Home Investigating the impact of wet rendering (solventless method) on PUFA-rich oil from catfish (Clarias magur) viscera
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Investigating the impact of wet rendering (solventless method) on PUFA-rich oil from catfish (Clarias magur) viscera

  • Jaydeep Dave , Ali Muhammed Moula Ali EMAIL logo , Nishant Kumar , Muralidharan Nagarajan , Marek Kieliszek and Sri Charan Bindu Bavisetty EMAIL logo
Published/Copyright: July 17, 2024
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Open Life Sciences
From the journal Open Life Sciences Volume 19 Issue 1

Abstract

Catfish (Clarias magur) is a popular freshwater fish food worldwide. The processing of this fish generates a significant amount of waste, mainly in the form of viscera, which constitutes around 10–12% of the fish’s total weight. This study was focused on extracting polyunsaturated fatty acid (PUFA)-rich oil from catfish viscera, aiming to enhance the extraction process and make the production of oil and handling of fish byproducts more cost-effective. The wet reduction method, a solvent-free approach, was used for extraction, with yield optimization done via the Box–Behnken design. The resulting oil was evaluated for its oxidative quality and chemical characteristics. The optimal conditions for the wet rendering process were as follows: viscera to water ratio, 1:0.5 (w/v); temperature, 90℃; and time, 20 min, yielding 12.40 g/100 g of oil. The oil extracted under optimal wet rendering conditions had quality and oxidative stability comparable to solvent extraction and fewer secondary oxidation compounds. This oil had a higher PUFA content, specifically a 4:1 ratio of omega 6 to omega 3. Such oil, derived from catfish viscera, is suitable for the food industry due to its solvent-free extraction method.

Keywords: catfish waste management; fish oil new resource; solvent-free extraction; polyunsaturated fatty acids; fatty acid analysis; volatile compounds

1 Introduction

Catfish is a freshwater fish popular among consumers worldwide, with a global production rate reaching 5.8 million metric tons/year in 2021 [1]. A massive increase in demand exists for catfish driven by the consumer’s need for sustainable sources of proteins and healthy lipids, low cost, and organoleptic properties [2]. Asia is the largest producer of catfish, and China is the topmost producer. The most farmed catfish for consumption are the channel catfish (Ictalurus punctatus) and the African catfish (Clarias gariepinus). The other type of catfish, Clarias magur, commonly found in South and Southeast Asia, is considered a delicacy in countries like Thailand, Indonesia, Vietnam, India, and Bangladesh [3,4]. C. magur is a relatively new species in aquaculture and has yet to be utilized by the processing industry [5]. Therefore, limited research has been conducted in developing value-added products from this fish or its processing byproducts.

Generally, fish processing generates significant quantities of waste (40–60%, based on wet weight) in the form of head, skin, scales, bones, trimmings, and viscera [6]. Typically, this waste is used to develop low-valued animal feed or fertilizers or discarded, which can contribute to environmental issues [7]. Fish processing waste, especially viscera, contains a significant amount of bioactive compounds, i.e., squalene, monounsaturated fatty acids (MUFA), and polyunsaturated fatty acids (PUFA) [8,9]. It has been reported that different species of catfish may contain 10–40% oil based on the weight of visceral biomass, with the composition including saturated and unsaturated fatty acids, omega 6, and omega 3 fatty acids [10]. Owning to their habitat in freshwater, catfish have lower levels of omega-3 fatty acids, i.e., eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA), as compared with marine fish oil [11]. While catfish still contain significant quantities of oleic acid, linoleic acid, and alpha-linolenic acid (ALA), which have been associated with numerous health benefits, including improved heart and brain health, maintained healthy skin, reduced inflammation, and lower cholesterol and triglycerides levels [12,13].

Typically, recovery of fish oil is done using various chemical solvents, enzymatic hydrolysis, supercritical fluid extraction, wet rendering, and cold pressing [14]. However, some of these methods are linked to several detriments. For instance, chemical solvent extraction can leave traces of hazardous chemicals and is not eco-friendly. Cold pressing, while solvent-free, yields very low amounts of oil. Enzymatic and supercritical fluid extraction, despite being effective, are associated with higher capital expenditures and are hence not cost-effective. Dry rendering, another solvent-free method, involves cooking the fish biomass at high temperatures without water, which can lead to the degradation of heat-sensitive nutrients and the development of off-flavors due to prolonged exposure to high heat [15]. Due to these considerations, the present research dwells on solvent-free, eco-friendly, and cost-effective methods for recovering fish oil [16,17]. In this regard, wet rendering is gaining interest, as it involves cooking fish biomass in water or steam. This method not only prevents the degradation of heat-sensitive nutrients but also minimizes the development of off-flavors, as the lower temperatures used in wet rendering are more gentle on the biomass. The oil released is easily separated from the water and other fish solids, making this process particularly efficient and environmentally friendly [18]. Taati et al. [19] found that wet rendering produced higher fish oil yields than other extraction methods, such as acid hydrolysis and solvent extraction.

Wet rendering is a simple method for fish oil extraction, but it produces relatively impure oil, which may contain residual water, protein, and contaminants. During wet rendering, several factors can affect the yield and quality of fish oil, including the ratio of water to biomass, cooking time, and temperature [20]. Studies show that varying temperatures and times significantly impact the extraction yields and quality of fish oil using the wet rendering process. For instance, using an autoclave at 121°C with a 30-min holding time optimized the oil extraction from yellowfin tuna heads, providing a balance in fatty acid content and minimizing oxidative damage [21]. Another study indicated that increasing extraction temperatures from 80 to 100°C increased oil yields from tilapia and mackerel viscera, with longer cooking times (up to 60 min) further enhancing oil recovery [22]. These variations underscore the necessity of optimizing temperature and time to maximize fish oil extraction efficiency and quality. Therefore, optimizing the cooking conditions can enhance the extraction process’s efficiency, cost-effectiveness, quality, and safety. Hence, this research aimed to optimize the wet rendering extraction to recover high-quality fish oil from catfish (C. magur) viscera. The effect of extraction on oil yield, quality, and oxidative stability was studied.

2 Materials and methods

2.1 Chemicals

Thiobarbituric acid (TBA), trichloroacetic acid (TCA), ammonium thiocyanate, ferrous chloride, and sodium hydroxide (NaOH) were purchased from Merck (Darmstadt, Germany). Solvents, including chloroform, methanol, and hexane, were obtained from Lab-Scan (Bangkok, Thailand). Supelco® 37 component FAME mix (purity 96 to 99%), 1,1,3,3-tetramethoxypropane (purity >98%), and ammonium thiocyanate (purity >98%) were procured from Sigma-Aldrich (St. Louis, MO, USA). Cumene hydroperoxide (C₆H₅C(CH₃)₂OOH) and 2-TBA (C4H4N2O2S) were obtained from Fluka Co. (Buchs, St. Gallen, Switzerland). The rest of the chemicals were obtained from Lab-Scan Ltd. (Bangkok, Thailand). All the chemicals used were of analytical grade.

2.2 Raw material

Freshly available catfish viscera (including liver and intestines) were procured from the Huatake fish market at Ladkrabang, Bangkok, Thailand. Approximately 2–3 kg of viscera was packed in polyethylene bags, placed into the polystyrene container containing ice, and transported to the faculty of the food industry, KMITL. Upon arrival, the viscera were minced using a meat grinder (Model: HR271331, Philips, Netherlands), and the minced visceral samples were used for further analysis.

2.3 Proximate composition

The proximate composition, moisture, proteins, fats, and ash were determined using the AOAC method to be 930.15, 923.03, 920.39, and 923.01, respectively [23,24].

2.4 Extraction of oil

A minced visceral sample (100 g) was used for oil extraction by wet rendering method (solvent-free extraction). The extraction conditions were optimized using the Box-Behnken design (BBD); the experimental parameters are mentioned in Table 1. Three independent variables were considered, i.e., viscera to water ratios (1:0.5, 1:1.25, and 1:2, w/v), cooking temperature (50, 85, and 120℃), and cooking time (5, 15, and 25 min). The wet rendering was performed as suggested by Dave et al. [25] with slight modifications. The extraction was carried out in a hot water bath for 50 and 85℃, and in an autoclave for 120℃. After the experimental run, the sample temperature was brought down to room temperature. The extraction mixtures were filtered using a muslin cloth and filtrated using Whatman no 1 filter paper. The filtrate was further subjected to centrifugation at 2,500 × g for 15 min, the pellets (cell debris) were discarded, and the liquid samples were transferred to a separating funnel to separate oil from an aqueous phase. The recovered oil was subjected to analysis, the excess fat was transferred to an amber bottle, and headspace was flushed using nitrogen gas and stored at −20℃. Figure 1 depicts the flowchart for the extraction process.

