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# Biopreservation of pre-processed fresh fish by bio-based coatings: A single strategy with multiple benefits towards waste prevention

Don Hettiarachchige Udana Eranda<sup>a</sup>, Manat Chaijan<sup>a</sup>, Ilke Uysal-Unalan<sup>b,c</sup>, Worawan Panpipat<sup>a</sup>, Azza Silotry Naik<sup>d</sup>, Amira Leila Dib<sup>e</sup>, Supatra Karnjanapratum<sup>f</sup> and Mohammed Gagaoua<sup>g,h</sup>*

## Abstract

The demand for pre-processed fresh fishery products is growing due to their convenience for subsequent processing and cooking. However, when improperly stored, the combined impacts of microbial deterioration and chemical reactions render them inedible, leading to significant food waste. To tackle this issue, several approaches have been proposed, among which bio-based edible coating provides a promising solution with a green approach. Edible coatings are formed of bio-based polymers, particularly using macromolecules such as proteins and polysaccharides that are biodegradable, edible, and can serve as carriers for naturally sourced active agents. Coatings made from a variety of proteins and polysaccharides with the main focus on gelatin and chitosan to preserve pre-processed fish products are discussed in this review along with their properties from the microbial, physicochemical, and sensory perspectives. Coating carriers and composite natural preservatives as well as formula optimization and investigation of coating mechanisms, challenges, and potential research prospects have been further reviewed. Overall, edible coatings with active substances can be used to efficiently preserve freshly pre-processed fish. The underlying mechanisms are multiple, and the applications were mainly for preventing or reducing moisture loss and purge accumulation, postponing microbial spoilage, and restricting the growth of pathogenic microorganisms, slowing and/or inhibiting lipid, protein, and pigment oxidation, and extending the shelf-life along with improving sensory properties. Edible coatings are viewed as ecologically friendly and sustainable packaging alternatives for the preservation of pre-processed fish products.

**Keywords:** Sustainable products; Bio-based polymers; Edible coatings; Fish products; Shelf-life; Sensory properties, Food wastage, Packaging.

<footnote><sup>a</sup> Food Technology and Innovation Research Center of Excellence, Division of Food Science and Innovation, Department of Food Industry, School of Agricultural Technology and Food Industry, Walailak University, Nakhon Si Thammarat, 80161, Thailand</footnote>

<footnote><sup>b</sup> Department of Food Science, Aarhus University, Agro Food Park 48, 8200, Aarhus N, Denmark</footnote>

<footnote><sup>c</sup> CiFOOD - Center for Innovative Food Research, Aarhus University, Agro Food Park, 48, 8200, Aarhus N, Denmark</footnote>

<footnote><sup>d</sup> School of Food Sciences and Environmental Health, Technological University Dublin, Grangegorman, D07 ADY7, Dublin, Ireland</footnote>

<footnote><sup>e</sup> Gestion de La Santé et Productions Animales Research Laboratory, Institut des Sciences Vétérinaires El-Khroub, Université Constantine 1 Frères Mentouri, Constantine, 25000, Algeria</footnote>

<footnote><sup>f</sup> Division of Marine Product Technology, Faculty of Agro-Industry, Chiang Mai University, Chiang Mai, Thailand</footnote>

<footnote><sup>g</sup> Food Quality and Sensory Science Department, Teagasc Ashtown Food Research Centre, Ashtown, Dublin 15, Ireland</footnote>

<footnote><sup>h</sup> PEGASE, INRAE, Institut Agro, 35590, Saint-Gilles, France</footnote>

<footnote>* Corresponding author: mohammed.gagaoua@inrae.fr</footnote>
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# 1. Introduction

Sustainable development is a global goal that requires an interdisciplinary approach to addressing current and future environmental, climate change, economic, and resource efficiency concerns. Food sustainability must be a global priority if we are to maintain our planet for future generations; nevertheless, this lofty target is difficult to achieve due to two fundamental difficulties in food handling, production, and storage, these being food loss and waste. Among food products of animal origin, fish is a perishable raw material that, more than other matrices, faces the issues of food waste and loss (up to 40% in some middle-low income countries) (Racioppo et al., 2021). Food loss is defined as a decrease in the quantity and quality of food because of the decisions made by food suppliers, excluding retail, providers, and consumers (FAO, 2019). The best example of food loss in the fishing industry is so-called low-value fish, which is frequently discarded since it is not of acceptable intrinsic quality for consumers. Fish loss transpires at every step of the value chain, from catch to final consumption. Loss is influenced by a number of factors, including the species of fish, the associated physical characteristics (i.e. composition, weight, and structure), the perceived value of the fish, the volumes handled, the level of seasonality, and the geographic location. Moreover, due to technical, financial, infrastructural, and managerial constraints, fish companies in developing countries are likely to incur greater fish losses than those in developed ones during loading and unloading, processing, storage, transportation, and marketing (De Silva, 2011). However, not all the seafood that is made accessible is consumed. Volumes of wastes are produced by discards, bycatch on board, and wastage ashore. The processing discards, such as skins, skulls, heads, viscera, fillet cut offs, spoilage discards, and others, account for up to 40% of the processed raw material (Venugopal & Sasidharan, 2021). In the past few decades, there has been a significant upsurge in close consideration surrounding the issue of food waste. This is mostly due to the numerous adverse effects of food waste and loss (FWL), as well as the growing recognition of the problem's global scale. Specifically, there is a heightened awareness of the associations between food waste and loss, food security, and climate change (Mokrane et al., 2023). Millions of people, including those in regions where malnutrition and micronutrient deficiencies are pervasive (FAO, 2011) are affected by the loss or nutritional degradation of highly nutritious materials. In fact, due to high ambient temperatures, lack of access to services, infrastructure, and basic technology, reliance on more traditional smoking and drying techniques for preservation, and absence of cooling (cold chain) facilities, spoilage and quality degradation of the product are more likely to occur in low-income economies (HLPE, 2014). Because of its significance in coastal and marine pollution, improper disposal of fishery waste in marine and coastal areas has received growing attention. Improved use of seafood waste can help meet the global demand for seafood, while avoiding the negative impacts of seafood harvesting and production.

The primary cause of fish quality degradation in fresh or minimally preserved fish is microbial decomposition, which in extreme circumstances can lead to 25%–30% loss of marketable fish (Mei et al., 2019; Nie et al., 2022; Tavares et al., 2021). Fresh fish has a distinct microflora, a relatively short shelf life, and its spoilage is caused by gram-negative aerobic bacteria such as *Pseudomonas* sp. and *Shewanella* sp. (Donna et al., 2007). Notably, some mesophilic and psychrotolerant bacteria, such as *Morganella psychrotolerans* and *Photobacterium* spp., can produce histamine in fish during storage by decarboxylating free histidine in the fish muscle, which can result in histamine fish poisoning (De Silva, 2011; Tsironi et al., 2008). In addition to microbiological problems, oxidative deterioration has a major detrimental effect on the quality of fish (Amit et al., 2017). Lipid oxidation
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can further degrade the flavor, color, and texture as well as a concurrent reduction in the nutritional attributes of the stored fish (Amit et al., 2017). As lipids spontaneously react with atmospheric oxygen, auto-oxidation of lipids is the most frequent form of oxidative deterioration in fish (Hassoun and Emir Çoban, 2017). As a result, fish tissues with high levels of unsaturated lipids are extremely vulnerable to oxidation and faster deterioration (Sahari et al., 2014). Another issue with dark-fleshed fish such as that of tuna and mackerel, is the discoloration caused by the redox instability of myoglobin when kept at a cold temperature, which can lead to sensory unacceptability (Nakazawa et al., 2020; Thiansilakul et al., 2011; Wongwichian et al., 2015). The brighter reddish hue of dark-fleshed fish meat is typically attributed to the presence of red pigment, primarily oxymyoglobin, which undergoes various biochemical changes during handling and storage to become brown metmyoglobin (Chaijan & Panpipat, 2017; Singh et al., 2021). Myoglobin has a close relationship with lipid oxidation which could further influence the deterioration in food (O'grady et al., 2001). Thus, both microbial and chemical actions can cause the deterioration in the quality of fish flesh during storage (Nawaz et al., 2020; Qiu et al., 2014; Singh et al., 2021).

The most popular method of delaying the microbiological and biochemical decomposition of fish is to lower the temperature with ice or mechanical refrigeration (Trigo et al., 2022). However, some disadvantages of cold storage include dehydration, textural hardness, nutritional loss, and decreased protein extractability. The shelf life of fresh fish fillets at 4 °C is 1–2 days. While being stored in the refrigerator, fish fillets will soon lose quality and become unfit for consumption in a very short time (Lahreche et al., 2020). Therefore, finding new approaches to lower quality loss and extend the shelf life of refrigerated fish fillets is of importance given the rising demand for refrigerated fresh fish fillets. Several processing technologies may be used to increase the raw fish's shelf life (Kulawik & Dordević, 2020), likely ultra-low temperature storage (Nakazawa et al., 2020), stepwise chilling (Park et al., 2021), modified atmospheric packaging (Abel et al., 2018), antimicrobial absorbent pads (Otoni et al., 2016), cold pasteurization/irradiation (Gautam & Venugopal, 2021) and the use of natural preservatives (Singh et al., 2021) or the combination of these techniques. In addition, recent EU regulations highlight the importance of limiting of the weight and volume of packaging to a minimum adequate amount without compromising the quality and safety of food products (Dörnyei et al., 2023; European Commission, 2020). Therefore, (active) bio-based coating as a part of food preservation and food packaging can be seen as a key strategy to cutting down the use of plastic by co-acting as a shelf-life extender together with primary plastic packaging. Moreover, edible coatings have been regarded as the optimal replacement for conventional packaging systems owing to its intrinsic properties, including water solubility, fats and oil resistance, flexibility, biodegradability, and superior barrier characteristics (Firouz et al., 2021; Syafiq et al., 2020). Edible coatings are thin layers made up of polysaccharides, proteins, lipids, bioactive substances, and composite materials. They are applied directly to the surface of food by either using the techniques of submerging or spraying (Baghi et al., 2022; Cheng et al., 2021). Edible coatings endure the capacity to serve as a vehicle for various bioactive substances, including natural antioxidants, phenolic compounds, and antimicrobials, and thus improve their efficacy in preserving food and impede microbial activity on the surface of the food (Pedreiro et al., 2021; Xu et al., 2022). There is a persistent need to develop economically feasible innovative methods for preserving the quality of fish and fish products, thus justifying the need for further research on coatings.

## 2. Spoilage mechanisms of fresh fish
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Fish are highly perishable due to their high moisture content (70%–80%), large amounts of small molecules, and neutral pH, which all provide ideal conditions for microbial and biochemical spoilage (Ojagh et al., 2010; Ramezani et al., 2015). Endogenous enzymatic reactions, oxidation, and microbial activities occur shortly after death in fish, causing changes in sensory and nutritional properties and limiting shelf-life (Karoui & Hassoun, 2017; Kostaki et al., 2009; Olatunde & Benjakul, 2018). In the dark-fleshed fish, the discoloration, formation of unpleasant odors and flavors, as well as the development of harmful molecules, are notable effects of lipid oxidation, with negative health and economic implications (Kuuliala et al., 2018; Mahmoud et al., 2016). Thus, any change in the initial condition of fish that results in an unpleasant odor, taste, appearance, or texture is referred to as spoilage. Fish spoilage can be classified into three distinct mechanisms, including microbial, autolytic, and oxidative spoilages.

## 2.1. Microbial spoilage

The biochemical composition of fish and seafood promotes bacterial growth, including Gram-positive and Gram-negative types that can survive in a wide temperature range. As a result, microbial spoilage is regarded as one of the primary causes of fish quality degradation, accounting for up to 25%–30% of such product loss (Mei et al., 2019). Microbial growth and metabolism generate biogenic amines like putrescine, histamine, and cadaverine, as well as organic acids, sulphides, alcohols, aldehydes, and ketones with irritating and undesirable off-flavors (Bulushi et al., 2009). Every fish species has a unique microflora that is determined by its microbial environment, raw materials, manufacturing parameters, and subsequent storage conditions, as well as the microorganisms' ability to withstand preservation conditions (Chaillou et al., 2015). For example, psychrotolerant Gram-negative bacteria, such as those belonging to the genera *Pseudomonas* and *Shewanella*, have been shown to be the most common spoilage bacteria in aerobically stored chilled fish. CO₂-tolerant microbes, such as *Photobacterium phosphoreum* and lactic acid bacteria, on the other hand, may dominate the microflora and lead to the spoilage of packed fish products (Chaillou et al., 2015). While fresh fish is normally contaminated with many microorganisms, only a small percentage of these microorganisms, known as specific spoilage organisms (SSO), cause spoilage. The SSO differ depending on the type of fish and the preservation conditions (Table 1). *Pseudomonas* was found to be the SSO for bighead carp (*Aristichthys nobilis*) fillets sprinkled with 2% salt, whereas *Aeromonas* was found to be the SSO for unsalted bighead during 4 °C storage (Hassoun & EmirÇoban, 2017; Liu et al., 2017). By assimilation of nonprotein-nitrogen in fish tissue, SSO have the potential to dominate and generate metabolites, resulting in unpleasant and undesirable off-flavors that directly impact the organoleptic properties of fish, leading to consumer rejection (Boziaris et al., 2013).

