It is important to point out that according to these results, EF1 showed less color change 7. These values are low compared to those reported by Rodríguez-Campos et al. Moreover, the results obtained in this study with EF1 at both temperatures were in the range 4.

In their study, the authors formulated avocado puree with ascorbic acid 0. Therefore, even though citric acid may decrease color change in avocados, it is not the main compound responsible for preventing color change in guacamole.

Table 2. At the beginning of the storage, phenolic compounds and antioxidant capacity presented values of Moreover, values of total phenolic compounds and antioxidant capacity were slightly higher than those reported by Rodríguez-Carpena et al.

However, no effect of storage conditions and the EF was observed. Therefore, it is possible to infer that the lesser effect of color change in guacamole added with EF1 was due to its antioxidant capacity, which also possesses a higher quantification among formulated EFs In this sense, under oxidative stress conditions, the antioxidant capacity of avocado pulp is mainly related to its tocopherol content γ- tocopherol mainly , which donated hydrogen to alyl or alkyl peroxy radicals of avocado pulp.

These tocopherols are stabilized through hydrogen donation by the ascorbic acid or flavonoids contained in the lemon-onion extract Basavegowda and Baek, ; Yin et al.

On the other hand, although citric acid is an antioxidant compound when it is added to the chitosan film formulation, it can act as a cross-linker giving stability to the EFs. In this sense, it is possible that increasing the amount of citric acid in the EF formulation reduces the action of lemon-onion extract, thus showing a reduction in the antioxidant capacity of EF Priyadarshi et al.

Figure 2. Total phenolic compounds A and antioxidant capacity B of guacamole with and without EF control. Bars indicate standard deviation. One of the main characteristics of EFs is protecting against contamination and microbial spoilage. In this sense, chitosan has been proven to reduce the microbial growth of several food products Abdel-Naeem et al.

Figure 3 shows the mesophylls and molds plus yeast growth in guacamole with and without EF. Moreover, results indicate the effect of the lemon-onion extract in maintaining or even reducing microbial load.

In this sense, thiosulfinates in onions may be responsible for the antimicrobial activity of EFs Loredana et al. Figure 3. Mesophylls A and molds plus yeasts B of guacamole with and without EF control.

The moisture content, tensile strength, extension, and WVP are in the range of those reported In this sense, Piccirilli et al. On the other hand, tensile strength significantly increased at 20°C, while a reduction in thickness was observed at the same temperature.

In this sense, an inverse relationship between the thickness and tensile strength of EF based on chitosan was reported Escamilla-García et al. Table 3. On the other hand, microbial load and antioxidant properties of EF remained constant during storage, regardless of the temperature, which is of paramount importance because it indicates that EF remained stable during storage and its safety and antioxidant properties did not change, even at the highest storage temperature.

In this sense, the antioxidant capacity of EF was corroborated in the color parameter of guacamole when EF1 was applied Table 4.

As observed, no significant effect of storage time and temperature was detected. Therefore, the developed EF fulfills several requirements due to its limited microbial spoilage and oxidative reaction, increasing the shelf life of guacamole Benbettaïeb et al. Table 4. The main characteristic of guacamole rejection by the consumer is likely to be its rapid color change caused by the polyphenol oxidase enzyme and its rancidity flavor caused by the oxidation of lipids.

As previously mentioned, a sensory acceptance study was carried out, evaluating the sensory properties and overall acceptance of control guacamole, guacamole with fresh EF, and guacamole with stored EF Figure 4.

After this time, EF and guacamole were mixed and used for sensory evaluation. As was observed, all sensory parameters evaluated were well-accepted by the consumers showing values between 5 and 6, which stood for I like and I like much, respectively.

Moreover, consumers did not perceive the addition of fresh and stored EF, regardless of the storage temperature. Similar results were obtained by Rodríguez-Campos et al. They informed values of 6 to 8 I like slightly to I like very much in a 9-hedonic scale of freshly formulated guacamole.

Probably, the adequate acceptance of the taste and flavor of the guacamole with EF obtained in this study was due to lemon-onion extract, two ingredients used for the formulation of guacamole. Moreover, according to Salgado-Cervantes et al. Figure 4.

Selected and applied EF on guacamole. In this study, EFs based on chitosan, glycerol, citric acid, and lemon-onion extract were formulated and evaluated in their capacity for maintaining the color and microbiological and antioxidant characteristics of guacamole.

Results indicated that EFs reduce the color change of guacamole, increase the antioxidant capacity, and reduce the microbial load of mesophylls and molds plus yeasts.

