Quality characteristics and antioxidant activities of rice cookies supplemented with red beet extracts

Abstract

In this study, the quality characteristics and antioxidant properties of red beet rice cookies were determined. Four different types of cookies were prepared using 0, 2.5, 5.0, and 10 g red beet extract. Consumer acceptability was evaluated after measuring the quality characteristics and antioxidant activity of the red beet cookie dough and cookies. The quality of red beet cookies varied depending on the amount of red beet extract added. The antioxidant properties of the cookies were correlated with the amount of red beet extract added to the cookie dough. The addition of 5.0% red beet extract to cookie dough increased its antioxidant effect without affecting consumer preference. Considering its nutritional value, consumer preferences, and antioxidant properties, this red beet cookie recipe can be a valuable resource for rice cookie production.

Supplementary Information

The online version contains supplementary material available at 10.1007/s10068-024-01805-8.

Introduction

The demand for natural foods is increasing because of health orientation and premiumization. In vivo, reactive oxygen species, such as superoxide radicals, alkyl radicals, hydroxyl radicals, hydrogen peroxide, and (alkyl) peroxyl radicals are produced by various physical and chemical factors. These free radicals are oxidized unsaturated fatty acids present in the in vivo membrane, which reduce membrane fluidity (Morita et al., 2019). These also impair the activity of enzymes and receptors, damage membrane proteins, inactivate cells, and contribute to aging and the development of various diseases. Accordingly, interest in the removal of free radicals is increasing and research is actively being conducted on the development of synthetic or natural antioxidants that enhance the antioxidant defense system or control reactive oxygen species (Nemati et al., 2024; Fomekong et al., 2024). Many synthetic antioxidants have been developed; however, they are known to cause side effects when consumed in large quantities. Therefore, safer and more powerful natural antioxidants must be developed crucially (Jang et al., 2015; Liu et al., 2013). Representative natural antioxidant compounds include vitamins, such as ascorbic acid and tocopherol; phenolic acids, including caffeic acid, chlorogenic acid, and ferulic acid; flavonoids, such as catechin, quercetin, and kaempferol; and carotenoids. (Lee et al., 2023; Liu et al., 2013).

Red beets are biannual crops of the family Chenopodiaceae that have been used for medicinal purposes since ancient Roman times. The plants have been traced to have originated in North Africa and spread through the Mediterranean Sea route, occupying the seashores of Asia and Europe (Neelwarne, 2012). It has relatively large roots, a red interior, and an epidermis, and is highly adaptable to soil; therefore, it can be cultivated in the spring, summer, and fall. In Korea, it is harvested in various locations including Jeju Island and Gangwon-do (Yoo and Ko, 2014). It is low in calories (42 kcal per 100 g) and rich in folic acid, which is recommended for pregnant women. It is also effective for anemia, blood purification, and red blood cell production. Furthermore, red beets are rich in fiber; therefore, they prevent constipation and are effective in treating cardiovascular disease and high blood pressure by reducing serum cholesterol (Babarykin et al., 2019). Betalains, flavonoids, polyphenols, and vitamins are excellent antioxidant sources (Vinson et al., 1998).

Rice has been used as a staple food in Korea for a long time (Kwak et al., 2017); however, its consumption has recently been gradually decreasing due to the westernization of dietary culture. Therefore research and development of various processed foods to promote rice consumption is crucially required. Rice contains various nutrients, such as starch, lipids, proteins, vitamin B, minerals, and fibers. Rice contains 7–8% protein (Chaudhari et al., 2018) and as it lacks the gluten protein of wheat flour, it does not cause irritable bowel or celiac disease (Fasano et al., 2012). Therefore, it is a good food ingredient to replace wheat flour when making bread, cookies, and noodles. Cookie, a type of confectionery, is consistently consumed as a favorite snack for all age groups. In response to the tendency of customers to purchase health-oriented products, research is being conducted on cookies that replace wheat flour with rice powder and to add functional ingredients, such as omija extract (Kim et al., 2015), watermelon rind waste (Naknaen et al., 2016), and pear peel powder (Nam et al., 2023). In previous studies, red beet powder cookies (Ingle et al., 2017) and beef sausage with red beet powder (El-Gharably and Ashoush, 2011) have been manufactured; however, no reports are available on the manufacturing and quality evaluation of rice cookies with red beet extract.