<table><caption>**Table 1.** Microorganisms and specific spoilage organisms (SSO) in fish and seafood products during storage.</caption><thead><tr><td>Specific spoilage organisms (SSO)</td><td>Fish product/seafood species</td><td>Packaging technology used, gas and water vapor barrier properties and storage conditions</td><td>Analysis method</td><td>References</td></tr></thead><tbody><tr><td>Acinetobacter johnsoni<br>Acinetobacter lwooffii<br>Shewanella baltica<br>Pseudomonas Fragi<br>Pseudomonas</td><td>Hybrid sturgeons (Acipenser schrenckii<br>female × Huso dauricus<br>males, females, 60 ± 5 kg)</td><td>Packaging technology used: wrapped in plastic bag, and packaged in a cardboard box with gel ice packed around the inner side</td><td>Plate colony-counting method<br>Plate count agar (PCA)<br>PCA plate method<br>MALDI Biotyper</td><td>Li et al. (2021)</td></tr></tbody></table>
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<table><thead><tr><th>Specific spoilage organisms (SSO)</th><th>Fish product/seafood species</th><th>Packaging technology used, gas and water vapor barrier properties and storage conditions</th><th>Analysis method</th><th>References</th></tr></thead><tbody><tr><td>koreensis<br>Pseudomonas<br>antarctica</td><td></td><td>Storage condition: kept in the cold storage (–40 °C) for 0.5–1 h</td><td>(MBT, Brucker<br>Daltonoics)</td><td></td></tr><tr><td>Photobacter<br>phosphoreum<br>Psychrobacter spp.<br>Moritella spp.<br>Carnobacterium spp.<br>Shewanella spp.<br>Vibrio spp.</td><td>Fresh hake fillets with<br>skin (Merluccius)</td><td>Packaging technology used:<br>Modified atmosphere packaging<br>Gaseous composition: 50 % CO<sub>2</sub>/50 % N<sub>2</sub> under gas/product<br>ratio of approximately 2:1<br>Storage temperature: stored at<br>four varied temperatures of 1,4,7<br>and 10 °C</td><td>Maxwell 16 Lev Blood<br>DNA Extraction Kit<br>(Promega, Madison,<br>USA), qPCR analysis,<br>high-throughput 16S<br>rRNA gene sequencing</td><td>Antunes-<br>Rohling et al.<br>(2019)</td></tr><tr><td>Lactobacillus algidus<br>Pseudomonas<br>fluorescens</td><td>fresh yellowfin tuna<br>fillets (Thunnus<br>albacares)</td><td>Packaging technology used:<br>Vacuum packaged<br>Film thickness: high barrier film<br>(90 µm in thickness)<br>Oxygen transmission rate at 23 °C<br>and 75 % Relative humidity:<br>1 cm<sup>3</sup> m<sup>-2</sup> 24 h<sup>-1</sup> atm<sup>-1</sup></td><td>Agar spread plate, PCR<br>amplification<br>16S rRNA sequencing</td><td>Jääskeläinen<br>et al. (2019)</td></tr><tr><td>Lactic acid bacteria<br>(LAB)<br>Pseudomonas spp.<br>B. thermosphacta</td><td>Atlantic cod (Gadus<br>morhua)</td><td>Packed under different<br>atmospheres gas-product ratio<br>2:1, tray sealer MECA 900<br>Multilayer packaging trays<br>(PP/EVOH/PP)<br>oxygen<br>transmission rate at 23 °C and 50<br>% RH: 0.03 cm<sup>3</sup>/tray *24h<br>Top film (PA/EVOH/PA/PP)<br>Oxygen transmission rate at<br>23 °C, 50 % RH and 1 atm:<br>6.57 cm<sup>3</sup>/m<sup>2</sup>*24h*atm</td><td>Total psychotropic<br>count (TPC) using<br>Marine agar<br>lactic acid bacteria<br>(LAB) count using<br>Man<br>Rogosa Sharpe agar or<br>modified MRS<br>Pour plate method</td><td>Kuuliala et al.<br>(2018)</td></tr><tr><td>Staphylococcus<br>aureus<br>Pseudomonas<br>fluorescence<br>Escherichia coli<br>Vibrio cholerae<br>Salmonella typhi<br>Aspergillus niger</td><td>Indian mackerel fish<br>(Rastrelliger<br>kanagurta)</td><td>Storage temperature: Stored in a<br>refrigerator at 4 °C</td><td>Pour plate method<br>(Serial dilution<br>technique)<br>Sterile Nutrient agar<br>and Sabouraud's<br>dextrose agar<br>Quebec colony<br>counter, Colony<br>morphology<br>Mobility tests, routine<br>mycological methods</td><td>Srinivasan<br>and Saranraj<br>(2017)</td></tr><tr><td>Pseudomonas spp.<br>Shewanella spp.<br>Janthinobacterium<br>spp.<br>Photobacterium spp.<br>Acinetobacter spp.</td><td>farmed Atlantic<br>salmon fillets</td><td>Packaging technology used:<br>Separate boxes (expanded<br>polystyrene; EPS boxes)<br>Storage condition: Stored on ice<br><1 °C (verified by the<br>temperature logging)</td><td>Iron agar medium<br>partial 16S rRNA gene<br>sequencing<br>Phylogenetic analysis by<br>Sequence Scanner<br>software 2 (Applied<br>biosystems)</td><td>Møretrø et al.<br>(2016)</td></tr><tr><td>Rhodanobacter<br>terrae<br>Brochothrix<br>thermosphacta<br>Pseudomonas spp.<br>Enterobacteriaceae<br>spp.<br>Lactic acid bacteria<br>(LAB)</td><td>tropical yellowfin<br>tuna (Thunnus<br>albacares)</td><td>First batch packaging and<br>storage condition: Vacuum-<br>packed individually in 80 µm<br>thickness plastic bags made from<br>polyamide and polypropylene<br>Gas permeability:<br>2.78 cm<sup>3</sup> m<sup>-2</sup>/day for water vapor<br>O<sub>2</sub> permeability:<br>19.95 cm<sup>3</sup>/m<sup>2</sup>/day</td><td>Plate count agar (PCA)<br>ELK and STAA<br>PCR amplification and<br>Bacterial 16S rRNA<br>gene amplification<br>barcode sequencing</td><td>Silbande et<br>al. (2016)</td></tr></tbody></table>
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<table><thead><tr><th>Specific spoilage organisms (SSO)</th><th>Fish product/seafood species</th><th>Packaging technology used, gas and water vapor barrier properties and storage conditions</th><th>Analysis method</th><th>References</th></tr></thead><tbody><tr><td></td><td></td><td>CO<sub>2</sub> permeability: 164.87 cm3/m2/day<br>Second batch packaging condition: Vacuum-packed in 80 µm thickness plastic bags used<br>Storage condition: Iced in a cool box by altering a fish layer with a layer of ice and stored in a cold room at 4 °C<br>Third batch storage condition: Placed in a film plastic tray. Packaging technology used: MAP (70 % CO<sub>2</sub> and 30 % O<sub>2</sub>)</td><td></td><td></td></tr><tr><td>Pseudomonas fluorescens<br>Psychrobacter spp.<br>Macrococcus caseolyticus<br>Pseudomonas fragi<br>Pseudomonas veroni<br>Carnobacterium maltaromaticum<br>Carnobacterium divergens<br>Vagococcus fluvialis</td><td>Sea bream fillet (Sparus aurata)</td><td>Packaging material used: Polystyrene boxes with melted ice.<br>Storage conditions: under air or MAP<br>MAP gas concentration: 60 % CO<sub>2</sub>/10 % O<sub>2</sub>/30 % N<sub>2</sub> (MAP)<br>MAP film material: BDF 8050 F packaged in insulated boxes with melted ice, stored in incubators operating at 0 °C and 5 °C</td><td>Cycloheximide (actidione) agar (STAA) tryptone soy agar (TSA)<br>Cephaloridine fucidin cetrimide (CFC) agar<br>Polymerase chain reaction method (PCR)<br>with 6S rRNA gene fragments amplification</td><td>Parlapani et al. (2014)</td></tr><tr><td>Photobacterium phosphoreum</td><td>Fillets of coalfish</td><td>Storage temperature: Stored at 10 °C or 5 °C<br>Gaseous composition: CO<sub>2</sub>/N<sub>2</sub> (60 %/40 %) (MAP)</td><td>Agar plates & tissues/Real time Polymerase Chain Reaction (PCR)/T-RFLP, amplification & sequencing</td><td>Rudi et al. (2004)</td></tr><tr><td>Brochothrix thermosphacta<br>Carnobacterium divergens<br>Carnobacterium piscicola</td><td>Fillets of salmon</td><td>Storage temperature: Stored at 10 °C or 5 °C<br>Gaseous composition: CO<sub>2</sub>/N<sub>2</sub> (60 %/40 %) (MAP)</td><td>Agar plates & tissues/Real time PCR/T-RFLP, amplification & sequencing</td><td></td></tr><tr><td>S. putrefaciens<br>Pseudoalteromonas spp.<br>Pseudomonas spp.</td><td>Squid (Todaropsis eblanae)</td><td>Storage medium: Ice<br>Storage method: Squid to ice ratio was approximately 2:1 (crushed freshwater ice), new ice was added periodically to maintain the ratio</td><td>Spread-plated chilled Iron Agar Lyngby (ADSA Micro), Iron agar with salt used</td><td>Paarup et al. (2002)</td></tr></tbody></table>

Abbreviations: TPC-total psychrotrophic count; LAB- lactic acidbacteria; PCA -plate count agar; MAP-modified-atmosphere packaging; PCR-polymerase Chain Reaction; T-RFLP- A terminal restriction fragment length polymorphism.

## 2.2. Autolytic spoilage

Autolysis, or protein degeneration caused by endogenous proteases, starts shortly after rigor mortis onset. This mechanism aids in the creation of an environment favorable to microbial growth (Olatunde & Benjakul, 2018). Even under refrigeration or freezing conditions, autolysis in fish continues at a very slow rate (Robertson, 2016). Protease and lipase enzymes, commonly found in
## Portails

* [Portails institutionnels](#)

* [HAL](#)

* [HAL SHS](#)

* [HAL Thèses](#)

* [MédiHAL](#)

## CCSD

* [CCSD](#)

* [Episciences](#)

* [Sciencesconf](#)
fish muscle and viscera, contribute to the postmortem deterioration of fish muscle and other tissues during storage and processing. In addition, proteases and lipases are responsible for changes in fish sensory characteristics (Engvang & Nielsen, 2001).

Protein degradation occurs rapidly in inappropriate storage. In fact, proteolytic products (free amino acids and peptides) may act as nutrients for microbial growth in conjunction with the production of biogenic amines, culminating in spoilage (Fraser & Sumar, 1998). The enzymes in the fish gut cause rapid protein decomposition, resulting in belly burst, a common occurrence in fish. Temperature and pH are the most important factors influencing the activity of proteases. When the pH is between alkaline and neutral, most proteases perform better (Ashie et al., 1996). Endogenous or microbial enzyme's function and metabolism raise the pH of stored seafood by converting trimethylamine oxide (TMAO) to TMA and other major volatile compounds (Olatunde & Benjakul, 2018). Lipase and phospholipase are two enzymes that can hydrolyze lipids in fish (Aryee et al., 2007). In the presence of water, triacylglycerol acyl hydrolases hydrolyze glycerides (mono, di, and tri) to glycerol and fatty acids (Fernandes, 2016; Olatunde & Benjakul, 2018). Fish and other seafood release free fatty acids (FFAs), which contribute to the development of fishy odors (i.e. off-odors). The fishy odor is caused by aldehydes, primarily polyunsaturated aldehydes (Maqsood et al., 2014). Furthermore, lipoxygenase, which is primarily found in the gills or skin, can cause oxidation in preserved fish, particularly when the fish is kept for an extended period (Fernandes, 2016). Table 2 lists various fish proteolytic and lipolytic enzymes and products, along with information on how they are controlled.

<table><caption>**Table 2.** Different enzymes and products of proteolysis and lipolysis in fish and their prevention strategies.</caption><thead><tr><td>Enzyme (s)</td><td>Substrate</td><td>Effect/final products</td><td>Prevention</td><td>References</td></tr></thead><tbody><tr><td>Acid phosphatase</td><td>Lyzozome</td><td>Lysosome disruption</td><td>Storage at higher pressure of 75 MPa/25 °C</td><td></td></tr><tr><td>Cathepsin D<br>Chymotrypsin, Trypsin,<br>Carboxy-peptidases</td><td>Proteins<br>Proteins, peptides</td><td>Disruption of lyzozomes<br>Belly-bursting</td><td>Store at 37 °C (residual activity 7 % & 21 %)<br>Problem increased with freezing/thawing or long-term chill storage</td><td>Fidalgo et al. (2020)</td></tr><tr><td>Cathepsins<br>Cathepsin B</td><td>Proteins, peptides<br>Proteins</td><td>Softening of tissue<br>Hydrolysis of tissue proteins</td><td>Avoid rough handling during storage<br>Stored at 25 °C or 37 °C (at AP; 42 and 3 %, respectively)</td><td>FAO (2005)<br>Fidalgo et al. (2020)</td></tr><tr><td>Calpain<br>Collagenases</td><td>Myofibrillar proteins<br>Connective tissue</td><td>Softening<br>Softening and gaping of tissue<br>Degradation of Type 5 collagen</td><td>Removal of calcium<br>Time and temperature of chilled Storage</td><td>FAO (2005)</td></tr><tr><td>Trimethylamine Oxide<br>(TMAO) demethylase</td><td>TMAO</td><td>Formaldehyde formation</td><td>Storage temperature less than -30 °C physical abuse, freeze/thawing</td><td>FAO (2005)</td></tr><tr><td>Glycolytic enzymes</td><td>Glycogen</td><td>Lactic acid production, resulting in pH drop</td><td>Avoid pre-rigor stress</td><td>FAO (2005)</td></tr><tr><td>Nucleotide breakdown<br>Enzymes</td><td>ATP, ADP, AMP,<br>IMP</td><td>Gradual production of hypoxanthine</td><td>Avoid pre-rigor stress and improved handling</td><td>FAO (2005)</td></tr><tr><td>Lipoxygenase (LOX)</td><td>Polyunsaturated fatty acids (PUFA)<br>Linoleic acid<br>Linolenic acids<br>Decosahexaenoic</td><td>13- and 9-hydroperoxides</td><td>Application of inhibitors Nordihydroguaiaretic acid (NDGA), esculetin (catechols), Butylated hydroxyanisole (BHA), Butylated</td><td>Banerjee et al. (2001)</td></tr></tbody></table>
<table><thead><tr><td>Enzyme (s)</td><td>Substrate</td><td>Effect/final products</td><td>Prevention</td><td>References</td></tr></thead><tbody><tr><td></td><td>acid<br>Arachidonic acid</td><td></td><td>hydroxytoluene (BHT),<br>Epicatechin gallate (ECG),<br>Epicatechin (EC), Potassium<br>cyanide (KCN)</td><td></td></tr><tr><td>Nucleodepolymerases<br>Ribonuclease<br>Acid deoxyribonuclease</td><td>RNA, DNA</td><td>Mononucleotides<br>enhance the flavor of fish<br>products</td><td>Blanching of fish in steam at<br>100 °C<br>Reducing the water activity by<br>drying</td><td>Mukundan<br>et al. (1986)</td></tr><tr><td>Lipase<br>Phosphorilipase</td><td>Lipids</td><td>Free fatty acids and<br>glycerol, lipolysis off<br>flavor, hydrolytic<br>rancidity and oxidative<br>rancidity</td><td>Blanching of fish in steam at<br>100 °C<br>Reducing the water activity by<br>drying</td><td>Mukundan<br>et al. (1986)</td></tr></tbody></table>

Abbreviations: AP: atmospheric pressure; TMAO- trimethylamine oxide; ATP- adenosine triphosphate; ADP-Adenosine diphosphate; AMP- adenosine monophosphate; IMP- inosine monophosphate; NDGA-nordihydroguaiaretic acid; BHA- butylated hydroxyanisole; BHT- butylated hydroxytoluene; ECG-epicatechin gallate; EC- epicatechin; KCN- potassium cyanide; PUFA- Polyunsaturated fatty acids; LOX-lipoxygenase.

