira, I. J. C., & De Carvalho, A. J. C. (2024). Electrical conductivity of nutrient solutions affects the growth, nutrient levels, and content and composition of essential oils of Acmella oleracea (L.) R. K. Jansen from southeastern Brazil. Journal of Agriculture and Food Research, 15, 100968. https://doi.org/10.1016/j.jafr.2024.100968 Ćavar Zeljković, S., Aucique-Perez, C. E., Štefelová, N., & De Diego, N. (2022). Optimizing growing conditions for hydroponic farming of selected medicinal and aromatic plants. Food Chemistry, 375, 131845. https://doi. org/10.1016/j.foodchem.2021.131845
48 INNOVATIVE STUDIES IN AGRICULTURE, FORESTRY AND AQUACULTURE – 2025 Chrysargyris, A., Maggini, R., Incrocci, L., Pardossi, A., & Tzortzakis, N. (2021). Copper Tolerance and Accumulation on Pelargonium graveolens L’Hér. Grown in Hydroponic Culture. Plants, 10(8), 1663. https://doi.org/10.3390/ plants10081663 Chutimanukul, P., Mosaleeyanon, K., Janta, S., Toojinda, T., Darwell, C. T., & Wanichananan, P. (2022). Physiological responses, yield and medicinal substance (andrographolide, AP1) accumulation of Andrographis paniculata (Burm. F) in response to plant density under controlled environmental conditions. PLOS ONE, 17(8), e0272520. https://doi.org/10.1371/journal.pone.0272520 Danova, K., & Pistelli, L. (2022). Plant Tissue Culture and Secondary Metabolites Production. Plants, 11(23), 3312. https://doi.org/10.3390/ plants11233312 Dsouza, A., Dixon, M., Shukla, M., & Graham, T. (2025). Harnessing controlled-environment systems for enhanced production of medicinal plants. Journal of Experimental Botany, 76(1), 76–93. https://doi.org/10.1093/jxb/ erae248 Fierascu, R. C., Fierascu, I., Baroi, A. M., & Ortan, A. (2021). Selected Aspects Related to Medicinal and Aromatic Plants as Alternative Sources of Bioactive Compounds. International Journal of Molecular Sciences, 22(4), 1521. https://doi.org/10.3390/ijms22041521 Giri, L., Angmo, J. C., Hussain, M., Singh, B., Bhatt, I. D., & Nautiyal, S. (2025). Hydroponic culture improves growth and secondary metabolite production in Rheum tibeticum , a near threatened species from the Ladakh Trans-Himalayan region of India. Plant Biosystems - An International Journal Dealing with All Aspects of Plant Biology, 159(2), 356–368. https://doi.org/10. 1080/11263504.2025.2468727 Hashim, M., Ahmad, B., Drouet, S., Hano, C., Abbasi, B. H., & Anjum, S. (2021). Comparative Effects of Different Light Sources on the Production of Key Secondary Metabolites in Plants In Vitro Cultures. Plants, 10(8), 1521. https://doi.org/10.3390/plants10081521 Hawrylak-Nowak, B., Dresler, S., Stasińska-Jakubas, M., Wójciak, M., Sowa, I., & Matraszek-Gawron, R. (2021). NaCl-Induced Elicitation Alters Physiology and Increases Accumulation of Phenolic Compounds in Melissa officinalis L. International Journal of Molecular Sciences, 22(13), 6844. https:// doi.org/10.3390/ijms22136844 Hazrati, S., Pignata, G., Casale, M., Binello, A., Cravotto, G., Devecchi, M., & Nicola, S. (2024). Impact of four hydroponic nutrient solutions and
MEDICINAL PLANT PHYTOCHEMICALS: SUSTAINABLE EXTRACTION . . . 55 2. Conventional and Green Extraction Techniques The recovery of bioactive compounds from medicinal plants represents a fundamental innovation process for the pharmaceutical and nutraceutical industries (Dincheva et al., 2025). Conventional extraction techniques such as maceration, percolation, reflux extraction, and Soxhlet extraction have long established industrial standards (Cao et al., 2025). However, these well established methodologies suffer from significant drawbacks, including high consumption of organic solvents, prolonged extraction times, and the application of elevated temperatures that may lead to the thermal degradation of bioactive constituents (Chaves et al., 2020). These inherent limitations of conventional methods have stimulated the search for more efficient and sensitive extraction strategies aimed at preserving the purity and biological stability of bioactive molecules (Olaniyan et al., 2025). As a result of these efforts, green extraction technologies have emerged as sustainable processes that minimize environmental impact while reducing energy consumption and employing non toxic alternative solvents (Mirzazadeh et al., 2024). The core philosophy of green extraction is to achieve high quality extracts while simultaneously preserving natural resources and ensuring food and pharmaceutical safety (Darwin et al., 2025). The transition from conventional techniques to these innovative approaches reflects the practical implementation of green chemistry principles at both laboratory and industrial scales in alignment with global sustainability goals (Martins et al., 2023). In this context, techniques such as pressurized hot water extraction and electrohydrodynamic methods demonstrate clear advantages over conventional counterparts, particularly in preserving thermosensitive antioxidant compounds (Bastos et al., 2025). Although the performance of green technologies varies depending on the methodology applied, techniques including microwave assisted extraction, ultrasound assisted extraction, and supercritical fluid extraction significantly enhance processing speed and extraction efficiency (Cao et al., 2025). For instance, a comparative study on anthocyanin extraction from red onion peel reported that high hydrostatic pressure assisted extraction achieved the highest efficiency, reaching 81.84% yield (Mirzazadeh et al., 2024). The advancement of these modern extraction techniques has progressed in parallel with the widespread adoption of advanced analytical tools such as high performance liquid chromatography and nuclear magnetic resonance spectroscopy, enabling more accurate characterization of bioactive compounds (El Allaoui et al., 2024). Nevertheless, it should be emphasized that no single technology offers a universal solution for industrial applications, as each method presents target
56 INNOVATIVE STUDIES IN AGRICULTURE, FORESTRY AND AQUACULTURE – 2025 compound specific advantages as well as economic and operational limitations (Usman et al., 2023). Beyond physical extraction techniques, the chemical nature of the solvents employed plays a strategic role in sustainability considerations (Herrero, 2024). Deep eutectic solvents and ionic liquids have gained increasing attention as alternatives to conventional volatile organic solvents due to their lower toxicity and higher selectivity (Bastos et al., 2025). In particular, responsive deep eutectic solvents that undergo phase separation in response to external stimuli such as temperature, pH, or carbon dioxide enable solvent recovery and reuse, thereby contributing directly to circular economy frameworks (Vicente et al., 2025). When combined with non thermal techniques such as pulsed electric fields, these innovative solvent systems form highly efficient extraction platforms that preserve compound integrity at an advanced level (Anwar, 2025). Recent studies increasingly emphasize that hybrid or integrated extraction strategies, which combine multiple techniques rather than relying on a single method, deliver superior extraction yields and biological activity (Sun et al., 2025). The optimization of these hybrid processes through artificial intelligence based models, including artificial neural networks and genetic algorithms, plays a critical role in resolving complex variable interactions and enhancing overall process sustainability (Anwar, 2025). Ultimately, these modern green technologies that are progressively replacing conventional methods provide an environmentally friendly and economically viable roadmap for maximizing the therapeutic potential of medicinal plants in industrial applications (Sun et al., 2025). 3. Advanced Green Extraction Technologies (UAE, MAE, SFE, SWE) The limitations of conventional extraction methods, including maceration, percolation, and Soxhlet extraction, such as high solvent consumption, long processing times, and the degradation of heat sensitive compounds, have driven the industry toward more sustainable alternatives (El Saadony et al., 2025). Advanced green extraction technologies aim to increase extraction efficiency while reducing environmental footprint, minimizing energy consumption, and producing high purity products free from toxic solvent residues (Cao et al., 2025). These innovative approaches represent a fundamental strategic transformation, particularly in the pharmaceutical and nutraceutical sectors, for the discovery and utilization of bioactive compounds (Dincheva et al., 2025).