Therefore, this study aimed to investigate the usability of red beet rice cookies by manufacturing highly functional red beet extract and comparing the quality characteristics and antioxidant capacities of different amounts of rice cookies.

Materials and methods

Chemical and reagent

The solvent for red beet rice cookie extract was diluted with 95% food-grade ethanol (Korea ethanol supplies company, Seoul, Korea). For the antioxidant activity experiment, DPPH from Alfa-Aesar (Ward Hill, MA, USA) and ABTS from Sigma (St. Louis, MO, USA) were used.

Preparation of red beet extract

The red beets used in this study were cultivated and harvested in Dangjin-si, Chungcheongnam-do, in December 2022, and purchased from the Green Farm Farming Association Corporation. The purchased red beets were washed, dried using a freeze dryer (FD8508, IlShinbiobase Co., Dongducheon, Korea), and ground (HR2041, PHILIPS, Seoul, Korea). Twenty grams of freeze-dried red beet powder was placed in a round flask, 200 ml of 95% food-grade ethanol was added, and the mixture was extracted in a 80 °C constant-temperature water tank (WEB-8, DAIHAN Scientific Co., Wonju, Korea) for 3 h using a reflux condenser. The extract was filtered using filter paper (Whatman No. 1) and completely concentrated using a rotary evaporator (Hei-VAP Value/Advantage; Heidolph, Schwabach, Germany). Three milliliters of bottled water was added and extracted. The concentrate frozen at –80 °C was used as a sample for sensory evaluation and analysis.

Cookie preparation

Dry basis soft rice flour (Daedoo Co., Gunsan, Korea), butter (Fonterra Limited, Seoul, Korea), small granulated sugar (CJ, Ulsan, Korea), egg (Heungsaeng Farm, Gyeongbuk, Korea), and baking powder (Bread Garden, Seongnam, Korea) were purchased from a commercial mart before experimenting and used as ingredients. Based on the results of several preliminary experiments, the rice cookies were prepared by supplementing with various amounts of red beet extract as follows: control, without red beet extract; RBE1, 2.5 g; RBE2, 5 g; RBE3, 10 g. The butter was left at room temperature for approximately 1 h and released using a hand mixer (DH-200; Guang Dong Xinbao Electrical Appliance Holdings Co., Foshan, China) at speed 1 for 1 min. Further, small granulated sugar was added and mixed in a cream state at speed 2 for 1 min. According to the ratios shown in Table 1, egg and red beet concentrates were added and kneaded at a rate of 2 for 1 min. After completing the dough by hand, thick rice and baking powders were added to a sieve once, refrigerated for 1 h, and pushed evenly to a thickness of 0.5 cm. This was molded using a cookie mold (4 cm in diameter) and baked in a preheated air fryer (MO22A7997CKR, Samsung Electronics Co., Suwon, Korea) at 180 °C for 4 min. Cookie samples were prepared as shown in Fig. S1, then cooled at room temperature for 1 h, stored in Ziploc bags, and used for the experiments.

Table 1.

Ingredient composition of cookies containing various amounts of red beet extract

Ingredients (g)	Red beet extract contents (g)
	Control	2.5 (RBE1)	5.0 (RBE2)	10 (RBE3)
Rice powder	100.0	100.0	100.0	100.0
Red beet extract	0.0	2.5	5.0	10.0
Butter	50.0	50.0	50.0	50.0
Sugar	50.0	50.0	50.0	50.0
Egg	20.0	20.0	20.0	20.0
Baking powder	2.0	2.0	2.0	2.0

Control, untreated; RBE1, rice cookie supplemented with 2.5 g red beet extract; RBE2, rice cookie supplemented with 5 g red beet extract; RBE3, rice cookie supplemented with 10 g red beet extract

Density and pH characteristics of red beet cookie dough

The pH of cookie dough was measured using a pH meter (A211, Thermo Scientific, MA, USA). Five grams of cookie dough was mixed with 50 ml distilled water for 5 min and filtered using a filter paper. Dough density was measured based on addition of 5 g of cookie dough to 30 ml of distilled water in a 50 ml mass cylinder. The difference between the volumes before and after the addition of cookie dough was measured. The density was calculated by dividing the weight of the cookie dough by the volume difference (g/mL).