## 2.3. Oxidative spoilage

Spoilage caused by lipid oxidation causes a variety of issues, including the development of off-flavors, changes in color and texture, and nutrient value alteration (Losada et al., 2007). Lipids can be oxidized in a variety of ways, including photo-oxidation, thermal oxidation, enzymatic oxidation, and auto-oxidation. The latter is defined as the spontaneous reaction of atmospheric oxygen with lipids (Hassoun & Emir Çoban, 2017). Typically, oxidation involves the reaction of oxygen with fatty acid double bonds. As a result, seafood with higher levels of polyunsaturated fatty acids (PUFAs) are more prone to oxidation. Other factors influencing the rate of oxidation include the availability of oxygen, light, the presence of metals, moisture, temperature, and the degree of unsaturation of the lipid (Domínguez et al., 2019; Maqsood, Benjakul, Abushelaibi, & Alam, 2014). Lipid oxidation occurs in three stages: initiation, propagation, and termination of free radicals. The initiation phase can begin with the abstraction of a hydrogen atom adjacent to a double bond in a fatty acid, which may be aided by light, heat, or metal ions to form a free radical. The free radicals react with oxygen to form peroxyl radicals, which then react with other lipid molecules to form hydroperoxides and a new free radical during the propagation phase. The process is terminated when free radicals associate to create non-radical products (Domínguez et al., 2019).

# 3. Edible coating as a biopreservation method for preprocessed fresh fish

## 3.1. General perspectives on edible coating

Edible coating has been extensively researched and proven to be an effective and green process for ensuring food quality and safety, including muscle foods, during refrigerated storage (Gagaoua et al., 2021). The implementation of edible films and coatings on the surface of foods aids in the control of moisture movement, lipid oxidation, microbial contamination, and enzyme activity, all of which are regarded as major contributors to food spoilage (Unalan et al., 2011, 2013). According to the largely increasing number of works in edible film and coating systems (Farris et al., 2014; Sapper & Chiralt, 2018; Suhag et al., 2020), bringing along their renewable resources and less independency to oil-
based conventional primary packaging, such systems can be recognized as “food and packaging waste reduction-focused sustainable system approach”. In particular, edible coating, is a thin layer of edible material obtained through the consecutive three stages; i) “wet process” in which biopolymer as a main matrix is dispersed or dissolved in a coating-forming solution, ii) the “deposition” in which coating solution is directly applied onto the surface of food via immersion or spraying techniques (Campos et al., 2011; Kang et al., 2013), and iii) “solvent evaporation” that takes place for drying of coating on the food surface.

Overall, the coating serves as a barrier against gases and water vapor, which reduces oxygen levels and helps to regulate microbial, enzyme, and oxidative reactions of the coated product during storage (Sapper & Chiralt, 2018). The composition of edible coatings affects their effectiveness and stability. Active compounds, mainly, proteins, bacteriocins, essential oils, and plant extracts are incorporated into coatings with the target of controlling or delaying active compound release towards food (Gagaoua et al., 2022). Consumer awareness for food safety and clean labels motivate the transition from synthetic to natural antimicrobial and antioxidant active compounds (Wong et al., 2020) in the fish preservation such as chilled fish fillets (Pan et al., 2021; Qiu et al., 2014). Antioxidants can also prevent oxidation by decreasing or delaying the production of free radicals, whereas antimicrobial compounds can inhibit or stop the proliferation of bacteria, hence boosting food safety (Wong et al., 2020).

Over the last decade, it has been demonstrated that a variety of edible coating materials, such as proteins (like gelatin and whey protein) and polysaccharides (like chitosan, alginate, and starch), can extend the shelf life of fishery products (Hosseini, Javidi, & Rezaei, 2016; Mohan et al., 2012; Neetoo et al., 2010; Yildiz & Yangilar, 2016), particularly in sea bass (Lateolabrax japonicas), rainbow trout (Oncorhynchus mykiss) (Andevari & Rezaei, 2011; Shokri et al., 2015), white leg shrimp (Litopenaeus vannamei) (Wang et al., 2015), silver pomfret (Pampus argentus) (Wu, Fu, et al., 2016), grass carp (Ctenopharyngodon idellus) (Yu, Li, et al., 2017), olive flounder (Paralichthys olivaceus) (Li et al., 2016), Asian sea bass (Lates calcarifer) (Chaijan et al., 2022), large yellow croaker (Larimichthys crocea) (Pei et al., 2022) and these studies are reported in Table 3 and Table 4 with relevant details.

<table><caption>**Table 3.** Different types of edible coating materials used for pre-processed fishery products under the dipping coating method.</caption><thead><tr><th>Fish/fishery product</th><th>Coating formulation</th><th>Results</th><th>References</th></tr></thead><tbody><tr><td>Mackerel (Auxis thazard)</td><td>Rice starch (RS)<br>Mon-pu (Glochidion wallichianum) leaf extract<br>(0.1 %,0.5 %,1.0 %)<br>RS<br>RS/MPE (0.02 %)<br>RS/MPE (0.1 %)<br>RS/MPE (0.5 %)<br>RS/MPE (1.0 %)</td><td>pH, TBARS, TVB-N, TMA, TVC, TPC↓<br>Heme protein degradation, Metmyoglobin↓<br>WHC, Liking score↑<br>Shelf-life (9 days) ↑</td><td>Chumsri et al. (2022)</td></tr><tr><td>Mackerel Tuna (Euthynnus affinis)</td><td>Xanthan gum (1.5 %)/Glycerol (0.5)/Ethanolic extract of Propolis (1 %)<br>Xanthan gum (1.5 %)/Glycerol (0.5)/Ethanolic extract of Propolis (2 %)<br>Xanthan gum/Ethanolic extract of Propolis (1:1)<br>Xanthan gum/Ethanolic extract of Propolis (1:2)</td><td>pH, Peroxide value, TBARS, TVB-N, K-value ↓<br>TVC, PTC,<br>Enterobacteriaceae, E. coli ↓<br>Pseudomonas fluorescens, Lactic acid bacteria ↓</td><td>Sheikha et al. (2022)</td></tr></tbody></table>
<table><thead><tr><th>Fish/fishery product</th><th>Coating formulation</th><th>Results</th><th>References</th></tr></thead><tbody><tr><td></td><td></td><td>Yeasts/molds ↓<br>Color, taste, odor↑<br>Shelf-life (7 days) ↑</td><td></td></tr><tr><td>Zander Fish Fillets<br>Sander lucioperca<br>(L.)</td><td>Xanthan gum (1 %)/Black tea extract (1 %)<br>Xanthan gum (1 %)/Black tea extract (2 %)<br>Xanthum gum (10 %)<br>Xanthum gum/Black tea extract (10:1)<br>Xanthum gum/Black tea extract (10:2)</td><td>pH, Cooking loss, TBA, FFA↓<br>Total bacterial count, Total yeast mold count, Total antioxidant activity↓<br>Physical quality, Shelf life (12 days) ↑</td><td>Anwar et al. (2022)</td></tr><tr><td>Silver pomfret<br>(Pampus argenteus)</td><td>Corn starch (2 % w/v) fumaric acid (0.5 % w/v)<br>Corn starch/fumaric acid (2:0.5)</td><td>pH, TVB-N, PV, TBA↓<br>Total mesophilic bacteria count↓<br>Total psychotropic bacteria count↓<br>Pseudomonas spp., Shewanella putrefaciens↓<br>Sensory Score ↑<br>Shelf-life (15 days) ↑</td><td>Remya et al. (2022)</td></tr><tr><td>Yellowfin Tuna<br>(Thunnus albacares)</td><td>Chitosan (1 %), Lemon peel extract (LPE) (1 %),<br>Chitosan (1 %)/LPE (1 %), Chitosan (2 %)/LPE (2 %)</td><td>pH, TVB-N, TMA-N, PV, TBARS, TPC↓<br>Sensory Score, Shelf-life (10–12 days) ↑</td><td>Sabu et al. (2020)</td></tr><tr><td>Blue tilapia<br>(Oreochromis aureus)</td><td>Fish gelatin (Gf-1.5 %), Kappa-carrageenan (Cr-0.75), pomegranate peels (PPE-0.5 %, 1.0 %, 1.5 %, 2.0 %)<br>Gf/Cr/PPE (1.5:0.75:0.5)<br>Gf/Cr/PPE (1.5:0.75:1.0)<br>Gf/Cr/PPE (1.5:0.75:1.5)<br>Gf/Cr/PPE (1.5:0.75:2)<br>Gf/Cr (1:2)</td><td>TVB-N, PV, TBARS↓<br>Microbial total count,<br>Aerobic count↓<br>Psychotropic count↓<br>Sensory Score, Shelf-life (10–15 days) ↑</td><td>Khojah (2020)</td></tr><tr><td>Shrimp (Litopenaeus vannamei)</td><td>Chitosan (1 %)/Fish gelatin (3 %)</td><td>pH, TVB-N, TBARS, K-Value, TVC↓<br>Color, Texture, Sensory value↑<br>Shelf-life (13 days) ↑</td><td>Farajzadeh et al. (2016)</td></tr><tr><td>Refrigerated Beluga sturegon (Huso)</td><td>Whey protein concentrate (8 %)/Glycerol (1 %)/Tween 80 (0.2 %)/Cinnamon essential Oil (1.5 %)</td><td>pH, TVB-N, PV, TBA, TVC↓<br>Sensory score↑<br>Shelf-life (16 days) ↑</td><td>Bahram et al. (2016)</td></tr><tr><td>Rohu (Labeo rohita)</td><td>whey proteins concentrate (8 % protein; w/w), Sorbitol, Glycerol<br>whey protein/Glycerol (1:2)<br>whey protein/sorbitol (1:2)<br>whey protein/glycerol/sorbitol (1:1:1)</td><td>pH, TBARS, POV, TVB-N,<br>Drip loss↓<br>Color, Thaw yield, Sensory score, texture ↑<br>Shelf life ↑</td><td>Khan et al. (2015)</td></tr><tr><td>Shrimp<br>(Parapenaeus longirostris)</td><td>Chitosan (1 %)/Garlic Oil (0 %, 0.5 %, 1.0 %, or 1.5 %)</td><td>pH, APC, TVB-N, PV, TBA, Weight Loss↓<br>Color, Shelf-life (11 days) ↑</td><td>Asik and Candogan (2014)</td></tr><tr><td>Atlantic salmon<br>(Salmo salar)</td><td>Whey protein concentrate (WPC) (8 % Protein w/w), WPC (1 %)/Glycerol (1 %), WPC (2 %)/Glycerol (1 %), WPC (1 %)/Sorbitol (1 %), WPC (2 %)/Sorbitol (1 %)</td><td>PV, TBARS, weight loss, drip loss, pH↓<br>Yield, sensory score, Shelf life↑</td><td>Rodriguez-Turienzo et al. (2011)</td></tr><tr><td>Refrigerated Bream<br>(Megalobrama amblycephala)</td><td>Sodium alginate (1.5 %)/Glycerin (10 %)<br>Sodium alginate (1.5 %)/Glycerin (10 %)/Vc antioxidant (5 %)</td><td>pH, TVB-N, TBA, TVC, Water loss, ↓<br>K-value, Sensory</td><td>Song et al. (2011)</td></tr></tbody></table>
<table><thead><tr><td>Fish/fishery product</td><td>Coating formulation</td><td>Results</td><td>References</td></tr></thead><tbody><tr><td></td><td>Sodium alginate (1.5 %)/Glycerin (10 %)/TP antioxidant (0.3 %)</td><td>characteristics↑<br>Shelf-life (12 days) ↑</td><td></td></tr></tbody></table>

Abbreviations: RS- Rice starch; MPE- Mon-pu (Glochidion wallichianum) leaf extract; TBARS- thiobarbituric acid reactive substances; TVB-N- total volatile basic nitrogen; TMA-trimethylamine; TVC- total volatile counts; TPC- total psychrophilic bacterial count; WHC- water holding capacity; PTC- psychotropic count; TBA-thiobarbituric acid; FFA-free fatty acids; PV-peroxide value; TMA-N- tri-methyl amino nitrogen; TPC- total aerobic plate count; POV- peroxide value; APC- aerobic Plate Count; Vc-purchased from (Biodee Biotechnology co., Ltd. Beijing, China) and added as an antioxidant; TP- purchased from (Keyi Chemical Co., Ltd., Zhengzhou City, China) and added as anti-oxidant.; LPE- Lemon peel extract; PPE-pomegranate peels; WPC- Whey protein concentrate.

