Effects of plant-origin binder sources in Asian catfish diets on growth, feed quality, blood chemistry, liver and gut health, and economic efficiency

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Effects of plant-origin binder sources in Asian catfish diets on growth, feed quality, blood chemistry, liver and gut health, and economic efficiency

Introduction

Aquaculture is one of the fastest-growing sectors in global food production and plays a crucial role in meeting the increasing demand for high-quality protein1,2. Asian catfish (Clarias batrachus) is a commercially significant species due to its adaptability to diverse environmental conditions, rapid growth rates, and strong consumer acceptance across multiple Asian countries3,4,5. As the aquaculture industry expands, optimising fish feed formulation is essential to enhance productivity and ensure economic sustainability. A key challenge in aquafeed production is maintaining pellet stability in water, which affects feed efficiency and environmental sustainability6. Commercial fish feeds typically incorporate micro-bound diets using binders to improve pellet integrity, minimizing nutrient leaching and disintegration in aquatic environments7,8,9. Pellet stability is influenced by ingredient composition, binder inclusion levels, and feed manufacturing processes10. In 2022, global compound feed production reached approximately 1,266.35 million tons11, with binders accounting for 2–3% of total production10.

Binders, available in liquid or solid powder forms, enhance feed structure by forming bridges, coatings, or films that improve inter-particle cohesion7,12,13. They play a crucial role in maintaining pellet integrity, reducing dust formation, and ensuring uniform nutrient distribution14,15. Among synthetic binders, carboxymethyl cellulose (CMC) is widely used in the aquafeed industry due to their superior binding properties and stability16,17. However, concerns regarding high import costs, supply chain dependency, sustainability, market volatility, and the negative economic impact of synthetic binders have spurred interest in natural alternatives18,19,20. The lack of an optimal, locally sourced binder and the limited availability of commercial alternatives present a significant challenge to the fish feed industry21. Consequently, natural plant-derived binders offer a promising, sustainable, and cost-effective alternative.

Plant-derived polysaccharides, composed of long chains of monosaccharide linked by glycosidic bonds, are considered environmentally sustainable and less likely to have adverse physiological effects on fish compared to synthetic binders22,23,24,25,26. In addition to their binding properties, natural polysaccharides exhibit biocompatibility, bioactivity, gel-forming capabilities, and prebiotic effects, potentially enhancing pellet quality, reducing feed wastage, and improving fish health, immunity, and physiological development26,27,28,29,30.

Previous studies have demonstrated the significant effects of dietary binder on growth performance, nutrient utilisation, feed stability, and health status in various aquaculture species, including common carp, Cyprinus carpio L.31, amberjack, Seriola dumerili32, Nile tilapia, Oreochromis niloticus33, Pacific white shrimp, Litopenaeus vannamei34,35, tongue sole, Cynoglossus semilaevis36, sea Cucumber, Apostichopus37, crayfish38, and largemouth bass, Micropterus salmoides39,40. However, research on the use of natural polysaccharide binders, specifically purple fingerling potato (Solanum tuberosum L.), taro root (Colocasia esculenta), and glutinous rice (Oryza sativa var. glutinosa), and their effects on the growth and health of aquaculture species remains limited. Purple fingerling potato, taro root, and glutinous rice were chosen as plant binders for the diet of Asian catfish (Clarias batrachus) because of their high starch content, strong binding capacity, and nutritional value41,42,43. In addition, they are cost-effective, easily accessible, and environmentally sustainable, making them viable alternatives to synthetic binders. Their capacity to enhance water stability in feed and improve fish health makes them viable alternatives for sustainable aquaculture practices. Thus, this study aims to evaluate the potential of natural plant-derived polysaccharide binders in aquafeed, focusing on their effects on feed stability, growth performance, health status, and economic efficiency in Asian catfish production.

Materials and methods

Animal ethics

The study was performed to a high standard (best practice) of experiment handling and approved by the Animal Ethic Committee of Sylhet Agricultural University with the approval code of SAU/291/083/24. This study also strictly adhered to the ARRIVE (Animal Research: Reporting of In Vivo Experiments) guidelines. All methods were performed in accordance with the relevant guidelines and regulations.

Fish husbandry and experimental conditions

A total of 1,000 Asian catfish (Clarias batrachus) fries (mean initial weight: 6.66 ± 0.01 g) were purchased from a commercial fish farm (New Desh Bandhu Fish Hatchery and Nursery, Mymensingh) and acclimatised in a 2,000 L aquarium for two weeks before the initiation of the feeding trial in the Aquaculture Laboratory, Faculty of Fisheries, Sylhet Agricultural University. During the acclimation period, the fish were fed a basal diet containing 32% crude protein and 6% crude lipid (Brand: ACI catfish feed, Bangladesh). Following acclimation, 360 fish were randomly selected, individually weighed, and assigned to treatment groups in triplicates. Each group was stocked in 90 L experimental tanks at a density of 30 fish per tank. During the trial, hydrological variables such as temperature, water pressure, pH, and dissolved oxygen (DO) were measured at 3 days interval using the YSI multiparameter probe (HI 9828, YSI Incorporation, Yellow Spring, USA). Additionally, HACH test kit (HI 28049, HACH, USA) was used to determine the water ammonia. Throughout the experimental period, water quality parameters were maintained at optimal levels, with a temperature of 28.91 ± 0.49 °C, water pressure of 752.21 ± 0.12 mm Hg, pH of 7.21 ± 0.13, DO of 5.18 ± 0.19 mg/L, and ammonia concentration maintained below 0.1 mg/L.