MEDICINAL PLANT PHYTOCHEMICALS: SUSTAINABLE EXTRACTION . . . 57 Among these modern techniques, microwave assisted extraction is one of the most prominent approaches. It rapidly heats and disrupts plant cell walls through ionic conduction and dipole rotation induced by microwave radiation, thereby accelerating the transfer of bioactive compounds into the solvent phase (Wang et al., 2025). Compared with conventional methods, microwave assisted extraction can reduce extraction time from 180 minutes to as little as 30 minutes while significantly lowering carbon emissions (Bodea et al., 2025). For example, in the recovery of polyphenols from Cinnamomum iners leaves, microwave assisted extraction processes optimized using response surface methodology were shown to produce extracts with high antioxidant capacity in substantially shorter times than conventional reflux extraction (Tanruean et al., 2025). Another widely applied green technology is ultrasound assisted extraction, which maximizes mass transfer by generating microfractures in plant tissues through acoustic cavitation phenomena (Cao et al., 2025). In recent years, ultrasound assisted extraction has been increasingly combined with environmentally friendly deep eutectic solvents to create synergistic effects in the extraction of polyphenolic compounds (Siddiqui et al., 2025). A comparative study conducted on feijoa flowers demonstrated that ultrasound assisted extraction achieved the highest yields for flavonoids such as quercetin and isoquercitrin, while the use of deep eutectic solvents showed superior selectivity for specific target compounds (Gil et al., 2023). Supercritical fluid extraction, developed in accordance with sustainability principles, enables the isolation of heat sensitive essential oils and lipids without leaving solvent residues, particularly through the use of supercritical carbon dioxide (Herzyk et al., 2024). Supercritical carbon dioxide extraction provides an environmentally friendly and commercially viable pathway for converting by products such as fruit seed wastes into value added oils (Nastić et al., 2025). Similarly, subcritical water extraction, also known as pressurized hot water extraction, modifies the dielectric constant of water through controlled temperature and pressure conditions, allowing the recovery of compounds with varying polarities using water as a single solvent (Aminzai et al., 2025). These approaches effectively preserve the structural integrity of thermosensitive antioxidants while contributing to circular economy strategies by enhancing the value of agro industrial residues (Bastos et al., 2025). Technological advances have also facilitated the development of hybrid extraction systems that combine the advantages of multiple methods. Techniques such as enzyme assisted microwave extraction and pulsed electric field treatments
58 INNOVATIVE STUDIES IN AGRICULTURE, FORESTRY AND AQUACULTURE – 2025 enhance extraction efficiency while preserving the biological activity of target compounds (Anwar, 2025). The use of bio based solvents and deep eutectic solvents in these hybrid systems further reinforces the green character of the extraction process by replacing petroleum derived toxic solvents (Usman et al., 2023). In addition, next generation approaches such as electrohydrodynamic methods enable the recovery of high quality antioxidants with substantially lower energy consumption (Bastos et al., 2025). In conclusion, the industrial applicability of advanced green extraction processes depends on factors including process optimization, economic feasibility, and life cycle assessment. Artificial intelligence models such as artificial neural networks and genetic algorithms play a critical role in optimizing complex extraction parameters and overcoming challenges associated with scale up (Anwar, 2025). The cost effectiveness and operational efficiency of these technologies are key determinants supporting the large scale utilization of valuable compounds derived from medicinal plants in the food, cosmetic, and pharmaceutical industries (Prado et al., 2017).
MEDICINAL PLANT PHYTOCHEMICALS: SUSTAINABLE EXTRACTION . . . 59 Table 1. Comparative overview of major green extraction technologies Extraction Technique Fundamental Principle Main Advantages Main Limitations Typical Target Compounds Industrial Maturity References MicrowaveAssisted Extraction (MAE) Rapid volumetric heating via microwaveinduced dipole rotation and ionic conduction Short extraction time, reduced solvent consumption, high extraction efficiency Limited penetration depth, risk of thermal degradation for highly sensitive compounds Polyphenols, flavonoids, alkaloids Pilot to semiindustrial Wang et al., 2025; Bodea et al., 2025; Cao et al., 2025 UltrasoundAssisted Extraction (UAE) Acoustic cavitation enhances cell wall disruption and mass transfer Low energy demand, mild conditions, compatibility with green solvents Scalability challenges, possible equipment erosion at high intensity Phenolics, flavonoids, carotenoids Pilot-scale Cao et al., 2025; Siddiqui et al., 2025; Gil et al., 2023 Supercritical Fluid Extraction (SFE–CO2) Supercritical CO₂ used as a tunable solvent under controlled pressure and temperature Solvent-free extracts, high selectivity, excellent for thermolabile compounds High capital cost, limited polarity range without co-solvents Essential oils, lipids, terpenoids Industrialscale Herzyk et al., 2024; Nastić et al., 2025; Prado et al., 2017 Subcritical/ Pressurized Hot Water Extraction (SWE/ PHWE) Polarity modulation of water under elevated temperature and pressure Non-toxic solvent, cost-effective, broad polarity range Elevated temperature may affect sensitive compounds Phenolic acids, flavonoids, sugars Pilot to industrial Bastos et al., 2025; Aminzai et al., 2025; Herrero, 2024
60 INNOVATIVE STUDIES IN AGRICULTURE, FORESTRY AND AQUACULTURE – 2025 The main green extraction techniques discussed in this section are comparatively summarized in Table 1 to facilitate a clearer evaluation of their technological characteristics and industrial applicability. Advanced green extraction technologies provide efficient and environmentally compatible pathways for recovering bioactive compounds from medicinal plants. Methods such as microwave assisted extraction, ultrasound assisted extraction, supercritical fluid extraction, and subcritical water extraction offer complementary advantages depending on compound properties and application goals. The integration of hybrid systems and data driven optimization strategies further enhances process efficiency and supports the translation of laboratory scale approaches into sustainable industrial applications. 4. Role of Ionic Liquids and Deep Eutectic Solvents in Sustainable Extraction The use of conventional organic solvents in the recovery of phytochemicals from medicinal plants is increasingly being abandoned due to concerns related to environmental pollution and potential risks to human health. In response to these challenges, ionic liquids and deep eutectic solvents have emerged as sustainable and environmentally friendly next generation alternatives, commonly described as designer solvents (Plastiras and Samanidou, 2022). These solvent systems offer unique advantages, including low vapor pressure, high thermal stability, and tunable solvation properties that can be adjusted according to the physicochemical characteristics of specific target compounds (Guazzelli et al., 2023). Ionic liquids are salts composed entirely of ions that remain in the liquid state at or near room temperature. This distinctive property enables their effective interaction with plant cell walls, facilitating cell disruption and the purification of structurally complex phytochemicals (Da Rocha Fernandes et al., 2025). In biotechnological applications, ionic liquids have demonstrated remarkable efficiency in selective separations. For example, studies focusing on the purification of folic acid have shown that specific ionic liquid combinations can achieve separation efficiencies as high as 99.56% (Blaga et al., 2023). In addition, specialized subclasses such as surface active ionic liquids exhibit significant potential in drug delivery applications by enhancing the solubility and oral bioavailability of poorly water soluble bioactive compounds (Sangiorgi et al., 2025). Deep eutectic solvents represent a further advancement in the sustainability hierarchy of green solvents. These systems are typically formed by combining