<table><caption>**Table 4.** Effect of fish gelatin dip coating method on quality and shelf life of pre-processed fishery products.</caption><thead><tr><td>Pre-processed fish product</td><td>Gelatin concentration (w/w %) or (w/v %) and active compound/concentration</td><td>Additional preservation</td><td>Results</td><td>References</td></tr></thead><tbody><tr><td>Common carp (Cypirnius carpio)</td><td>Furcellaran (3.5 %) (w/v): gelatin (3:7)</td><td>Thyme (5 %)<br>Rosemary (5 %)<br>Stored at 4 °C</td><td>TVC, pH, Biogenic amines↓<br>Sensory score, Shelf-life (12 days) ↑</td><td>Tkaczewska et al. (2023)</td></tr><tr><td>Smooth-hound shark (Mustelus)</td><td>Gelatin 3 % (w/v)<br>Moringa oleifera extract 20 µg/m (w/v)</td><td>Stored at 4 °C without packaging</td><td>pH, TVB-N, weight loss, MDA↓<br>Mesophilic, psychrophilic, lactic acid bacteria↓<br>H<sub>2</sub>S bacterial count↓<br>Sensory score, color, odor↓<br>Overall acceptability↑<br>Shelf-life (6 days)↑</td><td>Mezhoudi et al. (2022)</td></tr><tr><td>(Ctenopharyngodon idellus)</td><td>Gelatin (8 % w/v)</td><td>Ginger essential oil (0.5 % v/v)<br>Stored at 4 °C</td><td>Weight loss, texture, TVC↓<br>Volatile flavor ↓<br>Color, taste, shelf-life (8 days) ↑</td><td>Li et al. (2022)</td></tr><tr><td>Red Snappers</td><td>Gelatin (8g)</td><td>kaffir lime leaf essential oil (1 %, 1.5 %.2 % w/v)<br>Stored at room temperature</td><td>pH, Moisture, elasticity ↓<br>TPC, fat content ↓<br>Color, ash content, protein content ↑<br>Shelf-life ↑</td><td>Febriana et al. (2021)</td></tr><tr><td>Asian seabass (Lates calcarifer)</td><td>Gelatin 4 % (w/v)<br>Grape seed extract 0.5 % (w/v)</td><td>Vacuum pressure (10 s),<br>Stored at 4 °C</td><td>Drip loss, TVC, putrescine, cadaverine ↓<br>Color, Shelf-life (12 days)</td><td>Zhao et al. (2021)</td></tr><tr><td>Blue tilapia (Oreochromis aureus)</td><td>Fish gelatin 1.5 % (w/v)</td><td>K-Carrageenan 0.75 % (w/v)<br>Extract of pomegranate peel (0.5 %, 1.0 %, 1.5 % and 2.0 % w/v)<br>Stored at 4 ± 1 °C</td><td>Total aerobic count, psychotropic bacteria ↓<br>TVB-N, PV, TBARS ↓<br>Sensory score, Shelf-life (30 days) ↑</td><td>Khojah (2020)</td></tr><tr><td>Golden pompano (Trachinotus blochii)</td><td>Gelatin 2.5 % and 5 % (w/v)</td><td>Vacuum packaging stored at 4 °C</td><td>TVB-N, TBARS, TVC, pH, drip loss ↓<br>Freshness index, color, gel strength ↑<br>Shelf-life ↑</td><td>Wei et al. (2019)</td></tr></tbody></table>
<table><thead><tr><th>Pre-processed fish product</th><th>Gelatin concentration (w/w %) or (w/v %) and active compound/concentration</th><th>Additional preservation</th><th>Results</th><th>References</th></tr></thead><tbody><tr><td>Golden Pomfret</td><td>Gelatin (1.2 %, 2.4 %, 3.6 % w/w)</td><td>Tea Polyphenol (0.4 % w/v)<br>Stored at +4 °C in high quality polyethylene zip lock bags</td><td>Weight loss, pH, AMC ↓<br>Myofibril degradation ↓<br>PUFA oxidation ↓<br>TMA, TVB-N, TMAO ↓<br>Shelf life (17 days) ↑</td><td>Feng et al. (2016)</td></tr><tr><td>Rainbow trout (Oncorhynchus mykiss)</td><td>Cold water fish skin gelatin (4 % w/v)</td><td>Oregano essential oil (1.2 % w/v)<br>Stored at 4 °C</td><td>pH, TVB-N, PV, TBA, TVC ↓<br>Shelf-life (16 days) ↑</td><td>Hosseini et al. (2016)</td></tr><tr><td>Fresh white shrimp (Penaeus vannamei)</td><td>Catfish skin gelatin 5 % (w/w)</td><td>Potassium sorbate (2 %) + Sodium tripolyphosphate (2 %)<br>Glycerol (20 %)<br>Stored in ice (0 °C)</td><td>ABC, PBC ↓<br>No significant change in pH and springiness<br>Color, Shelf-life (10 days) ↑</td><td>Jiang et al. (2011)</td></tr><tr><td>Salmon (Onchorhynchus keta)</td><td>Gelatin (4g)</td><td>Glycerol (60 % w/v), Sorbitol (1 % w/v)<br>Stored at 5 ± 1 °C</td><td>Moisture, Volatile Basic Nitrogen ↓<br>Peroxide Value ↓ fatty acid composition ↓<br>Color, Shelf-life (12 days) ↑</td><td>Heu et al. (2010)</td></tr></tbody></table>

Abbreviations: TVC-total viable count; TVB-N-total volatile basic nitrogen; MDA-malondialdehyde; H₂S-dihydrogen monosulfide; TPC-total plate count; PV-peroxide value; TBARS-thiobarbituric acid reactive substances; AMC-aerobic mesophilic count; PUFA-polyunsaturated fatty acid; TMA-trimethylamine; TMAO-trimethylamine oxide; TBA-thiobarbituric acid; ABC-aerobic bacterial count; PBC-psychrophilic bacterial count.

## 3.2. Edible coating materials for fish products

Because of raw fish's initial high microbial load and the increasing global demand for safe, convenient, and environmentally sustainable seafood, research for green and efficient preservation methods or technologies necessitates the application and development of novel packaging solutions (Augusto et al., 2016). Increasing awareness and thereby demand of consumers towards natural, healthful and safe products is closely associated with implementation of edible coatings as a promising approach to preserve or improve fish quality without compromising safety. Food quality changes caused by moisture transfer, oxidation processes, volatile flavor loss, or microbial growth can be reduced or even prevented by selecting appropriate matrices (Tavassoli-Kafrani et al., 2016). In this part, biopolymers, particularly proteins and polysaccharides, that have been intensively used to create coating matrices for shelf life studies of preprocessed fishes have been discussed.

### 3.2.1. Gelatin and its potential applications in pre-processed fish

Protein-based materials are one of the most commonly used raw materials. The global gelatin market size is expected to reach USD 13.14 billion by 2030, as per the new report by Grand View Research, Inc. with an expansion of Compound Annual Growth Rate (CAGR) of 9.5 % from 2022 to 2030 (Jiang et al., 2023). Because it is a plentiful product, mammalian gelatin has received research attention as a biopolymer. However, because of mammalian gelatin allergies, its use can cause both ethical and health issues. Furthermore, the food industry's byproducts are being used to create more sustainable solutions. Fish gelatin has gained popularity as an alternative to mammalian gelatin due
to its excellent biocompatibility, biodegradability, non-toxicity, and film forming abilities (Etxabide et al., 2017). Thermal denaturation or partial hydrolysis of collagen can be used to extract gelatin from fish industry byproducts such as heads, skin, bones, fins, muscle pieces, scales, viscera, and others (Huang et al., 2017). The use of pretreatments, with either acid or alkali, to allow the swelling of collagen to increase gelatin extraction is associated with collagen thermal denaturation. Depending on the end goal or application, gelatin properties can be adjusted by varying the pH, temperature ranges, and time applied during both pre-treatments and the extraction process (Ahmad et al., 2017). Fish gelatin extraction procedure depicted in Fig. 1 (Montero & Acosta, 2020).

<figure><img src="image_1.png" /><figcaption>**Fig. 1.** Simplified procedure highlighting the steps for extracting fish gelatin from Tuna skin (Montero & Acosta, 2020).</figcaption></figure>

Another important consideration is the incorporation of natural bioactive compounds, which add functionality to coatings. Because of fish gelatin's excellent biocompatibility, fish gelatin-based coating can gain active functional properties including antioxidant, antimicrobial, and antifungal by the addition of bioactive compounds (Etxabide et al., 2017). The basic physicochemical properties of gelatin, such as solubility, composition parameters, color, transparency, absence of odor and taste, are the main attributes that best define its overall commercial quality. Likewise, gel strength (expressed as a normalized “bloom value”) and viscosity should be regarded as the most important physical properties because they influence gelatin quality and potential applications. These distinct properties can be listed in two groups: (i) properties related to surface behavior, such as protective colloidal function, emulsion and foam formation and stabilization, adhesion and cohesion, and film forming capacity, and (ii) properties related to gelling behavior, such as gel formation, thickening, texturizing, and water binding capacity (Schrieber & Gareis, 2007). Thus, gelatin seems to continue growing and create unique opportunities in a broad range of applications including but not limited to food, packaging and pharmaceutical (Galus & Kadzińska, 2015). In packaging industry, due to widely-explored and accepted film forming properties of gelatin, its packaging film implementation to protect food from dryness, light exposure, and/or oxygen exposure during its shelf life were largely investigated (Lu et al., 2022). Gelatin, due to its highly hygroscopic nature, tends to swell or dissolve
when in contact with the surface of foodstuffs with high moisture content. Several research studies have been conducted to assess the overall effect of adding various substances, such as crosslinkers, strengthening agents, plasticizers, or additives with antimicrobial or antioxidant properties, to gelatin-based products in order to improve the functional properties of gelatin and the shelf-life of food products (Mellinas et al., 2016; Ortiz-Zarama et al., 2016). These properties improve when the intermolecular forces of protein chains are reduced by the action of molecular structures that modify their hydrophilic character or promote the formation of strong covalent bonds in the film's protein network (Alfaro et al., 2014). Zhao et al. (2016) demonstrated the feasibility of using a natural extract as a new natural crosslinker for the modification of gelatin (type B, from bovine bone) through the formation of hydrogen bonds between water and free hydroxyl groups of amino or polyphenol groups. The results showed that adding this extract to gelatin significantly increased gel strength when compared to untreated gelatin (Zhao et al., 2016). Combining gelatin with different biopolymers, such as whey proteins (Fakhouri et al., 2015; Zhao et al., 2016), starch (Fakhouri et al., 2015; Zhao et al., 2016), chitosan (Benbettaïeb et al., 2016a, b) or pectin (Gupta et al., 2014) could be a good strategy for the development of films with desirable mechanical and water resistance properties.

Gelatin has been developed to preserve the quality and increase the shelf-life of fish products (Feng et al., 2016). Gelatin coatings, for example, have been developed to act as a barrier for water and oxygen, preventing contamination of the food surface (Chen et al., 2019), resulting in reduced water loss, lipid oxidation, and bacterial growth. Furthermore, gelatin enzymatic hydrolysate has been shown to have antioxidant properties (Hosseini, Rezaei, et al., 2016) and can be applied to formulate antioxidant coatings. However, there is a lack of studies on coating applications of gelatin for processed fish (Table 4, Table 5) and some promising findings have been briefly summarized below.

<table><caption>**Table 5.** Characteristics of gelatin-biopolymer mixtures in an edible coating in fishery products.</caption><thead><tr><td>Formulation</td><td>Physical/chemical/mechanical Characteristics</td><td>Biological characteristics</td><td>Application</td><td>References</td></tr></thead><tbody><tr><td>Gelatin, chitosan, gallic acid Clove oil</td><td>Prevent the reduction of lightness in the fillets TBARS, pH ↓ Free thiol value (39–40 nmol thiol/mg) Retard the lipid and protein oxidation</td><td>combination of gallic acid and clove oil further enhanced the antimicrobial effect Antioxidant activity ↑ Shelf-life (5 days) ↑</td><td>Edible coating for fresh salmon fillet</td><td>Xiong, et al. (2021)</td></tr><tr><td>Sturgeon gelatin Portulaca oleracea extract</td><td>Moisture (62 %), Protein (13 %) ↑ pH (5.6), TBARS (0.48) ↓</td><td>TVC (4.8 CFU g-1) ↓ Total mold (1.3 CFU/g) ↓ Shelf-life (30 days) ↑</td><td>Edible coating for tuna fillet Crucian carp sausages</td><td>Tanha et al. (2021)</td></tr><tr><td>Gelatin, Furcellaran green tea or puerh extract</td><td>TBARS (0.47), pH (5.63) ↓ TVB-N (10.15) ↓</td><td>not cause any significant inhibition of any of the studied groups of microorganisms (6 log CFU/g) Did not extend the shelf life</td><td>Edible coating for salmon sushi</td><td>Kulawik et al. (2019)</td></tr><tr><td>Gelatin Red Pitaya Peel Methanol Extract, ε-polylysin</td><td>Gelatin Red Pitaya Peel Methanol Extract, ε-polylysin</td><td>free amino acid content (31.78 mg/100g) ↓ Antioxidant activity (DPPH-38.54 %, ABTS – 0.39 mM) Muscle fiber is more regular, Muscle Fiber was not significantly ↓</td><td>Edible coating for cray fish (Procambarus clarkia)</td><td>Liu et al. (2019)</td></tr></tbody></table>
<table><thead><tr><th>Formulation</th><th>Physical/chemical/mechanical Characteristics</th><th>Biological characteristics</th><th>Application</th><th>References</th></tr></thead><tbody><tr><td></td><td></td><td>broken (SEM)<br>Shelf-life (7 days) ↑</td><td></td><td></td></tr><tr><td>Fish gelatin<br>Grape seed<br>extract<br>Vacuum<br>impregnation</td><td>hardness (1500/g), springiness (40 %) ↑<br>more organized (α & β) protein structures<br>higher stabilization of myofibrillar protein ATPase activity (0.24 µmol/mb/min) ↑</td><td>Myofibril degradation ↓<br>Myofibril length (4.22 µm) ↑<br>Sulfhydryl groups (2.29-4.45 mol/105g protein) ↓<br>Disulphide bonds (113.60 %) ↓<br>Protein oxidation ↓<br>Aerobic Psychrophilic count (1.19 log CFU/g) ↓<br>Shelf-life (12 days) ↑</td><td>Edible coating for chilled tilapia fillets</td><td>Zhao et al. (2019)</td></tr><tr><td>Fish skin gelatin<br>Hydrolysate<br>Fish gelatin</td><td>pH (8.31 ± 0.16) ↓<br>FFA (1.68 ± 0.00) ↓<br>TVB-N (24.49 ± 0.67) ↓<br>Lipid oxidation (0.54 ± 0.07) ↓</td><td>Total microbial count (6.74 ± 0.74) ↓<br>Psychrotrophic count (4.74 ± 0.384) ↓<br>Lactic acid bacterial count (4.31 ± 0.584) ↓<br>Effectively extend the shelf life by 9 days ↑</td><td>Active coating for whole shrimp (Penaeus metguiensis)</td><td>Kouhdasht and Nasab (2019)</td></tr><tr><td>Fish gelatin,<br>cucurmin and<br>βCD based<br>emulsion</td><td>Weight loss (10 %), TVB-N (6.56 mg/100 g) ↓<br>PV (0.9 mmol cumene hydroperoxide kg<sup>-1</sup>) ↓<br>TBA (0.75 mg MDA kg<sup>-1</sup>) ↓</td><td>Inhibit the degradation of high salt soluble protein (HSP), Free amino acids (FAA) (0.504 %) ↓ TVC (6.2 log10 CFU/g) ↓<br>Pseudomonas spp. (6 log10 CFU/g) ↓<br>H<sub>2</sub>S-producing bacteria (5.5 log10 CFU g<sup>-1</sup>) ↓<br>Shelf-life (9 days) ↑</td><td>Edible coating for grass calf (Ctenopharyngodon idellus)</td><td>Sun et al. (2018)</td></tr><tr><td>Thunnus obesus<br>skin gelatin<br>Codium spp. or<br>Fucus<br>vesiculosus<br>extract</td><td>pH (6.55 ± 0.01), TBA (4.2 + 1.12) ↓<br>TVB-N (26) ↓<br>WI (33.11 ± 3.09), AE (8.36) ↓</td><td>TVC (6.8 log10 CFU g<sup>-1</sup>) ↓<br>Antioxidant activity DPPH (2.56 ± 0.70) ↑<br>Total phenolic content (0.079 ± 0.015) ↑<br>Shelf-life (12 days) ↑</td><td>Edible coating for tuna fillet (Thunnus obesus)</td><td>Vala et al. (2017)</td></tr><tr><td>Gelatin<br>Orange leaf<br>essential oil<br>(Citrus sinensis<br>(L) Osbeck)</td><td>Surface with holes and porous morphology pH (8.51), TVB-N (41.08), TBA (1.03) ↓<br>PV (3.9) ↓</td><td>Total Viable count (5.9 log CFU/g) ↓<br>Psychrotrophic bacteria count (6.4 log CFU/g) ↓<br>DPPH radical scavenging activity (52 %) ↑<br>Effectively extend the shelf life by 14 days ↑</td><td>Edible coating for pink shrimp (Parapenaeus longirostris L. 1846)</td><td>Alparslan et al. (2016)</td></tr><tr><td>Gelatin<br>Benzoic acid</td><td>VBN (15 mg/100g) ↓<br>Color (5.94 + 1.82) ↑<br>Flavour (6.18 + 1.47) ↑</td><td>Aerobic plate count (6.5 x 10<sup>6</sup> CFU/g) ↓<br>Anerobic plate count (10<sup>6</sup> CFU/g) ↓<br>Antimicrobial gelatin coating effectively controls both aerobic and anaerobic microbial growth and Effectively extending the shelf life.</td><td>Edible coating for refrigerated tilapia fillet</td><td>Ou et al. (2002)</td></tr></tbody></table>