Preparation of polysaccharide binders

Purple fingerling potato (S. tuberosum L.), taro root (C. esculenta), and glutinous rice (O. sativa var. glutinosa) were sourced from the Mymensingh division in Bangladesh. The plant materials were thoroughly washed with deionised water to remove surface contaminants and impurities. The tuberous samples (purple fingerling potato and taro root) were peeled, sliced into uniform thin sections, and oven-dried at 50 °C until the moisture content was reduced to below 10%, ensuring prolonged stability and preventing microbial growth. The dried samples were then finely ground using a mechanical mill and sieved through a 60-mesh screen to achieve a uniform particle size distribution.

Glutinous rice was processed according the methods described by Tang, et al.44. Briefly, the rice grains were soaked in deionised water at room temperature for 12 h to soften the matrix and enhance polysaccharide release. The soaked grains were then drained, oven-dried at 50 °C, and ground into fine powder using a milling machine. All processed plant powders were stored in airtight plastic containers at 4 °C to prevent moisture absorption and maintain stability until further use.

Diet Preparation and feeding

Prior to feed formulation, the chemical composition of purple fingerling potato (D1), taro root (D2), and glutinous rice (D3) was analysed to assessed their nutritional profiles (Table 1). Four isoproteic diets (32% crude protein) were subsequently formulated, incorporating each plant-based binder at levels of 3%. The basal diet (D0) utilised carboxymethyl cellulose (CMC) as the standard binder. The experimental diets were prepared using Danish fish meal, soybean meal, wheat meal, de-oiled rice bran, oiled rice bran, flour, soybean oil, palm oil, vitamin, and mineral premix (Table 2). The feed mixture was processed through an extruder (Model: LM 40 floating fish feed machine; Manufacturer: Henan Lima Machinery Manufacture Co. Ltd., Zhengzhou, China) to produce 2 mm dietary pellets. During feed preparation, the extrusion process was conducted under controlled conditions: the feed mash was conditioned to a moisture content of 32%, with an extrusion temperature ranges from 110 to 120 °C and a barrel pressure of 0.4 MPa. The screw speed was maintained at 300 rpm, while the cutter speed operated at 1200 rpm. A circular die with 2 mm openings was used to produce uniform floating pellets. The prepared diets were oven-dried at 60 °C, cooled to room temperature, packaged in airtight plastic zipper bags, and stored at − 20 °C until use. Each experimental diet underwent to proximate composition analysis following the Official Methods of Analysis45with the results presented in Table 2. Fish were fed their respective experimental diets ad libitum twice daily at 9:15 AM and 5:15 PM for 90 days.

Table 1 Proximate composition of natural binders (% dry matter basis).

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Table 2 Feed formulation and proximate composition (% dry matter basis).

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Feed stability test

The feed diameter was measured using a digital Vernier caliper. In brief, a total of 50 feed pellets were randomly selected from each diets group and their diameters were recorded individually. The average diameter was then calculated and used as the representative value for each feed type. In addition, the feed stability parameters such as pellet durability index, water stability, floatability, and swelling percentage were calculated following the methods of Zulhisyam et al.46:

  1. i.

    Pellet durability index, PDI (%) = (Weight of residual feed pellets on the sieve after tumbling/ Initial weight of pellets) x 100.

  2. ii.

    Water stability (%) = (Total pellets weight after immersion/ Initial weight of pellets) x 100.

  3. iii.

    Floatability (%) = (Number of pellets floated after the immersion time/ Initial number of pellets) x 100.

  4. iv.

    Swelling (%) = (Mean dimension of swollen pellets/ Total dimension of dry pellets) x 100.

Attractability and palatability test

The attractability and palatability of each experimental diet were evaluated following the method described by Afrin, et al.47, with minor modifications. The experiment was conducted in four rectangular tanks (100 L capacity), each partitioned into two chambers using a net-covered wooden frame. A total of 10 randomly selected fish (average weight: 6.18 ± 0.09 g) from each dietary group were introduced into one chamber, while the respective experimental feed was placed in the adjacent chamber. Following a one-hour acclilmation period, the wooden partition was removed, allowing the fish to access the feeding chamber. The number of fish attracted to each diet was recorded over a10-minute observation period. This experiment was conducted in triplicate.

Palatability was assessed by collecting uneaten feed pellets after the feeding trial. The recovered pellets were oven-dried at 70 °C overnight and subsequently weighed. The experiment was repeated three times at the same time of day (9:30 AM) to ensure consistency. The attractability and palatability of the experimental diets were calculated using the following formulae:

  1. i.

    Attractability (%) = (Number of fish attracted in feeding zone/ Number of fish stocked) x 100.

  2. ii.

    Palatability = Feed intake (mg)/ Total fish biomass (g).

Growth performances and feed utilisation

After the 90-day feeding trial, fish from all treatment groups underwent a 24-hour fasting period before being anesthetised with MS-222 (0.1 g/L). Following anesthesia, individual fish weight, total biomass, and total feed consumption for each tank were recorded. Additionally, growth performance and feed utilisation indices were calculated using the formulae established by Kabir, et al.48 and Rahman, et al.49:

  1. i.