Table 1

Runs showing various factors of response surface methodology for oil extraction using the wet rendering method obtained through BBD

Run Factor A, viscera:water (w/v) Factor B, temperature (℃) Factor C, time (min)
1 1:1.25 85 15
2 1:1.25 85 15
3 1:2.00 85 5
4 1:2.00 50 15
5 1:0.50 85 25
6 1:1.25 120 5
7 1:2.00 85 25
8 1:1.25 85 15
9 1:0.50 85 5
10 1:1.25 120 25
11 1:1.25 50 5
12 1:0.50 50 15
13 1:0.50 120 15
14 1:1.25 50 25
15 1:2.00 120 15
Figure 1 Process flow diagram for oil extraction from minced visceral samples using a BBD for optimization.
Figure 1

Process flow diagram for oil extraction from minced visceral samples using a BBD for optimization.

The oils extracted using solvent methods, i.e., chloroform–methanol (2:1) mixture and hexane, were considered control [8].

2.5 Extraction yield

The obtained oil was used to determine the extraction yield. The extraction yield and oil recovery were calculated according to Dave et al. [25]. The yield in terms of g/100 g, wet basis, was calculated using the following formula:

(1) Yield ( g / 100 g ) = Weight of the obtained oil ( g ) Weight of the sample used for the extraction ( g ) × 100 .

The oil recovery in terms of percentage was calculated using the following formula:

(2) Oil recovery ( % ) = Weight of the obtained oil ( g ) Total oil present in the sample ( g ) × 100 .

2.6 Oil quality

2.6.1 Color

The color of the extracted oils was determined using a HunterLab colorimeter (LabScan® XE, Reston, VA, USA) and reported in the CIE color system (L*, a*, and b* representing lightness/brightness, redness/greenness, and yellowness/blueness, respectively) [17]. The corresponding values of a white standard were L* = 93.71, a* = −1.21, and b* = 0.48.

2.6.2 Acid value and free fatty acids (FFAs)

The acid value and FFA of the oils were estimated according to the method described by Chaijan et al. [26] with slight modifications. One gram of oil was dissolved in ten volumes of hexane. To this, two drops of phenolphthalein indicator were added and titrated against 0.02 N potassium hydroxide (KOH) solution until the pink color was obtained. The acid value was expressed as mg KOH consumed to neutralize the free acids in 1 g of oil and calculated using the following formula:

(3) Acid value = 56.1 × N × ( V s − V b ) W ,

where N is the normality of KOH, V s is the volume of KOH required for the sample, V b is the volume of KOH required for blank, and W is the weight of the sample (in g).

FFA was expressed as % oleic acid present in the oil sample, calculated using the following formula:

(4) FFA ( % ) = Acid value × M w of oleic acid × 10 M w of KOH × 1 , 000 ,

where M w is the molecular weight.

2.7 Oxidative stability of oil

2.7.1 Peroxide value (PV)

The PV of oil was determined spectrophotometrically, as described by Bruno et al. [27], using 20 mM ferric thiocyanate as a reducing agent. A standard curve was plotted using cumene hydroperoxide at 0–50 ppm concentration, and the PV was expressed as meq O2/kg of oil sample.

2.7.2 TBA reactive substances (TBARS)

TBARS analysis was performed as described by Bruno et al. [27] and Dave et al. [23], with slight modifications. Oil (10 mg) was thoroughly mixed with 2.5 mL of a TBA reagent (0.375% TBA, 15% TCA, and 0.25 M HCl). The mixture was incubated at 95℃ for 10 min. The reaction was stopped by cooling the tubes under running water and centrifuging at 3,600 × g for 20 min at 25℃. The upper layer was collected, and absorbance was measured at 532 nm. The TBARS content of the oil was measured using a standard curve plotted with 1,1,3,3-tetraethoxypropane and expressed as mg malonaldehyde (MDA)/kg oil.

2.8 Characterization of oil

To characterize the oil, four oil samples were selected as follows: 1. The sample was selected based on the highest yield with desirable oil quality (optimized condition), 2. Run 10 was selected as a negative control, which had lower oil quality (higher acid value, higher FFA, higher PV, and higher TBARS). The oil recovered via solvent extraction, i.e., 3. chloroform:methanol (2:1, v/v), and 4. hexane was taken as a positive control.

2.8.1 Fatty acid composition

Fatty acid methyl esters (FAMEs) were prepared according to the method described by Muhammed et al. [28]. The derivatized FAMEs (1 μL) were injected into a gas chromatography (GC) (7890B series; Agilent Technologies, Santa Clara, CA, USA) equipped with a flame ionization detector and an HP 88 capillary column (J & W Scientific Column from Agilent Technologies). The GC conditions were set according to the manufacturer's instructions. Peak identification was based on the retention time of the Supelco® external standard (37 Component FAME Mix). The identified peaks were integrated and calibrated against the standard curve using Open LAB CDS software (Chem Station edition; Agilent Technologies, Santa Clara, CA, USA) [23].

2.8.2 Volatile compounds

Volatile compounds were measured using a GC-mass spectrometer (GC-MS) (HP 5890 series II; Hewlett Packard, Atlanta, GA, USA) equipped with an HP 5972 quadrupole mass detector, as outlined by Moula Ali et al. [29]. The spectra were obtained at an ionization energy of 70 eV. Ionization was performed at 250°C in electron ionization (EI) mode. The acquisition was monitored at a scanning rate of 0.220 s/scan at 25–500 amu. The ChemStation Library was used to identify volatile compounds (Library No. Wiley 275. L). The volatile compounds are expressed in abundance based on the peak area [23].

2.9 Statistical analysis

All the experiments were conducted in triplicates. BBD with three factors was used, and a second-order polynomial model was studied to calculate the F-ratio, coefficient correlation, and lack-of-fit test. The optimal conditions were extracted using the numerical optimization toolbox of Design-Expert 12.0.3. Duncan’s multiple range test enumerated the significant difference between mean values for fatty acid compositions and volatile compounds.

3 Results and discussion

The proximate composition of catfish species C. magur viscera contained 54.67% ± 0.13% moisture, 12.21% ± 0.57% proteins, 24.35% ± 0.76% fat, and 1.86% ± 0.51% ash (wet weight basis). Similar catfish species, i.e., C. magur from Kenya, were reported to contain 38.20% fat, which was relatively higher than those from Thailand [30]. The difference in total fat could be possibly due to the difference in geographical distribution and fish feed [31].

3.1 Extraction of oil

3.1.1 Second-order polynomial distribution

Catfish visceral oil recovery optimization studies were carried out using BBD. After the experimental runs, the optimal conditions were developed using software, and the oil was extracted under optimal conditions and was used for further analysis (Table 2). The results for ANOVA are shown in Table 3. The F-values of each response revealed that the quadratic model was significant (p < 0.05). The R 2 value for each response under study was >0.95 (except for color a*, b*), suggesting that more than 95% of results matched the predicted values. According to Ferreira et al. [32], a model is adequate when R 2 > 0.95. This study’s lack of fit, which measures the model’s fitness, was insignificant (p > 0.05). Therefore, the number of experiments was sufficient to determine the effects of the independent variables on catfish viscera oil yield.