Abbreviations: CFU/g- Colony-forming unit per gram; TBA-thiobarbituric acid; MDA-g-1- malondialdehyde per unit mass; WI- whiteness index; DPPH- 2,2-diphenyl-1-picrylhydrazyl; ABTS- 2,2'-azino-bis (3-ethylbenzothiazoline-6-sulphonic acid; SEM-scanning electron microscopy; HSP- high salt soluble protein;
FAA- Free amino acids; TVC- Total viable counts; H₂S-Dihydrogen monosulfide; VBN-Volatile basic nitrogen; TVB-N- total volatile basic nitrogen; PV- peroxide value.

Catfish gelatin has been used to extend the shelf-life of fresh white shrimp (*Penaeus vannamei*) by acting as an antimicrobial coating (Jiang et al., 2010). Moreover, rheological, structural and preservation effect of *Hemiramphus far* skin gelatin has been studied on the smooth hound fillet (Abdelhedi et al., 2018). Preservation effect of fish gelatin coating enriched with Curcumin/β-cyclodextrin emulsion was investigated on grass carp (*Ctenopharyngodon idellus*) fillets (Sun et al., 2018). Another earlier study has been conducted to characterize a sprayable edible coating using salmon bone gelatin on Atlantic salmon fillets (Char et al., 2019) and shelf life extension of whole shrimp was studied with an application of active edible coating made up of fish skin gelatin hydrolysate (Kouhdasht and Nasab, 2019). In the implementation of active packaging approach into coatings, many recent researches highlighted the multi-functionality of (bio) active coatings on pre-processed fish (Mezhoudi et al., 2022; Zhao et al., 2021) including targeted and controlled delivery of bioactive compounds in addition to the functionality of traditional coatings. Recently, an active edible gelatin coating extracted from the skin of gray triggerfish (*Balistes capriscus*) was enriched with a leaf extract and applied to the fillet of the smooth-hound shark (*Mustelus*) (Mezhoudi et al., 2022).

3.2.2. Whey proteins and their potential applications in pre-processed fish

Whey proteins (WP) are the most economically and technologically interesting fraction of whey, accounting for nearly 15%–20% of total milk proteins (De Castro et al., 2017; Schmid & Müller, 2019). The WP composition varies according to cheese type, manufacturing process, coagulation method (acid or enzyme), milk origin (bovine, ovine, or caprine), feeding regime, and lactation stage. These differences have a significant impact on emulsification and other functional properties (Ramos et al., 2012; Yadav et al., 2015). The WP are heat labile, dephosphorylated, and less sensitive to calcium. They are compact secondary, tertiary, or quaternary globular molecules with various combinations of cross-sulfur bonds (Henriques et al., 2016).

Because of advancements in membrane and ion exchange techniques, the global production of WP
has increased dramatically, making it easier to recover WP with preferred and appropriate functional
properties (acid stability, gelation, film-forming efficiency, and aeration) and emulsification
characteristics (Perez-Gago & Krochta, 2002). The WP recovery processes such as ultrafiltration,
diafiltration, electrodialysis, gel filtration, ion-exchange chromatography, and reverse osmosis are
commonly used (Milani & Tirgarian, 2020). whey protein concentrates (WPC) are created by
selectively removing high molecular-weight fractions from whey permeate during the ultrafiltration
process (Bhattacharjee et al., 2015). The WPC is low in fat and cholesterol but high in bioactive
compounds when compared to other commercial whey formulations. Whey protein isolates (WPI)
are made by removing minerals and lactose from WPC through an additional ion-exchange
chromatographic step (Khwaldia et al., 2004). The WPI are high in protein but low in bioactive
compounds. Both of these fractions have these essential characteristics that make them suitable for
application in food: high amino acid and protein content; low calorie, fat, and sodium content; lack
of pathogens and toxic metabolites; biocompatibility and generally recognized safe status; ready
availability; and low price (Yadav et al., 2015). Using the spray-drying process, both WPC and WPI
fractions can be dehydrated and used in coating applications.
The WP film is primarily composed of a dry, highly interacting polymer network with a three-dimensional gel-like structure. Regardless of the film-formation techniques used, the final films can result in a spatially reorganized gel arrangement containing the additional film forming agents (Henriques et al., 2016). The WP films are made using the solvent casting method, in which the film solution is poured onto flat surfaces to form a dried gel, which is then used to wrap food products. In the case of WP coatings, the food products are typically immersed in the film-forming solution for a specific time period (30 s or 1 min) to ensure complete exposure of the food surface with good adherence and perfect integrity, and then air-dried (Fernandes et al., 2020; Ramos et al., 2012). The WP differs from other film-forming polymers in several ways, including its amphiphilic nature, conformation, denaturation, and electrostatic charges. Charge density and the hydrophilic-hydrophobic balance, for example, can modify the WP conformation, as these factors will eventually define the physical and mechanical properties of films/coatings (Perez-Gago & Krochta, 2002; Karaca et al., 2019). Because of its compact globular structure and small molecular size, native WP cannot be considered as a good adhesive polymer candidate. Under certain conditions, the globular structure can be reformed into relatively linear structures and then into irreversible aggregates via a thiol-disulfide exchange. Native WP can also form coherent films based on low-energy bonding such as electrostatic interactions, hydrogen bonding, or van der Waals force (Schmid & Müller, 2019).

The process of creating WP-based films and coatings can be broken down into several steps, which include dissolving WPC or WPI (5%-12%) in distilled water, adjusting the pH of the solution (either 7 or 8), and then heating (80-90 °C/10-30 min) for protein denaturation (Perez-Gago & Krochta, 2002). Other ingredients can be added before or after heating, depending on their compatibility. Heat-tolerant ingredients (prebiotics, starches, and blends) are added early in the process, while heat-sensitive ingredients (antioxidants, antimicrobial compounds, and probiotics) are added after heating (Fernandes et al., 2020).

Even though several heat treatment methods are available for gel formation, denaturation is the most commonly used technique for forming coherent films. The majority of the hydrophobic and sulfhydryl (SH) groups in native WP are buried in the center of the protein molecule. Denaturation exposes WP's functional and hydrophobic groups, resulting in the formation of a three-dimensional chemical network that promotes intermolecular disulphide (S-S) bonding and hydrophobic interactions during drying. Due to food intermolecular interactions, WP films produced without heat treatment easily break into tiny parts during drying (Ramos et al., 2012). At pH 6.0, the thermal denaturation process occurs in two steps. The first step occurs when β-lactoglobulin dimers dissociate into monomers at temperatures above 40 °C. Denaturation at temperatures above 65 °C unfolds the β-lactoglobulin molecule and exposes hydrophobic and thiol groups, which contribute to the formation of smaller aggregates with β-lactoglobulin or other thiol-containing proteins. These smaller aggregates then interact in a second step to form high-molecular-weight irreversible aggregates (Khwaldia et al., 2004; Schmid & Müller, 2019).

After forming a heat or cold-set gel, WP films are formed by dehydrating the gel. Drying under ambient conditions, usually between 21 and 23 °C and 50 % relative humidity (RH), is a common dehydration method. Nonetheless, when applying edible coatings to foods, the drying process must be handled carefully because quick drying changes the characteristics of the film, making it thinner, stiffer, and less flexible (Ramos et al., 2012). The drying method, and final film or coating properties are significantly affected by changes in drying rate, temperature, and humidity.
Casting, dipping, extrusion, enrobing, fluidization, foaming, spraying, and UV polymerization are
currently used to produce edible WP films/coatings (Ramos et al., 2012). Monitoring the processing
conditions is critical because changes in handling environments can alter the kinetics and reaction
mechanisms involved in film formation (Perez-Gago & Krochta, 2002; Ramos et al., 2012). To be
considered edible, the WP film-formation methods must continue to be suitable for holding foods
despite changes in pH, salt levels, heat, enzymatic variation, drying, solvent usage, and other
chemicals. Furthermore, plasticizers and other additives must work well with the biopolymer
(Henriques et al., 2016).

The WP can be used to coat fish fillets to overcome any inherent changes in quality during storage (Wang et al., 2019). Recent work demonstrated that WP coating fortified with crude ginger extract could extend the shelf-life of Asian sea bass fish by 15 days when compared to the control (8 days) (Chaijan et al., 2020). Furthermore, the same authors have lately addressed the new preservative approach of pre-treated fish with plasma activated water (PAW) before coating with WPI-crude ginger extract (Chaijan et al., 2022). The PAW-treatment demonstrated the potential to reduce the microbial load in fish steak, while the WPI coating acted as an oxygen barrier, limiting oxygen participation in the oxidation reactions caused by the early PAW treatment. The presence of phenolic compounds in the ginger extract has been proposed to further delay lipid oxidation through free radical scavengers and delaying the proliferation of spoilage microorganisms during fish storage. With this treatment, the shelf-life of Asian sea bass steak was extended to 30 days. In the case of salmon, which is high in fat, reduction in shelf life is due to unsaturated fatty acids that are easily oxidized. The WP coating on frozen king salmon reduced moisture loss by 42 %–65 % within the first three weeks of storage and slowed lipid oxidation as seen through a decreased peak peroxide value (Stuchell & Krochta, 1995). Coatings applied after freezing improved thaw yield, reduced drip loss, and altered color constraints in frozen and thawed fillets in a study comparing application of WP coating before and after freezing Atlantic salmon (Rodriguez-Turienzo et al., 2011). Another study demonstrated that ultrasound-treated WP coatings with or without microbial transglutaminase (TGase) addition delayed lipid oxidation in frozen Atlantic salmon as effectively as heat treated coatings containing TGase (Rodriguez-Turienzo, et al., 2013). Coatings made with 13 % WP were found to reduce lipid oxidation during frozen storage of gutted Kilka (Hasanzati Rostami et al., 2010).

3.2.3. Chitosan and its coating applications for pre-proceed fish

Chitosan, derived from the second most abundant biopolymer chitin which is found in the exoskeleton
of crustaceans such as crabs and shrimps, in fungal cell walls, and in other biological materials, has
been studied *in vitro* and in complex food matrices as an edible film and coating (Nanda et al., 2021).
In addition to its well-known and proven film-forming properties, the increased awareness of
upcycling of waste or by-products to value-added bio-based products, and research in food packaging
has intensified the focus on broadening the implementation of chitosan (Nanda et al., 2021). For
example, the upcycling of seafood wastes (Chakravarthy & Edwards, 2022) into chitosan could
contribute to a circular economy in multiple ways including reducing global seafood waste, reducing
loss of resources, and increasing waste-streamed bio-based and biodegradable coating materials, and
adding longer shelf-life to food products.

