Intestinal Enzymes Research Articles

Supplementation of Yellow River carp diet with lutein and ferrous fumarate: Growth performance, digestive enzyme activity, skin pigmentation, and intestinal microbiota

In this study, the effect of the diet with lutein and ferrous fumarate on Yellow River carp (Cyprinus carpio) was studied, aiming to evaluate skin pigmentation, intestinal digestive enzymes, intestinal microbial diversity, and growth performance. Three experimental diets, including a control group, a lutein group (150mg/kg lutein), and a lutein and ferrous fumarate mixture group (150mg/kg lutein and 100mg/kg ferrous fumarate), were designed. The carp (N=135; 25.0±2.0g) were fed with experimental diets for 42 days. The results showed that the intestosomatic index (ISI) and viscerosomatic index (VSI) of the carp fed with lutein and ferrous fumarate were increased, accompanied by significant changes in body color, with the higher value of blue (b), color difference (ΔE) and chroma (Ch) compared with control group (P <0.05). Meanwhile, the higher activity of amylase, lipase, and trypsin were observed in the mixture group (P<0.05). High-throughput sequencing and Venn diagrams revealed that lutein or ferrous fumarate has obvious effects on the intestinal microbiota community of carp. The abundance of Actinobacteria and Flavobacterium was significantly increased in the carp fed with the mixture group compared with the control group. In conclusion, the addition of lutein and ferrous fumarate to the feed can change the skin pigmentation and intestinal microbial composition of Yellow River carp, thereby enhancing the coloring effect and digestive function of the fish. These findings provide valuable insights for optimizing feed formulation and aquaculture management, which can contribute to the improvement of the quality and farming efficiency of Yellow River carp.

Photooxidation Cross-Linked, Glutaraldehyde Cross-Linked, or Enzyme and Hydrostatic Pressure Processed Decellularized Biomaterials for Cardiovascular Repair Do Not Affect Host Response in a Rat Right Ventricular Outflow Flow Tract Reconstruction (RVOT) Model.

Cardiovascular diseases (CVDs) were responsible for approximately 19 million deaths in 2020, marking an increase of 18.7% since 2010. Biological decellularized patches are common therapeutic solutions for CVD such as cardiac and valve defects. The preparation of biomaterials for cardiac patches involves two main processing methods: glutaraldehyde or photooxidation cross-linking (fixation) and noncross-linked (nonfixation) processing. Despite the variety of products available in the market, cardiac patches still suffer from significant limitations, failing to adequately mimic the properties of biological tissue and restore its function. This study assesses the impact of different processing methodologies on the biological and biomechanical outcomes of three commercially available cardiac patches (CorPatch, CardioCel, PhotoFix) and one newly developed decellularized cardiac patch (Adeka) when implanted as right ventricular outflow tract (RVOT) repair material on a rat model. Four different patches for cardiovascular repair were selected based on their processing approaches and included: photooxidation crosslinked (PhotoFix), glutaraldehyde crosslinked (CardioCel), noncross-linked small intestine submucosa (CorPatch) or enzyme, and hydrostatic pressure (Adeka) processed decellularized biomaterials. Structure and function were characterized prior to implantation via thickness mapping, cross-section morphology, 2D surface topography, 3D volume microstructure, biaxial testing, uniaxial tensile testing, ball burst, and suture retention. Their host-biomaterials response was assessed invivo using a relevant model for cardiovascular repair: a rat (RVOT) reconstruction with 8 and 16-week timepoints. Topological analysis showed that the crosslinked cardiac patches had a more homogeneous thickness distribution when compared to the noncrosslinked patches. This agreed with histological evaluation, where cross-linking processed materials better preserved collagen content than noncrosslinked patches who were also more delaminated. Biaxial data demonstrated that all patches, except CorPatch, recapitulated the anisotropic behavior of healthy left ventricle tissue. The Adeka patch in-plane mechanics at 16 weeks was the one who better resembled the mechanics of healthy cardiac tissue. All patches showed appropriate biocompatibility and function at 8- and 16-week timepoints for RVOT patching. This included echocardiographic assessment, biomechanics, macrophage infiltration and polarization, and angiogenesis. Consistently with a more porous laminae structure, explants histology showed higher cell infiltration in non-crosslinked Adeka when compared to the crosslinked PhotoFix. Overall, both invitro and invivo tests indicate that the material processing does not impact the function, biomechanical performance, and the host response of the patches that can be considered as equally effective as materials based cardiac repair solutions.