Determination of physical properties

Cookie spreadability, loss rate (cook loss), and hardness were measured from characterization of cookie quality factors (Nam et al., 2024). Spreadability was defined as the ratio of the thickness to the measured diameter. The diameter of the cookies was measured by dividing the length of six cookies in a line by six to produce the mean cookie diameter. Similarly, cookie thickness was measured by dividing the height of six cookies stacked in a column by six to produce the mean cookie thickness (height). Finally, spreadability was calculated by dividing the mean cookie diameter by the mean cookie thickness.

The loss rate was defined as the difference in weight before and after baking. The weight of a cookie before baking divided by the weight of the same cookie after baking was calculated and reported as the cookie loss rate (%).

Cookie hardness was measured using a texture analyzer (TA.XT Express v2.1, Stable Micro System, London, England). The highest peak in the produced graph was recorded as cookie hardness. The texture analyzer was set with test type = TPA, test speed at 30 mm/min, target value of g, and trigger force of 0.1 N. Prepared cookie samples were 40 mm in diameter and 50 mm thick for hardness measurements. A 5 mm cylinder probe was used for the hardness measurement.

Determination of proximate compositions

General ingredient analysis of the rice cookies was performed after making a powdery sample with a mortar. The moisture content was measured by heating at 105 °C for 5 min using a moisture content measuring device (MB95, OHAUS, Seoul, Korea), the reference content was measured according to 550 °C ashing method (FX-03: 3Lit, DAIHAN Scientific Co., Wonju, Korea), and the crude fat was measured according to the soxhlet method (WHM12295, DAIHAN Scientific Co., Wonju, Korea).

Preparation of red beet cookie extract for measuring antioxidant activity

The samples required for the antioxidant activity experiments were homogenized for 1 h using magnetic stirrers (MS-300HS, MISUNG Scientific Co., LTD, Korea) after adding 27 ml of 80% ethanol to 3 g of powdered rice cookies. Centrifugation (1248R, Labogene, Daejeon, Korea) was performed for 20 min at 4000 rpm 4 °C, and the supernatant taken and filtered using a filter paper was used as an extract (Nam et al., 2023).

Antioxidant properties of red beet cookies

DPPH and ABTS radical scavenging activities were measured to determine the antioxidant properties of red beet cookies. The DPPH radical scavenging activity method (Kim, 2023) using 96 well plate was used to measure the DPPH radical scavenging activity of rice cookies prepared according to the added amount of red beet extract (0, 2.5, 5.0, and 10 g). Two milliliters of extract and 1.5 ml of 0.2 mM DPPH solution were added to the test tube, mixed, and reacted at room temperature in the dark for 30 min, and the absorbance was measured at 517 nm using a microplate reader (SpectraMax M5, Molecular Devices, Sunnyvale, USA). The absorbance of the control, with 80% ethanol added instead of the extract, was measured three times.

The ABTS radical scavenging ability was measured using the method described by De Camargo et al. (De Camargo et al., 2014). Further, 7.4 mM ABTS solution and 2.6 mM potassium persulfate were mixed 1:1 (v/v) and diluted with 80% ethanol to obtain an absorbance value of 0.70 ± 0.05 at 734 nm after reaction at room temperature in the dark for 24 h. After mixing 450 μl of the ABTS solution and 50 μl of the sample solution and reacting at room temperature in the dark for 6 min, the absorbance was measured three times at 734 nm and calculated. The DPPH and ABTS radical scavenging activities were calculated as follows:

$$\text{Scavenging, activity}(\text{\%})=\frac{1-\text{cookie, extract, absorbance ,at,}517\text{nm}}{\text{control, absorbance, at,}517\text{nm}}\times 100$$

Consumer acceptance testing of red beet cookies

Sensory evaluation of red beet rice cookies was conducted by a panel of 30 college and graduate students from the Department of Food Science and Biotechnology at Sungshin Women's University after explaining the purpose and evaluation items of the experiment. Cookie samples of a certain size (5 mm in height and 40 mm in diameter) were prepared and provided in disposable paper cups. The samples were evaluated by determining the extract content in the following order: 0, 2.5, 5.0, and 10 g. During the evaluation, participants were asked to rinse their mouths with bottled water to reduce confusion among the samples. Color, appearance, aroma, texture, taste, and overall preference were measured on a 7-point scale. The highest score was seven for “like very much”, and the lowest was one for “dislike very much”. Approval for the use of human subjects was obtained from the Institutional Review Board (IRB) at Sungshin Women's University (IRB approval number SSWUIRB-2022-404).