Table 2

Yield, recovery, color, and quality parameters of catfish viscera oils recovered using the wet rendering method

Run Oil yield a Oil recovery (%) L * a * b * Δ E * Acid value b FFA c PV d TBARS e
1 11.25 ± 0.29 47.06 ± 0.44 62.17 ± 0.10 −1.91 ± 0.02 19.18 ± 0.07 36.70 ± 0.19 5.23 ± 0.01 0.26 ± 0.04 0.532 ± 0.002 0.45 ± 0.04
2 11.28 ± 0.32 47.12 ± 0.31 62.15 ± 0.12 −1.90 ± 0.01 19.12 ± 0.04 36.67 ± 0.17 5.26 ± 0.01 0.26 ± 0.04 0.527 ± 0.001 0.45 ± 0.04
3 6.41 ± 0.74 28.13 ± 1.41 65.11 ± 0.13 −1.95 ± 0.03 19.21 ± 0.04 34.23 ± 0.22 5.25 ± 0.25 0.26 ± 0.01 0.566 ± 0.004 0.36 ± 0.04
4 5.64 ± 0.29 17.04 ± 0.33 68.19 ± 0.22 −1.93 ± 0.04 19.38 ± 0.01 31.80 ± 0.03 4.95 ± 0.01 0.25 ± 0.05 0.426 ± 0.004 0.40 ± 0.03
5 11.37 ± 0.78 45.42 ± 0.97 61.44 ± 0.21 −1.86 ± 0.02 19.11 ± 0.05 37.29 ± 0.42 5.24 ± 0.27 0.26 ± 0.01 0.433 ± 0.002 0.45 ± 0.02
6 7.10 ± 0.33 29.6 ± 0.53 63.21 ± 0.13 −1.93 ± 0.03 19.11 ± 0.04 35.78 ± 0.27 5.41 ± 0.28 0.27 ± 0.01 0.410 ± 0.001 0.41 ± 0.02
7 8.87 ± 0.21 36.13 ± 0.63 63.23 ± 0.12 −1.88 ± 0.04 19.21 ± 0.02 35.81 ± 0.15 5.42 ± 0.18 0.27 ± 0.05 0.396 ± 0.002 0.32 ± 0.01
8 11.25 ± 0.62 47.06 ± 0.44 62.16 ± 0.14 −1.90 ± 0.02 19.21 ± 0.07 36.74 ± 0.19 5.21 ± 0.01 0.26 ± 0.04 0.536 ± 0.002 0.45 ± 0.04
9 7.23 ± 0.41 29.69 ± 0.57 67.23 ± 0.12 −1.91 ± 0.03 19.41 ± 0.04 34.63 ± 0.15 5.11 ± 0.28 0.26 ± 0.01 0.418 ± 0.001 0.34 ± 0.02
10 10.93 ± 0.27 44.88 ± 0.48 61.21 ± 0.14 −1.84 ± 0.04 19.21 ± 0.02 36.55 ± 0.32 5.44 ± 0.01 0.27 ± 0.04 0.395 ± 0.003 0.32 ± 0.03
11 3.68 ± 0.29 13.92 ± 0.22 66.24 ± 0.41 −1.91 ± 0.03 19.51 ± 0.02 33.46 ± 0.24 4.83 ± 0.07 0.24 ± 0.03 0.492 ± 0.003 0.31 ± 0.04
12 7.32 ± 0.58 29.28 ± 0.51 65.24 ± 0.11 −1.90 ± 0.02 19.25 ± 0.01 34.32 ± 0.31 4.83 ± 0.27 0.24 ± 0.01 0.459 ± 0.002 0.30 ± 0.03
13 11.52 ± 0.79 47.31 ± 1.19 61.21 ± 0.32 −1.90 ± 0.01 19.17 ± 0.02 37.52 ± 0.10 5.25 ± 0.26 0.26 ± 0.01 0.373 ± 0.001 0.51 ± 0.03
14 6.69 ± 0.31 28.41 ± 0.97 67.12 ± 0.14 −1.93 ± 0.04 19.35 ± 0.02 32.65 ± 0.13 5.25 ± 0.26 0.26 ± 0.01 0.377 ± 0.002 0.46 ± 0.03
15 9.17 ± 0.64 21.19 ± 0.33 64.11 ± 0.13 −1.95 ± 0.03 19.21 ± 0.04 35.07 ± 0.15 5.34 ± 0.01 0.27 ± 0.04 0.470 ± 0.003 0.31 ± 0.03
Optimum conditions 12.34 ± 0.34A 50.67 ± 0.81A 61.48 ± 0.33A −1.86 ± 0.03A 19.11 ± 0.02C 37.29 ± 0.25B 5.44 ± 0.21B 0.27 ± 0.02B 0.395 ± 0.001A 0.44 ± 0.02B
A 16.68 ± 0.51B 68.50 ± 0.73B 77.28 ± 0.04B −1.88 ± 0.02B 17.86 ± 0.03A 24.12 ± 0.13A 2.34 ± 0.21A 0.12 ± 0.02A 0.366 ± 0.001A 0.05 ± 0.02A
B 13.54 ± 0.39A 51.49 ± 0.61A 78.11 ± 0.10B −1.86 ± 0.03A 18.35 ± 0.01B 23.92 ± 0.05A 2.21 ± 0.18A 0.11 ± 0.01A 0.366 ± 0.002A 0.03 ± 0.01A

Solvent extraction methods were positive controls, i.e., hexane and chloroform:methanol.

aOil yield in terms of g/100 g, based on wet weight; bacid value in terms of mg KOH/g of oil; cFFAs in terms of % oleic acid; dPV in terms of meq O2/kg oil; eTBARS in terms of mg MDAEq/kg of oil.

Runs 1-15, oil extracted using the wet rendering method.

Optimum condition: viscera to water ratio, 1:0.5 (w/v); temperature, 90℃; and time, 20 min.

A: Oil extracted with chloroform:methanol (2:1, v/v).

B: Oil extracted with hexane.

Mean ± SD from triplicate determinations.

Different uppercase superscripts in the same column indicate significant differences (p < 0.05) between A, B, and the optimum condition.

Table 3

ANOVA and regression coefficients of the second-order polynomial model for response variables against yield, recovery, color, and quality parameters of catfish visceral oils

Source df p value F value
Oil yield Oil recovery L* Acid value FFA PV TBARS Oil yield Oil recovery L* Acid value FFA PV TBARS
Model 9 <0.0001 <0.0001 0.0060 0.0002 0.0093 0.0001 0.0207 100.31 100.29 12.73 50.07 10.48 64.99 7.29
A 1 0.0005 0.0005 0.0148 0.0027 0.8394 0.0015 0.0838 64.00 64.00 13.30 30.31 0.0456 40.01 4.64
B 1 <0.0001 <0.0001 0.0006 <0.0001 0.0007 0.0123 0.3572 272.35 272.26 59.01 269.40 55.25 14.64 1.03
C 1 <0.0001 <0.0001 0.0180 0.0006 0.0021 0.0001 0.0838 216.39 216.33 11.98 60.70 34.15 107.15 4.64
AB 1 0.2307 0.2306 0.5464 0.6778 0.9223 0.0012 0.0030 1.86 1.86 0.4180 0.1942 0.0105 43.93 28.91
AC 1 0.1017 0.1019 0.0870 0.5823 0.9713 0.0002 0.1057 4.01 4.00 4.51 0.3453 0.0014 88.98 3.89
BC 1 0.5255 0.5233 0.0474 0.0023 0.1269 0.0036 0.0077 0.4652 0.4704 6.84 32.83 3.35 26.40 18.50
A² 1 0.0023 0.0023 0.1318 0.0205 0.4719 0.0014 0.3369 32.34 32.31 3.24 11.16 0.6049 40.51 1.13
B² 1 <0.0001 <<0.0001 0.0173 0.0058 0.5230 <0.0001 0.2720 178.63 178.58 12.25 21.26 0.4712 175.57 1.52
C² 1 <0.0001 <0.0001 0.0665 0.0060 0.5992 0.0003 0.2187 175.97 176.00 5.47 20.83 0.3143 82.25 1.98
Lack of Fit 3 0.9986 0.9985 0.2306 0.3094 0.0534 0.1500 0.1010 0.0086 0.0087 3.49 2.38 17.90 5.83 9.06
R 2 0.9945 0.9945 0.9582 0.9227 0.935 0.9902 0.9902 0.9915
Adj. R 2 0.9846 0.9846 0.8829 0.7645 0.7520 0.9725 0.9725 0.9763
CV % 3.89 3.89 1.10 1.53 1.46 1.47 1.47 5.48

A (viscera: water, w/v), B (temperature), and C (time) represent independent variables.

p-values < 0.05 indicate that model terms are significant.

F-values of each response revealed that the quadratic model was significant (p < 0.05).

Lack of fit, which measures the model’s fitness, was insignificant (p > 0.05).

3.1.2 Optimization of extraction

A numerical optimization toolbox was used to optimize the wet rendering conditions. According to the second-order polynomial equation, the optimum conditions for oil yield included viscera to water ratio, 1:0.5 (w/v); temperature, 90℃; and time, 20 min, with a desirability of 0.842. Under these conditions, the predicted oil yield was 12.34 g/100 g. The results revealed that the experimental value for oil yield was 12.40 g/100 g, demonstrating the high desirability.