    Weight gain, WG (%) = [(Mean final wt. – Mean initial wt.)/ Mean initial wt.] x 100.

  2. ii.

    Total biomass gain, TBG (g) = Final biomass – Initial biomass.

  3. iii.

    Specific growth rate, SGR (%/ day) = [ln (Final wt. – ln (Initial wt.)/ Experimental days] x 100.

  4. iv.

    Survival rate, SR (%) = (Number of surviving fish/ Number of fish at the beginning of the trial) x 100.

  5. v.

    Feed conversion ratio, FCR = Total feed consumption/ Live weight gain.

  6. vi.

    Protein efficiency ratio, PER = Live weight gain/ Total protein consumption.

Proximate composition

The proximate composition of binder ingredients, experimental feeds, and whole body fish samples was analysed in triplicate following the standard procedures outlined by the AOAC45. Moisture content was determined by oven-drying samples at 105 °C until a constant weight was achieved with the weight loss used to calculate moisture levels. Ash content was measured by incinerating samples in a muffle furnace at 550 °C for 6 h. Crude lipid content was assessed using the Soxhlet solvent extraction method, while crude protein content was determined using the Kjeldahl method, applying a nitrogen-to-protein conversion factor of 6.25 (% N x 6.25).

Blood biochemical analysis

The hematological and biochemical indices of fish blood were analysed following the methodology described by Nandi, et al.Histological analysis of liver and gut samples

Histological analysis of the liver and distal intestine was conducted following standard histological procedures. Four fish per tank (n = 4) were randomly selected, anaesthetised with MS-222 (0.1 g/L water), and dissected to collect midgut and liver tissue samples. The excised tissues were immediately fixed in 10% neutral buffered formalin, dehydrated in a graded ethanol series, and cleared in xylene. The processed samples were then embedded in paraffin, sectioned at 5–7 μm) thickness, and mounted on glass slides. The sections were stained with hematoxylin and eosin (H & E) for histological examination. Microscopic analysis was conducted using a light microscopy (Leica DMIL-LED, Germany), and histological images were captured using Cellsens software (Cellsens, The Netherlands) for further histopathological evaluation.

Economic analysis

The cost of raw materials for Asian catfish feed formulation was calculated by summing the prices of individual ingredients. Farm-level economic efficiency was assessed using standard economic indices, following the methodologies described by Suma, et al.51 and Sezu, et al.52, using the following formulas:

  1. i.

    Total yield, TY (kg m-3) = Total biomass gain/ Tank volume.

  2. ii.

    Farm feed cost, FFC (US$/ kg) = Raw materials cost x FCR.

  3. iii.

    Farm revenue was calculated by an expected Asian catfish farm gate price (6.37$/ kg): FR (US$/ m3) = TY x 6.37.

  4. iv.

    Farm raw margin, FRM (US$/ m3) = [FR – (FFC x TY)]

  5. v.

    Return on investment, ROI (%) = [FRM/ (TY x FFC)] x 100.

Statistical analysis

All data were analysed using the Statistical Package for the Social Sciences (SPSS) version 26.0 (IBM, USA). A one-way analysis of variance (ANOVA) was performed to assess significant differences among the dietary treatment groups, followed by Duncan’s multiple range test (DMRT) for post-hoc comparisons. Statistical significance was set at p < 0.05. Results are presented as means ± standard deviation (SD).

Results

Feed stability parameters

Table 3 summaries the physical characteristics of the experimental diets. No significant differences (p > 0.05) were observed in feed diameter and water stability across the dietary treatments. However, the inclusion of different natural binders significantly influenced pellet durability index (PDI) and floatability (p < 0.05), with the D1 diet exhibiting the lowest values.

Figure 1 illustrates the swelling percentages of the experimental diets at 2, 6, and 10 min. The swelling rates at 2 and 6 min did not differ significantly (p > 0.05) among the diets. However, after 10 min of water exposure, significant differences were observed (p < 0.05), with D1 diet displaying the lowest swelling percentage (p < 0.05).

Table 3 Feed physical parameters.

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Fig. 1
figure 1

Swelling test of experimental diets formulated with different plant-based sources as binders. Diet groups: D0 (Control, binder carboxymethyl cellulose), D1 (Binder: purple fingerling potato), D2 (Binder: Taro root), D3 (Binder: Glutinous rice).

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Palatability of experimental diets

Figure 2 illustrates the palatability assessment of the experimental diets in Asian catfish over 5-minute feeding period. The results indicate that fish fed the D0 and D2 diets exhibited significantly higher palatability (p < 0.05) compared to other treatments. In contrast, fish fed the D1 and D3 diets demonstrated significantly lower (p < 0.05) palatability.

Fig. 2
figure 2

Palatability test of experimental diets. Diet groups: D0 (Control, binder carboxymethyl cellulose), D1 (Binder: purple fingerling potato), D2 (Binder: Taro root), D3 (Binder: Glutinous rice).

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Attractability of experimental diets

The attractability assessment of the control and experimental diets is presented in Fig. 3. Statistical analysis revealed that the D2 diet demonstrated significantly higher attractability (p < 0.05) compared to the other treatments. In contrast, the D3 diet showed no significant difference from the other diets. The D0 and D1 diets demonstrated significantly lower (p < 0.05) attractability than the other experimental diets.