Implementing chitosan produced from seafood waste as a preservative or as a quality enhanced
coating onto fish product is recognized as a sustainable approach. Soares et al. (2015) evidenced that
Atlantic salmon products coated with 1.5 % (w/v) chitosan performed better in terms of color stability and control of microbial contamination in frozen and thawed samples. Surimi gels made from African catfish (*Clarias gariepinus*) with 1.5 % chitosan, on the other hand, showed the greatest improvement in gel strength, whiteness, and water holding capacity. The lowest Total volatile basic nitrogen (TVB-N) value was obtained by incorporating 2.0 % (w/w) chitosan into gels. During the storage period, peroxide value (PV) and thiobarbituric acid (TBARS) increased at a slower rate in the chitosan-treated gels than in the control gels (Amiza & Kang, 2013). Bonilla et al. (2018) reported an 8-day increase in shelf-life in catfish fillets treated with a chitosan solution. Similarly, the chitosan and propolis extract, contains the major source of polyphenols (Ebadi et al., 2019), reduced the extent of lipid oxidation, as measured by TBA and free fatty acid (FFA), improving the TVB-N and pH values of *Nemipterus japonicus* compared to untreated samples, and thus extending the shelf-life of *N. japonicus* fillet by approximately 10 days (Ebadi et al., 2019). Fernandez-Saiz et al. (2013) investigated the biotic effectiveness of chitosan as an internal layer of fresh hake (*Merluccius*) and sole (*Solea*) fillet packaging. Under vacuum packaging conditions, the microbial species, primarily total aerobic mesophilic bacteria, hydrogen sulfide-producing bacteria, and *Pseudomonas*, were significantly inhibited by increasing the lag phase. Coatings not only promoted preservation but also had an effect on cooking characteristics as seen in fish fingers coated with chitosan and chitosan nanoparticles, resulting in the reduced oil uptake from 16.42 % to 4.56 % and increased moisture content from 34.61 % to 52.7 %. Essentially, the shelf-life increased by up to one month when compared to non-coated fish finger (Osheba et al., 2013). Jeon et al. (2002) found that chitosan coating reduced or prevented moisture loss, lipid oxidation, and microbial growth in herring and cod, resulting in a moderate to strong viscosity-dependent preservative effect in both fish. Interestingly, chitosan coating extracted from crab processing waste in cooked comminuted samples of herring flesh demonstrated lower peroxide values, TBA, and total volatile aldehydes than low viscosity chitosan (Kamil et al., 2002). The chitosan treatment preserved favorable sensory properties in sardine for a longer period, extended the lag phase, and reduced the formation of volatile bases and oxidation products. When compared to untreated samples, it also helped to maintain good water holding capacity, reduce drip loss, and improved the textural changes. The shelf-life of 1 % and 2 % chitosan treated samples was 8 and 10 days, respectively, compared to only 5 days for untreated sardine (Mohan et al., 2012). In another work, chitosan coating not only preserved the sensory quality of refrigerated grass carp (*Ctenopharyngodon idellus*) fillets but also significantly reduced microbial population, characteristic spoilage biogenic amines, TVB-N, increased physiochemical and sensory qualities, and extended shelf-life (Yu, Jiang, et al., 2017).

In addition to mono polymer chitosan systems, blending of chitosan with other type of biopolymer
has been proposed. Aleman et al. (2016) found that incorporating a chitosan-gelatin matrix into
Pacific white shrimp (*Litopenaeus vannamei*) cooking juice increased the lag phase of total viable
microorganisms and enterobacteria to 15 and 10 days, respectively, and significantly inhibited growth
of these microbial groups. The TVB-N and pH values decreased as well, and no S. aureus or lactic
acid bacteria were detected in any batches during storage. Feng et al. (2016) found that 0.4 % chitosan
coating blended with 7.2 % gelatin significantly reduced the deterioration of golden pomfret
(*Trachinotus blochii*) fillets during 17 days of cold storage. According to Jasour et al. (2015),
combining enzyme chitosan coating with endogenous enzyme lactoperoxidase (LPO) increased the
shelf-life of rainbow trout while significantly reducing the number of *Shewanella putrefaciens*,
*Pseudomonas fluorescens*, and psychrotrophic and mesophilic bacteria. When compared to control
samples, the coating treatments with chitosan and LPO increased the shelf-life of trout fillets by at
least 4 days. Similar to LPO, lysozyme obtained from hen egg white is one of mostly used enzyme for active packaging due to its generally recognized as safe (GRAS) status, and its stability and activity in food and packaging system at cold temperatures (Unalan et al., 2011, 2013). The use of chitosan-lysozyme coating has a high efficiency in inhibiting microorganism growth, lipid oxidation, and preserving sensorial quality, making it a promising method for preserving the quality and extending the shelf-life of refrigerated large yellow croaker (*Larimichthys crocea*) (Wu et al., 2018).

As discussed earlier, phenolic-rich natural plant extracts and phenolic compounds are of high interest in active film and coating systems. For instance, chitosan film wrapping with young apple polyphenols delayed the increase in microbial load, PV, thiobarbituric acid reactive substances (TBARS), TVB-N, and pH values caused by microbial growth and oxidation of lipids and proteins in grass carp fillets (GCF) and silver carp fillets during cold storage, along an increase of water holding capacity, functional properties of soluble myofibrillar protein, external acceptability, textural properties, and amino acids (Ramezani, Zarei, & Raminnejad, 2015; Sun et al., 2018; Yu, Jiang, Xu, & Xia, 2017). The acceptability was evaluated based on sensory evaluation system comprised of five indices, including odor, color, tissue elasticity and surface character. It was reported that the polyphenol-incorporated chitosan film could retard the spikes in colony forming units (CFU), PV, TBARS, TVB-N, pH, and *b*\* values caused by microbial reproduction and lipid and protein oxidation in GCF during cold storage (Sun et al., 2018). Yuan et al. (2016) discovered that during the later stage of iced storage, the melanosis and sensory scores, TVB-N, and total aerobic plate counts of white shrimp coated with chitosan in the combination of pomegranate peel extract (PPE) were lower than those treated with chitosan coating or PPE alone. Wang et al. (2015) demonstrated that 1 % chitosan coatings improved TVB-N amounts, K values, total viable count (TVC), hardness, and springiness of postharvest white leg shrimp stored at 4 °C for 10 days. Similarly, coating *P. japonicas* with gallic acid and chitosan inhibited microbial growth, protein decomposition, biogenic amine formation, lipid oxidation, nucleotide breakdown, and the preservation of better sensory characteristics during storage, resulting in a 6-day shelf-life extension (Wu, Li, et al., 2016). Since amines are synthesized at the end of a product's shelf life, their levels can be recognized as spoilage indices as compared to quality indices (Ozogul & Ozogul, 2006). A combination of chitosan (1.5 % w/v), tea polyphenol (0.2 % w/v), and rosemary extract (0.2 % w/v) effectively maintains good quality and extends the shelf-life of refrigerated *L. crocea* by 8–10 days compared to the untreated group (Li et al., 2012). Similarly, chitosan matrix incorporated propolis extract, contains the major source of polyphenols, reduced the extent of lipid oxidation, as measured by TBA and free fatty acid (FFA), improving the TVB-N and pH values of *Nemipterus japonicus* compared to untreated samples, and thus extending the shelf-life of *N. japonicus* fillet by approximately 10 days (Ebadi et al., 2019). Citric acid or licorice extract can significantly improve chitosan's preserving function by slowing lipid oxidation and inhibiting microbial growth, as reflected in TBA and TVC, respectively, and thus extend the shelf-life of Japanese sea bass fish fillets during refrigerated storage (Qiu et al., 2014). Cao et al. (2020) treated snakehead fish fillets with a chitosan solution containing chlorogenic acid (CGA). In their study, the lipid and protein oxidation were found to be inhibited by 2 % chitosan, 0.5 % CGA in 2 % chitosan, and 1.0 % CGA in 2 % chitosan coated fish fillet. All CGA/CS coatings caused a brown color and delayed the increase in pH. The combination of vanillin (2 mg/mL) and 1 % chitosan coating improves the sensory and chemical quality of turbot (*Scophthalmus maximus*) fillets and extends shelf life by 6–7 days (Li et al., 2020). Similarly, chitosan coating combined with sodium phytate, the sodium salt of phytic acid, effectively inhibited bacterial growth, increased pH and TVB-
N values, and decreased sensory properties of fresh Antarctic krill (Euphausia superba) during partially frozen storage (Liu et al., 2020).

As exemplified above, great efforts are being devoted to the development of chitosan films and coatings, however, high vapor permeability and water sorption of chitosan at the moist environments lowers its protection performance, therefore the combination of chitosan with essential oils which are concentrated hydrophobic liquids containing volatile aroma compounds, has deserved special attention in food packaging by developing an active coating layer with enhanced water vapor barrier performance. Essential oil is primarily composed of low-molecular weight aliphatic and aromatic secondary metabolite wherein terpenes (i.e., *p*-cymene, limonene, or pinene), terpenoids (i.e., thymol, carvacrol, or menthol), and phenylpropenes (i.e., eugenol, cinnamaldehyde, or vallinin) are key compounds (Kumar et al., 2020).

In Table 6, studies on chitosan coatings incorporated essential oils as preservation approach for pre-processed fish products have been summarized. Cinnamon oil containing chitosan active coating successfully inhibited lipid and microbial oxidation in refrigerated rainbow trout fillets and extended their shelf-life until the end of the storage period (Day 16) without significant loss of texture, odor, color, or overall acceptability, and without significant microbial growth, whereas control samples had a shelf-life of only 12 days (Ojagh et al., 2010). Rezaeifar et al. (2020) combined lemon verbena essential oil and extract with chitosan coating in rainbow trout meat, resulting in an enhancement of sensory characteristics. Chitosan coating (1.5 %) combined with lemon and thyme essential oils maintained the firmness, retarded lipid and protein oxidation, improved color and texture properties, reduced microorganism counts, and extended shelf-life of grass carp and European eel (Anguilla) fillets to 16 days (Cai et al., 2018; El-Obeid et al., 2018). Furthermore, the chitosan-citrus essential oil composite coating exhibits strong superoxide anion radical scavenging activity as well as hydroxyl radical scavenging activity with excellent antibacterial properties. It inhibits microbial growth more effectively, reduces lipid oxidation and peroxide production, and extends the shelf-life of Pacific mackerel by about 3 days (Li et al., 2019).

<table><caption>**Table 6.** The preservation effect of utilizing chitosan and essential oil-based edible coatings on pre-processed fishery products under the dip coating method.</caption><thead><tr><th>Fish/fishery product</th><th>Preservative formulation</th><th>Coating</th><th>Results</th><th>References</th></tr></thead><tbody><tr><td>Hairtail (Trichiurus haumela)</td><td>Chitosan 1 % (w/v)<br>Eugenol 11.61 % (w/w)</td><td></td><td>pH, TBA, TVB-N, TVC ↓<br>WHC, Sensory score ↑<br>Shelf-life (18 days) ↑</td><td>Liu et al. (2021)</td></tr><tr><td>Scomberoides commersonnianus</td><td>Chitosan 1 % (w/v)<br>Whey protein 1 % (w/v)<br>Tarragon Artemisia dracunculus essential oil (TEO)</td><td></td><td>pH, TVB-N, TBARS, FFA ↓<br>No difference in TMC and PTC<br>Sensory acceptance score ↑<br>Shelf-life (12 days) ↑</td><td>Farsanipour et al. (2020)</td></tr><tr><td>white shrimp (Litopenaeus vannamei)</td><td>Chitosan 1.5 % (w/v)<br>Fully deacetylated chitosan (FDCH) 1.0 %, 1.5 % (w/v)<br>Clove essential oil (CEO) 0.5 %, 0.25 % (w/v) kojic acid (0.5 %, 0.25 % (w/v))</td><td></td><td>pH, TVB-N, weight loss, TPC ↓<br>Hardness, Sensory score ↑<br>Shelf-life (15 days) ↑</td><td>Liu et al. (2020)</td></tr><tr><td>Rainbow trout (Oncorhynchus mykiss)</td><td>Chitosan 20 g/L (w/v)<br>Alginate 2 % (w/v)<br>Mentha Piperita essential oil</td><td></td><td>TBARS, TVBN ↓<br>TVC, TPC, LAB ↓<br>Shelf-life (12 days) ↑</td><td>Raeisi et al. (2020)</td></tr></tbody></table>
<table><thead><tr><th>Fish/fishery product</th><th>Preservative formulation</th><th>Coating</th><th>Results</th><th>References</th></tr></thead><tbody><tr><td></td><td>(MPEO) 0.2 % (w/v)<br>Artemisia dracunculus essential oil (ADEO) 0.2 % (w/v)<br>Zataria multiflora essential oil (ZMEO) 0.2 % (w/v)</td><td></td><td></td><td></td></tr><tr><td>Rainbow trout<br>(Oncorhynchus mykiss)</td><td>Chitosan 1 % (w/v)<br>Lemon verbena Essential oil 0.5 %, 1 %, 2 % (v/v)<br>Lemon verbena extract 0.5 %, 1 %, 2 % (v/v)</td><td></td><td>TBARS, PV, TVN, pH, TVC ↓<br>Psychrotrophic bacteria, H<sub>2</sub>S producing bacteria,<br>Enterobacteriaceae spp. ↓<br>Shelf-life (16 days) ↑</td><td>Rezaeifar et al. (2020)</td></tr><tr><td>frozen tambaqui fillet<br>(Colossoma macropomum)</td><td>Chitosan 2 % (w/v)<br>Clove essential oil 0.08 % and 0.16 % (w/v)</td><td></td><td>pH, Moisture, TBARS, CPB count ↓<br>Sensory score ↑<br>Shelf-life (120 days) ↑</td><td>Vieira et al. (2019)</td></tr><tr><td>Pacific mackerel<br>(Pneumatophorus japonicus)</td><td>Chitosan 1.5 % (w/v)<br>Citrus essential oil 1.5 % (w/v)</td><td></td><td>Drip loss, pH, TBARS, TVB-N, BA ↓<br>TVC, TPC ↓<br>Superoxide anion radical scavenging activity ↑<br>Hydroxyl radical scavenging activity ↑</td><td>Li et al. (2019)</td></tr><tr><td>Tilapia (Oreochromis niloticus)</td><td>Chitosan 2 % (w/v)<br>Carvacrol 0.125 %, 0.25 % (w/v)</td><td></td><td>TVBS, pH, WRC ↓<br>Total aerobic count, Total coliform count<br>V. parahaemolyticus, V. cholerae ↓</td><td>Chaparro-Hernandez et al. (2015)</td></tr><tr><td>Japanese sea bass<br>(Lateolabrax japonicas)</td><td>Chitosan 1.5 % (w/v) citric acid 0.6 % (v/v)<br>Citric acid 0.5 % (w/v)<br>Locorice extract 1 % (w/v)</td><td></td><td>pH, TVB-N, TBARS, TPC ↓<br>Sensory score ↑ shelf life (12 days) ↑</td><td>Qiu et al. (2014)</td></tr><tr><td>Large yellow croaker<br>(Pseudosciaena crocea) fillet</td><td>Chitosan 1.5 % (w/v)<br>Rosemary extract 0.2 % (w/v)<br>Tea polyphenol 0.2 % (w/v)</td><td></td><td>pH, PV, TBARS, TVC, TVB-N, K value ↓<br>Sensory score, shelf-life (8–10 days) ↑</td><td>Li et al. (2012)</td></tr></tbody></table>

Abbreviations: TBA-thiobarbituric acid; TVB-N- total volatile base and nitrogen; TVC- total viable count; WHC- water holding capacity; TBARS- thiobarbituric acid reactive substances; FFA-free fatty acids; TMC- total mesophilic count; PTC- psychrotrophic bacteria values; TPC- total aerobic plate counts; PV- peroxide value; TVN- total volatile nitrogen; CPB- culturable psychotropic bacteria; BA-biogenic amine; TPC- total psychrotrophic count; WRC- water retention capacity.