3.1.3 Effect of independent variables on the responses

3.1.3.1 Extraction yield

The extraction yield of oil and recovery of oil ranged from 3.39 to 12.28 g/100 g and 13.92 to 50.43%, respectively. This change in values of both parameters suggested that all three independent variables were at linear and quadratic levels (Table 3). The independent variable, viscera to water ratio (w/v), demonstrated that with an increase in water ratio, a significant (p < 0.05) decrease in extraction yield and recovery was observed (Table 4). Similarly, Mbatia et al. [33] reported that the oil yield decreased with an increase in water content, which might be due to the formation of emulsion resulting in the entrapment of oil. In contrast, the effect of temperature showed an increase in the extraction yield at a temperature from 50 to 100℃. Similarly, El-Rahman et al. [22] observed an increase in oil yields up to 100℃ during wet reduction of tilapia and mackerel viscera. Further increase in temperature from 100 to 120℃ resulted in a significant (p < 0.05) decrease (73%) in the extraction yield (Tables 2 and 4). Further, increasing the temperature to 121℃ might have led to higher protein denaturation exposing the hydrophobic domain, resulting in protein–lipid interaction and decreased yield [8]. A study on the effects of varying heat treatments on lipid removal during pelagic fishmeal production observed that an increase in temperature above 85℃ could affect the lipid extraction efficiency [34]. Time factor exhibited a linear correlation with oil recovery. The increase in the extraction time increased the oil yield and oil recovery. Kudre et al. [35] also observed that a more extended extraction period effectively increased the yield. This could be correlated with the higher breakdown of adipose tissues with respect to time and the release of more fat, thereby improving the recovery [20]. It was noted that when compared to the chloroform:methanol (2:1, v/v) extraction method, the optimal conditions had a relatively lower oil yield and recovery (2 and 7%, respectively) (Table 2). The results indicated that the values were comparable and not very low with the prospect of any extraction method compared to the standard solvent extraction method.

Table 4

Effect of individual independent variables on the oil yield, oil recovery, color, and quality parameters of the catfish visceral oil using the wet rendering method

Process variables Oil yield (g/100 g) Oil recovery (%) L* Acid value (mg KOH/g of oil) FFA (% oleic acid) PV (mg cumene hydroperoxide equivalent/kg of oil) TBARS (mg MDAEq/kg of oil)
A
B
C

A (viscera: water, w/v), B (temperature), and C (time) represent independent variables.

: increasing trend.

: decreasing trend.

: increasing followed by decreasing trend.

: Non-significant changes.

3.1.3.2 Color

The color values of oils from the solvent extraction and wet rendering processes are shown in Table 2. During the wet rendering, the L* (lightness) values ranged from 61.21 to 67.23. These values were significantly (p < 0.05) influenced by all three independent variables at a linear level, as shown in Table 3. When more water was used during extraction, the resulting oil had a lower lightness value (L*). It is unclear why this happened, but this could be due to the difference in the oil’s chemical composition. On the other hand, the lightness value decreased as the extraction process continued with an increase in temperature and time. Many researchers have suggested various explanations for this phenomenon. Aidos et al. [36] reported that protein and carbohydrates in viscera hydrolyzed to smaller fractions and mixed in oil to lower lightness. Another study by Grebenteuch et al. [37] reported that high temperatures could polymerize browning substances by condensation and dehydration of the secondary products from lipid oxidation during the wet rendering method. However, no significant difference was observed in other color parameters, including a* values (−1.83 to −1.99) and b* values (19.11 to 19.51), suggesting no difference in greenness and yellowness of the oil, irrespective of the extraction methods used.

3.1.3.3 Acid value and FFA

The acid value and FFA content are shown in Table 2. Among all the samples, the highest acid value of the wet rendering was from run 10 (5.44 mg KOH/g of oil), which was significantly higher than chloroform:methanol (56.98%) and hexane (59.37%) (p < 0.05). The optimal conditions exhibited lower acid values when compared to the chloroform:methanol and hexane method, i.e., 20.6 and 21.3%, respectively. The acid value is an index for the hydrolytic rancidity of fats and oils [38]. The increased acid value denotes a greater proportion of FFA, released due to hydrolysis from the glycerol backbone due to the extraction process, which is readily available for oxidation [39,40]. As determined, the FFA content was higher in the wet rendering from run 10, which was 20.8% higher than chloroform:methanol and 21.8% higher than hexane (p < 0.05). The acid value and FFA were significantly (p < 0.05) affected by all three independent variables at linear and quadratic levels (Table 3). It was observed that with an increase in water ratio, temperature, and time, there was a gradual increase in the acid value and FFA. This could be due to increased hydrolysis of fat and oil to release FFA [41,42]. Similar findings were observed by Suseno et al. [43] during the wet rendering of tilapia byproducts.

3.1.3.4 PV

Generally, fish oil contains a high amount of PUFA, which can react with atmospheric oxygen, especially during extraction, and produce primary and secondary oxidation products. The presence of heavy metals in unrefined fish oil can intensify these oxidation reactions, as these metals often act as catalysts, accelerating the formation of hydroperoxides and other oxidative products [44]. It was observed that the PV of both the solvent-extracted oils remained lower when compared to all the runs in the randomized design and optimal conditions. It was observed that the PV of both the solvent-extracted oils remained lower when compared to all the runs in randomized design and optimal conditions. The PV of optimal conditions was found to be 1.07 times higher than oil recovered through solvent extraction (p < 0.05). However, all oil samples showed the PV within the limit described by the Codex Alimentarius Commission, i.e., <5 meq O2/kg of fish oil [45]. Furthermore, it was also observed that the PV had a linear relation with the water ratio during the experiment. The higher water content has been reported to hydrolysis triglycerides and liberate FFAs, which can easily undergo oxidation reactions and contribute to PV [43]. Moreover, the temperature also shows a profound effect on PV in the wet rendering process at 50–80℃. With the increase in temperature, the thermal degradation and oxidation of lipids are higher, which can react with oxygen more effectively [46,47]. Further, with an increase in temperature (80–121℃), the PV reduced, but an increase in TBARS was observed. This could be due to the reduction of primary oxidation products and an increase in the formation of secondary oxidation products [48,49]. Charuwat et al. [50] documented that the hydroperoxide decomposition rate may increase at a higher temperature, producing secondary compounds (aldehyde and ketone). Probably, extraction time exhibited a similar pattern for PV and TBARS, i.e., from 5 to 15 min, a slight increase in PV was observed, and from 15 to 25 min, PV decreased (Table 4). This can be supported by the hydrolysis of triglyceride ester bonds occurring more at prolonged extraction time, elevating the FFA level, which reacts with oxygen and generates hydroperoxides [36]. Weber et al. [51] also observed decreased PV during boiling silver catfish fillets for 30 min. This could be due to prolonged cooking time, where the initial hydroperoxides might decompose into volatile and non-volatile products.

3.1.3.5 TBARS

The results for TBARS are presented in Tables 2 and 4. TBARS of oil extracted using the wet rendering method was higher, which was 8.8-fold higher than chloroform:methanol (2:1, v/v) and 14.66-fold higher than oil recovered through hexane (p < 0.05) under optimal conditions. These results correlated with PV, as discussed in the previous section. The high-value TBARS might be due to higher thermal degradation of fatty acids during wet rendering, resulting in more secondary oxidation products like MDA. These results were in line with that of volatile compounds detected in the oil samples. The results indicated that with an increase in temperature and time, a gradual increase in TBARS was observed (p < 0.05). In comparison, the water ratio did not affect the increasing trend of TBARS values. Weber et al. [51] also observed that thermal processing like boiling increases the TBARS of silver catfish fillets. Prolonged cooking may convert the primary oxidation products into secondary oxidation products, like aldehydes, i.e., MDA. Similar results were observed by Sajib et al. [52], where the TBARS increased during the prolonged cooking of herring fish fillets for 30 min to extract the fish oil.

3.2 Characterization

3.2.1 Fatty acid composition

The fatty acid compositions of all four oil samples selected for characterization are given in Table 5. Linoleic acid was the predominant fatty acid in all four samples, accounting for about 25% of the total fatty acids. The second-highest fatty acid found in all oil samples was oleic acid, accounting for about 21% of total fatty acids. Sathivel et al. [20] also observed the predominance of omega-6 and omega-9 in catfish oils. The other fatty acids were found to be in moderate or lower concentrations. In the present study, it was also noted that the total amount of PUFA was higher (32–63%) than the total amount of saturated fatty acid (p < 0.05). Among PUFA, EPA and DHA were found at lower concentrations in all samples. Sathivel et al. [53] observed that the long-chain omega-3 fatty acids in catfish oil were significantly less (ranging from 1.8 to 2.1 mg/g). Based on the type of extraction methods, it was observed that oil recovered through hexane had the highest content of PUFA followed by chloroform:methanol (2:1, v/v), optimal conditions, and Run 10 (p < 0.05). The thermal process during wet rendering profoundly affected the reduction of the PUFA [54], but the optimal conditions retained the PUFA to a considerable quantity. Similarly, the extraction can also affect the recovery of PUFA; a longer extraction time can degrade PUFA to a more significant extent [55]. The oil obtained under optimal conditions showed 27.40 μg/g of oil EPA and 30.80 μg/g of oil DHA, which was significantly (p < 0.05) higher than the oil recovered by Run 10 (13.32 μg/g EPA and 21.12 μg/g of DHA, respectively). This can also be due to the higher rate of oxidation in oil recovered through Run 10, thereby degrading these compounds to produce the secondary oxidation compounds. These results were in line with the findings of Dave et al. [25].

