Abstract
This study was conducted to determine the effect of replacing fishmeal (FM) with fermented soybean meal (FSBM) for 12 weeks on the growth performance, feed utilization, immunological parameters, antioxidant enzyme assays and lipid peroxidation, digestive enzymes, and histopathological analysis of juvenile Litopenaeus vannamei (L. vannamei). By substituting 0.0%, 20%, 30%, and 40% FSBM for fishmeal (w/w), four isonitrogenous diets were generated. A total of 300 juvenile L. vannamei (1.59 ± 0.01 g) were randomly allocated to the experimental fiber tanks at a rate of fifteen shrimp per tank, with three replicates for each treatment. Growth performance and feed utilization decline considerably (P < 0.05) with increasing amounts of FM replacement with FSBM in diets. In comparison to the juveniles fed the other experimental diets, the diet containing a moderate level of FM replacement (20% FSBM) considerably enhanced growth performance and feed consumption during the feeding trial. The 20% FSBM-fed group had the highest protein content. In contrast, raising FSBM levels significantly increased lipid content (P < 0.05) compared to the control. However, there were no statistically significant differences (P > 0.05) across FSBM treatments. Hemolymph plasma total protein (TP) concentration and lysozyme activity were substantially greater (P < 0.05) in 20% FSBM compared to 40% FSBM (P < 0.05). In addition, 20% FSBM exhibits a substantial (P < 0.05) increase in the activity of antioxidant enzymes (CAT SOD, GPX, and GR). In contrast, the control and 30% FSBM groups had considerably more lipid peroxidation markers (MDA) than the 20% and 40% FSBM groups. Hepatopancreas amylase activity was considerably elevated (P < 0.05) in the control group and with 40% FSBM. In addition, hepatopancreas and intestinal protease and lipase activity increased significantly by 20% FSBM. Considerably, more B cells were present in the 40% FSBM diet than in the control diet; however, they were significantly less prevalent in the 20% and 30% FSBM diets (P < 0.05).
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Introduction
Crustaceans are a common species that contribute significantly to the aquaculture industry. Because of high protein and minerals, it provides for healthy living, and shrimp has emerged as the favorite and fastest growing species in the crustacean aquaculture industry (Bondad-Reantaso et al. 2012). Pacific whiteleg shrimp (Litopenaeus vannamei; L. vannamei) is an important economic species in aquaculture (Pauly and Froese 2012). In 2020, whiteleg shrimp was ranked as the top-produced species with 5.81 Mmt (FAO 2022). The selection of shrimp species for culture is highly dependent on their ability to accept a wide range of feed formulations and ingredients (Chi et al. 2009). Despite its numerous advantages, the aquaculture industry faces numerous challenges; the most pressing of which is the demand for and supply of fishmeal (FM), the industry’s primary protein source in artificial aquatic and marine shrimp diets. FM has long been used in the aquafeed formulation as a source of high-quality protein with its high digestibility and good amino acid (AA) profile (Sookying et al. 2013). However, reliance on FM has been identified as a significant impediment to the aquaculture industry’s long-term development (Tacon and Metian 2008) because of its consistent price increase due to its limited supply and increased incorporation in livestock and aquaculture feed (Zhang et al. 2018). As a result, pursuing less expensive and more sustainable alternative protein sources of both animal and plant origin to reduce FM levels in aquafeed without impairing growth performance is a major ongoing global interest (Ding et al. 2015; Faggio et al. 2015; Dossou et al. 2018; Hoseinifar et al. 2011; Ishwarya et al. 2018; Nath et al. 2018). Over the last few decades, a wide range of protein ingredients has been studied as potential FM replacers, with plant proteins receiving the most attention as the most viable candidates due to their lower cost and abundant availability (Gatlin III et al. 2007; Dossou et al. 2018; Abd El-Naby et al. 2022). Plant protein sources such as soybean meal (SBM) are the primary options when replacing FM in aquafeed. However, SBM has several nutritional drawbacks, including the presence of anti-nutritional factors (ANFs) that reduce feed utilization, absorption, and feed conversion ratio (Kikuchi 1999), in addition to the amino acid imbalance and lower protein content when compared to FM. Thermal and mechanical processes, soaking, germination/malting, and fermentation have all been used to reduce the ANF content of SM and improve its nutrients, bioavailability, and nutritional value (Hotz and Gibson 2007). Among these methods, fermentation has been proposed as the most cost-effective for improving the nutritional quality of SBM not only through the biodegradation of ANFs (such as trypsin inhibitors, oligosaccharides, and phytic acid), proteins, and fibers but also through the production of probiotics and prebiotics, which may improve palatability, nutrient digestibility, and immune function (Hong et al. 2004). Fermented soybean meal (FSBM) was produced by fermenting SBM; many microorganisms were tested for SBM fermentation, and it was revealed that the nutritional value of the produced FSBM varies depending on the type of microorganism (Feizi et al. 2022). Thus, the goal of this study is to assess the effects of long-term feeding fermented soybean meal in different levels of replacement to the fish meal on juvenile L. vannamei. Growth performance, feed utilization, immunological parameters, antioxidant enzyme assays, lipid peroxidation, digestive enzymes, and histopathological analysis were measured for providing accurate evaluation.
Materials and methods
Ethical statement
The Animal Research and Ethics Committee of the National Institute of Oceanography and Fisheries in Suez, Egypt, approved the research work plan and permitted it to work with Litopenaeus vannamei post-larvae. All experiments were done by the committee’s rules.