### 3.2.4. Mode of action of chitosan

Several review papers have described the antibacterial mechanisms of chitosan, however, in the light of increased research efforts for chitosan, description needs an update with the most recent and relevant studies. Chitosan has antibacterial activity against both gram-positive and gram-negative bacteria. Chitosan's mechanism of action involves the modification of the cell envelope or the disruption of the integrity of the cytoplasmic membrane (Devlieghere et al., 2004; Hu & Ganzle, 2019). Chitosan exhibits antimicrobial activity only when its pH ranges between 6.2 and 7, and it can be dissolved in 1%–2% acetic acid concentration or as chitosan-based packaging film (Hu & Ganzle, 2019). However, chitosan's antimicrobial activity is influenced by several other factors, including molecular weight, degree of deacetylation, and degree of polymerization, as well as pH, as it is only soluble in acidic conditions (Ma et al., 2017; Qi et al., 2004). Chitosan with a higher degree of
deacetylation has a higher positively charged density, resulting in a stronger electrostatic interaction with the cell surface and increased antimicrobial activity (Chung et al., 2004; Younes et al., 2014).

Above all, when used in food preservation, chitosan can act in a variety of ways. According to Goy et al. (2009), chitosan can bind to bacterial DNA, inhibiting mRNA transcription and possibly interacting with microorganism surface molecules. However, the mechanism of DNA binding remains unknown due to chitosan's inability to reach targets within the cytoplasm; it could be hypothesized that it binds the bacterial membrane and disrupts it (Bowman & Leong, 2006; Ma et al., 2017). However, because of its larger size, chitosan is unable to cross the cell wall and interact directly with the membrane, more research is needed to understand this (Raafat et al., 2008).

Another mechanism that could alter the cell membrane is the polycationic nature of chitosan (Hu & Ganzle, 2019; Ma et al., 2017). Chitosan contains glucosamine amine groups ($NH_3^+$), and many microorganisms have negatively charged surface components; interaction of those compounds causes extensive alteration of the cell surface, resulting in leakage of intracellular substances and cell death. The lipopolysaccharide and teichoic acid found in gram-positive and gram-negative bacteria play a major role in binding chitosan to the cell membrane, altering, destabilizing, and ultimately disrupting cell wall dynamics (Ganan et al., 2009; Raafat et al., 2008). For example, 0.01–5 g/L chitosan increases *E. coli* and *Salmonella* sp. outer membrane permeabilization (Helander et al., 2001; Hu & Ganzle, 2019).

Chitosan may have antifungal properties similar to its antibacterial properties (Ma et al., 2017). Fungal and yeast strains such *Fusarium oxysporum*, *Botrytis cinerea*, *Rhizoctonia solani*, *Candida lambica*, and *Phomopsis asparagi* are inhibited by chitosan's antifungal potential. Chitosan and its derivative possess broad-spectrum antifungal activity and can inhibit the germination of sporangia, germination tubes, and mycelium development of pathogenic fungi (Qin et al., 2020). Further, through the action of plant host hydrolytic enzymes, fungal pathogens' cells are lysed following which chitosan can enter the nuclei of fungi and interfere with RNA and protein synthesis (Muzzarelli et al., 1986). Tayel et al. (2010) investigated the mode of action of chitosan with *Candida albicans* and discovered an interaction between chitosan and the microbe's cell, resulting in severe swelling and cell wall lysis. Furthermore, chitosan inhibited the growth of *Aspergillus flavus*, *Alternaria alternate*, *Fusarium solani*, and aflatoxin production in culture (Dutta et al., 2009; Reddy et al., 1997). Fungi are thought to be more responsive to chitosan than bacteria (Goy et al., 2009).

Chitosan's strong hydrogen-donating ability makes it a promising antioxidant for food preservation with enhanced scavenging activity. The antioxidant activities of chitosan are enhanced by oxidation products such as reactive oxygen species (ROS), superoxide anion radicals, hydroxyl radicals, and hydrogen peroxide. Aside from that, chitosan's low molecular weight and higher degree of quaternization have an influence in its antioxidant capacity (Vinsova & Vavrikova, 2011). Chitosan was investigated as a potential antioxidant in several studies, where the lipid peroxidation of phosphatidylcholine and linoleate liposomes was inhibited by chitosan scavenging of hydroxyl radicals (Feng et al., 2008).

It is also worth mentioning that Yen et al. (2008) reported that chitosan from crab shells with varying degrees of deacetylation had higher scavenging activities as well as high chelating abilities. Chitosan's high chelating capacity makes it useful as a food supplement (Vinsova & Vavrikova, 2011).
Consumer demand for synthetic antioxidant-free sea foods may encourage greater use of chitosan (Chiang et al., 2000), because chitosan significantly reduced lipid oxidation even after cooking. For example, viscous nature of chitosan has been found to protect cooked cod from oxidation (L'opez-Caballero et al., 2005; Shahidi et al., 2002). Similarly, chitosan combined with modified atmosphere packaging improved lipid stability in cold-storage lingcod (*Ophiodon elongates*) for 21 days (Duan et al., 2010). Another study showed chitosan enriched with cinnamon oil may delay peroxidation in rainbow trout (*Oncorhynchus mykiss*) after a 16-day storage period (Ojagh et al., 2010). A proposed antibacterial mechanism of chitosan is depicted in Fig. 2.

<figure><img src="image_2.png" /><figcaption>Fig. 2. Summary of the postulated mechanism of antibacterial activity of chitosan.</figcaption></figure>

4. Essential oils as preservative agents in edible coating for fresh pre-processed fish

Essential oils are complex mixtures of volatile organic compounds extracted from various plant parts such as leaves, barks, stems, roots, flowers, and fruits (Calo et al., 2015). Essential oils frequently have antimicrobial and antioxidant properties, and a growing demand for natural preservatives has led to the consideration and use of essential oils as possible chemical preservative substitutes (Jayasena & Jo, 2013). Oregano, rosemary, thyme, laurel, sage, cinnamon, clove, and basil are the most used essential oils in fish and fish products as antimicrobial and antioxidant compounds. Other essential oils, such as cumin and spearmint (Cai et al., 2015), *Zataria multiflora* Boiss (Emir Çoban & Tuna Keleştemur, 2017), horsemint (Heydari et al., 2015), orange, grapefruit, mandarin, and lemon (Durmus, 2020), turmeric, and lemongrass (Masniyom et al., 2012) are also explored by researchers for their taste enhancing as well as antimicrobial properties.

Essential oils can be used in a variety of ways in the fish processing industry, the most common being the direct application of essential oils or their compounds to fish and other seafoods. However, higher concentrations of essential oils may cause undesirable sensory characteristics on treated fish, reducing
the acceptability of fish and seafood products (Atares & Chiralt, 2016). To tackle the organoleptic challenges of direct application of essential oils onto fish and other seafoods, the incorporation of essential oils into active films and coatings for food protection has been proposed. In fact, several studies have been so far conducted to evaluate the effectiveness of essential oil added films and coatings in the packaging of fish and fisheries products. For example, on refrigerated bream fillets, a pectin coating enriched with clove essential oil at two concentrations (1 % and 1.5 %) was tested (Nisar et al., 2019). The results showed that this treatment reduced the rate of lipid oxidation and TVB-N, as well as the microbial growth, as measured by TVC, psychrophilic bacteria count, hydrogen sulfide-producing bacteria, lactic acid bacteria, *Pseudomonas*, and Enterobacteriaceae count, hence extending the shelf-life by at least 6 days. In another case, shrimp treated with gelatin coating enriched with orange leaf essential oil (2 %) improved sensory and microbial attributes while also extending shelf-life (14 days) compared to the control (6 days) (Alparslan et al., 2016).

Nanocomposite films based on soy protein isolate, montmorillonite, and clove essential oils were used to preserve bluefin tuna fillets during refrigerated storage, with films promoting a reduction in the microbial count and delaying lipid oxidation until 12 days (Echeverría et al., 2018). Ehsani et al. (2020) investigated the effect of chitosan, alginate, and gelatin-based films containing sage essential oil on fish burgers stored at 4 °C. The results showed that film wrapping significantly reduced the proliferation of microorganisms such as *Pseudomonas* sp. and *Shewanella* sp. Furthermore, Arfat et al. (2015) discovered that sea bass slices wrapped in fish protein/fish gelatin composite films infused with basil leaf essential oil had a shelf-life of 10–12 days compared to 6 days for the control.

An alternative strategy for incorporating essential oils into fish and other food products through micro- and nano-emulsions has recently been proposed as a promising alternative to direct essential oil addition. This technique is gaining popularity as a novel technique and as a model carrier for the transport of lipophilic substances such as essential oils due to its ease of preparation, small particle size, higher bioavailability, and kinetic stability (Ozogul et al., 2017). Recently, the effectiveness of nano-emulsions containing 4 % orange, grapefruit, mandarin, and lemon essential oils was assessed on the quality of rainbow trout fillets stored at 4 °C. When compared to the control group, the results showed that nano-emulsions based on essential oils eliminated fishy odor, improved organoleptic quality and delayed the growth of bacteria. Furthermore, the shelf-life of grapefruit and mandarin essential oils, lemon and orange essential oils treated and control samples were 16, 14, and 10 days, respectively (Durmus, 2020). Similarly, micro-emulsion-based lemon essential oil at two concentrations (0.3 % and 1 %) influenced the microbiological quality of salted sardines, with essential oil treated fish having significantly lower levels of Enterobacteriaceae, *Staphylococci*, and rod lactic acid bacteria than the control group (Alfonzo et al., 2017). Many volatile organic compounds belonging to monoterpene hydrocarbons, oxygenated monoterpenes, and sesquiterpene hydrocarbons may be responsible for the preservative functions of lemon essential oil. In fact, the antimicrobial activity of essential oils has been attributed to their main constituents, particularly phenolic constituents, as well as minor constituents found in oils (Bakkali et al., 2008).

The primary mode of action of essential oil components is targeting a broad spectrum of bacteria by disrupting their cell membrane and cytoplasm. They can also enter the bacterial cell's interior and inhibit its functional properties while also inducing cell morphology changes (Calo et al., 2015). Gram-positive bacteria are more sensitive to essential oils than Gram-negative bacteria (Trombetta et al., 2005); for example, essential oil from geraniol was found to be more active against
*Staphylococcus aureus* (Gram-positive) than *E. coli* (Gram-negative) (Zanetti et al., 2015). This is due to lipoteichoic acids in gram-positive bacteria cell membranes, which may facilitate the penetration of hydrophobic essential oil compounds, making them susceptible to essential oils. An extrinsic membrane surrounding the Gram-negative bacterial cell wall, on the other hand, slows the diffusion rate of those compounds through the lipopolysaccharide layer, conferring some resistance (Rodriguez-Garcia et al., 2016; Tongnuanchan & Benjakul, 2014).

The hydrophobicity of essential oils leads to interaction with the lipids of the bacterial cell membrane and mitochondria, making the cell more permeable. This interaction between cell membranes and essential oils inhibits both gram-positive and gram-negative bacterial growth (Calsamiglia et al., 2007; Friedly et al., 2009).

**5. Challenges and future directions**

Based on the literature reviewed above, bio-based edible coatings have emerged as a promising approach for preserving the quality and extending the shelf life of fresh fishery products during storage. However, most studies have solely focused on observing physicochemical changes, bacterial enumeration, and sensory evaluation, rather than attempting to understand the underlying mechanisms of edible coating's protective effect. To optimize its application in the preservation of fishery products, future studies on bio-based edible coating need to investigate the protective mechanisms in terms of film-forming characteristics, synergistic effects for microorganisms and oxidation control, flavor and sensory properties, and enzyme inhibition. Among the specific recommendations we propose.

* Dip-draining is currently the preferred method for fishery product. This method requires a long drain time for coatings containing chitosan and gelatin and may result in non-homogeneous coating. More research is needed to understand the factors affecting the thickness and uniformity of forming films, as well as to develop drying methods to control film formation and shorten drain time.

* Although many researchers studied the effects of bio-based edible coating on microbial succession through enumeration, only a few microorganisms were cultivated. During the spoilage of fishery products, only one or a few microorganisms were found dominant communities. As a result, monitoring the changes in the overall bacterial diversity of coated samples using high-throughput methods in the frame of foodomics would allow us to better explain the preservation mechanism(s) at the microbial level.

* Investigating the effects of bio-based edible coatings on the changes in taste and flavor precursors of fishery products during storage, with the goal of improved understanding of flavor and sensory quality retentions in coated products is required.

* Developing a better understanding of the synergistic effect among natural preservatives is needed to optimize the formulation of composite coatings according to the targeted effect as well as negligible negative impact on product sensory qualities, particularly avoiding color darkening and irritating odors from natural preservatives.
* Although edible coatings have shown the proof in reducing texture softening induced by endogenous enzymes during storage, further discoveries are needed in the enzyme inhibition mechanisms to slow this deterioration even further.