Table 5

Changes in the fatty acid composition of catfish viscera oils recovered using the wet rendering method compared to two solvent extraction methods

Fatty acids A B Optimal conditions* Run 10**
(μg/g of oil)
C14 32.61 ± 0.02a 32.61 ± 0.01a 32.62 ± 0.01a 31.82 ± 0.02b
C14:1 3.32 ± 0.00a 3.32 ± 0.01a 3.51 ± 0.02b ND
C15 4.61 ± 0.01c 4.51 ± 0.01b 4.61 ± 0.02c 4.22 ± 0.01a
C16 99.81 ± 0.21a 110.71 ± 0.32b 145.92 ± 0.29d 139.22 ± 0.30c
C16:1 (n7) 93.01 ± 0.31c 128.81 ± 0.23d 62.40 ± 0.24b 49.31 ± 0.21a
C17 4.12 ± 0.02c 4.22 ± 0.01d 4.01 ± 0.01b 3.71 ± 0.01a
C18 3.71 ± 0.02 3.72 ± 0.00b 3.61 ± 0.01a 3.61 ± 0.01a
C18:1 (n9) 249.80 ± 0.52c 256.81 ± 0.43d 229.90 ± 0.29b 201.22 ± 0.51a
C18:2 (n6) 305.11 ± 0.19c 324.32 ± 0.28d 262.90 ± 0.22b 212.51 ± 0.20a
C18:3 (n6) 0.72 ± 0.02b 0.81 ± 0.01c 0.61 ± 0.01a ND
C18:3 (n3) 11.21 ± 0.01c 13.62 ± 0.02d 9.12 ± 0.02b 6.21 ± 0.02a
C18:4 (n3) 6.21 ± 0.01c 6.21 ± 0.02c 6.01 ± 0.01b 4.71 ± 0.03a
C20 1.91 ± 0.03c 2.02 ± 0.01d 1.71 ± 0.02b 0.51 ± 0.03a
C20:1 (n9) 12.92 ± 0.02c 13.71 ± 0.04d 10.11 ± 0.04b 7.41 ± 0.05a
C20:2 (n6) 3.71 ± 0.01b 3.71 ± 0.00b 3.71 ± 0.01b 2.32 ± 0.02a
C20:3 (n6) 2.41 ± 0.02c 2.42 ± 0.01c 2.31 ± 0.01b 0.60 ± 0.02a
C20:4 (n6) 7.52 ± 0.02c 7.82 ± 0.02d 7.31 ± 0.03b 5.80 ± 0.01a
C20:3 (n3) 0.22 ± 0.00a 0.22 ± 0.01a 0.20 ± 0.00a ND
C20:4 (n3) 3.11 ± 0.03b 3.21 ± 0.02c 2.81 ± 0.02a ND
C20:5 (n3) 29.11 ± 0.13c 29.41 ± 0.18c 27.40 ± 0.15b 13.32 ± 0.10a
C22 0.71 ± 0.00a 0.71 ± 0.01a 0.71 ± 0.01a ND
C22:1 (n9) 2.12 ± 0.03c 2.22 ± 0.03d 1.92 ± 0.02b 0.51 ± 0.01a
C22:2 (n6) 6.82 ± 0.01c 7.11 ± 0.02d 6.02 ± 0.02b 4.20 ± 0.03a
C22:4 (n6) 1.21 ± 0.00b 1.31 ± 0.01c 1.22 ± 0.02b 0.41 ± 0.01a
C22:5 (n6) 2.41 ± 0.02b 2.41 ± 0.02b 2.51 ± 0.02c 1.31 ± 0.02a
C22:6 (n3) 33.21 ± 0.12c 34.91 ± 0.16d 30.80 ± 0.10b 21.12 ± 0.13a
C24:1 (n9) 0.91 ± 0.01c 0.82 ± 0.01b 0.71 ± 0.01a ND
Saturated fatty acids 150.72 ± 0.20c 161.71 ± 0.02d 196.62 ± 0.16b 183.00 ± 0.01a
MUFAs 358.51 ± 0.31c 402.32 ± 0.22d 305.01 ± 0.02b 258.51 ± 0.20a
PUFAs 412.82 ± 0.32c 506.31 ± 0.28d 362.51 ± 0.17b 272.41 ± 0.11a
Total fatty acids 922.21 ± 0.10a 1070.31 ± 0.12d 864.41 ± 0.10b 713.92 ± 0.25a
Omega 6:Omega 3 (Ʃn6:Ʃn3) 3.92:1 3.75:1 4:1 5:1

A: oil extracted with chloroform:methanol (2:1, v/v), B: oil extracted with hexane.

*Oil extracted under optimal conditions: viscera: water (w/v) = 1:0.5, extraction temperature = 90℃, and extraction time = 20 min.

**Negative control, viscera: water (w/v) = 1:1.25, extraction temperature = 120℃, and extraction time = 25 min.

Values are given as mean ± SD (n = 3).

Different lowercase superscripts in the same row denote significant differences (p < 0.05).

ND: not detectable.

The omega-6 to omega-3 ratio is important in fish oil because these are two essential fatty acids the human body cannot produce on its own and must be obtained through diet. While omega-6 and omega-3 fatty acids are important for maintaining optimal health, a proper balance between them is crucial for overall health and well-being. According to the FDA, the optimal ratio of omega-6 to omega-3 fatty acids should be around 4:1 or lower [56]. A higher ratio of omega-6 to omega-3 fatty acids is associated with an increased risk of chronic diseases, such as heart disease, arthritis, and certain cancers [57]. In this study, the oil recovered through solvent extraction and optimal conditions showed an omega-6 to omega-3 ratio of 3.75:1 to 4:1. However, the oil recovered through Run 10 showed 5:1 omega-6 to omega-3 ratio, which made the oil undesirable for consumption.

3.2.2 Volatile compounds

The volatile compounds in oils recovered through solvent and wet rendering methods are shown in Table 6. Aldehydes were found to be the most abundant volatile compounds in oils, including solvent extraction methods. Aldehydes have been widely used as indicators of lipid oxidation in various fats and oil-containing foods because they have low threshold values and mainly contribute to unpleasant odors [39]. The primary aldehydes identified in the present study were octanal, nonanal, pentanal, and hexanal, along with some minor aldehydes, i.e., 2,4-decadienal, and 17-octadecenal. The octanals are mainly produced due to omega-3 fatty acid oxidation, especially ALA [18]. The nonanal, pentanal, hexanal, and 2,4-decadienal are primarily derived from the oxidation of linoleic acid and oleic acid [58]. The oil extracted by solvent methods had fewer aldehydes than the wet rendering method. It can be noted that the temperature and time in the wet rendering process could affect the oxidation of oils. However, the oil obtained under optimal conditions had a low abundance of hexanal and 2,4-decadienal compared to Run 10.

Table 6

Changes in volatile compounds of catfish viscera oils recovered using the wet rendering method as compared to two solvent extraction methods

Volatile compounds A B Optimal conditions* Run 10**
Aldehydes
Hexanal ND ND 0.24b 0.49a
Nonanal ND ND 0.01b 0.05a
2-Octenal ND ND 0.02b 0.06a
2,4-Heptadienal ND ND 0.03b 0.08a
Benzaldehyde ND ND ND 0.07
2-Nonenal ND ND 0.03b 0.05a
2-Decenal ND ND ND 0.10
10-Undecenal ND ND ND 0.17
2-Dodecenal ND ND ND 0.10
2,4-Decadienal 0.04 ND 0.96b 1.12a
Hexadecenal ND ND ND 0.38
17-Octadecenal ND 0.02 0.13b 0.19a
Ketones
3,5-octadien-2-one ND ND ND 0.02
Hydrocarbons
Cyclohexene ND ND 0.02
Pentadecane ND ND ND 0.06
Hexadecane ND ND ND 0.02
Heptadecane ND ND ND 0.08
Others
Furan 2-pentyl ND ND ND 0.02

A: oil extracted with chloroform:methanol (2:1, v/v), B: oil extracted with hexane.