Solid-state fermentation of soybean meal preparation
Commercial soybean meal (SBM) was obtained from a company in Egypt’s Zagazig province and ground to a particle size of 500 m. The modified method of Yabaya et al. (2009) was used to perform fermented SBM. Briefly, 2 kg of SBM was combined with 1.1 L of distilled water (50% moisture), and 60.5 mg of commercial dry yeast Saccharomyces cerevisiae (S. cerevisiae) of cell density 3 × 106 cell g1 (Fermipan®, GB ingredients, China). In a Hobart food mixer, all the ingredients were homogenized for 15 min. The mixture was incubated for 48 h at 40 °C, the ideal growth temperature for S. cerevisiae, in a 10-L glass jar covered with aluminum foil. Ten grams of the solid-state fermented soybean meal (SSF-SBM) with yeast was sampled at 0, 12, 24, and 48 h of fermentation to assess the anti-nutritional factors (ANFs) and chemical composition content. Finally, the SSF-SBM was dried to a constant weight at 70 °C.
Diet formulation
A control diet with FM (FSBM0) as the primary source of protein and three experimental diets in which the FM in the control diet was replaced by FSBM at 20, 30, and 40% resulted in four isonitrogenous (41.2 g/kg crude protein and 8.2 g/kg isocaloric diets) (Table 1). The dry ingredients and fish oil were combined to make a stiff dough by adding water. Using a meat grinder, the mixtures were formed into 1.6-mm-diameter pellets. In a forced-air oven set at 40 °C, pellets were dried. Before use, fully dried diets were put in plastic bags and kept at 18 °C.
Amino acid analysis
Following procedures recommended by the Association of Official Analytical Chemists (AOAC 1990) and the protocol of Llames and Fontaine (1994), the amino acid profile of FSBM was determined using an amino acid analyzer (LA8080, Amino SAYA, Hitachi High-Tech, Japan).
Feeding trial
L. vannamei juveniles were obtained from the El-Sahaba hatchery in Damietta, Egypt, and transported to the National Institute of Oceanography and Fisheries’ Invertebrates Laboratory in Suez, Egypt. The juveniles (1.59 ± 0.01 g) were acclimated for 1 week before being distributed at random into fifteen fiberglass tanks (40 L volume). Compressed air was used to provide continuous aeration. For 12 weeks, shrimp juveniles were fed the four test diets in triplicate (0.0, 20%, 30%, and 40% FSBM, respectively) to apparent satiation twice daily. Dietary amounts were adjusted based on survival and body weight. Approximately 10% of the water volume in each aquarium was changed out daily with fresh aerated and filtered marine water after removing the accumulated wastes.
Physicochemical characteristics of water
Daily partial exchange of 10% of the water kept the water quality parameters within the acceptable ranges during the experimental trial. Dissolved oxygen levels were kept at 5.74 ± 0.50 mg/L using an oxygen meter (HANNA, HI 9146-04/10), pH was kept at 7.5 ± 0.3 using a pH meter (Adwa, AD-11), unionized ammonia concentration was kept at 0.22 ± 0.04 mg/L using DREL/2 HACH kits (HACH Co., Loveland, Co.), and salinity was kept at 30.56 ± 0.80 (Bioevopeak Co., Ltd., China), while the water temperature was maintained at a range of 28.52 ± 0.92 °C.
Growth performance and feed utilization
Weight in (g) and length in (cm) of each post larvae (PL) were measured at the start of the experiment and every 2 weeks for the next 12 weeks. All shrimp were weighed to determine their final weight at the end of the experiment, which was then compared to their initial weight on the first day. By counting the individuals in each aquarium, the survival rates of the PL were also estimated.
Feed and body composition chemical analysis
The standard methods of AOAC (2012) were used to determine the moisture, crude protein, crude lipid, and ash content of diets and shrimp samples. The dry matter of the samples was determined by drying them to a constant weight at 105 °C. Crude protein was calculated by multiplying nitrogen by 6.25 using the Kjeldahl method (Kjeltec TM8400, FOSS, Sweden). The Soxhlet method was used to determine crude lipid after diethyl ether extraction (Buchi 36680, Switzerland), after 16 h of combustion in a muffle furnace at 550 °C. According to Goering and van Soest (1970), the crude fiber was estimated. Gross energy was computed according to the standards of NRC (1993).
Immunological parameters
Hemolymph samples were taken from the ventral sinus of the recently molted shrimp using 1-mL syringes and a 27-G needle containing anticoagulant (1:1), according to El Asely et al. (2010). The molting stages were identified through periodic observation of the flock and were confirmed through microscopic examination of the epidermal retraction following Robertson et al. (1987).
Hemolymph plasma was obtained by centrifugation to the hemolymph-anticoagulant mix at (10,000 g/4 °C) for 5 min, according to Lamela et al. (2005). The obtained plasma was stored at − 20 °C.
The total protein of hemolymph plasma was determined following Bradford (1976) at 595 nm, where the bovine serum albumin was used as standard.
Lysozyme activity was determined through measurement of the decrease in the absorbance of lysis to the Micrococcus lysodeikticus cells (Sigma-Aldrich, Cat. no. LY0100) at a wavelength of 450 nm following the manufacturer instructions and the protocol of Sotelo-Mundo et al. (2003).
Antioxidant enzyme assays and lipid peroxidation
The shrimp were euthanized following the American Veterinary Medical Association (AVMA) guidelines for animal euthanasia (Leary et al. 2013). Shrimp were immersed in ice for 15 min till no motion was noticed. Hepatopancreas was dissected and kept in cold phosphate-buffered saline (PBS), and then, it was homogenized. Tissue homogenate was centrifuged at 12,000 × g for 12 min at 4 °C. The supernatant was removed and stored at − 75 °C. Catalase (CAT), superoxide dismutase (SOD), and glutathione peroxidase (Gpx) activities were measured photometrically using a microplate reader and the commercially available kits (Abcam; Catalase-ab83464, SOD-ab65354, GPx-ab219926) at the wavelength (OD 570 nm, OD 450 nm, and OD 405 nm) respectively, while GR activity was measured fluorometrically at Ex/Em = OD420/480 nm using kits (Abcam; GR-ab83461). Colorimetric lipid peroxidation assay kit (Abcam, ab118970) was used to detect the content of malondialdehyde (MDA) in the hepatopancreas tissue homogenate at OD = 532 nm.