Moreover, EF1 maintains the color characteristics of guacamole, and according to its composition, was well-accepted by the consumer, showing a similar acceptance to fresh guacamole. The raw data supporting the conclusions of this article will be made available by the authors, without undue reservation.

MM-S: Data curation, Writing — original draft. PH-C: Supervision, Visualization, Writing — original draft. CR-L: Methodology, Supervision, Writing — original draft. IR-L: Investigation, Methodology, Project administration, Writing — original draft. CO-V: Conceptualization, Formal analysis, Funding acquisition, Writing — original draft.

This study was financed by the Vicerrectoría de Investigación y Estudios de Posgrado of the Benemérita Universidad Autónoma de Puebla Project number: VIEP The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.

All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article, or claim that may be made by its manufacturer, is not guaranteed or endorsed by the publisher.

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Keywords: antimicrobial; antioxidant; chitosan; dairy; edible films; fruits; meat. Abstract Chitosan is the deacetylated form of chitin regarded as one of the most abundant polymers and due to its properties, both chitosan alone or in combination with bioactive substances for the production of biodegradable films and coatings is gaining attention in terms of applications in the food industry.

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The slurry was filtered and 5 mL of filtrate was added to a vial containing 0. After capping, the vials were heated for 10 min in a boiling water bath to develop the chromogen and then cooled to ambient temperature. Using freshly made, acidified 1,1,3,3-tetraethoxypropane, a standard curve 8—50 nmol of malondialdehyde MDA was prepared.

TMA content was determined using the method of Ward et al. After centrifuging the mixture at rpm, the supernatant was transferred to a mL test tube. After that, the mixture was rapidly agitated for 10 min while 1.

Anhydrous sodium sulfate 1. After mixing, 5 mL of 0. TVB measurement was conducted using the Kjeldahl method according to Cai et al. A total of 5 g of the minced tuna flesh sample and 50 mL of distilled water were mixed and agitated for 30 min at ambient temperature. The mixture was filtered followed by steam distillation.

An automated Kjeldahl system was used for the distillation process VELP Corporation, Italy. Coated and uncoated tuna filet samples were evaluated for their sensory characteristics color, odor, appearance, and overall acceptability during cold storage by 10 trained adult volunteers Rong et al.

A plate with three-digit numbers was used to assess the samples. Using a 9-point hedonic scale, the samples were judged on their sensory attributes, including color, odor, appearance, and overall acceptability 1 being extremely disliked and 9 being extremely liked.

All of the experiments were performed in triplicate, and the values were expressed as the means ± standard deviations SD. Data were analyzed by a two-way ANOVA. Moreover, the MetaboAnalyst software was used to create a 3D plot for the multivariate principal component analysis PCA , variable importance in projection VIP score, and clustering heatmap.

Because BAs are an important tool for evaluating the sanitary condition of foods, they can be utilized as indicators of food spoilage and, at high concentrations, can have toxicological consequences on consumers Saad et al.

Biogenic amine formation depends mainly on the decarboxylase enzyme that is secreted by spoilage or pathogenic bacteria Xia et al. Controlling pathogenic bacteria using natural antimicrobials and antioxidants reflects levels of BAs in fish which also represents a quality indicator of fish.

Edible coating fortified with plant extracts is one of the novel approaches to food safety and preservation that consumers seek in the food industry Sapper and Chiralt, In this study, chitosan edible coatings fortified with natural plants such as beetroot, curcumin, and garlic extracts were applied to chilled tuna filets.

The results in Figures 2A — C show the impacts of chitosan - bioactive coatings on histamine, putrescine, and cadaverine formation in tuna fish filets stored refrigerated at 4°C during storage for 14 days.

There were increases in the content of BAs histamine, putrescine, and cadaverine during the storage period in both uncoated control and coated tuna filets samples. However, the control sample showed a higher increase in BA values than coated samples. Among the treated samples, chitosan incorporated with curcumin CH-C showed the lowest increase in BA formation, followed by chitosan with garlic CH-G , chitosan with beetroot extract CH-B , and chitosan without extract addition CH.

Figure 2. Values are shown as the mean of triplicates ± SD. Each colored circle with a lowercase letter matches the same colored group at each time point treatment.

Histamine is an indicator of fish quality Mendes, Higher levels of histamine cause edema, urticaria, and anaphylactic shock which are life-threatening. Additionally, according to Egyptian standards [ Egyptian Organization for Standardization and Quality Control E.