*Oil extracted under optimal conditions: viscera: water (w/v) = 1:0.5, extraction temperature = 90℃, and extraction time = 20 min.

**Negative control, viscera: water (w/v) = 1:1.25, extraction temperature = 120℃, and extraction time = 25 min.

Values are expressed as abundance (×106).

Different lowercase superscripts in the same row denote significant differences (p < 0.05).

ND: not detectable.

The highest number of volatile compounds found in oil recovered through Run 10 followed by optimal conditions. In the present study, the 3,5-octadien-2-one was found only in oil recovered by Run 10. 3,5-octadien-2-one is the most critical ketone produced from the lipid oxidation of omega 3 PUFA Field [59]. Similarly, the oil recovered from Run 10 showed the presence of furan derivatives, i.e., furan 2-pentyl (Table 6). Furan 2-pentyl can be generated from the 12-hydroperoxide of EPA and 16-hydroperoxide of DHA Field [60].

4 Conclusions

The oil recovery from catfish viscera was successfully carried out, which contained PUFA, including oleic acid, linoleic acid, ALA, EPA, and DHA. BBD studies were conducted using three independent variables, i.e., viscera to water ratio (w/v), extraction temperature, and time. The selected variables have shown a significant effect on the quality attributes such as color, acid value, FFA, PV, and TBARS, which was also confirmed by fatty acid analysis and oil volatile compounds. The optimal conditions (viscera to water ratio, 1:0.5 (w/v); temperature, 90℃; and time, 20 min) was developed with a comparable oil yield and quality to conventional solvent extraction methods. Finally, this study concludes that the acquired conditions are very well suited to extract the oil from C. magur viscera for application in the food industry aimed at a solvent-free extraction process. Further studies on the partial purification of this extracted oil with storage stability and application will be carried out.

Acknowledgements

This work was supported by King Mongkut’s Institute of Technology Ladkrabang.

  1. Funding information: Authors state no funding involved.

  2. Author contributions: Jaydeep Dave: conducting research activities, investigating, writing – original draft preparation, and methodology. Ali Muhammed Moula Ali: work plan, designing the experiments, writing – original draft preparation, correcting and critically improvising the manuscript, and methodology. Nishant Kumar: writing – review and editing. Marek Kieliszek: writing – review and editing, investigating, source, and funding acquisition. Sri Charan Bindu Bavisetty: conducting lab experiments, documenting, and improvising the manuscript.

  3. Conflict of interest: Marek Kieliszek, who is the co-author of this article, is a current Editorial Board member of Open Life Sciences. This fact did not affect the peer-review process. Authors state no conflict of interest.

  4. Data availability statement: The datasets generated during and/or analyzed during the current study are available from the corresponding author upon reasonable request.

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Received: 2024-02-11
Revised: 2024-05-12
Accepted: 2024-06-03
Published Online: 2024-07-17

© 2024 the author(s), published by De Gruyter

This work is licensed under the Creative Commons Attribution 4.0 International License.

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  208. Retraction
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  210. Retraction to “A data mining technique for detecting malignant mesothelioma cancer using multiple regression analysis”
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https://doi.org/10.1515/biol-2022-0903
Keywords for this article
catfish waste management; fish oil new resource; solvent-free extraction; polyunsaturated fatty acids; fatty acid analysis; volatile compounds
Creative Commons
BY 4.0