Digestive enzyme assay
Hepatopancreas and intestines were weighed and homogenized in cold PBS. Tissue homogenates were centrifuged at 18,894 × g at 4 °C for 5 min. The supernatant was stored at − 20 °C until analysis. Amylase, lipase, and protease activities were determined using Abcam kits following the manufacturer’s instructions. Amylase activity was detected using colorimetric kits (ab102523) measured at OD = 405 nm. Lipase activity was assayed using kits (Abcam, ab102524), where lipase hydrolyzes a triglyceride substrate to form glycerol which is quantified enzymatically by monitoring a linked change in the absorbance of a probe (OD = 570nm). Lipase activity was calculated as nanomole of glycerol per milligram of protein. Protease activity was estimated by Protease Activity Assay Kit (fluorometric—green) (ab112152) with fluorescence intensity at Ex/Em = 490/525 nm.
Histopathological analysis
The hepatopancreas of three shrimp/replicate/group were dissected and then fixed for 24 h in Davidson’s fixative before being transferred to 70% ethanol for standard histological processing (Bell and Lightner 1988). The tissue sections (4–5m) were stained with hematoxylin-eosin, examined using a light microscope (Olympus CX 41, Japan), and photographed using an Olympus E-620 digital camera. Using Image J software equipped with a cell counter plugin, the number of R cells (Restzellen or resorptive cell) and B cells (Blasenzellen or blister cell) as well as tubule diameter were determined in 20 randomly selected tubules from each treatment (Romano et al. 2015).
Statistical analysis
SPSS software (ver. 17.0) was used to conduct the statistical analysis. All data were given as mean ± standard error (S.E.). Using one-way ANOVA and Tukey’s post hoc multiple comparisons, statistical significance was determined. P-values < 0.05 were deemed statistically significant.
Results
Essential amino acid content
In the current study, the partial replacement of FM with FSBM influenced the aa profile in the diet, with an improvement in the contents of some amino acids such as arginine, lysine, leucine, isoleucine, phenylalanine, threonine, and valine in shrimp diets and a decrease in the contents of some amino acids such as histidine and methionine (Table 2).
Growth performance and feed utilization
At the end of the feeding trial, the survival of L. vannamei juveniles was reduced with no significant difference between feeding groups. There was no significant difference in growth performance or feed utilization parameters between the four experimental groups (P > 0.05) (Table 3). The 20% FSBM diet had significantly improved growth performance in terms of FBW, BWG, RBWG, and SGR (P < 0.05), followed by the groups fed the 30, 0.0, and 40% FSBM diets, respectively, while there were no statistically significant differences in these parameters between the 30, 0.0, and 40% FSBM diets. The lowest was found at 40% FSBM.
As shown in Table 4, the low replacement level (20% FSBM) significantly increased feed intake, resulting in the greatest growth performance relative to the other replacements. In addition, 20% replacement exhibited the lowest FCR (P < 0.05) compared to those fed a 40% FSBM diet. Conversely, the low replacement level group (20% FSBM) significantly improved (P < 0.05) FER, PER, APU, and EU compared to the high replacement level group. Forty-percent FSBM diet produced the least amount of feed utilization.
Proximate body composition analysis
Table 5 presents the approximate composition of the entire body of L. vannamei juveniles. The present investigation revealed no statistically significant variations (P > 0.05) in the moisture content of L. vannamei shrimp across all dietary regimens, with moisture levels ranging from 75.64 to 76.46%. The group fed 20% FSBM had the highest protein content (21.99%), while the group fed 0% FSBM had the lowest protein level (20.36%). It was noticed that raising FSBM levels significantly increased lipid content (P < 0.05) compared to the control. However, there were no statistically significant changes (P > 0.05) among FSBM treatments. In contrast, as the amount of dietary FSBM increased from 0 to 40% FSBM, ash concentrations tended to decrease. The highest significant levels of ash content were recorded in the control diet (0.0% FSBM).
Immunological parameters
Hemolymph plasma TP concentration was substantially greater (P < 0.05) in 20% FSBM, followed by 30% FSBM and 0% FSBM, with 40% FSBM having the lowest concentration (Table 6). Similarly, lysozyme activity was greater in 20% FSBM than in other FSBM concentrations (Table 6).
Antioxidant enzyme assays and lipid peroxidation
The obtained results presented in Table 7 showed a significant increase (P < 0.05) in the activity of hepatopancreas catalase enzyme in 20% FSBM, and the same pattern was recorded in the other measured antioxidant enzymes (SOD, GPX, and GR). On the other hand, significantly low catalase activity was recorded in 30% FSBM, followed by control and 40% FSBM. In contrast, the content of lipid peroxidation marker (MDA) was significantly higher (P < 0.05) in control and 30% FSBM than in 20 and 40% FSBM levels.
Digestive enzyme assay
Hepatopancreas amylase activity was significantly high (P < 0.05) in 40% FSBM and control followed by 30% FSBM and 20% FSBM (Table 8). While protease activity showed a significant increase in 20% FSBM, its lowest activity was recorded in 40% FSBM. While protease activity showed a significant increase in the 20% FSBM diet, its lowest activity was recorded in the 40% FSBM diet. Contrarily, the values of lipase activity were significantly higher (P < 0.05) in 20% of FSBM, and the lowest activity was recorded in 40% of FSBM (Table 8).