Fig. 3
figure 3

Attractability test of experimental diets. Diet groups: D0 (Control, binder carboxymethyl cellulose), D1 (Binder: purple fingerling potato), D2 (Binder: Taro root), D3 (Binder: Glutinous rice).

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Growth performance

Table 4 demonstrates the growth performance of Asian catfish fed different experimental diets. The results indicated that FW (g), WG (%), TB (g), SGR (%/day), and PER were significantly higher (p < 0.05) in fish fed the D2 diet. In contrast, no significant differences were observed in these parameters in the D1 group. Fish fed the D1 and D2 diets exhibited significantly lower FCR (p < 0.05) compared to the other groups. However, SR was not significantly (p > 0.05) affected by the dietary treatments.

Table 4 Zootechnical performance of C. batrachus fed with different test diets for 90 days.

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Whole-body biochemical composition

Table 5 presents the proximate composition of the sampled fish. The results indicated that dietary treatments significantly (p < 0.05) affected crude protein and crude lipid levels. Fish fed the D2 diet exhibited significantly (p < 0.05) higher crude protein content and lower crude lipid levels compared to other treatments. However, no significant differences (p > 0.05) were observed in moisture and ash content among the groups.

Table 5 Whole-body proximate composition (% dry matter basis) of fish fed with different test diets for 90 days.

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Hematological parameters

Table 6 displays the haematological indices of Asian catfish subjected to different dietary treatments. The results indicated that the experimental diets significantly (p < 0.05) influenced most haematological parameters, except for monocytes (MON). Fish fed the D2 diet exhibited significantly (p < 0.05) higher levels of white blood cell (WBC), eosinophil (EOS), red blood cell (RBC), and platelet distribution width (PDW), while showing lower levels of mean corpuscular volume (MCV), mean corpuscular haemoglobin concentration (MCHC), red blood cell distribution width-coefficient of variance (RDW-CV), red blood cell distribution width-standard deviation (RDW-SD), platelets (PLT), and procalcitonin (PCT) compared to other dietary groups. In contrast, fish in the control group (D0) exhibited significantly (p < 0.05) lower levels of lymphocytes (LYM), EOS, basophils (BAS), RBC, hematocrit (HCT), and platelet distribution width (PDW). Additionally, fish fed the D3 diet showed a significant (p < 0.05) reduction in neutrophil (NEU), haemoglobin (HGB), and mean platelet volume (MPV).

Table 6 Hematological indices of experimental fish fed with different test diets for 90 days.

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Serum biochemical metrics

The serum biochemical parameters of the experimental fish are presented in Table 7. The results indicated that dietary treatments significantly (p < 0.05) influenced all measured biochemical indices. Fish fed the D2 diet exhibited significantly (p < 0.05) higher levels of glucose, serum glutamate pyruvate transaminase (SGPT), total protein, and globulin compared to other treatment groups. In contrast, fish supplemented with the D3 diet exhibited significantly (p < 0.05) elevated levels of creatinine, urea, and cholesterol, along with decreased glucose and alkaline phosphatase levels. Meanwhile, fish fed the basal diet (D0) had significantly (p < 0.05) higher albumin levels and lower concentrations of creatinine, bilirubin, urea, and SGOT. No significant differences in creatinine and bilirubin levels were observed in fish fed the D2 diet.

Table 7 Serum biochemical indices of experimental fish fed with different test diets for 90 days.

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Mid-intestine and liver histological analysis

Figure 4 illustrates the histological analysis of the distal intestine of Asian catfish following the experimental diets. The mid gut tissue of fish fed the D2 and D3 diets exhibited enhanced intestinal morphology, characterised by a higher density of villi and goblet cells, as well as an elongated lamina propria, compared to fish in the D0 and D1 groups. Additionally, the D2 diet group demonstrated enhanced stratum compactum bonding, despite structural variations in the tunica muscularis among dietary treatments.

Figure 5 represents the liver morphology of Asian catfish at the end of the feeding trial. Fish in the D1 and D2 treatment groups displayed a higher prevalence of intact nuclei and sinusoidal cytoplasm, with fewer vacuoles, compared to other groups. In contrast, fish fed the D0 and D3 diets exhibited increased vacuolar cytoplasm, larger vacuoles, and degenerative nucleus. However, erythrocyte distribution remained consistent across all dietary treatments.

Fig. 4
figure 4

Mid-gut histological investigation of Asian catfish fed with diets incorporated with different polysaccharides as binder for 90 days. The morphological variations were detected in LP: Lamina propria, GC: Goblet cell, LEM: Laminal epithelial mucosae, TM: Tunica muscularis, and SC: Stratum compactum. Magnification: 10X and Scale bar: 50 μm.

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Fig. 5
figure 5

Liver histological analysis of C. batrachus fed with diets incorporated with different polysaccharides as binder for 90 days. The histomorphological differences were examined in N: Nucleus, E: Erythrocyte, S: Sinusoid, and V: Vacuole. Magnification: 10X and Scale bar: 50 μm.

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Gut morphological measurements

Table 8 presents the intestinal morphological measurements of Asian catfish fed diets containing different binders. Although variations were observed in villi length, villi width, and crypt depth among the treatment groups (p < 0.05), these differences were not statistically significant.

Table 8 Morphological measurements of C. batrachus mid-gut post-feeding trial.