Impact of fermented bamboo powder on the morphology and physiology of the gastrointestinal tract in yellow-feather broiler chickens.

Bamboo powder, a novel ingredient, is gaining recognition for its potential as a dietary supplement in poultry feed. This study aimed to investigate the effects of fermented bamboo powder (FBP) on antioxidant status, gut hormone activities, intestinal digestive enzyme activities, gut morphological structure, gastrointestinal development, and the expression of nutritional transporter genes in dwarf yellow-feather broiler chickens. A total of 600 healthy 1-day-old chicks were allocated randomly into two groups, with 10 replicates per group and 30 chicks in each replicate. The control group was provided with a standard basal diet, whereas the experimental group received the same basal diet supplemented with 1.0, 2.0, 4.0, and 6.0 g/kg of fermented bamboo powder (FBP) in four phases: Phase I (days 1-22), Phase II (days 23-45), Phase III (days 46-60), and Phase IV (days 61-77). Phases I and II were categorized as the pretreatment period (days 0-45), while Phases III and IV represented the experimental period (days 46-77). Tissue samples were collected during Phase IV for further analysis. After 77 days of feeding, results revealed that FBP supplementation significantly enhanced the levels of gastrointestinal hormones (Glucagon-like peptide 1, Peptide YY, Cholecystokinin, and 5-hydroxytryptamine) in the duodenum, jejunum, and ileum. Similarly, the activities of digestive enzymes (protease, chymotrypsin, trypsin, and amylase) were significantly increased in the small intestine. It also improved gut morphology by increasing villus height, crypt depth, and goblet cell counts in the duodenum, jejunum, and ileum. Additionally, antioxidant enzyme activities (superoxide dismutase, glutathione peroxidase, and catalase) were significantly increased, while malondialdehyde content was significantly decreased in the jejunum. Additionally, FBP supplementation significantly enhanced gizzard development. Overall, FBP supplementation modulated gut hormones and enzymes, enhanced gut morphology and promoted antioxidant status and gene expression related to nutrient transport and antioxidant defenses in broiler chickens. These findings suggest that FBP has the potential as a beneficial dietary supplement in poultry feed.

Anticancer Attributes and Multifaceted Pharmacological Implications of Laetrile and Amygdalin.

Laetrile, known as vitamin B17, is often used interchangeably with amygdalin. Laetrile is a semi-synthesis product of amygdalin, whereas amygdalin is a naturally occurring substance in many plants. Both compounds have a nitrile functional group that, when activated by the intestinal enzyme β-glucosidases, releases hydrogen cyanide. The two compounds have been considered for a long time as alternative therapy for cancer treatment however, findings available in the literature are discordant on the real efficacy of laetrile/amygdalin for the treatment of cancer, often highlighting a negative benefit-risk ratio. In this regard, the study aimed to comprehensively analyze the scientific data on laetrile/amygdalin, with a special emphasis on their pharmacokinetics, underlying pharmacological properties, mode of action as a potent antitumor agent, and effect on human health. The results showed that there is no clear evidence on the efficacy of cancer therapy following laetrile/amygdalin administration, especially at the clinical trial level. However, the in vitro studies of the biological activity of these compounds showed positive effects related to their antifibrotic, anti-inflammatory, antiasthmatic, and immunoregulatory processes. Laetrile's mechanism of action closely resembles amygdalin, affecting cancer signaling pathways. However, due to its cyanide toxicity, it was banned by the food and drug administration (FDA) due to safety concerns. Despite not receiving permission from the FDA, laetrile emerged as an alternative therapy in the 1970s. Nonetheless, continuing research is investigating safer methods of activating Laetrile for targeted cancer treatment. This opens interesting prospects in using these compounds in alternative medical therapies, for which, however, further research is needed.

Effects of dietary Lactococcus lactis Z-2 on growth, host health and resistance to Aeromonas hydrophila in juvenile common carp (Cyprinus carpio L.).