MEDICINAL PLANT PHYTOCHEMICALS: SUSTAINABLE EXTRACTION . . . 61 two or more components, such as hydrogen bond donors and acceptors, resulting in mixtures with melting points substantially lower than those of the individual constituents (Płotka Wasylka et al., 2020). Compared with ionic liquids, deep eutectic solvents are generally more cost effective, simpler to synthesize, and often exhibit improved biodegradability (Cao et al., 2025). In particular, natural deep eutectic solvents prepared from plant derived components such as sugars, amino acids, and organic acids have gained increasing attention due to their ability to preserve bioactivity while minimizing toxicity during extraction processes (Bastos et al., 2025). In practical extraction applications, the performance of natural deep eutectic solvent systems is frequently enhanced through integration with advanced green technologies such as ultrasound assisted or microwave assisted extraction. For instance, the use of natural deep eutectic solvents in the extraction of rosmarinic acid from Thunbergia laurifolia resulted in improved extraction efficiency while maintaining the stability of the extract against oxidative and photolytic stress for up to ninety days (Kriengsaksri et al., 2025). Similarly, the application of deep eutectic solvents in the isolation of phytochemicals from Mangifera indica fruit and its processing by products has been shown to improve selectivity and enable the production of high quality raw materials for food and pharmaceutical applications (Rahman et al., 2025). Comparable outcomes have also been reported for choline chloride based deep eutectic solvents used in the extraction of carotenoids and phenolic compounds from Chlorella vulgaris biomass, where bioactivity was better preserved than with conventional solvent systems (Dardavila et al., 2023). Despite these advantages, the widespread industrial implementation of ionic liquids and deep eutectic solvents remains limited by challenges related to solvent recovery, recycling efficiency, and overall economic feasibility (Bastos et al., 2025). To address these limitations, the development of responsive deep eutectic solvents that undergo reversible phase separation in response to external stimuli such as temperature or pH has gained increasing attention. These systems offer promising opportunities for improving solvent recyclability and reducing process costs (Vicente et al., 2025). Furthermore, the application of artificial intelligence and machine learning models to predict solvent behavior and optimize extraction conditions has initiated a new phase in the rational design and efficient utilization of green solvent based extraction systems (Tian et al., 2025). In summary, ionic liquids and deep eutectic solvents provide sustainable and tunable alternatives to conventional organic solvents for phytochemical
62 INNOVATIVE STUDIES IN AGRICULTURE, FORESTRY AND AQUACULTURE – 2025 extraction. While ionic liquids offer high efficiency and selectivity, deep eutectic solvents and particularly natural deep eutectic solvents combine improved biocompatibility with economic advantages. Continued advances in solvent design and process optimization are expected to support their broader adoption in environmentally responsible extraction technologies. 5. Optimization Strategies for Phytochemical Recovery The recovery of bioactive compounds from medicinal plants is a critical process due to the high application value of these compounds in the food, pharmaceutical, and cosmetic industries (Cao et al., 2025). The limitations of conventional extraction methods, including low extraction efficiency and high solvent consumption, have increased the need for standardized and optimized approaches capable of enhancing both the quality and yield of bioactive constituents (El Saadony et al., 2025). In this context, modern optimization strategies aim to maximize extraction efficiency while minimizing environmental impact and preserving the biological integrity of target compounds (Abu Reidah, 2025). Response surface methodology forms the foundation of many optimization strategies and is widely applied to identify optimal extraction conditions by analyzing interactions among independent variables such as solvent ratio, temperature, and extraction time (Gupta et al., 2024). For example, in a study on hemp seed oil extraction, the application of a Box Behnken experimental design enabled optimization of ultrasound duration and temperature, resulting in an oil yield of 31.22% (Esmaeilzadeh Kenari and Dehghan, 2020). Similarly, optimization studies conducted on phytochemical recovery from Senna fistula using Design Expert software demonstrated that specific stirring speeds and extraction durations were critical for achieving optimal yield (Faboro et al., 2023). In addition, the selection of solvent polarity and extraction technique, including Soxhlet extraction, maceration, or ultrasound assisted extraction, has been shown to play a decisive role in determining phenolic content and antioxidant capacity in medicinal plants such as Mentha longifolia (Tourabi et al., 2025). However, due to the complexity of biological matrices, conventional statistical models may be insufficient in capturing nonlinear relationships among extraction parameters. In such cases, advanced computational approaches such as artificial neural networks are increasingly employed (Subramani et al., 2025). Compared with response surface methodology, artificial neural network
MEDICINAL PLANT PHYTOCHEMICALS: SUSTAINABLE EXTRACTION . . . 63 models provide higher predictive accuracy and sensitivity, particularly when applied to nonlinear data sets (Gupta et al., 2024). In the recovery of bioactive compounds from agro industrial residues, the integration of artificial neural networks with genetic algorithms has been shown to enhance extraction yield while simultaneously optimizing energy consumption, thereby offering a more sustainable processing pathway (Anwar, 2025). Furthermore, the combined use of radial basis function neural networks and genetic algorithms enables multi objective optimization and facilitates a more comprehensive understanding of complex extraction systems (Ma et al., 2023). Another innovative direction in optimization involves the application of hybrid extraction systems in combination with machine learning algorithms. In the isolation of bioactive compounds from Boerhavia diffusa, the combined use of response surface methodology, adaptive neuro fuzzy inference systems, and machine learning models enabled a deeper analysis of how extraction parameters influence bioactivity (Kabilan et al., 2024). When integrated with hybrid techniques such as enzyme assisted microwave extraction, these approaches achieve high extraction efficiency while maintaining compound integrity (Anwar, 2025). Since the choice and optimization of extraction techniques directly influence the phytochemical profile of the resulting extracts and consequently their therapeutic efficacy, these optimization strategies are of critical importance (Sun et al., 2025). Future oriented strategies increasingly focus on integrating omics technologies and network pharmacology approaches to tailor phytochemical profiles toward specific antibacterial or anti inflammatory targets (Abdallah et al., 2023). In addition, green nanotechnology applications, such as the biosynthesis of nanoparticles using plant extracts, require precise optimization of parameters including pH and metal ion concentration to ensure product stability and functional performance (Edo et al., 2025). Overall, optimized extraction protocols represent one of the most important tools supporting the economic and sustainable commercialization of plant derived bioactive compounds at an industrial scale (Usman et al., 2023). Overall, optimization strategies play a key role in improving phytochemical recovery while ensuring sustainability and product quality. Through the combined application of statistical approaches, artificial intelligence based techniques, and hybrid extraction systems, complex process parameters can be effectively managed. As a result, these strategies facilitate the scalable and economically feasible production of high value bioactive compounds from medicinal plants.
64 INNOVATIVE STUDIES IN AGRICULTURE, FORESTRY AND AQUACULTURE – 2025 6. Analytical Techniques for Phytochemical Characterization Accurate identification and characterization of bioactive compounds are essential for understanding the therapeutic potential of medicinal plants and for meeting industrial quality standards (El Allaoui et al., 2024). This process involves a multistep analytical approach that enables both qualitative and quantitative evaluation of complex phytochemical profiles, including phenolic compounds, alkaloids, saponins, and terpenoids present in plant extracts (Abdallah et al., 2023). Advances in modern analytical technologies have made it possible to detect even trace level constituents within plant matrices with high sensitivity, thereby providing a foundation for innovation in the pharmaceutical sector (Dincheva et al., 2025). At the initial stage of characterization, qualitative phytochemical screening methods are commonly employed to determine the presence of major compound classes such as flavonoids, steroids, glycosides, and polyphenols using specific chemical reagents and solvent systems (Rao et al., 2023). Thin layer chromatography remains a fundamental tool for the rapid, cost effective, and visual separation of complex mixtures, particularly through two dimensional applications. In addition to compound separation, thin layer chromatography offers important advantages in establishing chromatographic fingerprints and in determining the geographical origin of samples such as propolis (Cvetković et al., 2025). For advanced quantitative analysis and compound separation, high performance liquid chromatography and ultra performance liquid chromatography are among the most widely applied techniques (Sik et al., 2022). High performance liquid chromatography coupled with diode array detection provides high resolution data for mapping phenolic profiles in plant extracts and for correlating antioxidant capacity with chemical composition (Tourabi et al., 2025). These techniques are particularly critical for understanding the complex interactions between polyphenols and other food matrix components, including carbohydrates and proteins (Sik et al., 2022). In the analysis of volatile compounds and lipid based phytochemicals, gas chromatography coupled with mass spectrometry plays a prominent role. This technique enables precise identification of chemical constituents and structural elucidation through the use of mass spectral libraries. Gas chromatography mass spectrometry analyses form the basis for characterizing essential oils derived from plant by products and for predicting the biological activities of these compounds using in silico modeling approaches (Punetha and Vuppu, 2023).