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Economic analysis

Table 9 shows the economic analysis of Asian catfish production using diets formulated with different binders. The results indicated that farm economics varied significantly (p < 0.05) among the dietary treatments. The total yield (TY), farm revenue (FR), farm revenue margin (FRM), and return on investment (ROI) were significantly (p < 0.05) higher in the D2 group, though these values did not differ significantly from those observed in the D1 group. The feed formulation cost (FFC) was substantially reduced in diets incorporating natural polysaccharide binders. Additionally, ROI more than doubled across all experimental groups compared to the control.

Table 9 Farm economic analysis for different diet groups post-feeding trial.

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Discussion

Natural plant-derived polysaccharides hold significant potential as cost-effective and efficient binders in aquafeed formulations, offering enhanced feed stability. In this study, various polysaccharides were evaluated as alternative binding agents to replace the commercially used synthetic binder, carboxymethyl cellulose (CMC). The suitability and effectiveness of each binder were assessed based on their impact on growth performance, feed stability, biochemical composition, hematological parameters, gut and liver morphology, and economic viability in Clarias batrachus culture. The findings of this study identify the most suitable natural binder for application in aquafeed formulation within the aquaculture industry.

Maintaining the physical integrity of feed pellets is crucial to prevent breakage during production, storage, and distribution. Binders play a significant role in achieving a smoother pellet surface, reducing dust formation, and enhancing structural cohesion. In this study, water stability remained consistent across all dietary treatments and within an acceptable range, ensuring the preservation of feed nutritional value and water quality. According to Aksoy, et al.10 pellet water stability is influenced by dietary composition, manufacturing process, and binder type. The use of polysaccharides as binders in this study significantly affected the pellet durability index (PDI) and floatability. Specifically, the incorporation of purple fingerling potato (D1) resulted in reduced PDI and floatability, suggesting that this polysaccharide may have weakened pellet structural integrity and reduced buoyancy. This outcome could be attributed to the composition of purple fingerling potato, which possibly formed weaker gel structures or less cohesive bonds during the pelletization. In contrast, taro root (D2) and glutinous rice (D3) demonstrated superior binding properties, producing more durable and stable pellets. These findings are consistent with previous studies by Kannadhason, et al.53, Orire and Emine54, and Kiki Haetami55, which highlight the effectiveness of plant-derived polysaccharides in improving pellet stability and quality.

Pellet swelling is a critical parameter in assessing feed quality and stability52. In this study, no significant differences in swelling rates were observed among the diets at the 2- and 6-minute intervals, indicating similar initial water absorption across treatments. However, a significant difference emerged at the 10-minute mark, with the D1 diet exhibiting the lowest swelling percentage. This finding indicated that the purple fingerling potato-based diet absorbs less water over time, potentially due to its starch composition forming fewer stable gel structure. The reduced swelling capacity of the D1 diet may have contributed to its lower floatability and overall stability, as feeds with lower swelling rates tend to be less buoyant. Similar effects have been reported in previous research, where the incorporation of Abelmoschus esculentus leaves as a natural binder in Nile tilapia diets significantly influenced pellet swelling rates56.

The palatability of aquafeeds is mainly influenced by factors such as toxic components, nutritional composition, moisture content, and species-specific feeding behaviour57. In this study, the diet formulated with taro root (D2) exhibited significantly higher palatability, which may be attributed to its digestible starch content. Additionally, fish tend to prefer diets that resemble their natural feeding habits. As an omnivorous species, Asian catfish may have found to taro root, a plant-based ingredient, more familiar and appealing, leading to greater acceptance of the D2 diet. Previous research supports the role of polysaccharides in improving diet palatability. For instance58, reported that juvenile Cherax albidus fed a diet containing 5% polysaccharides exhibited enhanced feed acceptance. Similarly59, demonstrated that yellowtail kingfish diets incorporating 10% agar beads and alginate-chitosan resulted in improved palatability. In contrast, fish fed diets containing purple potato (D1) and glutinous rice (D) exhibited lower palatability, possibly due to the presence of anti-nutritional factors (ANF) that may have affected feed acceptance. Furthermore, the attractability assessment revealed that D2 diet achieved the highest attractability score (57.04%), suggesting that taro root have provide a mild, natural flavour and aroma that was more appealing to the fish than other binders60. Similar findings were reported by Afrin, et al.47 and Sezu, et al.52. Likewise61, demonstrated that supplementing Nile tilapia diets with 10% seaweed significantly enhanced feed attractibility. These findings underscore the importance of binder selection in optimizing both palatability and attractability in aquafeeds.

The present study demonstrated that the diet incorporating taro root (D2) significantly enhanced the zootechnical performance of Asian catfish compared to other dietary treatments. The synergistic composition of carbohydrates, fibre, antioxidants, and micronutrients in taro root provides a well-balanced source of energy and essential nutrients, supporting optimal fish growth. Previous research by Temesgen and Retta62 highlighted that taro contains higher levels of digestible protein and amino acids than other root crops, largely due to the presence of symbiotic soil bacteria in its roots and rhizomes. Similar findings have been reported in studies utilizing natural binders in aquafeed formulations32,33,63,64,65, further supporting the benefits of taro as functional feed ingredient. Conversely, fish fed the glutinous rice-treated diet (D3) exhibited the lowest growth performance and the highest FCR, indicating reduced efficiency in converting feed into biomass. This outcome could be attributed to the lower digestibility of glutinous rice, which likely restricted the availability of essential nutrients and energy for growth. Despite these variations in growth performance, the survival rate remained statistically similar across all treatments, indicating that the inclusion of different polysaccharide binders did not negatively affect fish survival. These findings highlight the potential of taro root as an effective natural binder that not only enhances feed stability but also improves growth performance in Clarias batrachus.

