Effects of hydroxyproline supplementation in low fish meal diets on collagen synthesis, myofiber development and muscular texture of juvenile Pacific white shrimp (Litopenaeus vannamei).

This experiment aimed to evaluate the impact of dietary hydroxyproline (Hyp) supplementation on the muscle quality of juvenile Pacific white shrimp (Litopenaeus vannamei) fed a low fishmeal diet. Six formulated diets included one high fishmeal (HF; 25% fishmeal content) and five low fishmeal diets (10% fishmeal content) with 0%, 0.2%, 0.4%, 0.6% and 0.8% Hyp (LF0, LF2, LF4, LF6 and LF8, respectively). Each diet was assigned to four replicates, and 40 shrimp (0.32 ± 0.00 g) per replicate were fed four times a day for 8 weeks. Dietary Hyp supplementation had little effects on growth performance, but increased the contents of Hyp, prolyl 4-hydroxylases (P4Hs), and collagen. The meat yield, springiness, hardness, chewiness, and cohesiveness of muscle were the highest in the LF4 group among the low fishmeal groups (P < 0.05). Cooking loss and freezing loss of muscle were the lowest in the LF4 group (P < 0.05). Dietary supplementation with 0.4% Hyp increased the myofiber density and decreased the myofiber diameter of muscle (P < 0.05). Supplementation of Hyp in the diet up-regulated the mRNA expression of smyhc5, smyhc15, col1a1, col1a2, igf-1f, tgf-ß and tor and down-regulated the mRNA expression of smyhc 1, smyhc 2, smyhc 6a (P < 0.05). Supplementation of Hyp in the diet up-regulated the protein expression of P-4E-BP1, P-AKT, AKT and P-AKT/AKT (P < 0.05). These results suggested that the addition of 0.4% Hyp to low fishmeal diets improved the muscle quality of L. vannamei.

Effects of dietary astaxanthin supplementation on growth performance, antioxidant status, immune response, and intestinal health of rainbow trout (Oncorhynchus mykiss).

A feeding trial was conducted to assess the impacts of dietary astaxanthin from wall-broken Haematococcus pluvialis (WBHPA) on the growth performance, antioxidant status, immune response, and intestinal health of rainbow trout (Oncorhynchus mykiss). Six experimental diets were formulated with various concentrations of WBHPA, ranging from 0 to 8.4 g/kg (containing 0 to 125 mg/kg astaxanthin). Each diet was fed to triplicate groups of rainbow trout (mean initial weight of 561 g) twice daily for 9 consecutive weeks. The survival rate and feed intake of fish exhibited no significant differences among the dietary groups (P > 0.05). Similarly, dietary inclusion of 25 to 100 mg/kg astaxanthin did not significantly affect the weight gain and daily growth coefficient (P > 0.05), but excessive inclusion of astaxanthin (125 mg/kg) slightly depressed these parameters (P < 0.05). Dietary inclusion of 25 to 50 mg/kg astaxanthin increased the activities of intestinal digestion and absorption enzymes (lipase, creatine kinase, and alkaline phosphatase), while the inclusion of 25 to 75 mg/kg astaxanthin improved the immune response of fish. Furthermore, regardless of inclusion level (25 to 125 mg/kg), dietary astaxanthin supplementation strengthened the intestinal mucosal barrier function and improved antioxidant activity, thereby promoting intestinal development. Conclusively, 25 to 75 mg/kg astaxanthin from WBHPA was recommended to be included in diets for rainbow trout.

Effects of replacing wheat bran with palm kernel cake or fermented palm kernel cake on the growth performance, intestinal microbiota and intestinal health of tilapia (GIFT, Oreochromis niloticus).

A nine-week feeding trial was conducted to evaluate the effects of replacing wheat bran (WB) with palm kernel cake (PKC) or fermented palm kernel cake (FPKC) on the growth performance, intestinal microbiota and intestinal health of genetically improved farmed tilapia (GIFT, Oreochromis niloticus) (initial weight 7.00 ± 0.01 g). Eleven isonitrogenous and isolipidic experimental diets were formulated by replacing 0, 20, 40, 60, 80, and 100% of dietary WB with PKC or FPKC. Replacement of WB with PKC concentrations up to 80% had no significant effect on the growth rate of tilapia or feed utilisation (p > 0.05). FPKC improved the growth performance of tilapia, with optimum growth achieved at 40% replacement level (p < 0.05). Complete replacement with PKC significantly decreased the activity of lipase and trypsin, and reduced the height of muscularis and the height of villus (p < 0.05). However, FPKC significantly increased amylase activity and villus height (p < 0.05). The apparent digestibility of dry matter and energy decreased linearly with increasing levels of PKC substitution, while FPKC showed the opposite trend (p < 0.05). PKC replacement of WB by 20% significantly reduced serum diamine oxidase activity and endothelin levels and increased intestinal tight junctions (p < 0.05). However, FPKC significantly decreased diamine oxidase activity and increased intestinal tight junctions (p < 0.05). PKC completely replaced WB, up-regulating the expression of pro-inflammatory factors (il-1ß) (p < 0.05). When 40% of WB was replaced with FPKC, the expression of pro-inflammatory factors (il-1ß and il-6) was decreased significantly (p < 0.05). Completely replacement of WB with PKC reduced the abundance of Firmicutes and Chloroflexi, while FPKC reduced the abundance of Fusobacteriota and increased the levels of Actinobacteriota. WB can be replaced with PKC up to 80% in tilapia feeds. However, the high percentage of gluten induced intestinal inflammation, impaired gut health, and reduced dietary nutrient utilisation and growth performance. Complete replacement of WB with FPKC promoted intestinal immunity. It also improved dietary nutrient utilisation and growth performance. However, the optimal growth was achieved at a 40% replacement level.