Lactococcus lactis Z-2 was previously isolated from common carp intestine. In order to investigate the effects of optimal dose of L. lactis Z-2 on growth, host health and disease resistance to Aeromonas hydrophila in common carp, five experimental diets, including without (CK and CK+ groups) or with 5 × 107 (group A), 5 × 108 (group B) and 5 × 109 CFU g-1 (group C) L. lactis Z-2, were evaluated for 8 weeks. Supplementation of L. lactis Z-2 increased final weights, weight gain, specific growth rate, condition factor and muscle crude protein content, while feed conversion ratio, muscle crude lipid and ash contents were lower than in the CK group (P < 0.05). Furthermore, L. lactis Z-2 also promoted intestinal digestive enzyme (lipase, amylase, protease) activities, improved intestinal morphology and upregulated gene (ZO-1, Occludin, Claudin) expression of intestinal barrier; stimulated secretion of proinflammatory (IL-6, IL-1β, TNF-α) and anti-inflammatory cytokines (TGF-β, IL-10) in serum; and boosted AKP, ACP, LZM and antioxidant enzyme (T-AOC, GSH-Px, T-SOD, CAT) activities of hepatopancreas (P < 0.05). Among them, the effect of group B was more prominent. After A. hydrophila infection, the CK+ group showed overexpression of immune indicators and decreased antioxidant capacity. Whereas dietary intervention with L. lactis Z-2 significantly reversed these changes. Moreover, the survival rate was significantly higher (100%) in group B than in the CK+ group (36.36%) after A. hydrophila infection (P < 0.05). Dietary L. lactis Z-2 improved the growth performance, immune response, antioxidant capacity, intestinal health and disease resistance of juvenile common carp. Ultimately, 5 × 108 CFU g-1 dietary L lactis Z-2 is recommended for the formulation of feed. © 2025 Society of Chemical Industry.

Study on the effects of Massa Medicata Fermentata with different formulations on the intestinal microbiota and enzyme activities in mice with spleen deficiency constipation.

This study aims to explore the therapeutic mechanism of Massa Medicata Fermentata (MMF) with different formulations on spleen deficiency constipation in mice by analyzing gastrointestinal hormones, D-xylose, intestinal microbiota, and intestinal enzyme activities. A spleen deficiency constipation model was established using an oral administration of Sennae Folium decoction combined with controlled diet and water intake. After successful model establishment, the mice with spleen deficiency constipation were treated with MMF S1, S2, S3. Following the intervention, serum samples from each group of mice were collected to measure VIP, 5-HT, and D-xylose. Additionally, small intestine contents were analyzed for intestinal enzyme activity and subjected to 16S rRNA high-throughput sequencing. Mice with spleen deficiency constipation showed significant decreases in body weight and fecal water content. In contrast, the body weight of the CS2 and CS3 groups returned to normal levels, and fecal water content in the CS2 and CS3 groups also returned to normal. The MMF S2 and S3 significantly increased protease and sucrase enzymes levels compared with CM group. Serum D-xylose levels were significantly reduced in the CM and CS2 group. VIP levels increased significantly in the CM group but decreased in the CS2 and CS3 groups. Additionally, 5-HT levels in the CM and CS1 groups decreased significantly, with the CS2 group returning to normal and the CS3 group showing significant increases. 16S rRNA sequencing analysis revealed that all three MMF formulations effectively restored the intestinal microbiota composition in mice. LEfSe analysis identified characteristic microbiota linked to different intervention groups. The CS3 group significantly upregulated the chloroalkane and chloroalkene degradation and vibrio cholerae pathogenic cycle pathways compared to the CM group. Candidatus_Arthromitus in the CS3 group and Psychrobacter in the CS2 group were positive and negative correlations with 5-HT and VIP, respectively. The three formulations of MMF significantly alleviated spleen deficiency constipation symptoms by modulating intestinal enzyme activities, D-xylose, VIP, and 5-HT levels, and restoring intestinal microbiota balance. Psychrobacter and Candidatus_Arthromitus were identified as potential biomarkers for the treatment of spleen deficiency constipation. Different formulations of MMF have different mechanisms of regulating constipation through intestinal microbiota.

Mechanistic insight into human milk extracellular vesicle-intestinal barrier interactions.

Human milk extracellular vesicles (EVs) are crucial mother-to-baby messengers that transfer biological signals. These EVs are reported to survive digestion and transport across the intestine. The mechanisms of interaction between human milk EVs and the intestinal mucosa, including epithelial uptake remain unclear. Here, we studied the interaction of human milk EVs with the gut barrier components, including intestinal biofluids, enzymes, mucus and epithelium. Additionally, we probed the endocytic mechanisms mediating the EV intestinal uptake. Finally, using proteomic analysis, we determined the existence and identification of proteins enriched in the EV fraction transported across the intestinal epithelium. We show that human milk EVs are largely stable in the biochemical gut barriers and demonstrate high mucus diffusivity. EVs show a high level of epithelial cell uptake (∼70%) and efficient transport across Caco-2 monolayers. Whilst cell uptake of EVs was mediated by multiple routes, none of the pathway-specific inhibitors inhibited their epithelial translocation. Proteomic analysis of EVs transported across Caco-2 monolayers identified 14 enriched EV proteins that may facilitate intestinal transport. These findings significantly expand our understanding of the interactions between human milk EVs and the gut barriers, including their intestinal uptake.