Articles in the same Issue

  1. Biomedical Sciences
  2. Constitutive and evoked release of ATP in adult mouse olfactory epithelium
  3. LARP1 knockdown inhibits cultured gastric carcinoma cell cycle progression and metastatic behavior
  4. PEGylated porcine–human recombinant uricase: A novel fusion protein with improved efficacy and safety for the treatment of hyperuricemia and renal complications
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  7. Brucella infection combined with Nocardia infection: A case report and literature review
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  9. Ang-1, Ang-2, and Tie2 are diagnostic biomarkers for Henoch-Schönlein purpura and pediatric-onset systemic lupus erythematous
  10. PTTG1 induces pancreatic cancer cell proliferation and promotes aerobic glycolysis by regulating c-myc
  11. Role of serum B-cell-activating factor and interleukin-17 as biomarkers in the classification of interstitial pneumonia with autoimmune features
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  17. Aquaporin-8 promotes human dermal fibroblasts to counteract hydrogen peroxide-induced oxidative damage: A novel target for management of skin aging
  18. Current research and evidence gaps on placental development in iron deficiency anemia
  19. Single-nucleotide polymorphism rs2910829 in PDE4D is related to stroke susceptibility in Chinese populations: The results of a meta-analysis
  20. Pheochromocytoma-induced myocardial infarction: A case report
  21. Kaempferol regulates apoptosis and migration of neural stem cells to attenuate cerebral infarction by O‐GlcNAcylation of β-catenin
  22. Sirtuin 5 regulates acute myeloid leukemia cell viability and apoptosis by succinylation modification of glycine decarboxylase
  23. Apigenin 7-glucoside impedes hypoxia-induced malignant phenotypes of cervical cancer cells in a p16-dependent manner
  24. KAT2A changes the function of endometrial stromal cells via regulating the succinylation of ENO1
  25. Current state of research on copper complexes in the treatment of breast cancer
  26. Exploring antioxidant strategies in the pathogenesis of ALS
  27. Helicobacter pylori causes gastric dysbacteriosis in chronic gastritis patients
  28. IL-33/soluble ST2 axis is associated with radiation-induced cardiac injury
  29. The predictive value of serum NLR, SII, and OPNI for lymph node metastasis in breast cancer patients with internal mammary lymph nodes after thoracoscopic surgery
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  31. P2X7 receptor: A receptor closely linked with sepsis-associated encephalopathy
  32. Probiotics for inflammatory bowel disease: Is there sufficient evidence?
  33. Identification of KDM4C as a gene conferring drug resistance in multiple myeloma
  34. Microbial perspective on the skin–gut axis and atopic dermatitis
  35. Thymosin α1 combined with XELOX improves immune function and reduces serum tumor markers in colorectal cancer patients after radical surgery
  36. Highly specific vaginal microbiome signature for gynecological cancers
  37. Sample size estimation for AQP4-IgG seropositive optic neuritis: Retinal damage detection by optical coherence tomography
  38. The effects of SDF-1 combined application with VEGF on femoral distraction osteogenesis in rats
  39. Fabrication and characterization of gold nanoparticles using alginate: In vitro and in vivo assessment of its administration effects with swimming exercise on diabetic rats
  40. Mitigating digestive disorders: Action mechanisms of Mediterranean herbal active compounds
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  45. Application of metagenomic next-generation sequencing technology in the etiological diagnosis of peritoneal dialysis-associated peritonitis
  46. Clinical diagnosis, prevention, and treatment of neurodyspepsia syndrome using intelligent medicine
  47. Case report: Successful bronchoscopic interventional treatment of endobronchial leiomyomas
  48. Preliminary investigation into the genetic etiology of short stature in children through whole exon sequencing of the core family
  49. Cystic adenomyoma of the uterus: Case report and literature review
  50. Mesoporous silica nanoparticles as a drug delivery mechanism
  51. Dynamic changes in autophagy activity in different degrees of pulmonary fibrosis in mice
  52. Vitamin D deficiency and inflammatory markers in type 2 diabetes: Big data insights
  53. Lactate-induced IGF1R protein lactylation promotes proliferation and metabolic reprogramming of lung cancer cells
  54. Meta-analysis on the efficacy of allogeneic hematopoietic stem cell transplantation to treat malignant lymphoma
  55. Mitochondrial DNA drives neuroinflammation through the cGAS-IFN signaling pathway in the spinal cord of neuropathic pain mice
  56. Application value of artificial intelligence algorithm-based magnetic resonance multi-sequence imaging in staging diagnosis of cervical cancer
  57. Embedded monitoring system and teaching of artificial intelligence online drug component recognition
  58. Investigation into the association of FNDC1 and ADAMTS12 gene expression with plumage coloration in Muscovy ducks
  59. Yak meat content in feed and its impact on the growth of rats
  60. A rare case of Richter transformation with breast involvement: A case report and literature review
  61. First report of Nocardia wallacei infection in an immunocompetent patient in Zhejiang province
  62. Rhodococcus equi and Brucella pulmonary mass in immunocompetent: A case report and literature review
  63. Downregulation of RIP3 ameliorates the left ventricular mechanics and function after myocardial infarction via modulating NF-κB/NLRP3 pathway
  64. Evaluation of the role of some non-enzymatic antioxidants among Iraqi patients with non-alcoholic fatty liver disease
  65. The role of Phafin proteins in cell signaling pathways and diseases
  66. Ten-year anemia as initial manifestation of Castleman disease in the abdominal cavity: A case report
  67. Coexistence of hereditary spherocytosis with SPTB P.Trp1150 gene variant and Gilbert syndrome: A case report and literature review
  68. Utilization of convolutional neural networks to analyze microscopic images for high-throughput screening of mesenchymal stem cells
  69. Exploratory evaluation supported by experimental and modeling approaches of Inula viscosa root extract as a potent corrosion inhibitor for mild steel in a 1 M HCl solution
  70. Imaging manifestations of ductal adenoma of the breast: A case report
  71. Gut microbiota and sleep: Interaction mechanisms and therapeutic prospects
  72. Isomangiferin promotes the migration and osteogenic differentiation of rat bone marrow mesenchymal stem cells
  73. Prognostic value and microenvironmental crosstalk of exosome-related signatures in human epidermal growth factor receptor 2 positive breast cancer
  74. Circular RNAs as potential biomarkers for male severe sepsis
  75. Knockdown of Stanniocalcin-1 inhibits growth and glycolysis in oral squamous cell carcinoma cells
  76. The expression and biological role of complement C1s in esophageal squamous cell carcinoma
  77. A novel GNAS mutation in pseudohypoparathyroidism type 1a with articular flexion deformity: A case report
  78. Predictive value of serum magnesium levels for prognosis in patients with non-small cell lung cancer undergoing EGFR-TKI therapy
  79. HSPB1 alleviates acute-on-chronic liver failure via the P53/Bax pathway
  80. IgG4-related disease complicated by PLA2R-associated membranous nephropathy: A case report
  81. Baculovirus-mediated endostatin and angiostatin activation of autophagy through the AMPK/AKT/mTOR pathway inhibits angiogenesis in hepatocellular carcinoma
  82. Metformin mitigates osteoarthritis progression by modulating the PI3K/AKT/mTOR signaling pathway and enhancing chondrocyte autophagy
  83. Evaluation of the activity of antimicrobial peptides against bacterial vaginosis
  84. Atypical presentation of γ/δ mycosis fungoides with an unusual phenotype and SOCS1 mutation
  85. Analysis of the microecological mechanism of diabetic kidney disease based on the theory of “gut–kidney axis”: A systematic review
  86. Omega-3 fatty acids prevent gestational diabetes mellitus via modulation of lipid metabolism
  87. Refractory hypertension complicated with Turner syndrome: A case report
  88. Interaction of ncRNAs and the PI3K/AKT/mTOR pathway: Implications for osteosarcoma
  89. Association of low attenuation area scores with pulmonary function and clinical prognosis in patients with chronic obstructive pulmonary disease
  90. Long non-coding RNAs in bone formation: Key regulators and therapeutic prospects
  91. The deubiquitinating enzyme USP35 regulates the stability of NRF2 protein
  92. Neutrophil-to-lymphocyte ratio and platelet-to-lymphocyte ratio as potential diagnostic markers for rebleeding in patients with esophagogastric variceal bleeding
  93. G protein-coupled receptor 1 participating in the mechanism of mediating gestational diabetes mellitus by phosphorylating the AKT pathway
  94. LL37-mtDNA regulates viability, apoptosis, inflammation, and autophagy in lipopolysaccharide-treated RLE-6TN cells by targeting Hsp90aa1
  95. The analgesic effect of paeoniflorin: A focused review
  96. Chemical composition’s effect on Solanum nigrum Linn.’s antioxidant capacity and erythrocyte protection: Bioactive components and molecular docking analysis
  97. Knockdown of HCK promotes HREC cell viability and inner blood–retinal barrier integrity by regulating the AMPK signaling pathway
  98. The role of rapamycin in the PINK1/Parkin signaling pathway in mitophagy in podocytes
  99. Laryngeal non-Hodgkin lymphoma: Report of four cases and review of the literature
  100. Clinical value of macrogenome next-generation sequencing on infections
  101. Overview of dendritic cells and related pathways in autoimmune uveitis
  102. TAK-242 alleviates diabetic cardiomyopathy via inhibiting pyroptosis and TLR4/CaMKII/NLRP3 pathway
  103. Hypomethylation in promoters of PGC-1α involved in exercise-driven skeletal muscular alterations in old age
  104. Profile and antimicrobial susceptibility patterns of bacteria isolated from effluents of Kolladiba and Debark hospitals
  105. The expression and clinical significance of syncytin-1 in serum exosomes of hepatocellular carcinoma patients
  106. A histomorphometric study to evaluate the therapeutic effects of biosynthesized silver nanoparticles on the kidneys infected with Plasmodium chabaudi
  107. PGRMC1 and PAQR4 are promising molecular targets for a rare subtype of ovarian cancer
  108. Analysis of MDA, SOD, TAOC, MNCV, SNCV, and TSS scores in patients with diabetes peripheral neuropathy
  109. SLIT3 deficiency promotes non-small cell lung cancer progression by modulating UBE2C/WNT signaling
  110. The relationship between TMCO1 and CALR in the pathological characteristics of prostate cancer and its effect on the metastasis of prostate cancer cells
  111. Heterogeneous nuclear ribonucleoprotein K is a potential target for enhancing the chemosensitivity of nasopharyngeal carcinoma
  112. PHB2 alleviates retinal pigment epithelium cell fibrosis by suppressing the AGE–RAGE pathway
  113. Anti-γ-aminobutyric acid-B receptor autoimmune encephalitis with syncope as the initial symptom: Case report and literature review
  114. Comparative analysis of chloroplast genome of Lonicera japonica cv. Damaohua