High replacement of soybean meal by different types of rapeseed meal is detrimental to rainbow trout (Oncorhynchus mykiss) growth, antioxidant capacity, non-specific immunity and Aeromonas hydrophila tolerance.

A 12-week feeding trial was conducted to evaluate the effects of replacing soybean meal with different types of rapeseed meal (RSM; Chinese 95-type (oil press model) rapeseed meal [C95RM], Chinese 200-type rapeseed meal [C200RM], cold pressed rapeseed cake [CPRC], Indian rapeseed meal [IRM] and Canadian rapeseed meal [CRM]) on growth, antioxidant capacity, non-specific immunity and Aeromonas hydrophila infection tolerance in 990 fingering (average weight 12.77 ± 0.01 g) rainbow trout (Oncorhynchus mykiss). A basal diet was prepared using fishmeal and soybean meal as the main protein sources, the other 10 diets were formulated with five types of RSM at 20% (C95RM20, C200RM20, CPRC20, IRM20, CRM20) or 35% (C95RM35, C200RM35, CPRC35, IRM35, CRM35) inclusion levels to replace iso-nitrogenous soybean meal. Regardless of the RSM source, dietary inclusion of 20% RSM significantly reduced the weight gain rate (WGR) and digestive enzymes activities (except C200RM20) of fish, but increased the blood urea nitrogen (BUN) and hepatic malondialdehyde (MDA) content (except CRM20). Fish fed with CPRC20 and IRM20 exhibited relatively higher plasma cortisol and MDA content, but lower content/activities of triiodothyronine (T3), thyroxine (T4) and glutathione peroxidase (GPx) in plasma, lysozyme (LZM) and complement 3 (C3) in serum, catalase (CAT) in liver, and respiratory burst activity (RBA) of head kidney macrophages. The intestinal and hepatic tissues fed with 20% RSM were damaged to some extent, with the CPRC20 and IRM20 groups being the most severely affected. Regardless of the RSM source, dietary inclusion of 35% RSM significantly decreased WGR and digestive enzymes activities, but significantly increased plasma BUN and MDA content. The fish fed with CPRC35 and IRM35 exhibited relatively higher plasma cortisol, MDA, serum triglyceride, BUN content, but lower content/activities of T3, T4, C3, and LZM in serum, CAT, peroxidase and GPx in plasma, CAT in liver, RBA and phagocytic activity of head kidney macrophage. The hepatic and intestinal tissues damage was the worst in the IRM35 group among the 35% RSM inclusion groups. These results indicate that including ≥20% RSM in the diet, regardless of the source, reduced the growth, antioxidant capacity, immunity, and survival to Aeromonas hydrophila infection in rainbow trout.

Effects of dietary citrus pulp level on the growth and intestinal health of largemouth bass (Micropterus salmoides).

BACKGROUND: Citrus pulp (CP) is rich in pectin, and studies have shown that pectin possesses antioxidant, anti-inflammatory, and gut microbiota-regulating properties. However, the application of CP in aquafeed is limited. In this study, the effect of dietary inclusion of CP on the intestinal health of largemouth bass (Micropterus salmoides) was investigated. Juveniles of similar size (6.95 ± 0.07 g) were fed isonitrogenous and isoenergetic diets containing different levels of CP (0%, 3%, 6%, 9%, 12%, or 15%) for 58 days. RESULTS: As the level of CP in the feed for largemouth bass increased, the fish's growth performance and intestinal health initially improved and then declined. Adding low doses of CP (≤9%) to the feed had no significant impact on the growth performance of large-mouth black bass, whereas high doses of CP (>9%) significantly reduced their growth performance. Adding 6%, 9%, or 12% of CP to that feed enhanced the expression of genes related to tight junctions, anti-inflammatory activity, anti-apoptotic activity, and antioxidant activity in the intestines of largemouth bass. It reduced intestinal inflammation and improved intestinal nutrient absorption, intestinal mucosal barrier function, and intestinal antioxidant capacity. Moreover, it improved the α-diversity, structure, and function of the intestinal flora. The addition of 6% CP had the most beneficial effect on the intestinal health of largemouth bass. On the other hand, the addition of 15% CP had adverse effects on the intestinal antioxidant capacity and intestinal mucosal barrier function of largemouth bass. CONCLUSION: Adding 6-9% CP to the feed for largemouth bass can improve their intestinal health without having a significant impact on their growth performance. CP could serve as a novel prebiotic and immunostimulant ingredient in aquafeed. © 2023 Society of Chemical Industry.