Statistical analysis

The experimental results were subjected to one-way analysis of variance (ANOVA) using SPSS Ver 26 for Windows. To test the significance of differences in each section, a comparative analysis was performed using Duncan’s multiple range test at p < 0.05 level.

Results and discussion

Dough pH and density quality characteristics

The density and pH of the rice cookies, measured according to the amount of red beet extract added (0, 2.5, 5.0, and 10 g) are shown in Table 2. The pH of the control group without red beet extract was highest at 7.29 ± 0.02 and the pH of the group with red beet extract was 6.84 ± 0.04–7.04 ± 0.03, which significantly decreased as the amount of red beet extract increased (p < 0.05). pH affects the flavor and color of cookies. The pH value decreased in omija cookies (Kim et al., 2015), American cookies with coffee extract residues (Han and Lee, 2021), and eggplant flour addition in cookie (Soares et al., 2022), which is consistent with the results of the present study. Red beets contain large amounts of antioxidants (Babarykin et al., 2019) and are assumed to influence the pH changes in rice cookies.

Table 2.

Density and pH of rice cookies containing various contents of red beet extract

	Density	pH
Control	2.17 ± 0.29 ns*	7.29 ± 0.02a**
RBE1	1.50 ± 0.50	7.04 ± 0.03b
RBE2	2.00 ± 0.00	7.03 ± 0.05b
RBE3	1.67 ± 0.58	6.84 ± 0.04c

All values are expressed as mean ± SD (n = 3)

*ns: not significant

**Different letters (a–c) within the same column indicate significant differences (p < 0.05)

Lower density means greater crispiness and superior texture. Dough density relies upon the type of fat and oil used in the formulation. The less aerated dough is denser than aerated dough bringing about stiffer dough (Devi and Khatkar, 2018). No significant differences were observed between the densities of the control rice cookies and the density based on the amount of red beet extract added. These results are similar with those of studies on cookies prepared using sweet potato flour (Jemziya and Mahendran, 2017).

Physical properties of cookies

Table 3 shows the spread factor, loss rate (%), and hardness (%) of rice cookies prepared with different amounts of red beet extract (0, 2.5, 5.0, and 10 g). The spreadability of the control group was the lowest at 5.54 ± 0.07 and the spreadability of the additive group was 5.62 ± 0.08–5.74 ± 0.15, which was higher than the control group; however, no significant difference was observed. Spreadability is affected by factors, such as the viscosity of the dough, auxiliary ingredients, moisture content, and protein content. Generally, the greater the spreadability, the better will be the cookie quality (Ikuomola et al., 2017). In the case of cookies containing tannia cocoyam, the spreadability index of the additional group cookies was greater than that of the control group. The addition of cocoyam was reported to increase the moisture content in the dough, delaying the drying time in the oven, and as a result, the time for spreadability to stop was prolonged, increasing the spreadability index (Charushika et al., 2024). This supports the results of this study, in which the moisture content of the red beet extract-supplemented group was higher than that of the control group, resulting in increased spreadability.

Table 3.

Spread factor and loss rates of rice cookies with various contents of red beet extract

	Spread factor	Loss rates (%)
Control	5.54 ± 0.07 ns*	13.78 ± 0.12a**
RBE1	5.62 ± 0.08	12.78 ± 0.96ab
RBE2	5.74 ± 0.15	12.53 ± 1.34ab
RBE3	5.72 ± 0.13	11.62 ± 0.87b

All values are expressed as mean ± SD (n = 3)

*ns: not significant

**Different letters (a and b) within the same column indicate significant differences (p < 0.05)

The loss rate was highest in the control group at 13.78 ± 0.12 and in the addition group it decreased significantly from 12.78 ± 0.96 to 11.62 ± 0.87 (p < 0.05). In a study on cookies with added persimmon leaf powder, the cookie dough underwent a physicochemical reaction with the added ingredients to form bound water, and as the amount of additives increased, the amount of bound water also increased, and the loss rate decreased (Lim and Lee, 2016). Therefore, it is assumed that as the amount of red beet extract increases, the amount of bound water also increases, and the loss rate decreases. A study on rice cookies with wheat-barley flour blends also reported that the loss rate decreased as the amount of added auxiliary ingredients increased, showing a trend similar to that observed in this study (Sharma and Gujral, 2014).