On the other hand, intestinal amylase activity was significantly high (P < 0.05) in 0.0% FSBM, while 20% FSBM was the lowest (Table 8). Protease activity showed a significant increase in 20% FSBM, and its lowest activity was recorded at 40% FSBM. Contrarily, the values of lipase activity were significantly higher (P < 0.05) in 20% of FSBM, and the lowest activity was recorded in 40% of FSBM (Table 8).
Gross pathology
Gross pathology revealed that shrimp given 30 and 40% FSBM exhibited a pale white distal hepatopancreas and a slightly shrunken stomach (Fig. 1a, b).
Histology of hepatopancreas
Figure 2a–d depicts the hepatopancreatic tubules of shrimp fed the control and experimental diets. Considerably, more B cells were present in the 40% FSBM diet than in the control diet; however, they were significantly less prevalent in the 20% and 30% FSBM diets (P < 0.05). In addition, 30% of FSBM-fed shrimp had considerably more R cells than the others. In terms of hepatopancreatic tubule diameters, the 30% and 40% FSBM diets had considerably larger tubules than the other diets (P < 0.05). The predominance of B cells and R cells in tubules, as well as the tubule diameter, is shown in Table 9.
Discussion
Plant proteins especially soya bean (SB) could be a promising source of protein in aquatic animal feed; however, they are high in cellulose, which is difficult for fish and other monogastric animals to digest (Gatlin III et al. 2007). Researchers are working on bioprocessing SB in other products such as soybean meal (SBM), soy protein isolate (SPI), fermented soybean meal (FSBM), and soybean protein concentrate (SPC) in shrimp diets in order to reduce ANFs (Abdul Kader et al. 2012; Gamboa-delgado et al. 2013). In addition, fermentation of SB has been reported to increase its protein content (Teng et al. 2012), promote antibacterial and antioxidant activities (He et al. 2013; Akbari and Wu 2015), and diminish immunoglobulin E immunological activity (Song et al. 2008).
The findings of this study indicate that 20% of FM can be substituted with FSBM without harming the health of shrimp. In addition, juvenile L. vannamei given 20% FBSM displayed enhanced growth performance and feed utilization, despite the fact that the amino acid composition of all experimental groups was identical. In addition, this improvement may be attributable to the improved lipid digestibility of FSBM (Refstie et al. 2005) and the reduction of anti-nutritional components during fermentation (Su et al. 2018). In contrast, the growth of shrimp fed a diet containing 40% FSBM was diminished. This decline may be attributable to the presence of non-digestible oligosaccharides, decreased protein digestibility, or a nutritional imbalance (Sharawy et al. 2016).
Our study’s growth performance results were comparable to those of earlier research conducted on other crustacean species. According to the findings of Ding et al. (2015), the optimal growth performance of M. nipponense was achieved when 25% of the FM was substituted with FSM. Research on F. indicus suggests that replacing up to 28.57% of FM with FSM is significantly more economical (Sharawy et al. 2016). Shao et al. (2018) observed that a meal with a moderate FSM replacement level (20% of FM protein) efficiently boosted the growth of juvenile white shrimp and that a replacement level of up to 40% had no influence on shrimp growth performance.
In the present investigation, there were no statistically significant differences (P > 0.05) in the moisture content of L. vannamei shrimp between diets, while the addition of S. cerevisiae increased the protein content of FSBM-fed groups significantly. The same rise in body lipid content was observed with increased dietary FSBM, which included more carbohydrates than FM (Makkar et al. 2007). According to Kaushik et al. (2004), one of the major factors leading to increased lipid retention is an increase in dietary plant protein, which is associated with an increase in hepatic lipogenic enzyme activity, imbalances in dietary amino acid content, and higher whole-body lipid levels in sea bass and salmonids. In addition, Sharawy et al. (2016) reported that there were statistically significant variations (P < 0.05) in the protein, dry matter, lipid, and ash content of shrimp fed different experimental diets compared to control diets containing 0.0% protein (FSBM). In our study, ash contents tended to decrease as FSBM levels increased from 0 to 40% of the meal.
Hemolymph metabolites serve as physiological, nutritional, and immunological stress indicators in crustaceans. In addition, it has been used to evaluate the nutritional health of shrimp, whose blood protein and glucose levels are very sensitive to the protein content of their diet (Rosas et al. 2001). In the current study, the inclusion of FSBM in the diet had a significant effect on the hemolymph total protein (TP) content, with the maximum TP concentration seen in groups fed 20% FSBM, followed by 30% FSBM. According to Shiu et al. (2015), the increased protein content of soya bean meal after fermentation may account for the observed results. On the other hand, the decreased TP concentration in 40% FSBM is due to the detrimental effect of high soya bean levels on the digestibility, absorption, and utilization of dietary protein (Gilani et al. 2012).
With its antibacterial effectiveness against bacterial infection, lysozyme activity is one of the most important indicators of shrimp immunity (Kaizu et al. 2011). The addition of S. cerevisiae to ferment SBM was also beneficial in regulating serum lysozyme activity. The results demonstrated a considerable increase in the lysozyme activity of 20% of FSBM-fed groups. The process involved in boosting the immune system of shrimp hinges on the protein recognition pattern of the circulating sugars which evoke the immune cells (Vargas-Albores and Yepiz-Plascencia 2000). It is assumed that the yeast harboring β-glucan effectively stimulated lysozyme synthesis. In contrast to what was expected, the lysozyme activity decreased as the concentration of FM-replacement FSBM increased. This was postulated as a result of the fatigue of lysozyme-producing cells from long-term exposure to the triggering agents, recommending the use of a low dose for a brief period of time for an effective response (El-Barbary et al. 2021; Babu et al. 2013; El Asely et al. 2011).