Dietary Malondialdehyde Damage to the Growth Performance and Digestive Function of Hybrid Grouper (Epinephelus fuscoguttatus♀ × E. lanceolatu♂).

Malondialdehyde (MDA) is the dominant component of lipid peroxidation products. Improper storage and transportation can elevate the lipid deterioration MDA content of diets to values that are unsafe for aquatic animals and even hazardous to human health. The study aimed to investigate the effect of dietary MDA on growth performance and digestive function of hybrid grouper (Epinephelus fuscoguttatus♀ × E. lanceolatu♂). Six isoproteic and isolipidic diets were formulated to contain 0.03, 1.11, 2.21, 4.43, 8.86 and 17.72 mg/kg MDA, respectively. The study shows that the increased dietary MDA content linearly reduced the growth rate, feed utilization, body index and body lipid content of hybrid grouper, while the low dose of dietary MDA (≤2.21 mg/kg) created no difference. Similarly, dietary MDA inclusion linearly depressed the activities of intestinal digestive and absorptive enzymes as well as antioxidant enzymes, enhanced the serum diamine oxidase activity, endotoxin level and intestinal MDA content. A high dose of MDA (≥4.43 mg/kg) generally impaired the gastric and intestinal mucosa, up-regulated the relative expression of Kelch-like ECH-associated protein 1 but down-regulated the relative expression of nuclear factor erythroid 2-related factor 2 in hindgut. In conclusion, the effect of MDA on hybrid grouper showed a dose-dependent effect in this study. A low dose of dietary MDA had limited effects on growth performance and intestinal health of hybrid grouper, while a high concentration damaged the gastrointestinal structure, depressed the intestinal digestive and antioxidant functions, and thereby impaired the growth and health of hybrid grouper.

Excessive dietary soluble arabinoxylan impairs the intestinal physical and immunological barriers via activating MAPK/NF-κB signaling pathway in rainbow trout (Oncorhynchus mykiss).

Arabinoxylan (AX) has been deemed as an antinutritional factor, but limited information has addressed the effects of dietary AX on intestinal health of fish. The present study investigated the effects of dietary AX on intestinal mucosal physical and immunological barriers of rainbow trout (Oncorhynchus mykiss). Five isoproteic and isolipidic experimental diets (AXE, AX0, AX2.5, AX5 and AX10) were formulated to contain 0.03% arabinoxylanase as well as 0%, 2.5%, 5% and 10% AX, respectively. Each diet was randomly distributed to triplicate groups of 35 juvenile (average weight 3.14 ± 0.02 g) per tank in a rearing system maintained at 17 ± 1 °C for 9 weeks. Dietary AX supplementation regardless of inclusion levels significantly (P < 0.05) depressed the growth performance and feed utilization. The plasma endothelin-1 and d-lactic acid contents as well as diamino oxidase activity were significantly higher in fish fed diet AX10 compared to fish fed diet AX0. Dietary inclusion of 5-10% AX resulted in decreased intestinal villus height, goblet cell number and desmosome density, increased crypt depth, short and irregular microvilli, widened intercellular space; down-regulated the mRNA levels of occludin in hindgut, claudin3 and ZO-1 in foregut and midgut, but up-regulated the mRNA levels of claudin12 and claudin15 in midgut as well as claudin23 in foregut, midgut and hindgut. Furthermore, dietary 5-10% AX supplementation decreased the midgut and hindgut complement 3, complement 4 and sIgT contents as well as the midgut IgM and hindgut IL-10 contents. Conversely, the hindgut TNF-α and IL-6 contents increased with the rising dietary AX level. RT-qPCR demonstrated that the pro-inflammatory cytokines (IL-1ß, IL-6, IL-8, IL-12ß, IFN-γ, and TNF-α) and pIgR mRNA levels in midgut and hindgut were up-regulated by dietary AX inclusion of 5-10% AX. Meanwhile, the mRNA levels of p38 MAPK, IκBα, and NF-κB p65 in midgut and hindgut raised gradually with the increasing dietary AX content. The Western blot results showed that the protein expression levels of p38 MAPK and NF-κB generally increased with the rising dietary AX content. Dietary treatment with 0.03% arabinoxylanase did not affect the growth performance and intestinal health of rainbow trout (P > 0.05). In conclusion, excessive dietary AX inclusion (5-10%) increased the intestinal permeability and induced the intestinal inflammatory response via activating MAPK/NF-κB signaling pathway, and ultimately damaged the intestinal barrier function of rainbow trout.

Potential mechanisms of different methylation degrees of pectin driving intestinal microbiota and their metabolites to modulate intestinal health of Micropterus salmoides.