The hardness of the cookies was the highest in the control group at 14.73 ± 1.68. A significant difference was observed in the strength factor, which decreased as the amount of red beet extract added increased. These results show that as the amount of sub-materials increases, the hardness decreases because they interfere with gluten formation, which is consistent with research on cookies from composite flours of watermelon seed, cassava and wheat (Ubbor and Akobundu, 2009) and with Tenebrio molitor, Protaetia brevitarsis, and Gryllus bimaculatus powder (Jang et al., 2022).

Proximate compositions

The results of the analysis of the general ingredients of cookies manufactured using different amounts of red beet extract (0, 2.5, 5.0, and 10 g) are presented in Table 4. The moisture content (%) of rice cookies with red beet extract increased significantly compared to that of the control group, 3.47 ± 0.06 and tended to decrease to 2.23 ± 0.21 in 10% rice cookies (p < 0.05). It was reported that moisture content in cookies increased when it was prepared by adding banana flour (Agama-Acevedo et al., 2012), barley flour (Gupta et al., 2011). Additionally, the moisture content of rice cookies was lower than that of wheat cookies (Chung et al., 2014). With the rice flour substitution in this study, the cookies might have increased heterogeneity by the changes in the components such as proteins, starches and fibers, which might contribute to the change in moisture retention during baking (Chung et al., 2014).

Table 4.

Water content, crude ash, and crude fat of rice cookies with various contents of red beet extract

	Moisture content (%)	Crude ash (%)	Crude fat (%)
Control	3.47 ± 0.06b*	0.85 ± 0.03c	27.75 ± 1.58 ns**
RBE1	3.83 ± 0.11ab	0.94 ± 0.04b	25.41 ± 3.08
RBE2	3.96 ± 0.32a	0.97 ± 0.01b	25.66 ± 0.02
RBE3	2.23 ± 0.21c	1.20 ± 0.00a	29.47 ± 2.81

All values are expressed as mean ± SD (n = 3)

*Different letters (a–c) within the same column indicate significant differences (p < 0.05)

**ns: not significant

The ash content (%) of rice cookies significantly increased from 0.85 ± 0.03 to 1.20 ± 0.00 as the content of red beet extract increased compared to that of the control group, 0.85 ± 0.03 (p < 0.05). These results were consistent with those of a previous study on the physicochemical and sensory characteristics of cookies containing malted barley bran (Ikuomola et al., 2017).

The crude fat content (%) increased as the red beet extract content increased; however, the difference was not significant. All cookies tended to have a high fat content of more than 25%, which may be because of the butter (100 g), added to prepare the cookies. This is consistent with previous research on the quality characteristics and antioxidant activity of wheat-sesame flour cookies (Olagunju and Ifesan, 2013).

Free radical-scavenging activity is the most widely used method for measuring the antioxidant ability of red beet cookies. It shows the electron-donating ability of antioxidant substances and is used to inhibit the aging caused by free radicals in the human body (Biesalski, 2002). The results of measuring the DPPH and ABTS radical scavenging activities (%) of rice cookies are shown in Table 5. DPPH radical scavenging activity was measured as the degree to which purple DPPH was reduced and discolored when it reacted with antioxidant substances (Gülçin et al., 2005). Compared to the control group measurement result of 19.91 ± 0.72, rice cookies with red beet extract showed higher DPPH radical scavenging activity. Particularly, 10% rice cookies showed the highest score of 74.48 ± 0.23 and significantly increased as the amount of the added red beet extract increased (p < 0.05). According to a study using cake flour, the DPPH radical scavenging activity increased as wheat flour was replaced with rice flour (Wanyo et al., 2009). Accordingly, rice cookies are assumed to exhibit a relatively higher antioxidant capacity than wheat cookies.

Table 5.