MEDICINAL PLANT PHYTOCHEMICALS: SUSTAINABLE EXTRACTION . . . 71 production. Exhaustive, the convergence of green chemistry, technological innovation, and optimization frameworks provides a robust foundation for the scalable and responsible industrial exploitation of plant based bioactive compounds. References Abdallah, E. M., Alhatlani, B. Y., De Paula Menezes, R., & Martins, C. H. G. (2023). Back to Nature: Medicinal Plants as Promising Sources for Antibacterial Drugs in the Post-Antibiotic Era. Plants, 12(17), 3077. https://doi. org/10.3390/plants12173077 Abu-Reidah, I. M. (2025). Special Issue on “Phytochemicals: Extraction, Optimization, Identification, Biological Activities, and Applications in the Food, Nutraceutical, and Pharmaceutical Industries.” Processes, 13(5), 1390. https:// doi.org/10.3390/pr13051390 Ahmad, M., Tahir, M., Hong, Z., Zia, M. A., Rafeeq, H., Ahmad, M. S., Rehman, S. U., & Sun, J. (2025). Plant and marine-derived natural products: Sustainable pathways for future drug discovery and therapeutic development. Frontiers in Pharmacology, 15, 1497668. https://doi.org/10.3389/ fphar.2024.1497668 Ali, S. S., Al-Tohamy, R., Al-Zahrani, M., Badr, A., & Sun, J. (2025). Essential oils and plant-derived bioactive compounds: A comprehensive review of their therapeutic potential, mechanisms of action, and advances in extraction technologies. Phytochemistry Reviews. https://doi.org/10.1007/s11101-02510123-8 Aminzai, M. T., Yabalak, E., Akay, S., & Kayan, B. (2025). Recent developments in subcritical water extraction of industrially important bioactive substances from plants, microorganisms, and organic wastes. Biomass Conversion and Biorefinery, 15(12), 17927–17949. https://doi.org/10.1007/ s13399-024-06392-6 Angioletti Decker, B. L., Fonteles, T. V., Fernandes, F. A. N., & Rodrigues, S. (2026). Green extraction technologies for valorising Brazilian agri-food waste. Sustainable Food Technology, 10.1039.D5FB00323G. https://doi.org/10.1039/ D5FB00323G Ansari, P., Reberio, A. D., Ansari, N. J., Kumar, S., Khan, J. T., Chowdhury, S., Abd El-Mordy, F. M., Hannan, J. M. A., Flatt, P. R., Abdel-Wahab, Y. H. A., & Seidel, V. (2025). Therapeutic Potential of Medicinal Plants and Their Phytoconstituents in Diabetes, Cancer, Infections, Cardiovascular Diseases,
72 INNOVATIVE STUDIES IN AGRICULTURE, FORESTRY AND AQUACULTURE – 2025 Inflammation and Gastrointestinal Disorders. Biomedicines, 13(2), 454. https:// doi.org/10.3390/biomedicines13020454 Anwar, M. M. J. (2025). Advances in Green Technologies for Bioactive Extraction and Valorization of Agro-Waste in Food and Nutraceutical Industries. Haya: The Saudi Journal of Life Sciences, 10(05), 184–195. https://doi. org/10.36348/sjls.2025.v10i05.005 Ashraf, S. A., Elkhalifa, A. E. O., Siddiqui, A. J., Haque, A., & Mahmood, D. (2023). Phytochemicals as Immunomodulators, Nutraceuticals, and Pharma Foods. In M. Adnan, M. Patel, & M. Snoussi, Ethnobotany and Ethnopharmacology of Medicinal and Aromatic Plants (1st ed., pp. 143–163). CRC Press. https://doi.org/10.1201/b22842-9 Banerjee, S., Haldar, S., Bhattacharya, M., Kadeppagari, R.-K., Patil, A. G., Pradeep, B., & G., I. (2026). Phytochemical characterization: Analytical tools and techniques. In Phytoceuticals in Food for Health and Wellness (pp. 55–68). Elsevier. https://doi.org/10.1016/B978-0-443-26494-8.00011-2 Bastos, K. V. L. D. S., De Souza, A. B., Tomé, A. C., & Souza, F. D. M. (2025). New Strategies for the Extraction of Antioxidants from Fruits and Their By-Products: A Systematic Review. Plants, 14(5), 755. https://doi.org/10.3390/ plants14050755 Bernykov, V., Bilousko, T., Bilousko, R., Pitel, N., & Starovoyit, V. (2025). Green Technologies and Their Contribution to a Sustainable Global Economy in 2020-2025: A Literature Review. Futurity Economics&Law, 5(1), 161–178. https://doi.org/10.57125/FEL.2025.03.25.09 Blaga, A. C., Dragoi, E. N., Tucaliuc, A., Kloetzer, L., & Cascaval, D. (2023). Folic Acid Ionic-Liquids-Based Separation: Extraction and Modelling. Molecules, 28(8), 3339. https://doi.org/10.3390/molecules28083339 Bodea, I. M., Garre Pérez, A., Cătunescu, G. M., & Palop, A. (2025). A Review on Microwave and Ultrasound-Assisted Extractions of Essential Oil from Orange Peel Waste. Food and Bioprocess Technology, 18(8), 7060–7082. https://doi.org/10.1007/s11947-025-03882-x Cao, S., Liang, J., Chen, M., Xu, C., Wang, X., Qiu, L., Zhao, X., & Hu, W. (2025). Comparative analysis of extraction technologies for plant extracts and absolutes. Frontiers in Chemistry, 13, 1536590. https://doi.org/10.3389/ fchem.2025.1536590 Chaachouay, N., & Zidane, L. (2024). Plant-Derived Natural Products: A Source for Drug Discovery and Development. Drugs and Drug Candidates, 3(1), 184–207. https://doi.org/10.3390/ddc3010011
MEDICINAL PLANT PHYTOCHEMICALS: SUSTAINABLE EXTRACTION . . . 73 Chaves, J. O., De Souza, M. C., Da Silva, L. C., Lachos-Perez, D., TorresMayanga, P. C., Machado, A. P. D. F., Forster-Carneiro, T., Vázquez-Espinosa, M., González-de-Peredo, A. V., Barbero, G. F., & Rostagno, M. A. (2020). Extraction of Flavonoids From Natural Sources Using Modern Techniques. Frontiers in Chemistry, 8, 507887. https://doi.org/10.3389/fchem.2020.507887 Coyle, D. (2024). Everything Everywhere All At Once: Competition policy and industrial policy choices in an era of structural change. Oxford Review of Economic Policy, 40(4), 718–728. https://doi.org/10.1093/oxrep/grae040 Cvetković, D., Somogyi Škoc, M., Meštrović, E., & Meštrović, I. R. (2025). Rapid Chromatographic and Spectroscopic Analysis of Extracted Raw Propolis. Molecules, 30(24), 4729. https://doi.org/10.3390/molecules30244729 Da Rocha Fernandes, E., Barros, J. H. T., Guarda, P. M., & Guarda, E. A. (2025). The Use of Ionic Liquids and Deep Eutectic Solvents in the Extraction of Phytochemicals with Bioactive Properties: A Review. Food Biophysics, 20(4), 158. https://doi.org/10.1007/s11483-025-10037-0 Dardavila, M. M., Pappou, S., Savvidou, M. G., Louli, V., Katapodis, P., Stamatis, H., Magoulas, K., & Voutsas, E. (2023). Extraction of Bioactive Compounds from C. vulgaris Biomass Using Deep Eutectic Solvents. Molecules, 28(1), 415. https://doi.org/10.3390/molecules28010415 Dari, D. N., da Silva Vieira, R., Lima Júnior, A. M. B., Salamba, M. Q., Da Silva, J. L., Araújo, S. F. A., de Matos Filho, J. R., Da Silva, L. F., Melo, R. L. F., & dos Santos, J. C. S. (2025). Economic Challenges and Future Perspectives for Industrial Scale Biocrude Production. In M. Aslam, S. Mishra, J. A. Aburto Anell, & A. K. Sarma (Eds.), Biocrude Oil Biorefinery: An Emerging Biorefining Approach (pp. 531–556). Springer Nature Switzerland. https://doi. org/10.1007/978-3-031-85036-3_20 Darwin, R., Valmon, R., Chithanna, S., Galla, S. H., Syed, S. H., Mohathasim Billah, A. A., Kumar Reddy, K. T., & Arjun, U. V. N. V. (2025). Sustainable Extraction and Purification of Phytochemicals: A Review of Green Solvents and Techniques. Chemical Methodologies, Online First. https://doi. org/10.48309/chemm.2025.504050.1892 Dasiewicz, J., Wronka, A., & Kowaluk, G. (2025). From Extraction to Valorization: Unlocking the Potential of Bark-Derived Extraction Residues for Sustainable Material Development. Molecules, 30(23), 4537. https://doi. org/10.3390/molecules30234537 Dincheva, I., Badjakov, I., & Galunska, B. (2025). New Insights in the Research on Bioactive Compounds from Plant Origins with Nutraceutical