Four diets containing 8 % cellulose, low methyl-esterified pectin (LMP), high methyl-esterified pectin (HMP) and MMP (half LMP and half HMP) were designed to evaluate the potential mechanisms by which different esterification degrees of pectin drive intestinal microbiota and their metabolites modulating the intestinal health of Micropterus salmoides. The results showed that both dietary LMP and HMP consistently upregulated intestinal zonula occludens protein 1 (Zo-1), Caludin-1, and Caludin-4, and downregulated intestinal tumor necrosis factor-alpha (TNF-α), interleukin-8 (IL-8), and interleukin-1 beta (IL-1ß) gene expression (P < 0.05). Dietary HMP separately upregulated intestinal Occludin, nuclear factor erythroid2-related factor 2 (Nrf2), B-cell lymphoma-2 (Bcl-2), and Bcl-2 associated agonist of cell death (BAD) gene expression, as well as the digesta propionate content, OTUs, Sobs, Shannon, Chao, and ACE indices (P < 0.05), whereas dietary LMP decreased digesta arginine, 4-aminobutyric, L-tyrosine, and phenylalanine contents (P < 0.05). Moreover, dietary HMP decreased plasma lipopolysaccharide and d-lactic acid contents and increased intestinal superoxide dismutase and glutathione peroxidase activities and immunoglobulin (Ig) receptor and IgM levels (P < 0.05). Collectively, dietary HMP improves intestinal health by increasing intestinal flora α-diversity and enhancing intestinal mechanical barrier, anti-inflammatory, antioxidant, and immune functions. On the contrary, the interference of dietary LMP with butyrate, tyrosine, arginine, and 4-aminobutyric acid metabolism is the main reason for its detrimental effects on intestinal health.

The effects of sodium butyrate (NaB) combination with soy saponin dietary supplementation on the growth parameters, intestinal performance and immune-related genes expression of hybrid grouper (Epinephelus fuscoguttatus♀ × E. lanceolatus♂).

Soy saponins are generally known to have negative effects on growth and the intestines of aquatic animals, and appropriate levels of sodium butyrate (NaB) may provide some mitigating effects. We investigated the effects of low and high levels of soy saponin and the protective effects of NaB (based on high level of soy saponin) on growth, serum cytokines, distal intestinal histopathology, and inflammation in hybrid grouper (Epinephelus fuscoguttatus ♀ × E. lanceolatus ♂). The experiment included four groups: fishmeal group (FM, 0.00% saponin and 0.00% NaB), low saponin group (SL, 0.30% saponin and 0.00% NaB), high saponin group (SH, 1.50% saponin and 0.00% NaB) and high saponin with NaB group (SH-NaB, 1.50% saponin and 0.13% NaB). The results showed compared to FM, the final body weight (FBW) and weight gain (WG) were significantly higher and lower in SL and SH, respectively (P < 0.05). Compared to SH, the FBW and WG were significant higher in SH-NaB (P < 0.05). In the serum, compared to FM, the interferon γ (IFN-γ) and interleukin-1ß (IL-1ß) levels in SH were significantly increased (P < 0.05). Compared to SH, the IFN-γ level was significantly decreased in SH-NaB (P < 0.05). In the distal intestine, based on Alcian Blue-Periodic Acid-Schiff (AB-PAS) observation, the goblet cell/µm was significantly increased and decreased in the SL and SH, respectively, compared to FM. The intestinal diameter/plica height ratio in the SH was significantly higher than those in the FM, SL and SH-NaB (P < 0.05). The NO and ONOO- levels in the SH were significantly higher than that in FM and SL (P < 0.05). At the transcriptional level in the distal intestine, compared to FM, the mRNA levels of tumor necrosis factor (tnfα), il1ß, interleukin-8 (il8) and ifnγ were significantly up-regulated in the SH (P < 0.05). Compared to the SH, tnfα, il8 and ifnγ were significantly down-regulated in the SH-NaB (P < 0.05). Compared to the FM, the mRNA levels of claudin3, claudin15, zo2 and zo3 were significantly up-regulated in the SL (P < 0.05). The mRNA levels of occludin, claudin3, claudin12, claudin15, zo1, zo2 and zo3 were significantly down-regulated in the SH compared to the FM (P < 0.05). Additionally, compared to the SH, the mRNA levels of occludin, claudin3, claudin12, claudin15, zo1, zo2 and zo3 were significantly up-regulated in the SH-NaB (P < 0.05). After the 7-day Vibrio parahaemolyticus challenge test, the survival was significantly higher and lower in the SL and SH, respectively, compared to FM (P < 0.05). Overall, low and high levels of soy saponins had positive and negative effects on growth, disease resistance, serum cytokines, and distal intestinal development and anti-inflammation, respectively, in hybrid grouper. NaB effectively increased disease resistance and improved distal intestinal inflammation in hybrid grouper, but the effects of NaB were mainly observed in improving distal intestinal tight junctions.

Effects of Dietary Lysine Level on Growth Performance and Protein Metabolism in Juvenile Leopard Coral Grouper (Plectropomus leopardus).