DPPH and ABTS radical scavenging activity of rice cookies with various contents of red beet extract

	DPPH radical scavenging activity (%)	ABTS radical scavenging activity (%)
Control	19.91 ± 0.72d*	14.77 ± 0.16d
RBE1	70.33 ± 0.84c	31.77 ± 0.40c
RBE2	71.52 ± 0.02b	43.23 ± 0.49b
RBE3	74.48 ± 0.23a	88.16 ± 0.33a

All values are expressed as mean ± SD (n = 3)

*Different letters (a–d) within the same column indicate significant differences (p < 0.05)

ABTS radical scavenging activity was measured based on the principle that the blue-green color of the ABTS radical, which is expressed by the reaction between ABTS and potassium persulfate, is discolored by antioxidant substances (Re et al., 1999). The control group’s 14.77 ± 0.16 and the added group’s 31.77 ± 0.40–88.16 ± 0.33 significantly increased as the amount of the added red beet extract increased (p < 0.05). These results are consistent with those of the reports that the DPPH and ABTS radical scavenging activities of cookies supplemented with oleaster flour (Sahan et al., 2019) and date seed powder (Najjar et al., 2022) increased as the concentration of additives increased.

Sensory evaluation

The sensory profiles of the cookie samples were evaluated in terms of color, appearance, flavor, texture, taste, and overall acceptability (Table 6). For color, the 10% rice cookie was the highest at 5.77 ± 1.30 and for appearance and flavor, the 5% rice cookie was rated the highest at 5.97 ± 1.10 and 5.83 ± 0.99; however, no significant difference was observed. Regarding texture, the 10% rice cookie showed significantly the highest preference at 5.93 ± 1.26, whereas the control group was rated the lowest at 4.87 ± 1.66 (p < 0.05). In terms of taste, the 5% rice cookie had a significantly higher score of 5.93 ± 0.98 compared to the control group (p < 0.05). The overall acceptability results also showed that the 5% rice cookie received a significantly higher score of 5.80 ± 1.13 (p < 0.05). In this study, rice cookies containing 5–10% red beet extract showed higher values than the control group for all sensory evaluation items. Accordingly, it was determined that the taste of the user could be satisfied if cookies were manufactured by adding 5%–10% red beet extract to rice flour.

Table 6.

Consumer preference test of rice cookies with various contents of red beet extract

	Color	Appearance	Flavor	Texture	Taste	Overall acceptability
Control	5.47 ± 1.68 ns*	5.37 ± 1.63 ns	5.20 ± 1.65 ns	4.87 ± 1.66c**	5.03 ± 1.67b	5.00 ± 1.66b
RBE1	5.17 ± 1.44	5.30 ± 1.62	5.47 ± 1.14	5.20 ± 1.30bc	5.23 ± 1.36ab	5.10 ± 1.35ab
RBE2	5.50 ± 1.38	5.97 ± 1.10	5.83 ± 0.99	5.80 ± 1.06ab	5.93 ± 0.98a	5.80 ± 1.13a
RBE3	5.77 ± 1.30	5.90 ± 0.96	5.73 ± 1.08	5.93 ± 1.26a	5.73 ± 1.23ab	5.57 ± 1.17ab

All values are expressed as mean ± SD (n = 3)

*ns: not significant

**Different letters (a–c) within the same column indicate significant differences (p < 0.05)

This study investigated the potential of using red beet as a functional ingredient and evaluated the effect of adding red beet extract in varying amounts (2.5 g, 5 g, and 10 g) to develop the most suitable cookie for consumer preferences. There was no significant difference in the density of the control rice cookie compared to cookies with varying amounts of red beet extract. However, the pH decreased significantly with increasing red beet extract. While spreadability values were higher in the red beet extract groups, the difference was not significant. The loss rate decreased significantly as the amount of red beet extract increased. Moisture content significantly increased with the addition of red beet extract, though it tended to decrease at 10%. The crude fat content increased with red beet extract, but the difference was not significant. Notably, both DPPH and ABTS radical scavenging activities showed a significant increase as red beet extract levels rose, highlighting the antioxidant benefits of the extract. In the sensory test, no significant differences were observed in color, appearance, or aroma, but cookies with 5% and 10% red beet extract had higher scores in texture, taste, and overall preference.

The addition of red beet extract to cookie formulations can enhance antioxidant properties without negatively affecting consumer acceptance. This strategy not only promotes the development of functional foods but can also be directly applied to the production of value-added products in the food industry.

Supplementary Information

Below is the link to the electronic supplementary material.

Funding

This study was supported by Korea Institute of Marine Science & Technology Promotion (KIMST) funded by the Ministry of Oceans and Fisheries (RS-2024-00404977).

Declarations

Conflict of interest

There are no conflicts of interest to declare.