The inclusion of different polysaccharides into the diets of Asian catfish significantly influenced CP and lipid content in the treated groups. Notably, fish fed polysaccharide-enriched diets exhibited higher CP levels than those in the control group. Previous studies by Gao, et al.66 and Mohammadi, et al.67 have shown that polysaccharide-based binders exert synergistic effects that enhance CP accumulation in fish. Additionally, polysaccharides may reduce protein catabolism, thereby allowing more dietary protein to be utilized for growth and physiological maintenance. This mechanism likely contributed to the increased CP levels observed in the treatment groups, suggesting an improvement in protein synthesis and retention in Clarias batrachus. Similar trends have been reported in other species, including seabass fed chitosan-supplemented diets63 and rainbow trout supplemented with non-starch polysaccharides68, both of which showed elevated CP levels compared to the control group. Furthermore, a slight reduction in crude lipid content was observed in fish fed the D2 and D3 diets compared to other treatments. The decline in lipid content, couple with increased CP levels, may have enhanced the overall nutritional quality by of the fish, making them more desirable for human consumption69. These changes in protein and lipid composition could be attributed to the nutritional properties and bioactive compounds present in the different polysaccharides used as feed binders. Similar modifications in nutrient composition have been reported in previous studies70,71. Despite these differences, moisture and ash content remained unchanged across all treatments, indicating that polysaccharide supplementation did not significantly affect water retention or mineral composition in the fish.

Hematological parameters are critical indicators of the physiological condition and overall health status of cultured aquatic species51,72. In this study, the inclusion of different polysaccharide binders significantly influenced the hematological profiles of Asian catfish. The highest white blood cell (WBC) count was observed in fish fed the D2 diet, suggesting enhanced immune function, potentially due to the bioactive compounds in taro root. Moreover, diets containing natural polysaccharide binders resulted in significantly elevated neutrophils (NEU), basophils (BAS), lymphocytes (LYM), and eosinophils (EOS) compared to the CMC-based control diet. These findings align with previous, such as73, who reported that varying concentrations of aloe vera polysaccharides significantly affected hematological parameters in African catfish. Similarly, Hayati and Prihanto74 demonstrated that polysaccharide supplementation at levels of 0, 0.5, 1, and 1.5 g/kg had no detrimental effects on the hematological indices of Pangasius pangasius.

The NEU and EOS play vital roles in combating infections, while BAS are involved in mediating allergic responses. LYM, on the other hand, contributed to targeted, long-term immunity through adaptive immune mechanisms52. In the present study, RBC counts were significantly higher in fish fed the D2 diet, indicating an enhanced oxygen-carrying capacity. This finding aligns with a previous study by75, which reported increased RBC counts in red tilapia supplemented with 30% polysaccharides. Fish fed the glutinous rice-based diet (D3) exhibited a significantly higher platelet count, which may be attributed to the influence of polysaccharides on blood clotting mechanisms. Other hematological parameters, including MCV, MPV, MCHC, RDW-CV, RDW-SD, PDW, and PCT, varied across treatments but showed no statistically significant differences. These fluctuations may reflect subtle physiological variations among treatment groups. Similar observations have been reported in previous studies evaluating fish fed diets with varying levels of polysaccharides73,75,76,77,78.

Different polysaccharides significantly affected the serum biochemical parameters of Asian catfish in the present study. The highest glucose level was recorded in fish fed the taro root-based diet (D2), which may attributed to the influence of taro root on carbohydrate metabolism, resulting in elevated blood glucose levels. According to Abdul Kari, et al.79, glucose concentration serves as a sensitive indicator of physiological stress in fish. Similar findings were reported by80, who demonstrated that varying inclusion levels of Coriolus versicolor polysaccharide affected glucose levels in Crucian carp. Conversely, fish fed the glutinous rice diet (D3) exhibited significantly higher creatinine and urea levels, indicating increased protein catabolism or potential alterations in renal function. In contrast, fish fed the D2 diet showed reduced bilirubin levels, indicative of improved liver function, aligning with previous studies49,81. Furthermore, the D2 group exhibited significantly higher total protein and globulin, suggesting enhanced immune function and protein synthesis, potentially due to the bioactive compounds present in taro root. Similar trends have been reported in Asian seabass fed diets supplemented with mannan oligosaccharides, where no adverse effects on serum biochemistry were observed70, as well as in red tilapia fed polysaccharide-enriched diets (Abdelrhman, et al.75). Interestingly, the D3 group showed lower levels of SGPT and SGOT, which are key biomarkers of liver function. Elevated levels of these enzymes typically indicate liver damage, metabolic disturbances, or physiological stress82. Comparable findings were reported in spotted seabass, where varying levels of Astragalus membranaceus polysaccharides influenced albumin, alkaline phosphatase, and cholesterol levels83. Collectively, these findings imply that dietary polysaccharides distinctly affect liver function, protein metabolism, and stress responses, with taro root and glutinous rice exhibiting the most pronounced effects.