An 8-week feeding trial was conducted to evaluate the effects of dietary lysine level on growth performance and protein metabolism of juvenile leopard coral grouper (Plectropomus leopardus) and thereby obtained the optimal dietary lysine requirement of P. leopardus. Six isoproteic and isolipidic experimental diets were formulated to contain 1.10%, 1.69%, 2.30%, 3.08%, 3.56%, and 4.36% lysine of diets, respectively. Each diet was assigned at random to triplicate groups of 25 juveniles (initial mean weight is 10.57 g) per tank in a flow-through mariculture system maintained at 27-30°C. Dietary inclusion of 2.30-3.08% lysine improved the weight gain rate (WGR) and specific growth rate and decreased the feed conversion ratio (FCR) of juveniles (P < 0.05). The intestinal digestive enzyme (trypsin, amylase, and lipase) activities were overall enhanced by dietary inclusion of 3.08-3.56% lysine (P < 0.05). The mammalian target of rapamycin (mTOR) signaling pathway was activated in fish fed diets with 1.69-2.30% lysine by upregulating the relative expression levels of hepatic TOR and S6K1 (p70 ribosomal protein S6 kinase 1) but downregulating the relative expression level of hepatic 4E-BP2 (eIF4E-binding protein 2). Conversely, the amino acid response signaling pathway was inhibited in fish fed diet with 2.30% lysine by downregulating the relative expression levels of hepatic GCN2 (general control nondepressible 2), ATF3 (activating transcription factor 3), ATF4a (activating transcription factor 4a), and ATF4b (activating transcription factor 4b). Additionally, dietary 1.69-3.08% lysine enhanced the plasma total protein level and hepatic lysine α-ketoglutarate reductase activity but depressed the blood urea nitrogen level and hepatic adenosine monophosphate deaminase activity (P < 0.05). Moreover, dietary 3.08% lysine increased the contents of whole-body crude protein and total amino acids, while 1.69%-4.36% lysine depressed the whole-body lipid content (P < 0.05). These results indicated that optimal dietary lysine increased the digestive enzyme activities, promoted protein synthesis but depressed protein degradation, and thereby improved the growth performance of P. leopardus. Based on the second-order polynomial model, the optimal lysine requirement of juvenile P. leopardus for WGR, FCR, and lysine deposition was 2.60%-2.97% of diets (4.91%-5.60% of dietary protein).

Evaluation of Cottonseed Meal as an Alternative to Fish Meal in Diet for Juvenile Asian Red-Tailed Catfish Hemibagrus wyckioides.

A 10-week trial was performed to investigate the effects of replacing fishmeal with cottonseed meal (CSM) on the growth rate, protein metabolism, and antioxidant response of Asian red-tailed catfish Hemibagrus wyckioides. Five isonitrogenous and isocaloric diets (C0, C8.5, C17.2, C25.7, and C34.4) were prepared to contain 0%, 8.5%, 17.2%, 25.7%, and 34.4% CSM replacing fishmeal, respectively. The weight gain, daily growth coefficient, pepsin, and intestinal amylase activities initially increased and then decreased with the raising dietary CSM levels; the highest values were observed in the C17.2 group (P < 0.05). However, feed cost exhibited the opposite trend. With the increasing dietary CSM levels, the protein efficiency ratio and intestinal trypsin activity decreased but feed conversion rate increased gradually; while no differences were observed among the C0, C8.5, and C17.2 groups (P > 0.05). Dietary CSM inclusion regardless of levels increased the plasma growth hormone level as well as hepatic aspartate aminotransferase (AST) and γ-glutamyl transpeptidase activities but decreased the plasma glutamate dehydrogenase and AST activities (P < 0.05). With the increasing dietary CSM levels, the plasma alkaline phosphatase (AKP) and hepatic superoxide dismutase activities decreased but malondialdehyde content increased gradually, while no differences were observed among the C0, C8.5, and C17.2 groups (P > 0.05). The plasma immunoglobulin M content and hepatic glutathione reductase activity initially increased but then decreased with the raising dietary CSM levels; the highest values were found in the C17.2 group. These results indicated that dietary CSM inclusion level up to 17.2% improved the growth rate, feed cost, digestive enzyme activity, and protein metabolism without compromising antioxidant capacity of H. wyckioide, whereas these parameters were depressed by further inclusion of CSM. CSM is a potentially cost-effective alternative plant protein source in diet of H. wyckioide.

Dietary Histamine Impairs the Digestive Physiology Function and Muscle Quality of Hybrid Grouper (Epinephelus fuscoguttatus♀ × Epinephelus lanceolatus♂).