In addition to exogenous sources of reactive oxygen species (ROS), regular cellular metabolism generates electrons that can alter the membrane protein structure, lipids, cell division, and apoptosis signaling pathway (Redza-Dutordoir and Averill-Bates 2016; Bauer and Bauer 1999). Antioxidants’ function in cells is to maintain balance and scavenge excess reactive oxygen species (ROS) to mitigate their corrosive effect (Kurutas 2015).
In the present study, hepatopancreas CAT, SOD, Gpx, and GR activities increased significantly in the group fed 20% FSBM, indicating that the 20% substituted FSBM meal had a greater anti-oxidative effect than fish fed FM and the other two concentrations. Ding et al. (2015) detected a drop in CAT, SOD, and GSH-PX activities with increased FSM content in the diet of Macrobrachium nipponense. The results obtained were almost identical to those reported by Ding et al. which suggested that the anti-oxidative capacity of shrimp was compromised by the substitution of fishmeal. Xu et al. (2008) found that fish CAT activity fell dramatically from 30 to 20% when fishmeal was substituted. Despite the fact that Daiyong et al. (2009) discovered that the CAT activity of shrimp was unaffected, the SOD activity declined dramatically when fishmeal was reduced from 25 to 20%. In addition to the enhanced flavonoid content created during soybean fermentation, tiny peptides, organic acids, and probiotics are also produced (Mukherjee et al. 2016). Saccharomyces cerevisiae produces vitamins and other metabolites that serve as exogenous antioxidant sources (Farid et al. 2019).
The hepatopancreatic MDA level of shrimp given 20% FSBM was much lower than that of shrimp fed the FM and other diets, demonstrating that the dietary replacement of FM with FSBM did not stimulate oxidative stress and was even successful at decreasing it. This could be attributable to the FSBM’s high isoflavonoid content, which can neutralize free radicals and prevent lipid peroxidation (Yoon and Park 2014).
The hepatopancreas is responsible for the generation and release of digestive enzymes, the absorption of nutrients, and the mobilization and transport of nutrients such as lipids, glycogen, minerals, and organic compounds to muscle and other tissues in response to growth and reproductive needs (Ceccaldi 1989). The hepatopancreas secretes enormous quantities of digestive enzymes, such as amylases and proteases (Gamboa-delgado et al. 2003). Dietary content has a significant influence on digestive enzyme production and activity (Le Moullac et al. 1997; Guzman et al. 2001). In the present investigation, the substitution of fish meal with FSBM had a substantial effect on the activity of digestive enzymes; amylase activity was significantly higher in 40% FSBM and control diets than in other diets, indicating a higher carbohydrate content.
Interestingly, the digestive enzyme concentration in the intestine was substantially identical to that of the hepatopancreas, corroborating the findings of Córdova-Murueta et al. (2003).
The hepatopancreas is the most essential digestive organ in shrimp. Histologically, it consists of four distinct cell types contained within blind-ending tubules. E cells differentiate into R cells (nutrient absorption and storage), F cells (production of digestive enzymes), and B cells (presumed to be secretory in function) at the apex of the tubules (Gopinath and Paul Raj 2009).
In the present study, shrimp fed 30% partial fish meal replacement had significantly more R cells than other groups (P < 0.05); this increase in R cells, which are responsible for lipid storage in the hepatopancreas gland, may be indicative of an increase in energy reserve in the hepatopancreas as a result of the treatment. The B cells are large cells responsible for enzyme storage, and this study revealed that their prevalence was significantly higher in the 40% partial fish meal replacement group than in the control group but significantly lower in the 20% and 30% partial fish meal replacement groups (P < 0.05). In previous research, hypertrophied B cells were identified in shrimp fed moderate dosages of the mycotoxin deoxynivalenol (DON), indicating oxidative stress in shrimp (Xie et al. 2018). Also, the increased B-cell prevalence in L. vannamei has been observed to increase at low salinities, suggesting that this is a response to the increased nutrient use required for higher osmoregulatory functions (Li et al. 2008). Similar to our findings, Romano et al. (2015) concluded that while the prevalence of R cells was significantly higher in shrimp fed organic acid–blended diets, indicating greater energy reserves, the prevalence of B cells, which are primarily responsible for the secretion of digestive enzymes, was significantly lower. The 30% and 40% partial fish meal replacement groups had substantially larger hepatopancreatic tubule diameters than the other groups (P < 0.05). The increase in hepatopancreatic tubule diameter may be correlated with the greater prevalence of R cells and the resulting fat storage inside them (Johnston et al. 2003; Simon and James 2007; Pourmozaffar et al. 2019), which may be associated with the gross pathology picture of the hepatopancreas with the white fuzzy zone.
Conclusion
In conclusion, the current study demonstrates that 20% replacement of FM with FSBM improved growth performance, feed utilization, immunological parameters, antioxidant enzymes assays, lipid peroxidation, digestive enzymes, and histopathological analysis in juvenile L. vannamei, despite the fact that there was no difference in the amino acid content between the experimental groups. This improvement may also be attributable to the increased lipid digestibility of FSBM and the decrease in anti-nutritional components during fermentation.
Data availability
Data of the present article will be available on request.