The intestines of omnivorous fish are highly responsive to dietary modifications and are characterised by an increased surface area that facilitates efficient nutrient absorption84. In the present study, improved gut morphology was observed in Asian catfish fed diets containing D2 (taro root) and D3 (glutinous rice), as evidenced by an increased number of villi and goblet cells, along with enhanced bonding in the stratum compactum. These improvements may be attributed to the antioxidant, anti-inflammatory, and prebiotic properties of the polysaccharides present in these ingredients. Polysaccharides are known to support intestinal integrity by stimulating mucus secretion via goblet cells, which plays a critical role in protecting the intestinal lining and facilitating nutrient uptake85. Jutfelt86 emphasised that goblet cells are essential for producing mucus that forms a protective barrier on the distal intestine, shielding the epithelium from toxins, pathogens, and mechanical stress. Moreover, an increased number of villi enhances the absorptive surface are of the intestine, while stronger bonding in the stratum compactum suggests a more robust intestinal architecture that supports overall gut health. These findings are consistent with previous studies reporting that the beneficial effects of dietary polysaccharides on the intestinal morphology in various aquaculture species, such as Nile tilapia87, banana shrimp88, rainbow trout68, largemouth bass89, and spotted seabass90.

No statistically significant differences were observed in intestinal morphological measurements among the dietary treatment groups. However, fish fed the D3 diet exhibited the greatest villi length and width, along with the lowest crypt depth. Increased villi length and width enhance the absorptive surface area of the intestine, potentially improving the efficiency of nutrient digestion and assimilation49,91. Additionally, the reduced crypt depth observed in the D3 group may reflect improved gut maturation and a lower rate of intestinal epithelial cell turnover, which is often associated with better digestive efficiency and overall intestinal health92. These morphological trends suggest that glutinous rice may contribute positively to intestinal function, even in the absence of statistically significant differences.

The condition of the liver services as a key indicators of a fish’s nutritional status, given its central role in nutrient metabolism, digestion, and overall physiological regulation93. In the present study, histological analysis revealed that fish fed the control diet exhibited increased vacuolar cytoplasm and degenerative nuclei, which are indicative of hepatic stress. The presence of large vacuoles typically reflects lipid accumulation, while degenerative nuclei suggest cellular damage or metabolic dysfunction. In contrast, fish fed the D1 and D2 diets showed improved liver histoarchitecture, characterised by a greater presence of intact nuclei, healthy cytoplasm, and increased erythrocyte distribution. These observations suggest that the inclusion of polysaccharides in the diet may help mitigate the adverse effects of dietary stress on liver function. Specifically, plant-derived polysaccharides appear to support hepatic health by enhancing metabolic efficiency, improving nutrient utilisation, and reducing inflammation and oxidative stress. The favourable liver histological profiles observed in this study are consistent with findings reported in previous research63,88,94, further supporting the beneficial role of polysaccharide supplementation in promoting liver health in aquaculture species.

The farm economic analysis revealed that substituting CMC with natural polysaccharides as binders in Asian catfish diets had a significant positive impact on TY, FR, FRM, and ROI. Diets incorporating purple fingerling potato (D1) and taro root (D2) showed notable improvements in these economic indicators compared to other experimental feeds. The use of locally available and cost-effective natural polysaccharides significantly reduced FFC, while simultaneously enhancing FR, FRM, and ROI. In contrast, the control diet containing CMC exhibited lower economic efficiency, primally due to the higher costs associated with imported binders and their limited availability in local markets. These findings underscore the economic advantage of incorporating natural, plant-derived binders in aquafeed formulations.

This study presents valuable insights into the potential of plant-based binders in the diet formulation for C. batrachus; however, several limitations acknowledged and addressed in future research. One key limitation is the variability in the biochemical composition of different plant-based binders, which may lead to inconsistencies in feed stability and nutrient availability. Such variability could influence the effectiveness of the binders and compromise feed performance. Additionally, some plant-based binders contain may contain ANF that interfere with nutrient absorption and reduce overall feed efficiency. Although the present study considered these factors, future research could explore strategies such as enzyme supplementation or fermentation techniques to enhance nutrient bioavailability and mitigate the effects of ANFs. Another limitation is that feed stability was evaluated under controlled laboratory conditions, which may not fully represent the dynamics of real-world aquaculture environment. Factors such as water temperature fluctuations, pH variability, and microbial activity in natural or commercial systems could influence feed integrity and nutrient leaching, warranting further evaluation under field conditions. Moreover, this study primarily focused on short-term growth performance and health indicators, which may not capture the long-term impacts of plant-based binders on fish immunity, gut microbiota, and overall physiological well-being. Longitudinal studies are needed to better understand these broader impacts. Lastly, while the economic analysis showed promising results, the large-scale applicability of plant-based binders requires further assessment. Consideration such as market price fluctuations, scalability of production, and farmer acceptance could influence the widespread adoption of these alternative binders in commercial aquaculture operations.