Effect of Spirulina Suplemantation to Canola Meal-Containing Goldfish (Carrasius auratus) Diets on Growth Performance, Nutrient Digestibility and Digestive Enzyme Activity

The aim of this study was to determine the effects of spirulina added to diets containing canola meal in juvenile goldfish (Carrasius auratus) on growth, feed utilization, nutrient digestibility and intestinal digestive enzyme activity. In the experiment planned as 4 groups and 3 replications, fish with an average weight of 2.3 ± 0.9 g were placed in 12 glass aquaria with a capacity of 60 liters (60x30x35) with 20 fish per aquarium. Diets were prepared to contain 30% canola meal. 10%, 20% and 30% spirulina was added to the diets except the control diet, respectively. Fish were fed twice a day until satiated for 7 weeks. At the end of the experiment, it was determined that diets containing spirulina significantly (p

Development and Evaluation of Non-Antibiotic Growth Promoters for Food Animals.

The widespread utilization of antibiotic growth promoters (AGPs) boosts the growth rate of food animals and enhances human living standards. Nevertheless, it is accompanied by escalating antibiotic resistance. Consequently, there is an urgent demand to develop novel alternatives to growth promoters. The objective of this study was to develop a non-antibiotic growth promoter (NAGP) for augmenting the growth rate of food animals. The growth-promoting effect of plant-derived NAGPs was assessed in mice and broiler chickens, and its growth-promoting mechanism was initially investigated. The results reveal that a combination of hawthorn (also known as shanzha) and astragalus (also known as huangqi) extracts (SQ) enhanced the growth rate of mice both in vivo and in vitro, attributed to their significant capacity to promote muscle growth and improve immunity (p < 0.05). The composite super energy extract M (CSEE-M), further optimized on the basis of SQ, significantly improved growth performance and feed conversion ratio, and elevated the activity of intestinal digestive enzymes (p < 0.05) in both mice and broilers and reshaped the gut microbiota of broilers. The addition of 0.5% CSEE-M to broiler drinking water significantly increased muscle content and improved carcass quality (p < 0.05). In conclusion, both SQ and CSEE-M hold great promise as NAGPs and serve as effective substitutes to AGPs. This research not only furnishes new solutions for the misuse of antibiotics but presents a fresh perspective for the development of growth promoters.

Effects of Bacillus subtilis KG109 on Growth Performance, Carcass Quality, Serum Indicators, Intestinal Morphology, and Digestive Enzymes in Broilers.

The purpose of this experiment is to investigate how different doses of Bacillus subtilis KG109 powder affect the growth performance, carcass quality, serum biochemical indexes, serum antioxidant and immunological index, intestinal morphology, and digestive enzyme activity of broilers. Four hundred chicks of a similar weight (1 day old) are randomly assigned to four groups of five replicates of 20 chicks each (half males and half females). The control group is fed a basal ration, and the experimental groups T1, T2, and T3 are supplemented with 6.0 × 108 CFU/kg, 1.2 × 109 CFU/kg, and 1.8 × 109 CFU/kg of Bacillus subtilis KG109 bacterial powder, respectively, in the basal ration. The feeding cycle is 52 d. Compared with the control group, Bacillus subtilis KG109 powder (1) increases the broiler feed conversion ratio (FCR) (p < 0.05), (2) improves the carcass quality (slaughter rate, cooking loss, L* and b* values) (p < 0.05), (3) enhances the serum biochemical indexes (alanine transaminase (ALT), alkaline phosphatase (ALP), aspartate transaminase (AST), albumin (ALB), and triglycerides (TG)) (p < 0.05), (4) improves the serum antioxidant capacity (total an-tioxidant capacity (T-AOC), total superoxide dismutase (T-SOD), and glutathione peroxidase (GSH-PX)) and immunoglobulins (lg A, lg G, lg M) (p < 0.05), (5) improves the intestinal morphology (villus height and villus height to crypt depth (VCR)) (p < 0.05), and (6) increases the intestinal digestive enzyme activities (amylase, protease, and lipase) (p < 0.05). In summary, adding Bacillus subtilis KG109 to broiler diets can result in a significant decrease in broilers' FCR, an increase in their slaughtering rate, a decrease in their serum ALT, ALP, and AST activities, an increase in their serum TG content, an improvement of their immune and antioxidant capacity, an improvement of their intestinal morphology, and an improvement of their intestinal digestive enzyme activity. It is recommended to add 1.8 × 109 CFU/kg of bacteria.