References
Abd El-Naby AS, El Asely AM, Hussein MN, Fawzy RM, Abdel-Tawwab M (2022) Stimulatory effects of dietary chia (Salvia hispanica) seeds on performance, antioxidant-immune indices, histopathological architecture, and disease resistance of Nile tilapia. Aquac 563:738889
Abdul Kader M et al (2012) Can fermented soybean meal and squid by-product blend be used as fishmeal replacements for Japanese flounder (Paralichthys olivaceus). Aquac Res 43(10):1427–1438
Akbari A, Wu J (2015) An integrated method of isolating napin and cruciferin from defatted canola meal. LWT-Food Sci Technol 64(1):308–315
AOAC (1990) Association of Official Analytical Chemists (AOAC), 1990. In: Official methods of analysis of the Association of Official Analytical Chemists, 15th edn. AOAC, Washington, DC
AOAC (2012) Official methods for analysis, 19th edn. Association of Official Analytical Chemists, Washington, DC
Babu DT, Antony SP, Joseph SP, Bright AR, Philip R (2013) Marine yeast Candida aquaetextoris S527 as a potential immunostimulant in black tiger shrimp Penaeus monodon. J Invertebr Pathol 112(3):243–252
Bauer V, Bauer F (1999) Reactive oxygen species as mediators of tissue protection and injury. Gen Physiol Biophys 18:7–14
Bell TA, Lightner DV (1988) A handbook of normal penaeid shrimp. World Aquaculture Society, Baton Rouge, LA
Bondad-Reantaso MG, Subasinghe RP, Josupeit H, Cai J, Zhou X (2012) The role of crustacean fisheries and aquaculture in global food security: past, present and future. J Invertebr Pathol 110(2):158–165
Bradford M (1976) A rapid and sensitive method for the quantification of microgram quantities of protein utilizing the principle of protein dye binding. Anal Biochem 72:248–253
Ceccaldi H (1989) Anatomy and physiology of digestive tract of Crustaceans Decapods reared in aquaculture. In: Advances in Tropical Aquaculture, Workshop at Tahiti, French Polynesia
Chi S et al (2009) Growth and feed efficiency of juvenile shrimp Litopenaeus vannamei fed formulated diets containing different levels of poultry by-product meal. J Ocean Univ China 8(4):399–403
Córdova-Murueta JH, Garcıa-Carreno FL, de los A Navarrete-del M (2003) Digestive enzymes present in crustacean feces as a tool for biochemical, physiological, and ecological studies. J Exp Mar Biol Ecol 297(1):43–56
Daiyong W, Yuantu Y, Baotong Z (2009) Effects of cotton seed meal and rapeseed meal on growth performance, non-specific immune indexes and body compositions of Litopenaeus vannamei. China Feed 23:12
Ding Z, Zhang Y, Ye J, Du Z, Kong Y (2015) An evaluation of replacing fish meal with fermented soybean meal in the diet of Macrobrachium nipponense: growth, nonspecific immunity, and resistance to Aeromonas hydrophila. Fish Shellfish Immunol 44(1):295–301
Dossou S et al (2018) Effect of partial replacement of fish meal by fermented rapeseed meal on growth, immune response and oxidative condition of red sea bream juvenile, Pagrus major. Aquac 490:228–235
El Asely AM, Shaheen AA, Abbass AA, Inada M, Okugawa S, Tachikawa Y, Itami T (2011) Changes in the immune-related gene expression of kuruma shrimp Marsupenaeus japonicus in response to dietary inclusion of macrophage activating Chinese herbs (MACH). In: Proceedings of the 4th Global Fisheries and Aquaculture Research Conference, the Egyptian International Center for Agriculture. Massive Conferences and Trade Fairs, Giza, Egypt, 3-5 October 2011, pp 295–303
El Asely AM, Shaheen AA, Abbass AA et al (2010) Immunomodulatory effect of plant-mixed feed in kuruma shrimp, Marsupenaeus japonicus, and its protective efficacy against white spot syndrome virus infection. J Fish Dis 33:859–863
El-Barbary Y, Gaafar A, Younes A, El-Ashram A (2021) The influence of continuous and intermittent Bacillus subtilis AQUA-GROW® application on the white leg shrimp, Litopenaeus vannamei, immune-related genes. Egypt J Aquat Biol Fish 25(3):241–261
Faggio C, Fazio F, Marafioti S, Arfuso F, Piccione G (2015) Oral administration of gum arabic: effects on haematological parameters and oxidative stress markers in Mugil cephalus. Iran J Fish Sci 14(1):60–72
FAO (2022) The state of world fisheries and aquaculture 2022. In: Towards Blue Transformation. FAO, Rome. https://doi.org/10.4060/cc0461en
Farid F, Sideeq O, Khan F, Niaz K (2019) Saccharomyces cerevisiae. In: Nonvitamin and nonmineral nutritional supplements. Academic Press, pp 501–508 44349
Feizi LK, Seifdavati J, Rafiee H, Rezazadeh F, Meléndez JH, Molina OM et al (2022) Biotechnological valorization of fermented soybean meal for sustainable ruminant and non-ruminant feeding: modulating ruminal fermentation, gut or ruminal microflora, immune system, and growth performance. Biomass Conv Bioref. https://doi.org/10.1007/s13399-022-02971-7
Gamboa-delgado J, Molina-poveda C, Cahu C (2003) Digestive enzyme activity and food ingesta in juvenile shrimp Litopenaeus vannamei (Boone, 1931) as a function of body weight. Aquac Res 34(15):1403–1411
Gamboa-delgado J, Rojas-Casas MG, Nieto-López MG, Cruz-Suárez LE (2013) Simultaneous estimation of the nutritional contribution of fish meal, soy protein isolate and corn gluten to the growth of Pacific white shrimp (Litopenaeus vannamei) using dual stable isotope analysis. Aquac 380:33–40