A dearth of study has also been conducted on the development of novel synergistic gelling systems. In turn, it is possible to conduct such a comparative investigation. The majority of research on edible coating applications has been conducted in the laboratory, resulting in a paucity of actual applications. Future applied research ought to focus on edible coatings given that they can extend food's shelf life. To eliminate these concerns, coating application methods can be modified to include a recycling process that does not waste an excessive amount of coating solution, reduce the number of microorganisms in the solution during recycling, develop spraying techniques for uneven surfaces, design industrial-sized vacuum tanks, and so on.

This permits novel applications of food contact films and coatings on a large scale. The development of edible films and coatings spanning two or more nanomaterials is anticipated to be a future trend in edible packaging. These products are anticipated to have improved gas barrier properties, superior product stability, unique color and flavor, and a greater nutritional value. New approaches for modulating mechanical properties and gas transport must be investigated in order to maintain food quality. By altering their properties in response to environmental factors such as relative humidity and temperature, edible films and coatings must assist food products in adapting to their environments. To effectively regulate the passage of oxygen, carbon dioxide, and water vapor in a system, nanomaterials must be meticulously evaluated.

The use of gelatin-based edible films and coatings is an exciting new route for developing new food packaging materials. Because of gelatin's hygroscopic properties, some research studies have been conducted to evaluate the overall effect of adding different substances such as crosslinkers, strengthening agents, plasticizers, or additives with antimicrobial or antioxidant properties to gelatin-based products in order to improve the functional properties of gelatin-based edible films and the shelf-life of food products. A growing number of publications have reported the development of gelatin-based films for meat applications as coatings to reduce lipid oxidation-induced color deterioration from red to brown. In the case of fish products, various studies have focused on the use of gelatin in conjunction with other biopolymers or active additives to protect fresh fish from cooking processes and microbial/oxidation deterioration. Extensive research on new methods for gelatin-based film formation is still required to improve final properties and expand the applications.

The WP and active ingredients are being incorporated into edible films/coatings, which can be used in a variety of food products. Depending on the purpose, product, nature of the film, type of active ingredient, and various inclusions, numerous combinations could be used on an industrial scale. We described several characteristics of edible WP films/coatings as novel packaging materials in this review. There are numerous promising approaches for improving food quality, extending shelf life, and ensuring safety, maintaining functionality, and reducing environmental impacts. Furthermore, these films and coatings can be used as separate pouches of homogeneous substances and active ingredient carriers. In food packaging, the formulation of a biopolymer coating using WP-based plastic films can replace existing synthetic oxygen-barrier layers, such as ethylene vinyl alcohol (EVOH). Whey is not a direct food competitor because it is a byproduct of cheese production and in terms of intrinsic edibility and biodegradability, WP-based films/coatings outperform synthetic
plastics. The WP films/coatings described in this review demonstrated exceptional optical and barrier
properties that outperformed existing biopolymers. In this regard, WP coatings have been
demonstrated to be effective gas barriers capable of acting as vehicles for a variety of compounds
such as antioxidants, antimicrobials, or various nutrients, though their mechanical properties need to
be improved. Thus, chemical, physical, or enzymatic protein cross-linking (Di Pierro et al., 2018), as
well as other ways such as blending and nanotechnology, are potential methods for improving the
tensile strength and elongation at break characteristics of WP films. The greatest commercial potential
for WP films/coatings, in our opinion, includes its use for a delivery vehicle of functional and
bioactive compounds in fresh fish and seafood products, thereby extending shelf life and improving
safety, nutrition, and sensory qualities. The WP-based multilayer laminates have been successfully
validated for the storage of a variety of food products. This novel WP coating can be detached,
allowing multilayer films to be reused. Due to the use of natural by-products from the food industry
as raw materials, WP-based packaging ideas may play beneficial roles in sustainability due to the
possibility of recycling materials rather than incineration, as done in synthetic laminates. However,
in the face of industrial setbacks, cost-effectiveness will always be a driving force in current and
future WP processing developments. The industrial application of this new technology is still
dependent on additional scientific research to identify the mechanism of film formation to improve
the performance of both the product and the process. Furthermore, consumer studies and long-term
toxicity assessments must be investigated before gaining a significant market share. On the other side
the exorbitant cost of equipment for novel technologies used in food preservation such as plasma or
irradiation is not associated with coating technology.

Due to its nontoxic, antibacterial, and antifungal qualities, chitosan has garnered considerable interest. Because of this, chitosan is regarded as an ideal material for the development of food grade films/coatings. Due to the protonation of the amino groups, chitosan has a positive charge and is soluble at pH levels below 6.5. Chitosan can be shaped into numerous forms, including films, gels, and nanoparticles (Koc et al., 2020). Wang, Liao, et al. (2019) stated that it is among the most researched polysaccharides for the production of packaging film due to its exceptional film-forming capacity. Gelatin is commonly used as a composite component with chitosan to sidestep problems associated with using coating made from individual biopolymers (Wang et al., 2021). United States Food and Drug Administration (FDA) has classified chitosan as GRAS (GRAS notification number GRN 170) (FDA, 2005), in 2007 under the European union regulation (No. 749/2012) and China national standards (GB 29941–2013) (Nair et al., 2020). When incorporating food-grade functional materials within the limits specified by the regulations, chitosan-gelatin composite films could have improved physical and chemical properties without compromising the safety and edibility, indicating an excellent packaging potential (Wang et al., 2021). The acute and subacute toxicity of two chitosan derivatives, produced by a depolymerization procedure, was assessed by orally administering them to a mouse model (Punarvasu & Prashanth, 2023). The findings indicate that administering a dosage of chitosan derivatives up to 5000 mg/kg bw did not result in fatalities or any other proof of toxicity. Hence, the LD50 value of both the chitosan derivatives exceeds 5000 mg/kg bw. Furthermore, in the subacute toxicity assessment, the chitosan derivatives exhibited no discernible destruction following daily oral doses of 1000 and 2000 mg/kg bw. In an overview, the chitosan derivatives (LMWC and SAMC) that underwent investigation showed no signs of toxicity in mice. There were no instances of fatalities or alterations in general behavior observed during both single and repeated dose administration studies. Therefore, these derivatives appear to be safe for use in biomedical and food applications (Punarvasu & Prashanth, 2023).
Multiple investigations have demonstrated that the incorporation of various active chemicals into chitosan films results in intriguing modifications to their mechanical, barrier, and functional properties. J.P. Verwijs, a frozen fish vendor based in the Netherlands, recently switched to organic biodegradable packaging for its diverse range of seafood items (Kearns, 2019). J.P. Verwijs asserted that the design of the fresh packaging was influenced by the principles of the circular economy, a business strategy that values sustainability, transparency, simplicity, and excellence. By adopting its novel biodegradable packaging solution, developed from agricultural waste, the supplier can effectively diminish its plastic waste production and substantially reduce its carbon dioxide emissions. The packaging is fully compliant with the European Food Safety Authority (EFSA) and FDA regulations, as well as Europe's biodegradable and compostable standards EN 13432/14995. It is also an innovative biodegradable approach. The presence of phenolic compounds has been demonstrated to improve both mechanical and barrier characteristics. Chitosan-based edible films containing antioxidant properties were fabricated by adding *Zataria multiflora* Boiss essential oil (ZEO) at concentrations of 5 and 10 g/L, as well as grape seed extract (GSE) at a concentration of 10 g/L, both on its own and in combination. Except for the 10 g/L GSE +10 g/L ZEO film, all other films showed reduced strength and elongation values. The inclusion of only 10 g/L ZEO in the GSE film enhanced the water vapor transmission rate of chitosan films (Moradi et al., 2012). Talón and co-workers investigated the antioxidant properties and physical characteristics of various polymeric matrices composed of chitosan and starch (Talón et al., 2017). These matrices were modified by including a thyme extract (TE) that is abundant in polyphenols, along with the inclusion of tannic acid (TA) as a cross-linking agent. The integration of TE resulted in modifications to the microstructural and physical properties of the films, contingent upon the specific matrix employed. Thyme extract was found to be most effective in improving the properties of chitosan-based films. This is attributed to the cross-linking activity between the polyphenols and chitosan, resulting in enhanced tensile response characterized by increased resistance at break and improved stiffness (Talón et al., 2017). As for the functional qualities of the films, the addition of essential oils, phenolic compounds, and fruit extracts considerably enhanced their antibacterial and antioxidant capacities. To build chitosan-based packaging that can meet practical criteria and compete with petroleum-based packaging, however, considerably more research is required. It is being proposed to conduct further investigations (1) on chitosan and its combination with other materials to suit practical needs, (2) Conduct research on chitosan films and determine the potential risks associated with their use, (3) Determine interaction of types of foods with chitosan packaging (Florez et al., 2022).

Sustainable development is an overarching goal that mandates an interdisciplinary approach to resolve the societal challenge of climate change, environment, resource efficiency, and raw materials. In this context, the first step in closing the waste-to-consumption loop in accordance with the seventh main objective of the circular economy that is the valorization of abundant and readily available bio-waste with a high potential for manufacturing products with added value. Biomass leftovers can serve as a viable substitute for fossil fuel sources in the production of plastic packaging for food. Substituting biomass residues for fossil fuel sources in plastic manufacture has two primary benefits: severing the connection between food packaging and fossil fuels, and utilizing agricultural by-products to minimize demand for plastic derived from fossil fuel-based sources. Although bioplastic food packaging holds promise, it confronts numerous hurdles such as exorbitant production expenses, inadequate policy and standard support, and a scarcity of composting facilities (Symons, 2022). Plastic derived from secondary biomass resources, such as by-products, effectively confronts numerous issues associated with society and the environment. This includes the reduction of
greenhouse gas emissions, without compromising food security by substituting food crops. In along with substituting fossil fuel sources, the potential benefits of recycling or composting at the end of a product's life cycle enhance the environmental and social benefits. Efforts have been undertaken to measure both the cost and the benefits of shifting to biomass plastics, such as the economic gains resulting from declined greenhouse gas emissions and reduced plastic waste in marine ecosystems. However, these studies are broad in scope and prone to uncertainty. Several studies have utilized the life cycle analysis methodology to calculate these figures. However, these studies fail to comprehensively account for all the costs and benefits, as they do not incorporate various end-of-life alternatives. The financial expenditures linked to the adoption of biomass plastics are substantial and encompass elevated costs of using biomass sources, expenses for research and development, and shortfalls in economies of scale. These hurdles are substantial and necessitate additional investigation, beginning with the establishment of a baseline for the costs of packaging made from fossil fuels. This will help identify the cost benchmarks that biomass plastics must meet. Moreover, a further investigation is necessary to determine the complete environmental and economic implications of various end-of-life circumstances, as well as the potential advantages and disadvantages of waste systems that incorporate composting infrastructure (Symons, 2022).

The future development of bioplastic production will be strongly influenced by the fluctuation of conventional plastic costs. In addition, the development is impacted by other factors such as technological advancements, economies of scale, and raw material costs (Döhler et al., 2022). By increasing the scale of bioplastics manufacturing, organizations can leverage cost efficiencies by distributing current fixed expenses over a larger amount, resulting in higher output at reduced average costs per unit. At present, the production volume of bioplastics remains fairly modest. Simultaneously, the expenses associated with constructing structures and acquiring equipment constitute a substantial portion of the overall manufacturing costs (Manandhar & Shah, 2020). Simultaneously, the expenses associated with constructing buildings and acquiring equipment, known as fixed capital expenditures, constitute a substantial portion of the overall production costs (Manandhar & Shah, 2020). This implies a notable potential for untapped economies of scale. The cost progression in increasingly sophisticated biotechnologies demonstrates the actualization of these capabilities as time passes (de Jong et al., 2017; Skovsgaard & Jacobsen, 2017).

The enormous quantities of waste generated during the refining of seafood can be effectively managed to produce renewable and biodegradable raw materials (Caba et al., 2019). This management incorporates the use of environmentally benign and cost-effective extraction techniques to ensure that innovative biorefinery practices designed to add value to byproducts contribute to the sustainable development of materials. Currently, chitosan and fish gelatin production has been scaled up, and these materials are commercially available. Utilizing side streams, co-products, or even seafood processing waste (specially the crustacean shell fish processing waste) can support the concept of a circular bio-economy, provide more sustainable solutions, and maximize the use of our natural resources. Considering the annual amounts produced and the rising demand for biodegradable films/coatings in the food and agriculture industries, the processing or fabrication of animal-based products generates a substantial amount of potential feedstock.

# 6. Conclusion
Bio-based edible coatings are gaining momentum in the sustainable and circular ecosystem. This review has covered many aspects of coating preservation approach for fishery products, such as coating formulations and carriers, including those formulated with natural additive compounds, coating application methods and their benefits and challenges, and protection capacity and mechanism as determined by microbial, physicochemical, and sensorial evaluations. More discoveries and deep understanding are very critical and essential to engineering and designing fish product-specific bio-based coating formulations with recycling in mind, without the need for complex processing aids.

**CRediT authorship contribution statement**

**Don Hettiarachchige Udana Eranda:** Writing – original draft, Investigation, Data curation, Conceptualization.
**Manat Chaijan:** Writing – review & editing, Supervision, Investigation, Conceptualization.
**Ike Uysal-Unalan:** Writing – review & editing, Investigation, Data curation.
**Worawan Panpipat:** Writing – review & editing.
**Azza Silotry Naik:** Writing – review & editing.
**Amira Leila Dib:** Writing – review & editing.
**Supatra Karnjanapratum:** Writing – review & editing, Supervision.
**Mohammed Gagaoua:** Writing – review & editing, Writing – original draft, Supervision, Investigation, Data curation, Conceptualization.

**Declaration of competing interest**

All authors declare that they have no conflicts of interest.

**Acknowledgements**

This work has been supported by Walailak University Ph.D. Scholarships for High Potential Candidates to Enroll in Doctoral Programs (Contract No. HP018/2021). Also, this research was financially supported by Walailak University Graduate Research Fund (Contract No. CGS-RF-2023/07).

**Data availability**

No data was used for the research described in the article.

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