An 8-week feeding experiment was conducted to investigate the effect of dietary histamine on growth performance, digestive physiology function and muscle quality in a hybrid grouper (Epinephelus fuscoguttatus♀ × Epinephelus lanceolatus♂). Seven isoproteic (50%) and isolipidic (11%) diets were prepared with various histamine inclusion levels of 0, 30, 60, 120, 240, 480 and 960 mg/kg in diets (actual contents were 72.33, 99.56, 138.60, 225.35, 404.12, 662.12 and 1245.38 mg/kg), respectively. Each diet was randomly assigned to triplicates of 30 juveniles (average body weight 14.78 g) per tank in a flow-through mariculture system. The increase in the dietary histamine level up to 1245.38 mg/kg made no significant difference on the growth rate and feed utilization of the grouper. However, the increased histamine content linearly decreased the activities of digestive enzymes, while no differences were observed in groups with low levels of histamine (≤404.12 mg/kg). Similarly, high levels of histamine (≥404.12 mg/kg) significantly damaged the gastric and intestinal mucosa, disrupted the intestinal tight junction structure, and raised the serum diamine oxidase activity and endotoxin level. Meanwhile, high doses of histamine (≥662.12 mg/kg) significantly reduced the activities of antioxidant enzymes, upregulated the relative expression of Kelch-like ECH-associated protein 1, and hardened and yellowed the dorsal muscle of grouper. These results showed that dietary histamine was detrimental to the digestive physiology function and muscle quality of the grouper, although it did compromise its growth performance.

Soybean protein concentrate causes enteritis in juvenile pearl gentian groupers (Epinephelus fuscoguttatus ♀ × Epinephelus lanceolatus ♂).

Due to diminishing fish meal (FM) supplies, superb protein (PRO) sources are needed for aquaculture, such as soy-based PRO. However, these can cause enteritis and even intestinal injury in fish when used at high proportions in feed. This research examines the effects of substituting soybean protein concentrate (SPC) for FM on the growth performance and intestinal balance of pearl gentian groupers and investigates the mechanism of SPC-induced enteritis. Experimental fish (n = 720) were fed 1 of 3 following diets: (1) a 50% FM diet (control), (2) a diet with 20% of the FM substituted with SPC (group SPC20), and (3) a 40% SPC-substituted diet (SPC40). Fish were fed for 10 wk iso-nitrogenous (50% PRO) and iso-lipidic (10% lipid) diets. Groups SPC20 and SPC40 showed significantly lower developmental performance and intestinal structures than control. Group SPC40 had significantly higher expressions of pro-inflammatory-related genes, such as interleukin 1ß (IL1ß), IL12, IL17 and tumor necrosis factor α and significantly lower expressions of anti-inflammatory-related genes, such as IL5, IL10 and transforming growth factor ß1. Biochemical and 16S high-throughput sequencing showed that the abundance and functions of intestinal flora in group SPC40 were significantly affected (P < 0.05), and there were significant correlations between operational taxonomic unit abundance variations and inflammatory gene expressions at genus level (P < 0.05). The second- and third-generation full-length transcriptome sequence was used to analyze the mechanism of SPC-induced enteritis in pearl gentian groupers, which showed that enteritis induced by SPC may be caused by disturbances to intestinal immune function induced by an imbalance in intestinal nutrition and metabolism, such as the intestinal immunity network for IgA production pathway. However, it remains unclear as to which intestinal immune or nutritional imbalance is most important in enteritis development. This study provides a basis for further research into soy PRO-related enteritis in fish.

Feeding Rainbow Trout with Different Types of Non-Starch Polysaccharides: Impacts on Serum Metabolome and Gut Microbiota.

A 70-day feeding trial investigated the effects of dietary inclusion of different types of non-starch polysaccharides (NSPs) on gut microbiota and serum metabolome of rainbow trout. Four practical feeds (42% crude protein, 17% crude lipid) were prepared with 8% insoluble NSP (INSP, cellulose), 16.8% soluble NSP (SNSP, composed of 1.12% ß-glucan, 1.28% mannan, 4.8% arabinoxylan, and 9.6% pectin), 24.8% NSPs (8% INSP + 16.8% SNSP), or no NSPs inclusion, respectively. Dietary NSPs inclusion had no significant influence on the Shannon, Simpson, ACE, and Chao1 indices of gut microbiota but induced a significant increase in the abundance of Pseudomonas aeruginosa and Photobacterium kishitanii, and a decrease in Firmicutes and Alistipes finegoldii. Besides, dietary SNSP upregulated the carnitine synthesis metabolic pathway. Our data suggest that dietary NSPs are detrimental to gut microbiota homeostasis and the health of rainbow trout, and dietary SNSP exhibit a stronger ability to interfere with physiological metabolism of rainbow trout than INSP. Therefore, the physiological effects of dietary NSPs, especially SNSP, should be carefully considered when designing the commercial feed formulations of rainbow trout.

High dietary non-starch polysaccharides detrimental to nutrient digestibility, digestive enzyme activity, growth performance, and intestinal morphology in largemouth bass, Micropterus salmoides.