Gatlin DM III, Barrows FT, Brown P, Dabrowski K, Gaylord TG, Hardy RW et al (2007) Expanding the utilization of sustainable plant products in aquafeeds: a review. Aquac Res 38(6):551–579
Gilani GS, Xiao CW, Cockell KA (2012) Impact of antinutritional factors in food proteins on the digestibility of protein and the bioavailability of amino acids and on protein quality. Br J Nutr 108(S2):S315–S332
Goering HK, van Soest PJ (1970) Forage fiber analyses (apparatus, reagents, procedures, and some applications). US Agricultural Research Service
Gopinath R, Paul Raj R (2009) Histological alterations in the hepatopancreas of Penaeus monodon Fabricius (1798) given aflatoxin B1-incorporated diets. Aquac Res 40(11):1235–1242
Guzman C, Gaxiola G, Rosas C, Torre-Blanco A (2001) The effect of dietary protein and total energy content on digestive enzyme activities, growth and survival of Litopenaeus setiferus (Linnaeus 1767) postlarvae. Aquac Nutr 7:113–122
He G, Sui J, Du J, Lin J (2013) Characteristics and antioxidant capacities of five hawthorn wines fermented by different wine yeasts. J Inst Brew 119(4):321–327
Hong K-J, Lee C-H, Kim SW (2004) Aspergillus oryzae GB-107 fermentation improves nutritional quality of food soybeans and feed soybean meals. J Med Food 7(4):430–435
Hoseinifar SH, Mirvaghefi A, Merrifield DL (2011) The effects of dietary inactive brewer’s yeast Saccharomyces cerevisiae var. ellipsoideus on the growth, physiological responses and gut microbiota of juvenile beluga (Huso huso). Aquac 318(1–2):90–94
Hotz C, Gibson RS (2007) Traditional food-processing and preparation practices to enhance the bioavailability of micronutrients in plant-based diets. J Nutr 137(4):1097–1100
Ishwarya R, Vaseeharan B, Jayakumar R, Ramasubramanian V, Govindarajan M, Alharbi NS, Benelli G (2018) Bio-mining drugs from the sea: high antibiofilm properties of haemocyanin purified from the haemolymph of flower crab Portunus pelagicus (L.) (Decapoda: Portunidae). Aquac 489:130–140
Johnston DJ, Calvert KA, Crear BJ, Carter CG (2003) Dietary carbohydrate/lipid ratios and nutritional condition in juvenile southern rock lobster, Jasus edwardsii. Aquac 220(1-4):667–682
Kaizu A, Fagutao FF, Kondo H, Aoki T, Hirono I (2011) Functional analysis of C-type lysozyme in penaeid shrimp. J Biol Chem 286(52):44344–44349
Kaushik SJ, Coves D, Dutto G, Blanc D (2004) Almost total replacement of fish meal by plant protein sources in the diet of a marine teleost, the European seabass, Dicentrarchus labrax. Aquac 230(1-4):391–404
Kikuchi K (1999) Use of defatted soybean meal as a substitute for fish meal in diets of Japanese flounder (Paralichthys olivaceus). Aquac 179(1-4):3–11
Kurutas EB (2015) The importance of antioxidants which play the role in cellular response against oxidative/nitrosative stress: current state. Nutr J 15(1):1–22
Lamela REL, Silveira Coffigny R, Quintana YC, Martínez M (2005) Phenoloxidase and peroxidase activity in the shrimp Litopenaeus schmitti, Pérez-Farfante and Kensley (1997) exposed to low salinity. Aquac Res 36(13):1293–1297
Le Moullac G, Klein B, Sellos D, Van Wormhoudt A (1997) Adaptation of trypsin, chymotrypsin and a-amylase to casein level and protein source in Penaeus vannamei (Crustacea Decapoda). J Exp Mar Biol Ecol 208:107–125
Leary S, Underwood W, Anthony R, Cartner S, Corey D, Grandin T, Greenacre C, Gwaltney-Brant S, McCrackin MA, Meyer R et al (2013) AVMA guidelines for the euthanasia of animals. American Veterinary Medical Association, Schaumburg, IL
Li E, Chen L, Zeng C, Yu N, Xiong Z, Chen X, Qin JG (2008) Comparison of digestive and antioxidant enzymes activities, haemolymph oxyhemocyanin contents and hepatopancreas histology of white shrimp, Litopenaeus vannamei, at various salinities. Aquac 274(1):80–86
Llames CR, Fontaine J (1994) Determination of amino acids in feeds: collaborative study. J AOAC Int 77(6):1362–1402
Makkar HPS, Francis G, Becker K (2007) Bioactivity of phytochemicals in some lesser-known plants and their effects and potential applications in livestock and aquaculture production systems. Animal 1(9):1371–1391
Molina-Poveda C, Lucas M, Jover M (2015) Utilization of corn gluten meal as a protein source in the diet of white shrimp Litopenaeus vannamei. Aquac Nutr 21:824–834
Mukherjee R, Chakraborty R, Dutta A (2016) Role of fermentation in improving nutritional quality of soybean meal a review. Asian Australas J Anim Sci 29(11):1523
Nath M, Bhatt D, Bhatt MD, Prasad R, Tuteja N (2018) Microbe-mediated enhancement of nitrogen and phosphorus content for crop improvement. In: Crop improvement through microbial biotechnology. Elsevier, pp 293–304
National Research Council (1993) Nutrient requirements of fish. National Academy Press, Washington DC
Pauly D, Froese R (2012) Comments on FAO’s state of fisheries and aquaculture, or ‘SOFIA 2010. Mar Policy 36(3):746–752
Pourmozaffar S, Hajimoradloo A, Paknejad H, Rameshi H (2019) Effect of dietary supplementation with apple cider vinegar and propionic acid on hemolymph chemistry, intestinal microbiota and histological structure of hepatopancreas in white shrimp, Litopenaeus vannamei. Fish Shellfish Immunol 86:900–905
Redza-Dutordoir M, Averill-Bates DA (2016) Activation of apoptosis signaling pathways by reactive oxygen species. Biochimica et Biophysica Acta (BBA)-Molecular. Cell Res 1863(12):2977–2992