74 INNOVATIVE STUDIES IN AGRICULTURE, FORESTRY AND AQUACULTURE – 2025 and Pharmaceutical Potential II. Plants, 14(4), 500. https://doi.org/10.3390/ plants14040500 Edo, G. I., Mafe, A. N., Ali, A. B. M., Akpoghelie, P. O., Yousif, E., Isoje, E. F., Igbuku, U. A., Ismael, S. A., Essaghah, A. E. A., Ahmed, D. S., Ozsahin, D. U., Umar, H., & Alamiery, A. A. (2025). Green Biosynthesis of Nanoparticles Using Plant Extracts: Mechanisms, Advances, Challenges, and Applications. BioNanoScience, 15(2), 267. https://doi.org/10.1007/s12668-025-01883-w El Allaoui, H., El Ahmadi, K., El Abdouni, A., Dira, I., El Bastrioui, M., Bouhrim, M., Eto, B., Shahat, A., Herqash, R., & Haboubi, K. (2024). Trends and Insights in Medicinal Plant Extract Research: A Ten-Year Bibliometric and Visualization Study. Horticulturae, 10(11), 1163. https://doi.org/10.3390/ horticulturae10111163 El-Saadony, M. T., Saad, A. M., Mohammed, D. M., Alkafaas, S. S., Abd El-Mageed, T. A., Fahmy, M. A., Ezzat Ahmed, A., Algopishi, U. B., Abu-Elsaoud, A. M., Mosa, W. F. A., AbuQamar, S. F., & El-Tarabily, K. A. (2025). Plant bioactive compounds: Extraction, biological activities, immunological, nutritional aspects, food application, and human health benefits—A comprehensive review. Frontiers in Nutrition, 12, 1659743. https:// doi.org/10.3389/fnut.2025.1659743 Esmaeilzadeh Kenari, R., & Dehghan, B. (2020). Optimization of ultrasound‐assisted solvent extraction of hemp ( Cannabis sativa L.) seed oil using RSM: Evaluation of oxidative stability and physicochemical properties of oil. Food Science & Nutrition, 8(9), 4976–4986. https://doi.org/10.1002/ fsn3.1796 Faboro, E. O., Adekunle, D. O., Obisesan, I. A., & Oyinlola, T. A. (2023). Optimization of extraction conditions for phytochemicals from Senna fistula using cheminformatics. SN Applied Sciences, 5(8), 209. https://doi.org/10.1007/ s42452-023-05421-9 Gil, K. A., Jokić, S., Cikoš, A.-M., Banožić, M., Jakovljević Kovač, M., Fais, A., & Tuberoso, C. I. G. (2023). Comparison of Different Green Extraction Techniques Used for the Extraction of Targeted Flavonoids from Edible Feijoa (Acca sellowiana (O.Berg) Burret) Flowers. Plants, 12(7), 1461. https://doi. org/10.3390/plants12071461 Guazzelli, L., Marrucho, I. M., Mu, T., & Banerjee, T. (2023). Editorial: Ionic liquids and deep eutectic solvents: Two contrasting options or opposite sides of the same coin? Frontiers in Chemistry, 11, 1169688. https://doi. org/10.3389/fchem.2023.1169688
MEDICINAL PLANT PHYTOCHEMICALS: SUSTAINABLE EXTRACTION . . . 75 Gupta, R., Singh, A., Nema, P. K., Roy, T., Kumar, S., & Kumar, A. P. (2024). Implementation of RSM and ANN Optimization Approach for Natural Deep Eutectic Solvents-Based Extraction of Bioactive Compounds from Orange Peel. ACS Omega, 9(32), 34880–34892. https://doi.org/10.1021/acsomega.4c04468 Hasnain, A., Naqvi, S. A. H., Ayesha, S. I., Khalid, F., Ellahi, M., Iqbal, S., Hassan, M. Z., Abbas, A., Adamski, R., Markowska, D., Baazeem, A., Mustafa, G., Moustafa, M., Hasan, M. E., & Abdelhamid, M. M. A. (2022). Plants in vitro propagation with its applications in food, pharmaceuticals and cosmetic industries; current scenario and future approaches. Frontiers in Plant Science, 13, 1009395. https://doi.org/10.3389/fpls.2022.1009395 Herrero, M. (2024). Towards green extraction of bioactive natural compounds. Analytical and Bioanalytical Chemistry, 416(9), 2039–2047. https://doi.org/10.1007/s00216-023-04969-0 Herzyk, F., Piłakowska-Pietras, D., & Korzeniowska, M. (2024). Supercritical Extraction Techniques for Obtaining Biologically Active Substances from a Variety of Plant Byproducts. Foods, 13(11), 1713. https://doi. org/10.3390/foods13111713 Kabilan, S. J., Sivakumar, O., Sumanth, G. B., Kannan, S., Kunjiappan, S., & Sundar, K. (2024). Optimization and analysis of ultrasound-assisted solvent extraction of bioactive compounds from Boerhavia diffusa Linn. Using RSM, ANFIS and machine learning algorithm. Journal of Food Measurement and Characterization, 18(6), 4204–4220. https://doi.org/10.1007/s11694-02402487-w Kawatra, A., Gupta, S., Dhankhar, R., Singh, P., & Gulati, P. (2022). Application of Phytochemicals in Therapeutic, Food, Flavor, and Cosmetic Industries. In M. K. Swamy & A. Kumar (Eds.), Phytochemical Genomics (pp. 85–108). Springer Nature Singapore. https://doi.org/10.1007/978-981-19-57796_4 Kriengsaksri, K., Thongphichai, W., Uttarawichien, T., Khoochonthara, J., Towiwat, P., & Sukrong, S. (2025). Optimization of Natural Deep Eutectic Solvent-Assisted Extraction of Rosmarinic Acid from Thunbergia laurifolia Lindl. And Evaluation of Antioxidant Activity. Molecules, 30(24), 4795. https:// doi.org/10.3390/molecules30244795 Kumar, M. (2024). ADVANCEMENTS IN ANALYTICAL TECHNIQUES FOR THE IDENTIFICATION OF PHYTOCONSTITUENTS IN HERBAL MEDICINES. International Journal of Pharmaceutical Science and Medicine, 2(4), 94–101. https://doi.org/10.70199/IJPSM.2.3.94-101
76 INNOVATIVE STUDIES IN AGRICULTURE, FORESTRY AND AQUACULTURE – 2025 Lackner, M., & Besharati, M. (2025). Agricultural Waste: Challenges and Solutions, a Review. Waste, 3(2), 18. https://doi.org/10.3390/waste3020018 Lamponi, S., Barletta, R., & Santucci, A. (2025). Agricultural and AgroIndustrial Residues as Sustainable Sources of Next-Generation Biomedical Materials: Advances, Challenges, and Perspectives. Life, 15(12), 1908. https:// doi.org/10.3390/life15121908 Liu, X., & Meng, H. (2021). Consideration for the scale‐up manufacture of nanotherapeutics—A critical step for technology transfer. VIEW, 2(5), 20200190. https://doi.org/10.1002/VIW.20200190 Ma, J., Yao, J., Ren, X., Dong, Y., Song, R., Zhong, X., Zheng, Y., Shan, D., Lv, F., Li, X., Deng, Q., He, Y., Yuan, R., & She, G. (2023). Machine learning-assisted data-driven optimization and understanding of the multiple stage process for extraction of polysaccharides and secondary metabolites from natural products. Green Chemistry, 25(8), 3057–3068. https://doi.org/10.1039/ D2GC04574E Maranghi, S., Parisi, M. L., Basosi, R., & Sinicropi, A. (2020). LCA as a Support Tool for the Evaluation of Industrial Scale-Up. In S. Maranghi & C. Brondi (Eds.), Life Cycle Assessment in the Chemical Product Chain (pp. 125–143). Springer International Publishing. https://doi.org/10.1007/978-3030-34424-5_6 Martins, R., Barbosa, A., Advinha, B., Sales, H., Pontes, R., & Nunes, J. (2023). Green Extraction Techniques of Bioactive Compounds: A State-of-theArt Review. Processes, 11(8), 2255. https://doi.org/10.3390/pr11082255 Masyita, A., Hardinasinta, G., Astuti, A. D., Firdayani, F., Mayasari, D., Hori, A., Nisha, I. N. A., Nainu, F., & Kuraishi, T. (2025). Natural pigments: Innovative extraction technologies and their potential application in health and food industries. Frontiers in Pharmacology, 15, 1507108. https://doi. org/10.3389/fphar.2024.1507108 Mirzazadeh, N., Bagheri, H., Mirzazadeh, M., Soleimanimehr, S., Rasi, F., & Akhavan‐Mahdavi, S. (2024). Comparison of different green extraction methods used for the extraction of anthocyanin from red onion skin. Food Science & Nutrition, 12(10), 7347–7357. https://doi.org/10.1002/ fsn3.4354 Mykhailenko, O., Jalil, B., McGaw, L. J., Echeverría, J., Takubessi, M., & Heinrich, M. (2025). Climate change and the sustainable use of medicinal plants: A call for “new” research strategies. Frontiers in Pharmacology, 15, 1496792. https://doi.org/10.3389/fphar.2024.1496792
MEDICINAL PLANT PHYTOCHEMICALS: SUSTAINABLE EXTRACTION . . . 77 Nastić, N., Mazumder, J. A., & Banat, F. (2025). Supercritical CO2 extraction of oil from fruit seed by-product: Advances, challenges, and pathways to commercial viability. Critical Reviews in Food Science and Nutrition, 1–18. https://doi.org/10.1080/10408398.2025.2527946 Olaniyan, M. F., Olaniyi, O. D., Odegbemi, F., Olaniyan, T. B., & Odegbemi, O. B. (2025). Isolation and purification techniques for bioactive compounds from Nigerian medicinal plants and their therapeutic applications. Discover Chemistry, 2(1), 13. https://doi.org/10.1007/s44371-025-00098-y Patel, P., Pang, Y. L. J., Choi, W. J., & Wong, A. (2025). Protein Extraction and Isolation from Legumes and Algae: An Industry Primer. Food and Bioprocess Technology, 18(10), 8380–8408. https://doi.org/10.1007/s11947-025-03958-8 Plastiras, O.-E., & Samanidou, V. (2022). Applications of Deep Eutectic Solvents in Sample Preparation and Extraction of Organic Molecules. Molecules, 27(22), 7699. https://doi.org/10.3390/molecules27227699 Płotka-Wasylka, J., De La Guardia, M., Andruch, V., & Vilková, M. (2020). Deep eutectic solvents vs ionic liquids: Similarities and differences. Microchemical Journal, 159, 105539. https://doi.org/10.1016/j. microc.2020.105539 Prado, J. M., Veggi, P. C., & Meireles, M. A. A. (2017). Scale-Up Issues and Cost of Manufacturing Bioactive Compounds by Supercritical Fluid Extraction and Ultrasound Assisted Extraction. In G. V. Barbosa-Cánovas, G. María Pastore, K. Candoğan, I. G. Medina Meza, S. Caetano Da Silva Lannes, K. Buckle, R. Y. Yada, & A. Rosenthal (Eds.), Global Food Security and Wellness (pp. 377–433). Springer New York. https://doi.org/10.1007/978-1-4939-6496-3_20 Punetha, S., & Vuppu, S. (2023). GC–MS, FTIR and physico-chemical analysis of phytochemicals from Vellore floral waste and its in-silico studies. Chemical Papers, 77(11), 6873–6905. https://doi.org/10.1007/s11696-02302984-0 Rahman, A. M. A., Bakar, A. R. A., Yee, A. Q., Zainudin, M. A. M., Daud, N. M. A. N., Gunny, A. A. N., Sarip, M. S. M., Peron, R. V., & Khairuddin, N. H. (2025). A review on the role of deep eutectic solvents in mango ( Mangifera indica ) extraction. RSC Advances, 15(6), 4296–4321. https://doi.org/10.1039/ D5RA00097A Rao, A., Kumari, S., Laura, J. S., & Dhania, G. (2023). Qualitative Phytochemical Screening of Medicinal Plants Using Different Solvent Extracts. Oriental Journal Of Chemistry, 39(3), 621–626. https://doi.org/10.13005/ ojc/390312