  115. Human umbilical cord mesenchymal stem cells regulate glutathione metabolism depending on the ERK–Nrf2–HO-1 signal pathway to repair phosphoramide mustard-induced ovarian cancer cells
  116. Electroacupuncture on GB acupoints improves osteoporosis via the estradiol–PI3K–Akt signaling pathway
  117. Renalase protects against podocyte injury by inhibiting oxidative stress and apoptosis in diabetic nephropathy
  118. Review: Dicranostigma leptopodum: A peculiar plant of Papaveraceae
  119. Combination effect of flavonoids attenuates lung cancer cell proliferation by inhibiting the STAT3 and FAK signaling pathway
  120. Renal microangiopathy and immune complex glomerulonephritis induced by anti-tumour agents: A case report
  121. Correlation analysis of AVPR1a and AVPR2 with abnormal water and sodium and potassium metabolism in rats
  122. Gastrointestinal health anti-diarrheal mixture relieves spleen deficiency-induced diarrhea through regulating gut microbiota
  123. Myriad factors and pathways influencing tumor radiotherapy resistance
  124. Exploring the effects of culture conditions on Yapsin (YPS) gene expression in Nakaseomyces glabratus
  125. Screening of prognostic core genes based on cell–cell interaction in the peripheral blood of patients with sepsis
  126. Coagulation factor II thrombin receptor as a promising biomarker in breast cancer management
  127. Ileocecal mucinous carcinoma misdiagnosed as incarcerated hernia: A case report
  128. Methyltransferase like 13 promotes malignant behaviors of bladder cancer cells through targeting PI3K/ATK signaling pathway
  129. The debate between electricity and heat, efficacy and safety of irreversible electroporation and radiofrequency ablation in the treatment of liver cancer: A meta-analysis
  130. ZAG promotes colorectal cancer cell proliferation and epithelial–mesenchymal transition by promoting lipid synthesis
  131. Baicalein inhibits NLRP3 inflammasome activation and mitigates placental inflammation and oxidative stress in gestational diabetes mellitus
  132. Impact of SWCNT-conjugated senna leaf extract on breast cancer cells: A potential apoptotic therapeutic strategy
  133. MFAP5 inhibits the malignant progression of endometrial cancer cells in vitro
  134. Major ozonated autohemotherapy promoted functional recovery following spinal cord injury in adult rats via the inhibition of oxidative stress and inflammation
  135. Axodendritic targeting of TAU and MAP2 and microtubule polarization in iPSC-derived versus SH-SY5Y-derived human neurons
  136. Differential expression of phosphoinositide 3-kinase/protein kinase B and Toll-like receptor/nuclear factor kappa B signaling pathways in experimental obesity Wistar rat model
  137. The therapeutic potential of targeting Oncostatin M and the interleukin-6 family in retinal diseases: A comprehensive review
  138. BA inhibits LPS-stimulated inflammatory response and apoptosis in human middle ear epithelial cells by regulating the Nf-Kb/Iκbα axis
  139. Role of circRMRP and circRPL27 in chronic obstructive pulmonary disease
  140. Investigating the role of hyperexpressed HCN1 in inducing myocardial infarction through activation of the NF-κB signaling pathway
  141. Characterization of phenolic compounds and evaluation of anti-diabetic potential in Cannabis sativa L. seeds: In vivo, in vitro, and in silico studies
  142. Quantitative immunohistochemistry analysis of breast Ki67 based on artificial intelligence
  143. Ecology and Environmental Science
  144. Screening of different growth conditions of Bacillus subtilis isolated from membrane-less microbial fuel cell toward antimicrobial activity profiling
  145. Degradation of a mixture of 13 polycyclic aromatic hydrocarbons by commercial effective microorganisms
  146. Evaluation of the impact of two citrus plants on the variation of Panonychus citri (Acari: Tetranychidae) and beneficial phytoseiid mites
  147. Prediction of present and future distribution areas of Juniperus drupacea Labill and determination of ethnobotany properties in Antalya Province, Türkiye
  148. Population genetics of Todarodes pacificus (Cephalopoda: Ommastrephidae) in the northwest Pacific Ocean via GBS sequencing
  149. A comparative analysis of dendrometric, macromorphological, and micromorphological characteristics of Pistacia atlantica subsp. atlantica and Pistacia terebinthus in the middle Atlas region of Morocco
  150. Macrofungal sporocarp community in the lichen Scots pine forests
  151. Assessing the proximate compositions of indigenous forage species in Yemen’s pastoral rangelands
  152. Food Science
  153. Gut microbiota changes associated with low-carbohydrate diet intervention for obesity
  154. Reexamination of Aspergillus cristatus phylogeny in dark tea: Characteristics of the mitochondrial genome
  155. Differences in the flavonoid composition of the leaves, fruits, and branches of mulberry are distinguished based on a plant metabolomics approach
  156. Investigating the impact of wet rendering (solventless method) on PUFA-rich oil from catfish (Clarias magur) viscera
  157. Non-linear associations between cardiovascular metabolic indices and metabolic-associated fatty liver disease: A cross-sectional study in the US population (2017–2020)
  158. Knockdown of USP7 alleviates atherosclerosis in ApoE-deficient mice by regulating EZH2 expression
  159. Utility of dairy microbiome as a tool for authentication and traceability
  160. Agriculture
  161. Enhancing faba bean (Vicia faba L.) productivity through establishing the area-specific fertilizer rate recommendation in southwest Ethiopia
  162. Impact of novel herbicide based on synthetic auxins and ALS inhibitor on weed control
  163. Perspectives of pteridophytes microbiome for bioremediation in agricultural applications
  164. Fertilizer application parameters for drip-irrigated peanut based on the fertilizer effect function established from a “3414” field trial
  165. Improving the productivity and profitability of maize (Zea mays L.) using optimum blended inorganic fertilization
  166. Application of leaf multispectral analyzer in comparison to hyperspectral device to assess the diversity of spectral reflectance indices in wheat genotypes
  167. Animal Sciences
  168. Knockdown of ANP32E inhibits colorectal cancer cell growth and glycolysis by regulating the AKT/mTOR pathway
  169. Development of a detection chip for major pathogenic drug-resistant genes and drug targets in bovine respiratory system diseases
  170. Exploration of the genetic influence of MYOT and MB genes on the plumage coloration of Muscovy ducks
  171. Transcriptome analysis of adipose tissue in grazing cattle: Identifying key regulators of fat metabolism
  172. Comparison of nutritional value of the wild and cultivated spiny loaches at three growth stages
  173. Transcriptomic analysis of liver immune response in Chinese spiny frog (Quasipaa spinosa) infected with Proteus mirabilis
  174. Disruption of BCAA degradation is a critical characteristic of diabetic cardiomyopathy revealed by integrated transcriptome and metabolome analysis
  175. Plant Sciences
  176. Effect of long-term in-row branch covering on soil microorganisms in pear orchards
  177. Photosynthetic physiological characteristics, growth performance, and element concentrations reveal the calcicole–calcifuge behaviors of three Camellia species
  178. Transcriptome analysis reveals the mechanism of NaHCO3 promoting tobacco leaf maturation
  179. Bioinformatics, expression analysis, and functional verification of allene oxide synthase gene HvnAOS1 and HvnAOS2 in qingke
  180. Water, nitrogen, and phosphorus coupling improves gray jujube fruit quality and yield
  181. Improving grape fruit quality through soil conditioner: Insights from RNA-seq analysis of Cabernet Sauvignon roots
  182. Role of Embinin in the reabsorption of nucleus pulposus in lumbar disc herniation: Promotion of nucleus pulposus neovascularization and apoptosis of nucleus pulposus cells
  183. Revealing the effects of amino acid, organic acid, and phytohormones on the germination of tomato seeds under salinity stress
  184. Combined effects of nitrogen fertilizer and biochar on the growth, yield, and quality of pepper
  185. Comprehensive phytochemical and toxicological analysis of Chenopodium ambrosioides (L.) fractions
  186. Impact of “3414” fertilization on the yield and quality of greenhouse tomatoes
  187. Exploring the coupling mode of water and fertilizer for improving growth, fruit quality, and yield of the pear in the arid region
  188. Metagenomic analysis of endophytic bacteria in seed potato (Solanum tuberosum)
  189. Antibacterial, antifungal, and phytochemical properties of Salsola kali ethanolic extract
  190. Exploring the hepatoprotective properties of citronellol: In vitro and in silico studies on ethanol-induced damage in HepG2 cells
  191. Enhanced osmotic dehydration of watermelon rind using honey–sucrose solutions: A study on pre-treatment efficacy and mass transfer kinetics
  192. Effects of exogenous 2,4-epibrassinolide on photosynthetic traits of 53 cowpea varieties under NaCl stress
  193. Comparative transcriptome analysis of maize (Zea mays L.) seedlings in response to copper stress
  194. An optimization method for measuring the stomata in cassava (Manihot esculenta Crantz) under multiple abiotic stresses
  195. Fosinopril inhibits Ang II-induced VSMC proliferation, phenotype transformation, migration, and oxidative stress through the TGF-β1/Smad signaling pathway
  196. Antioxidant and antimicrobial activities of Salsola imbricata methanolic extract and its phytochemical characterization
  197. Bioengineering and Biotechnology
  198. Absorbable calcium and phosphorus bioactive membranes promote bone marrow mesenchymal stem cells osteogenic differentiation for bone regeneration
  199. New advances in protein engineering for industrial applications: Key takeaways
  200. An overview of the production and use of Bacillus thuringiensis toxin
  201. Research progress of nanoparticles in diagnosis and treatment of hepatocellular carcinoma
  202. Bioelectrochemical biosensors for water quality assessment and wastewater monitoring
  203. PEI/MMNs@LNA-542 nanoparticles alleviate ICU-acquired weakness through targeted autophagy inhibition and mitochondrial protection
  204. Unleashing of cytotoxic effects of thymoquinone-bovine serum albumin nanoparticles on A549 lung cancer cells
  205. Erratum
  206. Erratum to “Investigating the association between dietary patterns and glycemic control among children and adolescents with T1DM”
  207. Erratum to “Activation of hypermethylated P2RY1 mitigates gastric cancer by promoting apoptosis and inhibiting proliferation”
  208. Retraction
  209. Retraction to “MiR-223-3p regulates cell viability, migration, invasion, and apoptosis of non-small cell lung cancer cells by targeting RHOB”
  210. Retraction to “A data mining technique for detecting malignant mesothelioma cancer using multiple regression analysis”
  211. Special Issue on Advances in Neurodegenerative Disease Research and Treatment
  212. Transplantation of human neural stem cell prevents symptomatic motor behavior disability in a rat model of Parkinson’s disease
  213. Special Issue on Multi-omics
  214. Inflammasome complex genes with clinical relevance suggest potential as therapeutic targets for anti-tumor drugs in clear cell renal cell carcinoma
  215. Gastroesophageal varices in primary biliary cholangitis with anti-centromere antibody positivity: Early onset?
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