Future research should focus on optimizing the processing techniques of plant-based binders in C. batrachus diets through approaches such as enzymatic hydrolysis, fermentation, and extrusion. These methods can enhance the functional properties of binders and reduce the presence of ANFs, thereby improving nutrient bioavailability and feed performance. However, the present study did not evaluate digestive enzyme activities related to carbohydrate breakdown (e.g., amylase, maltase, sucrose) or measure digestibility-related indicators such as fecal consistency, which could have provided deeper mechanistic insights into nutrient assimilation and feed functionality. Comparative studies with conventional binders, such as wheat gluten and gelatin, are essential to evaluate their relative efficiency in terms of water stability, digestibility, and cost-effectiveness. Additionally, investigations into nutrient leaching and its subsequent impact on water quality are essential for advancing environmentally sustainable aquaculture practices. Further research should also explore the effects of plant-based binders on gut microbiota and immune responses using advanced molecular tools such as metagenomics and transcriptomics. These techniques can provide deeper insights into the long-term implications of dietary binders on fish health and physiological functions. Large-scale field trials in commercial aquaculture systems are necessary to validate laboratory findings under real-world farming conditions. Moreover, conducting a life cycle assessment (LCA) to compare the environmental footprint of plant-based binders with conventional alternatives may provide critical data on their sustainability and resources efficiency. Addressing these research gaps will contribute to the development of more sustainable, nutrionally efficient, and economically viable feeding strategies for C. batrachus, ultimately promoting the broader application of plant-based binders in aquafeed formulations.

Conclusion

The use of natural polysaccharides, particularly taro root (D2), as an alternative binder in Asian catfish diets significantly improved growth performance, feed utilisation, whole-body composition, hematological and biochemical profiles, intestinal and liver health, as well as overall farm economics. This study highlights the potential of plant-based binders to enhance both fish health and production efficiency while reducing reliance on synthetic additives. Furthermore, the development of novel binders from natural sources provides aquafeed manufacturers with viable, cost-effective, and sustainable alternatives for aquafeed formulation, supporting the advancement of environmentally responsible aquaculture practices.

Data availability

The datasets used and/or analysed during the current study are available from the corresponding author upon reasonable request.

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Acknowledgements

This research article is a collaboration between Universiti Malaysia Kelantan, Chiang Mai University, and Sylhet Agricultural University. This collaboration is part of the planning by the Advanced Livestock and Aquaculture Research Group – ALAReG under the Faculty of Agro-Based Industry, Universiti Malaysia Kelantan, Jeli Campus.

Funding

The primary source of funding for this research project was provided by University Grants Commission and Sylhet Agricultural University Research System (Ref. No. UGC-SAURES: Agri-lifescience-139-2021-2022) and Universiti Malaysia Kelantan Rising Star Research Project under grant number R/STA/A0700/01083A/007/2024/01327. This research was partially supported by Chiang Mai University.

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Authors and Affiliations

  1. Department of Aquaculture, Faculty of Fisheries, Sylhet Agricultural University, Sylhet, 3100, Bangladesh

    Muhammad Anamul Kabir, Sanzida Haque, Shishir Kumar Nandi, Tanwi Dey & Md. Sakhawat Hossain

  2. Advanced Livestock and Aquaculture Research Group, Faculty of Agro- Based Industry, Universiti Malaysia Kelantan, Jeli Campus, Jeli, 17600, Malaysia

    Muhammad Anamul Kabir, Zulhisyam Abdul Kari & Nor Dini Rusli

  3. Faculty of Environmental Agricultural Sciences, Fish Research Centre, Arish University, El-Arish, Egypt

    El-Sayed Hemdan Eissa

  4. Department of Chemical Pathology, School of Medical Sciences, Universiti Sains Malaysia, Health Campus, Kubang Kerian, Kelantan, Malaysia

    Martina Irwan Khoo

  5. Department of Agricultural Sciences, Faculty of Agro-Based Industry, Universiti Malaysia Kelantan, Jeli Campus, Jeli, 17600, Malaysia

    Zulhisyam Abdul Kari & Nor Dini Rusli

  6. Department of Animal and Aquatic Sciences, Faculty of Agriculture, Chiang Mai University, Chiang Mai, 50200, Thailand

    Hien Van Doan

  7. Functional Feed Innovation Centre (FuncFeed), Faculty of Agriculture, Chiang Mai University, Chiang Mai, 50200, Thailand

    Hien Van Doan

Authors

  1. Muhammad Anamul Kabir
  2. Sanzida Haque
  3. Shishir Kumar Nandi
  4. Tanwi Dey
  5. Md. Sakhawat Hossain
  6. El-Sayed Hemdan Eissa
  7. Martina Irwan Khoo
  8. Zulhisyam Abdul Kari
  9. Nor Dini Rusli
  10. Hien Van Doan

Contributions

M.A.K: Conceptualisation, experimental design, project investigation, supervision, writing-review and editing the draft, and funding acquisition; S.H: Original draft preparation, formal analysis, visualization, methodology, writing and editing; S.K.N: Formal analysis, writing-review and editing the draft; Z.A.K and H.V.D: Validation, supervision, writing-review and editing the draft, and funding acquisition; T.D, E.S.H.E, M.I.K, N.D.R and M.S.H: Writing-review and editing the draft.

Corresponding authors

Correspondence to Zulhisyam Abdul Kari or Hien Van Doan.

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Kabir, M.A., Haque, S., Nandi, S.K. et al. Effects of plant-origin binder sources in Asian catfish diets on growth, feed quality, blood chemistry, liver and gut health, and economic efficiency. Sci Rep 15, 29953 (2025). https://doi.org/10.1038/s41598-025-16117-y

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