An 8-weeks feeding trial was carried out to evaluate the effects of different levels of dietary non-starch polysaccharide on the growth, apparent nutrient digestibility, intestinal development, and morphology of largemouth bass (Micropterus salmoides). Seven isoproteic and isolipidic experimental diets were formulated (crude protein 47.00%, crude lipid 12.50%), containing 0, 3, 6, 9, 12, 15, and 18% non-starch polysaccharides (NSPs) (named Control, NSPs3, NSPs6, NSPs9, NSPs12, NSPs15, and NSPs18), respectively. Dietary inclusion of NSPs below 9% showed no negative impacts on fish growth and feed utilization efficiency, whereas dietary NSPs inclusion level above 9% decreased weight gain rate, specific growth rate, protein efficiency, protein deposition rate, apparent digestibility of dry matter and protein, and were accompanied by a reduction in intestinal protease, Na+/K+-ATPase and alkaline phosphatase activity and an increase in feed intake and feed coefficient. The activity of lipase was significantly decreased when dietary inclusion of 15 and 18% NSPs. Moreover, the lipid deposition rate and the apparent digestibility of lipids were significantly decreased since dietary inclusion of 9% NSPs. Dietary inclusion of NSPs above 12% significantly up-regulated intestinal GLP-2 gene's expression, and was accompanied by significant changes in hindgut morphology, including increases in villus length and width, muscularis thickness and number of goblet cell, as well as a decrease in crypt depth. Additionally, dietary inclusion of NSPs above 3% significantly increased intestinal length index, and the viserosomatic index was significantly increased when dietary NSPs exceeded 15%. The linear regression analysis based on weight gain rate and feed coefficient showed that the appropriate dietary NSPs level of juvenile largemouth bass should not above 5.51%. In conclusion, high dietary NSPs adversely affects digestive enzyme activity and intestinal morphology, which in turn reduced the apparent digestibility of dietary nutrients and growth of juvenile largemouth bass.

High Dietary Histamine Induces Digestive Tract Oxidative Damage in Juvenile Striped Catfish (Pangasianodon hypophthalmus).

A 56-day feeding trial investigated the effects of dietary histamine on the antioxidant capacity, gastric and intestinal barrier functions, and growth performance of striped catfish (Pangasianodon hypophthalmus). Seven isonitrogenous (34.0% crude protein) and isolipidic (10.5% crude lipid) diets were formulated with supplemental 0, 15, 30, 60, 120, 240, and 480 mg/kg of histamine, named H0, H15, H30, H60, H120, H240, and H480 group, respectively. Results showed that the weight gain rate, specific growth rate, relative intestinal length in the H240 and H480 groups, and the condition factors in the H480 group were significantly lower than those in the H0 group. Intestinal total antioxidant capacity, peroxidase, catalase, superoxide dismutase, glutathione peroxidase, and glutathione reductase activities in the H480 group were significantly lower than those in the H0 group, whereas intestinal malondialdehyde content exhibited the opposite trend. Intestinal complement 3, complement 4, immunoglobulin M, and Recombinant Mucin 2 in the H480 group were significantly lower than those in the H0 group, in contrast to intestinal lipopolysaccharide content. Intestinal IL-10 gene expression in the H480 group was significantly lower than that in the H0 group, whereas the TNF-α, IL-1, IL-6, and IL-8 gene expression exhibited opposite results. Scanning and transmission electron microscopic observation of the gastrointestinal tract revealed severe damage to the gastric mucosa and intestinal epithelium in the H480 group. The abundance of Treponema in the histamine groups was significantly higher than that in the H0 group. These results indicated that high dietary histamine decreases intestinal immunity and antioxidant capacity, inducing digestive tract oxidative damage and ultimately decreasing the growth of striped catfish.

Assessing effects of guar gum viscosity on the growth, intestinal flora, and intestinal health of Micropterus salmoides.

A 56-day feeding trial was conducted to assess the effects of different viscous guar gum on the growth, intestinal flora, and intestinal health of Micropterus salmoides. Four practical diets with 42.5 % crude protein and 13.7 % crude lipid were formulated to contain 8 % cellulose and three different viscosities (2500, 5200, and 6000 mPa·s) of guar gum. Dietary guar gum inhibits fish growth and feed utilization, decreases the α-diversity of the intestinal flora, and negatively alters the intestinal flora structure and metabolite composition. High viscous guar gum down-regulated the intestinal tight junction, anti-inflammatory, and anti-apoptotic related gene's expression, decreased digesta butyrate/histamine ratio; and increased the abundance of Plesiomonas shigelloides. These results suggest that dietary guar gum adversely affects intestinal health by disrupting intestinal flora structure and metabolite composition, and that viscosity should be considered when using guar gum as a binder in aquafeeds.

Different Types of Non-Starch Polysaccharides Alter the Growth, Intestinal Flora and Serum Metabolite Profile of Grass Carp, Ctenopharyngodon idella.

Dietary non-starch polysaccharides (NSPs) broadly influence fish intestinal flora and physiological metabolism, but limited information is available on grass carp (Ctenopharyngodon idella). This study investigated the effects of different types of NSPs on the growth, nutrient metabolism status, gut microbiota, and serum metabolome of grass carp. Fish were fed with diets containing 4.4% insoluble NSPs (INSP), 9.24% soluble NSPs (SNSP), 13.64% NSPs (4.4% INSP + 9.24% SNSP, NSP) and non NSPs (FM), respectively, for 9 weeks. Results showed that dietary SNSP decreased protein efficiency ratio and serum protein content, but increased feed coefficient ratio, feed intake, plasma blood urea nitrogen content, and plasma aspartate aminotransferase activity (AST); conversely, dietary INSP decreased plasma AST activity. Dietary INSP and SNSP increased serum free cholesterol content. Dietary NSPs altered the abundance of dominant bacteria and serum metabolite profiles. The differential metabolites between groups were significantly enriched in amino acid synthesis and metabolic pathways. In conclusion, dietary INSP exhibited a growth-promoting effect compared to SNSP. Dietary INSP is beneficial for improving nutrient metabolism and intestinal health. Moreover, dietary NSPs may regulate the physiological metabolism and feeding behavior of grass carp by altering amino acid synthesis and metabolism.