Refstie S, Sahlström S, Bråthen E, Baeverfjord G, Krogedal P (2005) Lactic acid fermentation eliminates indigestible carbohydrates and antinutritional factors in soybean meal for Atlantic salmon (Salmo salar). Aquac 246(1-4):331–345
Robertson L, Bray W, Leung-Trujillo J, Lawrence A (1987) Practical molt staging of Penaeus setiferus and Penaeus stylirostris. J World Aquacult Soc 18(3):180–185
Romano N, Koh CB, Ng WK (2015) Dietary microencapsulated organic acids blend enhances growth, phosphorus utilization, immune response, hepatopancreatic integrity and resistance against Vibrio harveyi in white shrimp, Litopenaeus vannamei. Aquac 435:228–236
Rosas C, Cuzon G, Gaxiola G, Le Priol Y, Pascual C, Rossignyol J, Van Wormhoudt A (2001) Metabolism and growth of juveniles of Litopenaeus vannamei: effect of salinity and dietary carbohydrate levels. J Exp Mar Biol Ecol 259(1):1–22
Shao J, Zhao W, Liu X, Wang L (2018) Growth performance, digestive enzymes, and TOR signaling pathway of Litopenaeus vannamei are not significantly affected by dietary protein hydrolysates in practical conditions. Front Physiol 9:998
Sharawy Z, Abbas EM, Desouky MG, Kato M (2016) Descriptive analysis and molecular identification of the green tiger shrimp Penaeus semisulcatus (De Haan, 1844) in Suez Gulf, Red Sea. Int J Fish Aquat Stud 4(5):426–432
Shiu YL, Hsieh SL, Guei WC, Tsai YT, Chiu CH, Liu CH (2015) Using Bacillus subtilis E20-fermented soybean meal as replacement for fish meal in the diet of orange-spotted grouper (E pinephelus coioides, Hamilton). Aquac Res 46(6):1403–1416
Simon CJ, James PJ (2007) The effect of different holding systems and diets on the performance of spiny lobster juveniles, Jasus edwardsii (Hutton, 1875). Aquac 266(1-4):166–178
Song YS et al (2008) Immunoreactivity reduction of soybean meal by fermentation, effect on amino acid composition and antigenicity of commercial soy products. Food Chem 108(2):571–581
Sookying D, Davis DA, Da Silva F SD (2013) A review of the development and application of soybean-based diets for Pacific white shrimp Litopenaeus vannamei. Aquac Nutr 19(4):441–448
Sotelo-Mundo RR, Islas-Osuna MA, De-la-Re-Vega E, Hernández-López J, Vargas-Albores F, Yepiz-Plascencia G (2003) cDNA cloning of the lysozyme of the white shrimp Penaeus vannamei. Fish Shellfish Immunol 15(4):325–331
Su LW, Cheng YH, Hsiao FSH, Han JC, Yu YH (2018) Optimization of mixed solid-state fermentation of soybean meal by species and. Pol J Microbiol 67(3):297–305
Tacon AGJ, Metian M (2008) Global overview on the use of fish meal and fish oil in industrially compounded aquafeeds: trends and future prospects. Aquac 285(1–4):146–158
Teng D, Gao M, Yang Y, Liu B, Tian Z, Wang J (2012) Bio-modification of soybean meal with Bacillus subtilis or Aspergillus oryzae. Biocatal Agric Biotechnol 1(1):32–38
Vargas-Albores F, Yepiz-Plascencia G (2000) Beta glucan binding protein and its role in shrimp immune response. Aquac 191(1-3):13–21
Xie S, Zheng L, Wan M, Niu J, Liu Y, Tian L (2018) Effect of deoxynivalenol on growth performance, histological morphology, anti-oxidative ability and immune response of juvenile Pacific white shrimp, Litopenaeus vannamei. Fish Shellfish Immunol 82:442–452
Xu PL, Guo YK, Bai JG, Shang L, Wang XJ (2008) Effects of long-term chilling on ultrastructure and antioxidant activity in leaves of two cucumber cultivars under low light. Physiol Plant 132(4):467–478
Yabaya A, Akinyanju JA, Jatau ED (2009) Yeast enrichment of soybean cake. World J Dairy Food Sci 4(2):141–144
Yoon GA, Park S (2014) Antioxidant action of soy isoflavones on oxidative stress and antioxidant enzyme activities in exercised rats. Nutr Res Pract 8(6):618–624
Zhang C, Rahimnejad S, Wang YR, Lu K, Song K, Wang L, Mai K (2018) Substituting fish meal with soybean meal in diets for Japanese seabass (Lateolabrax japonicus): effects on growth, digestive enzymes activity, gut histology, and expression of gut inflammatory and transporter genes. Aquac 483:173–182
Acknowledgements
This research work was supported by the Department of Aquatic Animal Medicine, Faculty of Veterinary Medicine, Benha University, Egypt.
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Asmaa S. Abd El-Naby: conceptualization, writing (original draft), methodology and growth and feed utilization analysis, and final draft preparation. Eid, A. E: supervision. Alkhateib Y. Gaafar: histopathology analysis and conceptualization. Zaki Sharawy: investigation and supervision. Mohamed S. El-sharawy: writing (original draft), methodology statistical analysis, and data tabulation final draft revision. Khattaby A. A: methodology and final revision. Amel M. El Asely: conceptualization, immune and antioxidant methodology, data curation, investigation, and final draft review and editing.
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Abd El-Naby, A.S., Eid, A.E., Gaafar, A.Y. et al. Overall evaluation of the replacement of fermented soybean to fish meal in juvenile white shrimp, Litopenaeus vannamei diet: growth, health status, and hepatopancreas histomorphology. Aquacult Int 32, 1665–1683 (2024). https://doi.org/10.1007/s10499-023-01234-0
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DOI: https://doi.org/10.1007/s10499-023-01234-0