78 INNOVATIVE STUDIES IN AGRICULTURE, FORESTRY AND AQUACULTURE – 2025 Râpă, M., Darie-Niță, R. N., & Coman, G. (2024). Valorization of Fruit and Vegetable Waste into Sustainable and Value-Added Materials. Waste, 2(3), 258–278. https://doi.org/10.3390/waste2030015 Rodríguez-Negrete, E. V., Morales-González, Á., Madrigal-Santillán, E. O., Sánchez-Reyes, K., Álvarez-González, I., Madrigal-Bujaidar, E., ValadezVega, C., Chamorro-Cevallos, G., Garcia-Melo, L. F., & Morales-González, J. A. (2024). Phytochemicals and Their Usefulness in the Maintenance of Health. Plants, 13(4), 523. https://doi.org/10.3390/plants13040523 Sangiorgi, S., Albertini, B., Bertoni, S., & Passerini, N. (2025). An Overview on the Role of Ionic Liquids and Deep Eutectic Solvents in Oral Pharmaceuticals. Pharmaceutics, 17(3), 300. https://doi.org/10.3390/pharmaceutics17030300 Shahzadi, S., Fatima, S., Ul Ain, Q., Shafiq, Z., & Janjua, M. R. S. A. (2025). A review on green synthesis of silver nanoparticles (SNPs) using plant extracts: A multifaceted approach in photocatalysis, environmental remediation, and biomedicine. RSC Advances, 15(5), 3858–3903. https://doi.org/10.1039/ D4RA07519F Siddiqui, S. A., Ali Redha, A., Salauddin, M., Harahap, I. A., & Rupasinghe, H. P. V. (2025). Factors Affecting the Extraction of (Poly)Phenols from Natural Resources Using Deep Eutectic Solvents Combined with Ultrasound-Assisted Extraction. Critical Reviews in Analytical Chemistry, 55(1), 139–160. https:// doi.org/10.1080/10408347.2023.2266846 Sik, B., Székelyhidi, R., Lakatos, E., Kapcsándi, V., & Ajtony, Z. (2022). Analytical procedures for determination of phenolics active herbal ingredients in fortified functional foods: An overview. European Food Research and Technology, 248(2), 329–344. https://doi.org/10.1007/s00217-021-03908-6 Singh, J., Rasane, P., Kaur, R., Kaur, H., Garg, R., Kaur, S., Ercisli, S., Choudhary, R., Skrovankova, S., & Mlcek, J. (2023). Valorization of grape (Vitis vinifera) leaves for bioactive compounds: Novel green extraction technologies and food-pharma applications. Frontiers in Chemistry, 11, 1290619. https://doi. org/10.3389/fchem.2023.1290619 Skov, K. B., Portillo-Perez, G. A., & Martinez, M. M. (2025). Deep eutectic solvent-assisted starch acetylation within stale bread particles to improve water resistance. International Journal of Biological Macromolecules, 288, 138603. https://doi.org/10.1016/j.ijbiomac.2024.138603 Subramani, V., Tomer, V., Balamurali, G., & Mansingh, P. (2025). Artificial neural network in optimization of bioactive compound extraction: Recent trends and performance comparison with response surface methodology. Analytical Sciences, 41(2), 101–117. https://doi.org/10.1007/s44211-024-00681-w
MEDICINAL PLANT PHYTOCHEMICALS: SUSTAINABLE EXTRACTION . . . 79 Sun, S., Yu, Y., Jo, Y., Han, J. H., Xue, Y., Cho, M., Bae, S.-J., Ryu, D., Park, W., Ha, K.-T., & Zhuang, S. (2025). Impact of extraction techniques on phytochemical composition and bioactivity of natural product mixtures. Frontiers in Pharmacology, 16, 1615338. https://doi.org/10.3389/fphar.2025.1615338 Tanruean, K., Luangkamin, S., Srisurat, T., Bunmusik, W., & Suttiarporn, P. (2025). Optimization of Microwave-Assisted Extraction Process for Production of Polyphenol-Rich Crude Extract from Cinnamomum iners Leaves. Applied Sciences, 15(3), 1265. https://doi.org/10.3390/app15031265 Teixeira, N. (2025). Circular economy perspectives: Challenges, innovations, and sustainable futures. Discover Sustainability, 6(1), 738. https:// doi.org/10.1007/s43621-025-01606-x Tian, Y., Zhang, H., Qiao, Y., Yang, H., Liu, Y., & Ji, X. (2025). Intelligent prediction of ionic liquids and deep eutectic solvents by machine learning. Chinese Journal of Chemical Engineering, 84, 227–243. https://doi. org/10.1016/j.cjche.2025.06.006 Tourabi, M., Faiz, K., Ezzouggari, R., Louasté, B., Merzouki, M., Dauelbait, M., Bourhia, M., Almaary, K. S., Siddique, F., Lyoussi, B., & Derwich, E. (2025). Optimization of extraction process and solvent polarities to enhance the recovery of phytochemical compounds, nutritional content, and biofunctional properties of Mentha longifolia L. extracts. Bioresources and Bioprocessing, 12(1), 24. https://doi.org/10.1186/s40643-025-00859-8 Usman, M., Nakagawa, M., & Cheng, S. (2023). Emerging Trends in Green Extraction Techniques for Bioactive Natural Products. Processes, 11(12), 3444. https://doi.org/10.3390/pr11123444 Uti, D. E., Atangwho, I. J., Alum, E. U., Egba, S. I., Ugwu, O. P.-C., & Ikechukwu, G. C. (2025). Natural Antidiabetic Agents: Current Evidence and Development Pathways from Medicinal Plants to Clinical use. Natural Product Communications, 20(3), 1934578X251323393. https://doi. org/10.1177/1934578X251323393 Veera, S., Chirumamilla, P., Dharavath, S. B., & Taduri, S. (2025). Detection of chemical compounds by GC–MS and evaluation of antioxidant, anti-inflammatory, antibacterial and anti-cancer potential in methanolic extracts of Corallocarpus epigaeus (Rottler) Hook. F. Vegetos. https://doi.org/10.1007/ s42535-025-01564-9 Vicente, F. A., Tkalec, N., & Likozar, B. (2025). Responsive deep eutectic solvents: Mechanisms, applications and their role in sustainable chemistry. Chemical Communications, 61(6), 1002–1013. https://doi.org/10.1039/ D4CC05157B
80 INNOVATIVE STUDIES IN AGRICULTURE, FORESTRY AND AQUACULTURE – 2025 Wang, C., Wei, X., Zhong, L., Chan, C.-L., Li, H., & Sun, H. (2025). MetalBased Approaches for the Fight against Antimicrobial Resistance: Mechanisms, Opportunities, and Challenges. Journal of the American Chemical Society, 147(15), 12361–12380. https://doi.org/10.1021/jacs.4c16035 Yadav, S., Malik, K., Moore, J. M., Kamboj, B. R., Malik, S., Malik, V. K., Arya, S., Singh, K., Mahanta, S., & Bishnoi, D. K. (2024). Valorisation of Agri-Food Waste for Bioactive Compounds: Recent Trends and Future Sustainable Challenges. Molecules, 29(9), 2055. https://doi.org/10.3390/ molecules29092055 Zhang, Z., & Zhou, Y. (2025). Economic growth vs environmental sustainability: Examining resource extraction in the World’s largest economies. Frontiers in Environmental Science, 13, 1586619. https://doi.org/10.3389/ fenvs.2025.1586619 Zhao, B.-N., Xie, Z.-Y., Liu, J.-N., Chen, X.-R., Wang, X.-X., Li, J.-Y., Zhang, R., & Si, C. (2025). Concentration-dependent effects of fermented spent coffee grounds and contrasting effects of earthworms on growth and phytochemicals in medicinal plant Glechoma longituba. PLOS One, 20(12), e0339185. https://doi.org/10.1371/journal.pone.0339185
THE IMPORTANCE OF SILAGE CORN (ZEA MAYS L.) PLANT . . . 87 album. Three types of management measures stand out: cultural practices (rotation of crops, suitable tillage, and compact sowing) (Liebman and Davis, 2000), mechanical (hoeing, cover crop) measures, and chemical control. While there is widespread utilization of herbicides laced with atrazine, mesotrione, and nicosulfuron, there is an increase in atrazine resistances that calls for integral strategies (Knezevic et al., 2009; Heap, 2024). The disease pathogens include fungal, bacterial, and viral pathogens, while mycotoxin producers in this group include Fusarium spp. as a significant risk (Munkvold and Desjardins, 1997). The key disease pathogens include root and stalk rot (Fusarium graminearum, F. verticillioides), northern leaf blight (Exserohilum turcicum), corn gall (Ustilago maydis), and Maize dwarf mosaic virus. In this regard, weed and disease control in silage corn should not be dependent simply upon chemicals alone; IPM measures must be adopted so as to secure yield, quality, sound soils, as well as sustainability of production over time. In this regard, technical empowerment of producers as well as repeated field-level surveillance are paramount. 3. Influencing Factors of Silage Corn’s Yield and Quality 3.1.SilagePreparationandHarvesting Al Silage corn production targets high-energy, stable, and digestible forage. This goal is grounded in proper management of harvesting time, harvesting approach, and silage-making (Extension PSU, 2023). Optimal harvesting time is when the plant is at 30–35% dry matter and ½-¾ milk-yielding line; early harvesting will lead to moisture loss, while late harvesting will lead to low digestibility (AgReliant Genetics, 2022; EgeFarm, 2022; Penn State Extension, 2023). Digestibility improves by 8–15 mm chopping length, 15–20 cm cutting, and proper adjustments of crushing grain systems (Åkesson et al., 2022; Çiftliğim Agromarket, 2019). Quality silage results in proper maintenance of anaerbiosis, proper compression, and fermentation of at least 3-6 weeks (AHDB, n.d.; Penn State Extension, 2023). Milk-stage and dough-stage harvesting maintains forage quality as well as performance by animals, as it incorporates physiological maturity signs (Masseeds, 2023; da Silva et al., 2025; Karnatam et al., 2023; Bayer, 2024). 3.2.ShapingandChoppingPhase All plant parts at harvesting are chopped by forage harvesters to 4-20 mm in diameter (Lactanet, 2018; USU Extension, 2021). The chopping length is