In this study, histamine did not exceed the acute reference dose in all studied samples during the 14 days of storage. Cadaverine levels in fish are considered an index for spoilage Al Bulushi et al. In this study, tyramine was not detected in the investigated tuna samples.

Chitosan bioactive coatings greatly reduced putrescine and cadaverine contents in treated tuna samples. According to research by Hao et al. Additionally, chitosan monomers preserve two hydroxyls and an amino group, which can react with free radicals.

As a result, they have the scavenging ability Abd El-Hack et al. The pH values of the treatments ranged from 6. The pH of the control sample was increased significantly compared to the treated samples.

The control samples showed an initial value of 6. Among the treated samples, the CH-C treatment showed the lowest increase in pH, followed by the CH-G, CH-B, and CH treatments, which were the highest ones.

The same pattern in pH rise is consistent with the findings of Hassanzadeh et al. These elevations in pH levels may be caused by the bacteria which produce alkaline substances such as trimethylamine and ammonia Hassanzadeh et al.

The low pH in chitosan-bioactive coated tuna filet samples during storage might also be attributed to their antioxidant and antibacterial properties and the acidic pH of chitosan Hassanzadeh et al.

Figure 3. Impacts of chitosan bioactive coating fortified with natural extracts on pH A , TBA B , TVB C , and TMA D values in chilled tuna filets. Each colored circle with lower case letter matches the same colored group at each time point treatment. Up until the end of the storage, changes in the TBARS index generally revealed significantly higher levels in all of this study's treatments Figure 3B.

After 14 days of refrigeration, the control sample's TBA level of 0. The lowest alteration in TBA was noticed in the CH-C, CH-G, and CH-B treatments compared to chitosan only CH. The TBA values were 8. The chitosan-treated samples incorporated with the curcumin, garlic, and beetroot extracts performed better in lipid oxidation reduction than the CH treatment.

According to the Egyptian standard EOSQC, , 4. On the 6th day of storage, the control sample with a TBA content of 4. On the 14th day end of storage period , however, TBA levels in all treatments were higher than the permitted range, and the chitosan-coated treatment incorporated with curcumin CH-C with a TBA content of 4.

Phenolic substances stabilize hydroperoxides to stop their oxidation, further deterioration, and the production of molecules such as malondialdehyde Hernández-Hernández et al. According to a study by Mahdavi et al. Additionally, Qin et al. Mehrabi et al. pogonosperma extract resulted in a reduction in the rate of TBA alterations.

Several volatile compounds, including ammonia, methylamine, dimethylamine, trimethylamine, and other compounds formed during storage of meat in refrigerated conditions as a result of microbial activity, are included in TVB, one of the most significant indications of fresh meat detection Anderson, ; Rodríguez et al.

The lowest alteration in TVB values was noticed in chitosan-coated samples containing curcumin, garlic, and beetroot extracts compared to chitosan only. TVB level of 3. On the 6th day of storage, the control sample with a TVB level of The chitosan-coated treatment containing curcumin CH-C , with a TVB concentration of Uncoated and coated tuna samples showed a similar way in TMA value alterations, where clear increases were noticed during refrigerated storage Figure 3D.

The lowest alteration in TMA values was observed in chitosan-coated samples containing curcumin, garlic, and beetroot extracts compared to chitosan only.

TVB level of 1. The refrigerated storage of treated Indian white prawn Fenneropenaeus indicus revealed an increase in TMA, according to the finding of Bindu et al. TMA level of 1.

On the 6th day of storage, the control sample's TVB level of However, at the end of storage day 14 , TMA concentrations in all treatments were higher than the standard permitted range, and curcumin-containing treatment CH-C with a TVB content of 9.

However, Li et al. It can be concluded that among the treated samples, chitosan incorporated with curcumin CH-C demonstrated a remarkable superiority in all the studied parameters followed by chitosan with garlic CH-G extract and then chitosan with beetroot extract CH-B.

The results of the sensory evaluation of tuna filet samples are presented in Figure 4. Sensory scores exhibited a significant reduction in the color, odor, appearance, and overall acceptability of C and CH treatments during storage.

It has been proven that when fish spoils, it develops flavors that are strongly fishy, rancid, and putrid. As a result, the sensory score of 4 was utilized as the upper limit of acceptability in the current investigation Fan et al.

Figure 4. Impact of chitosan bioactive coating fortified with natural extracts sensory attributes of chilled tuna filets including color A , odor B , appearance C , and overall acceptability D. Values are shown as the mean of triplicates.