Vitamin D3 deficiency induced intestinal inflammatory response of turbot through nuclear factor-κB/inflammasome pathway, accompanied by the mutually exclusive apoptosis and autophagy.

Vitamin D3 (VD3) participated widely in the nuclear factor-κB (NF-κB)-mediated inflammation, apoptosis, and autophagy through the vitamin D receptor (VDR). However, the molecular mechanisms remain not understood in teleost. The present study investigated the functions of VD3/VDR on intestinal inflammation, autophagy, and apoptosis of turbot in vivo and in vitro. Triple replicates of 30 fish were fed with each of three diets with graded levels of 32.0 (D0), 1012.6 (D1), and 3978.2 (D2) IU/kg VD3. Obvious intestinal enteritis was observed in the D0 group and followed with dysfunction of intestinal mucosal barriers. The intestinal inflammatory response induced by VD3 deficiency was regulated by the NF-κB/inflammasome signalling. The promotion of intestinal apoptosis and suppression of intestinal autophagy were also observed in the D0 group. Similarly, VD3 deficiency in vitro induced more intense inflammation regulated by NF-κB/inflammasome signalling. The mutually exclusive apoptosis and autophagy were also observed in the group without 1,25(OH)2D3 in vitro, accompanied by similar changes in apoptosis and autophagy increased apoptosis. The gene expression of VDRs was significantly increased with the increasing VD3 supplementation both in vivo and in vitro. Moreover, VDR knockdown in turbot resulted in intestinal inflammation, and this process relied on the activation of inflammasome mediated by NF-κB signalling. Simultaneously, intestinal apoptosis was promoted, whereas intestinal autophagy was inhibited. In conclusion, VD3 deficiency could induce intestinal inflammation via activation of the NF-κB/inflammasome pathway, intestinal apoptosis, and autophagy formed a mutually exclusive relation in teleost. And VDR is the critical molecule in those processes.

Effects of Different Viscous Guar Gums on Growth, Apparent Nutrient Digestibility, Intestinal Development and Morphology in Juvenile Largemouth Bass, Micropterus salmoides.

An 8-week feeding trial was conducted to investigate the effects of different viscous guar gums on the growth performance, apparent nutrient digestibility, intestinal development and morphology of juvenile largemouth bass. Four isoproteic and isolipidic diets (crude protein 42.5%, crude lipid 13.7%) were formulated to contain 8% cellulose (Control group), 8% low viscous guar gum with 2,500 mPa s (Lvs-GG group), 8% medium viscous guar gum with 5,200 mPa s (Mvs-GG group) and 8% high viscous guar gum with 6,000 mPa s (Hvs-GG group), respectively. Each diet was fed to quadruplicate groups of 40 fish (6.00 ± 0.01 g) per repetition. Dietary guar gum inclusion significantly decreased the weight gain rate, specific growth rate, protein efficiency ratio, protein productive value and lipid deposition rate, and these parameters decreased considerably with increasing guar gum viscous and were lowest in the Hvs-GG group. Dietary guar gum inclusion significantly decreased the apparent digestibility of dry matter, crude protein and crude lipid, and these parameters decreased considerably with increasing guar gum viscous and were lowest in the Hvs-GG group. Intestinal protease, lipase and creatine kinase activities in the guar gum groups were significantly lower than those in the control group, and intestinal protease and lipase activities decreased considerably with increased guar gum viscous. Intestinal alkaline phosphatase activity in the Hvs-GG group and intestinal Na+/K+-ATPase activity in the Mvs-GG and Hvs-GG groups were significantly lower than those in the Lvs-GG and control groups. Serum high-density lipoprotein, total cholesterol and triglyceride concentrations and superoxide dismutase activity in the guar gum groups were significantly lower than those in the control group. Intestinal villus height and muscular thickness in the guar gum groups were considerably higher than those in the control group, whereas the goblet cell relative number in the Mvs-GG and Hvs-GG groups and the microvillus height in the Lvs-GG and Hvs-GG groups were significantly lower than those in the control group. The expression level of IGF-1 in the guar gum groups and the expression level of GLP-2 in the Mvs-GG and Hvs-GG groups were significantly higher than those in the control group. These results indicated that guar gum diets adversely affected intestinal morphology, decreased intestinal digestive and absorptive enzyme activities, and caused poor nutrient digestibility and growth performance in juvenile largemouth bass. Moreover, the adverse effects of guar gum are closely related to its viscous, and high viscous guar gum produces more extreme negative impacts on juvenile largemouth bass.