88 INNOVATIVE STUDIES IN AGRICULTURE, FORESTRY AND AQUACULTURE – 2025 critical in compressibility and efficiency in fermentation of materials. Short chopping creates gas, while long chopping creates spoilage by exposing it to oxygen (MSU Extension, 2018; USU Extension, 2021). The research shows that in the 10-20 mm category, there is regulated fermentation that also involves taking care of milk yield while sustaining the health of animals by achieving proper fiber in the rumen (USU Extension, 2021; Aquilian et al., 2020). 3.3.SilageProductionProcessandFermentation It should be promptly put into silos or plastic bags, and in layers, it should be compacted without oxygen (Penn State Extension, 2023). In anaerobic conditions, carbohydrate-fermenting lactic acid bacteria decrease pH by 4.0– 4.2; this action is done in 3–6 weeks, and the silage stabilizes (Extension PSU, 2023; AgReliant Genetics, 2022). Silage will keep under proper conditions for 8–12 months, yet it will develop mold, mycotoxin, as well as nutrient loss, if it is not used after opening promptly. Proper harvest time, proper chop length, effective compaction, and sanitary anaerobic conditions are a must for good silage (Extension PSU, 2023). 3.4.StorageConditions Conditions in which silage is kept are key to maintaining its quality and nutritious contents. The timely displacement of oxygen and the creation of an anaerobic environment are paramount in reducing the growth of unwanted microorganisms, such as mold and yeast (Kurt and Tan, 2019). While silo types (pit, reinforced concrete, horizontal, tower) vary with operating conditions, principles of close compaction and even filling are paramount in all types of silos (Sezen et al., 2018). The use of premium plastic cover is imperative in blocking air entry, and in controlling temperature and humidity fluctuations by suitable drainage and waterproofing (Aydın and Geren, 2021). Immediate response is prudent in countering anticipated leaks or infestations by mold, made easy by frequent checks. Optimal storage conditions optimize lactic acid bacteria activity, stabilize fermentation activity, and ensure long-term feed conservation (Soundharrajan et al., 2025). In turn, silage quality, animal health, and operating efficiency are maintained (Wilkinson and Chamberlain, 2016; Guo et al., 2022). 4. Silage Corn Through Sustainable Agricultural Lenses Silage corn (Zea mays L.) is a critical forage commodity under sustainable farming systems because of high biomass yield, high nutrient content, and wide
THE IMPORTANCE OF SILAGE CORN (ZEA MAYS L.) PLANT . . . 89 adaptability. The crop plays a great role in tackling feed deficiencies in response to climate change, due to its effective use of water and nitrogen associated with its C4 photosynthetic activity (Hatfield and Dold, 2019). Cultural practices such as rotation, reduced tillage, and precision fertilizer practices that focus on building soils’ health reduce environmental impacts while enhancing economic sustainability (Liu et al., 2020; Aydın and Geren, 2021). Efficient harvesting, silage handling practices also enhance feed quality as well as resource use efficiency, hence enhancing sustainability in crop production (Wilkinson and Rinne, 2018). 5. Strategic Value and Regions of Application in Animals 5.1. Its Function in Ruminant Nutrition and its Role in Dietary Formulations Tricalcium phosphate Silage corn possesses significant strategic value in ruminant nutrition due to its high concentration of energy as well as balanced nutrient composition. When subjected to fermentation and stored in appropriate storage conditions, its incorporation into feeding programs triggers higher weight gain and milk production. The presence of high starch trait increases starch digestion by ruminal microorganisms, thereby improving feed efficiency (Kurt and Tan, 2019). The former is corroborated by increased milk yield and component parts in dairy cattle, as well as higher daily weight gain and carcass production in beef cattle (Geren and Aydın, 2020). Furthermore, good-quality silages with balanced NDF and ADF compositions increase rumination and maintain pH stability by controlling rumen activity (Kılıç et al., 2018). Consequently, feed utilization efficiency is improved while risks of metabolic disorder are minimized. In summary, silage corn improves not only production outcomes in ruminant nutrition but also enhances aspects of animal welfare, cost efficiency, and sustainability. 5.2.ImpactuponAnimalProductQualityandAnimalProductivity Silage corn, with its high energy and digestibility properties, is a staple roughage in ruminant nutrition, directly affecting animal product quality and productivity. Silage obtained under appropriate harvesting, slaughtering, and fermentation conditions improves milk yield and milk components, while increasing live weight gain and carcass yield in meat production (Dellait, 2020;
90 INNOVATIVE STUDIES IN AGRICULTURE, FORESTRY AND AQUACULTURE – 2025 Åkesson et al., 2022). Grain crushing and harvesting at optimal dry matter levels enhance feed utilization by increasing starch digestibility (Aquilian et al., 2020). Indeed, the use of high-quality silage is reported to increase feed conversion efficiency by up to 10% and reduce production costs (Wilkinson & Rinne, 2018). Therefore, scientifically based silage management is strategically important for both crop yield and the sustainability of animal production. 6. Conclusions and Recommendations Silage corn (Zea mays L.) is a priority crop to supply sustainable forage requirements of contemporary livestock farming by virtue of its high biomass potential, nutrient concentration, and digestibility (Han et al., 2021). However, effective exploitation of this potential is contingent upon precise estimation of agroecological needs and effective exploitation of suitable farming practices. Suitable variety selection and planting methods, in addition to climate, soil texture, irrigation and fertilizer practices, remain as key determinants of both yield and quality (McWilliams et al., 2001; Kurt and Tan, 2019; Aydın and Geren, 2021). In addition, harvesting time, silage production, and storage practices remain paramount in terms of feed value conservation (AgReliant Genetics, 2022; Penn State Extension, 2023; Soundharrajan et al., 2025). From a sustainable farming standpoint, silage corn brings forth, in addition to high yield and high-quality feed production; So too, it excels in contributing towards sustaining a healthy root zone, enhanced water use efficiency, and ecological equilibrium (Liu et al., 2020; Hatfield and Dold, 2019). The adoption of scientifically structured production as well as integrated management practices inures towards both animal products quality and productivity as well as sustaining economically and environmentally friendly farming practices (Guo et al., 2022). Hence, silage corn farming will remain at the forefront of animal husbandry and sustainable farming practices in the future. RESOURCES Adana Tarım Rehberi. (n.d.). Mısır yetiştiriciliğinde sulama ve bakım önerileri. Adana İl Tarım Müdürlüğü Yayınları. AgReliant Genetics. (2022). Corn silage harvest recommendations. AgReliant Genetics. AHDB. (n.d.). Making better silage: Best practice guide. Agriculture and Horticulture Development Board.
THE IMPORTANCE OF SILAGE CORN (ZEA MAYS L.) PLANT . . . 91 Aquilian, L., Smith, R., & Johnson, P. (2020). Optimal chop length and its effect on rumen function and dairy cow performance. Journal of Animal Nutrition, 106(4), 1123–1134. Åkesson, M., Olsson, K., & Gustavsson, A. M. (2022). Effect of kernel processing and chop length on silage quality and digestibility in dairy cows. Journal of Dairy Science, 105(3), 2561–2573. Aydın, R., & Geren, H. (2021). Effects of crop management practices on sustainable silage maize production in Turkey. Turkish Journal of Field Crops, 26(2), 221–229. Bayer. (2024). Ensiling maize: Harvest timing and quality management. Bayer Crop Science. Bayer Crop Science. (2021). Corn irrigation and water management guide. Bayer AG. Bayer Crop Science. (2023). Corn nutrition and fertilization guide. Bayer AG. Bayram, G., Yılmaz, S., & Kara, B. (2017). İkinci ürün silajlık mısırda farklı ekim şekilleri ve bitki sıklıklarının verim ve kaliteye etkileri. Uludağ Üniversitesi Ziraat Fakültesi Dergisi, 31(1), 57–66. Bozkurt, H., & Kaya, M. (2018). Tohum yatağı hazırlığının mısırda çıkış ve verim üzerine etkileri. Tarla Bitkileri Merkez Araştırma Enstitüsü Dergisi, 27(1), 35–41. Çakır, R., Yıldız, H., & Öztürk, A. (2017). Silajlık mısırda morfolojik özellikler ve kalite parametreleri üzerine bir değerlendirme. Anadolu Tarım Bilimleri Dergisi, 32(1), 55–63. Çiftliğim Agromarket. (2019). Mısır silajında doğru biçim teknikleri. Çiftliğim Tarım Yayınları. da Silva, T. C., Ribeiro, G. O., & Santos, F. A. (2025). Harvest maturity effects on maize silage nutritive value and animal performance. Animal Feed Science and Technology, 310, 115646. Dekalb. (2023). Mısır yetiştiriciliğinde gübreleme önerileri. Dekalb Türkiye. Dědina, M., Jevič, P., Čermák, P., Moudrý, J., Mukosha, C. E., Lošák, T., Hrušovský, T., & Watzlová, E. (2024). Environmental life cycle assessment of silage maize in relation to regenerative agriculture. Sustainability, 16(2), 481. Dellait, H. (2020). Corn silage management for optimal yield and nutritive value. Journal of Animal Science and Biotechnology, 11(3), 245–252.