After 6 days, the C and CH samples were no longer acceptable; however, the CH-B sample remained in good and acceptable form after 10 days, and the CH-G and CH-C samples were still in good and acceptable condition for up to 14 days end of the storage period. A multivariate analysis principal component analysis, PCA was used to reveal the relationship between the various treatments and variables.

All variables were included in the three principal dimensional components PC1, PC2, and PC3 , which together accounted for Most of the examined variables were distinguished by PC1, and hence revealed the greater portion of variance Furthermore, the PCA showed that the CH and CH-B treated samples were clustered together on the upper part of the gel.

Similarly, CH-C and CH-G treated samples were clustered together on the bottom of the gel, and both were segregated from the control samples. Figure 5. Principal components analysis PCA and data clustering analysis of control Cont and chitosan-treated samples CH, CH-B, CH-C, and CH-G.

A 3D score plot of PCA for discerning the five treatments. Percentage values specified on the axes depict the contribution rate of PC1 B Variable importance in projection VIP : the colored boxes on the right display the relative concentrations of the relevant measured parameters in each study group, while, the contribution intensity is illustrated with colors ranging from highest red to lowest blue.

C All datasets can be visually represented intuitively thanks to hierarchical clustering heatmap. With varied averages in rows and various treatment sets in columns, the concentration levels are indicated by each colored cell on the map.

On the gradation scale, dark red is the highest value and blue is the lowest. The variable importance in projection VIP revealed the BAs putrescine, histamine, and cadaverine , and biochemical quality indicators pH, TVB, TBA, and TMA and sensory overall acceptability were the most important variables that affected the quality and shelf life of tuna filet samples Figure 5B.

The clustering heatmap shown in Figure 5C presents a clear overview of all the datasets and highlights the distinct changes in concentration values of all tested parameters in response to effective treatments when compared to other groups.

These findings revealed that the CH-C and CH-G showed more improvements in the overall characteristics of tuna filet samples than other groups. The results showed that coating tuna filets with chitosan incorporated with beetroot, garlic, and curcumin extracts resulted in a reduction of BA formation in the treated samples.

Furthermore, decreases in pH, TBA index, TMA, and TVB were observed during the storage period. Beetroot, garlic, and curcumin extracts could be used in pharmaceutical and food applications as replacers for synthetic antioxidants and chemical preservatives to slow down lipid oxidation and prevent the growth of microorganisms due to their potential antioxidant activity and high concentrations of phenolic compounds.

The raw data supporting the conclusions of this article will be made available by the authors, without undue reservation. The study protocol was approved by the Ethics Committee of the Faculty of Veterinary Medicine, Benha University Approval no. BUFVTM Conceptualization: RE, SSI, EA-E, AA, and MM.

Methodology: RE, SSI, EA-E, AA, AE-S, and MM. Software: RE, SSI, LŞ, RA, and AE-S. Validation: MA, SFI, RA, and AE-S. Formal analysis: RE, SSI, AA, AR, LŞ, and MM. Investigation: RE, SSI, AR, AE-S, and MM. Resources: AA, SFI, FI, and LŞ.

Data curation: RE, SSI, AA, MA, RA, and AE-S. Writing—review and editing: RE, AA, AR, RA, and FI. Visualization: SFI, MA, RA, and FI. Supervision: AA and AR. Project administration: RE.

Funding acquisition: SFI and FI. All authors contributed to the article and approved the submitted version. This research was funded by the Princess Nourah bint Abdulrahman University Researchers Supporting Project number PNURSPR , Princess Nourah bint Abdulrahman University, Riyadh, Saudi Arabia; and the project 6PFE of the University of Life Sciences King Mihai I from Timisoara and Research Institute for Biosecurity and Bioengineering from Timisoara.

The authors appreciate all support provided by the Center of Excellence in Screening Environmental Contaminants CESEC , Benha University, Egypt. The authors also gratefully thank the resources provided by the Princess Nourah bint Abdulrahman University Researchers Supporting Project number PNURSPR , Princess Nourah bint Abdulrahman University, Riyadh, Saudi Arabia.

Moreover, this article is published from the project 6PFE of the University of Life Sciences King Mihai I from Timisoara and Research Institute for Biosecurity and Bioengineering from Timisoara. The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.

All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers.

Any product that may be evaluated in this article, or claim that may be made by its manufacturer, is not guaranteed or endorsed by the publisher. Abd El-Hack, M.

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