92 INNOVATIVE STUDIES IN AGRICULTURE, FORESTRY AND AQUACULTURE – 2025 EgeFarm. (2022). Silajlık mısır hasadı ve depolama teknikleri. EgeFarm Tarım Yayınları. Extension PSU. (2023). Corn silage harvesting and storage. Penn State Extension. FAO. (1992). Crop water requirements – FAO Irrigation and Drainage Paper 24. Food and Agriculture Organization of the United Nations. FarmProgress. (2023). Critical water stages in corn development. Farm Progress Media. Fuksa, P., Hakl, J., & Konečná, J. (2023). Effect of seeding rate on yield and nutritive value of silage maize. Agronomy, 13(2), 421. Geerts, S., & Raes, D. (2009). Deficit irrigation as an on-farm strategy to maximize crop water productivity in dry areas. Agricultural Water Management, 96(9), 1275–1284. Golden Harvest. (2023). Corn growth stages and water needs. Golden Harvest Seeds. Gram, M., Jensen, L. S., & Thomsen, I. K. (2024). Optimizing split nitrogen applications to improve maize yield and nitrogen use efficiency. Nutrient Cycling in Agroecosystems, 118(1), 23–35. Guo, X., Li, J., & Zhang, Y. (2022). Advances in silage fermentation and preservation: Impacts on quality and animal performance. Animal Feed Science and Technology, 287, 115260. Haifa Group. (2016). Water management in maize production. Haifa Chemicals Ltd. Han, H., Zhou, H., & Wang, C. (2021). Maize silage utilization in ruminant nutrition: A global perspective. Journal of Integrative Agriculture, 20(10), 2424–2436. Hatfield, J. L., & Dold, C. (2019). Water-use efficiency: Advances and challenges in a changing climate. Frontiers in Plant Science, 10, 103. Heap, I. (2024). The international herbicide-resistant weed database. WeedScience.org. Kalaycı, R., Demirtaş, A., & Akman, H. (2016). Farklı ekim yöntemlerinde silajlık mısırda verim ve kalite parametreleri. Türk Tarım ve Doğa Bilimleri Dergisi, 3(4), 278–285. Karnatam, S., Singh, R., & Sharma, P. (2023). Effect of harvest stage on yield and nutritive value of maize silage. Forage Research, 49(2), 85–92. Karnatam, K. S., Mythri, B., Un Nisa, W., Sharma, H., Meena, T. K., Rana, P., Vikal, Y., Gowda, M., Dhillon, B. D., & Sandhu, S. (2023). Silage maize as a
THE IMPORTANCE OF SILAGE CORN (ZEA MAYS L.) PLANT . . . 93 potent candidate for sustainable animal husbandry development—perspectives and strategies for genetic enhancement. Frontiers in Genetics, 14, 1150132. Kaya, Y., Demir, E., & Öztürk, A. (2020). Mısır tarımında mekanizasyon uygulamalarının verim ve iş gücü üzerine etkileri. Tarım Makinaları Bilimi Dergisi, 16(2), 101–109. Kim, J. (2023). Climate change and maize production: Agronomic perspectives on yield stability and adaptation. Journal of Crop Science and Biotechnology, 26(4), 321–333. Kitaw, G., Terefe, G., Faji, M., Mengistu, G., Dejene, M., et al. (2024). Effect of maize (Zea mays L.) genotypes, harvesting stages and ensiling periods on fermentation and nutritional value of silage. Grass Research, 4, e009. Kılıç, H., & Özkan, U. (2020). Tohum ekim derinliğinin mısırda çimlenme ve çıkış üzerine etkileri. KSÜ Tarım ve Doğa Dergisi, 23(4), 755–762. Knezevic, S. Z., Streibig, J. C., & Ritz, C. (2009). Utilizing dose–response curves to determine synergism for herbicide mixtures. Weed Technology, 17(2), 242–251. https://doi.org/10.1614/0890-037X(2003)017[0242:UDRCTD]2.0 .CO;2 Kurt, O., & Tan, M. (2019). FAO olgunlaşma gruplarına göre silajlık mısır çeşitlerinin değerlendirilmesi. Atatürk Üniversitesi Ziraat Fakültesi Dergisi, 50(1), 25–34. Kurt, O., & Tan, M. (2019). Silaj teknolojisi ve kaba yem üretiminde önemi. Atatürk Üniversitesi Ziraat Fakültesi Dergisi, 50(1), 45–53. Lactanet. (2018). Corn silage: Harvest and processing guide. Lactanet Canada. Laskari, K., Papadopoulos, I., & Katsoulas, N. (2022). Impact of water stress on maize productivity and environmental costs. Agronomy, 12(4), 812. Liebman, M., & Davis, A. S. (2000). Integration of soil, crop and weed management in low-external-input farming systems. Weed Research, 40(1), 27-47. Limagrain Türkiye. (2021). Silajlık mısır üretim rehberi. Limagrain Tohumculuk. Liu, X., Zhang, Y., & Wang, J. (2020). Sustainable maize production through conservation tillage and nutrient management: A review. Agriculture, Ecosystems & Environment, 296, 106924. Liu, Y., Chen, X., & Wang, H. (2021). Split nitrogen application improves yield and nitrogen recovery efficiency in maize. Field Crops Research, 270, 108229.
94 INNOVATIVE STUDIES IN AGRICULTURE, FORESTRY AND AQUACULTURE – 2025 Liu, Y., Zhang, H., & Wang, X. (2023). Smart irrigation management in maize under water scarcity conditions. Field Crops Research, 295, 108874. Masat, F., İkikat Tümer, S., & Gürün, B. (2025). Silajlık mısırda farklı sulama seviyelerinin verim ve kaliteye etkileri. Kahramanmaraş Sütçü İmam Üniversitesi Tarım ve Doğa Dergisi, 27(1), 55–64. Masseeds. (2023). Determining the right harvest stage for silage corn. Masseeds. McWilliams, D. A., Berglund, D. R., & Endres, G. J. (2001). Corn growth and management quick guide (Extension Bulletin A-1173). North Dakota State University Extension Service. MSU Extension. (2018). Corn silage harvest and storage management. Michigan State University Extension. Munkvold, G. P., & Desjardins, A. E. (1997). Fumonisins in maize: Can we reduce their occurrence? Plant Disease, 81(6), 556–565. Netafim. (2018). Drip irrigation for maize: Solutions for water efficiency. Netafim Ltd. Öktem, A., & Oral, E. (2020). Silajlık mısırda toprak işleme teknikleri ve verim ilişkisi. Harran Tarım ve Gıda Bilimleri Dergisi, 24(3), 299–307. Öktem, A., Öktem, A. G., & Çelik, M. (2003). Effect of deficit irrigation on yield and water use efficiency of silage maize in southeastern Turkey. Agricultural Water Management, 61(1), 63–74. Özarslan, C. (2018). Mısır tarımında farklı sulama yöntemlerinin karşılaştırılması. Ankara Üniversitesi Ziraat Fakültesi Yayınları. Öztürk, A., Kaya, Y., & Yıldırım, T. (2008). Silajlık mısırda farklı ekim yoğunluklarının verim ve kaliteye etkisi. Tarım Bilimleri Dergisi, 14(2), 112– 118. PDA. (2015). Nutrient management for maize. Potash Development Association. Penn State Extension. (2023). Harvesting corn silage at the right moisture. Penn State College of Agricultural Sciences. Qian, Y. (2023). Optimizing the growth of silage maize by adjusting planting density and nitrogen application. Agronomy, 13(11), 2785. https://doi. org/10.3390/agronomy13112785 Sade, B., & Sezer, İ. (2010). Mısırda bitki sıklığının verim ve kalite unsurlarına etkisi. Anadolu Tarım Bilimleri Dergisi, 25(1), 55–62. Sezen, Y., Acar, R., & Tansı, V. (2018). Silage storage systems and their effects on feed quality. Çukurova University Journal of Agricultural Faculty, 33(2), 15–24.
THE IMPORTANCE OF SILAGE CORN (ZEA MAYS L.) PLANT . . . 95 Şahin, M., Yıldırım, A., & Öner, F. (2018). Düşük sıcaklık stresinin mısırda çimlenme ve erken dönem gelişimi üzerine etkileri. Anadolu Tarım Bilimleri Dergisi, 33(2), 201–210. Sharma, H., et al. (2023). Silage maize as a potent candidate for sustainable animal husbandry development—perspectives and strategies for genetic enhancement [Review]. Frontiers in Genetics. Soundharrajan, I., Kim, D., & Choi, K. (2025). Role of lactic acid bacteria in improving silage quality under climate stress conditions. Frontiers in Microbiology, 16, 1458. Tarım ve Orman Bakanlığı. (2012). Mısır yetiştiriciliği gübreleme rehberi. T.C. Tarım ve Orman Bakanlığı Yayınları. Tarım ve Orman Bakanlığı. (2013). Toprak verimliliği ve gübreleme teknik talimatları. T.C. Tarım ve Orman Bakanlığı Yayınları. Tarım ve Orman Bakanlığı. (2019). Mısır yetiştiriciliği sulama rehberi. T.C. Tarım ve Orman Bakanlığı Yayınları. Tarım ve Orman Bakanlığı. (2021). Mısır yetiştiriciliği teknik talimatları. T.C. Tarım ve Orman Bakanlığı Yayınları. Tarım ve Orman Bakanlığı. (n.d.). Mısır tarımı teknik talimatları. T.C. Tarım ve Orman Bakanlığı Yayınları. USU Extension. (2021). Silage production and chop length recommendations. Utah State University Extension. https://extension.usu.edu USDA. (2023). Irrigation scheduling and water use efficiency in corn. United States Department of Agriculture. Van Donk, S. J., Martin, D. L., & Irmak, S. (2013). Crop water use and irrigation management in maize. Applied Engineering in Agriculture, 29(3), 389–397. https://doi.org/10.13031/aea.29.9812 Wang, L., & Li, Z. (2023). Nitrogen losses and management strategies in maize-based systems. Soil & Tillage Research, 227, 105656. Wilkinson, J. M., & Chamberlain, A. T. (2016). Feeding the dairy cow. Chalcombe Publications. Wilkinson, J. M., & Rinne, M. (2018). Highlights of progress in silage conservation and future perspectives. Grass and Forage Science, 73(1), 40–52. https://doi.org/10.1111/gfs.12327 Yılmaz, İ., & Demir, E. (2021). Farklı ekim yöntemlerinin silajlık mısırda verim ve kaliteye etkileri. Tekirdağ Ziraat Fakültesi Dergisi, 18(3), 345–352. Zhang, Q., Liu, H., & Zhao, Y. (2021). Late-season nitrogen application improves grain filling and forage quality in maize. Plant and Soil, 465(1–2), 213–227.
96 INNOVATIVE STUDIES IN AGRICULTURE, FORESTRY AND AQUACULTURE – 2025 Zhang, Q., Liu, H., & Zhao, Y. (2021). Supplemental irrigation mitigates heat stress impacts on maize yield and water productivity. Agricultural Water Management, 245, 106593. Zhou, T., Wang, X., & Chen, Y. (2021). Foliar micronutrient applications enhance maize yield and photosynthetic efficiency. Journal of Plant Nutrition, 44(15), 2249–2262. Zimdahl, R. L. (2004). Weed–crop competition: A review (2nd ed.). Ames, IA: Blackwell Publishing. Zimdahl, R. L. (2018). Fundamentals of weed science (5th ed.). Academic Press.