Effect of High-glucose Incubation on the Antioxidant System in Rat Gastrointestinal Tract Wall Tissue

Abstract Background: Hyperglycemic metabolic disorders such as diabetes can impair gastrointestinal (GI) physiological functions leading to multiple digestive manifestations. Oxidative stress which is an imbalance between reactive oxygen species production and antioxidants is contributed to GI complications that occur during hyperglycemia. Aim: To investigate the effect of high-glucose concentration on the antioxidant enzymes in the GI tract. Methods: Small intestine and colon tissues extracted from rats were incubated in a high-glucose medium for 3 hours. Following tissue homogenization, antioxidant enzyme activity and expression were evaluated. Results: Catalase (CAT) activity was increased in the small intestine (1742 ± 113.1–2265 ± 242.4 [mU/mL]) and decreased in the colon (3791 ± 516.2–1532 ± 292.9). Total antioxidant capacity was decreased in the small intestine (10.1 ± 1.83–9.048 ± 0.441 [nmole/μl]) and increased in the colon (8.114 ± 0.9–11.01 ± 0.99). Messenger RNA (mRNA) expression of antioxidant enzymes in the small intestine was increased (CAT: 0.03941 ± 0.0041–0.1917 ± 0.0165, glutathione peroxidase 1 [GPx1]: 1.156 ± 0.0855–16.24 ± 1.618, glutathione reductase [GR]: 0.0413 ± 0.0014-0.1549 ± 0.0145, superoxide dismutase 1 [SOD1]: 1.03 ± 0.1095–8.52 ± 0.471, SOD2: 0.00106 ± 3.559e-005–0.0028 ± 0.00052, SOD3: 0.0352 ± 0.0044–0.0493 ± 0.0223). mRNA expression of antioxidant enzymes in the colon was decreased (CAT: 0.02148 ± 0.0032–0.01057 ± 0.0014, GPx1: 0.48 ± 0.146–0.1090 ± 0.0209, GPx4: 0.2391 ± 0.063–0.01671 ± 0.0019, GR: 0.0393 ± 0.0031–0.0093 ± 0.0014, SOD1: 0.389 ± 0.1159–0.088 ± 0.0251, SOD2: 0.000934 ± 0.00020–0.000233 ± 2.39023e-05, SOD3: 0.0114 ± 0.00107–0.0017 ± 0.000176). Conclusion: Most of the results indicate a state of oxidative stress in the GI tract mediated by the exposure to high glucose level. Diabetic GI complications could be reversed using specific modalities that act to increase the antioxidant capacity.

Effects of Malondialdehyde on Growth Performance, Gastrointestinal Health, and Muscle Quality of Striped Catfish (Pangasianodon hypophthalmus).

Malondialdehyde (MDA) is a reactive carbonyl compound produced through lipid peroxidation during feed storage, which poses a significant threat to fish health. This study aimed to evaluate the effects of dietary MDA on the growth rate, gastrointestinal health, and muscle quality of striped catfish (Pangasianodon hypophthalmus). A basal diet (M0) containing 34% crude protein and 10.5% crude lipid was formulated. Each group was sprayed with malondialdehyde solution (0, 5, 10, 20, 40, and 80 mg/kg, on dietary crude lipid basis; 0, 0.53, 1.07, 2.13, 4.26, and 8.52 mg/kg, on dietary basis) before feeding, respectively. Each diet was randomly assigned to triplicates of 30 striped catfish (initial weight 31.38 g) per net cage. After 8 weeks, dietary inclusion of MDA regardless of level significantly depressed the growth rate and feed utilization. The extent of structural damage to the gastrointestinal tract increased progressively with increasing dietary MDA levels. The extent of damage to the intestinal biological barrier (intestinal microbial structure), chemical barrier (trypsin, lipase, amylase, and maltase activity), physical barrier (zonula occludent-2, occludin, claudin 7α, and claudin 12 relative expression), and immune barrier (contents of complement 4, complement 3, immunoglobulin M, and lysozyme activity) was dose-related to dietary MDA. Moreover, a linear decline in the activities of intestinal antioxidant enzymes (catalas, superoxide dismutase, et al.) and anti-inflammatory factor (transforming growth factor beta1, interleukin 10) relative expression was noted alongside an increase in dietary MDA content. In contrast, the relative expression levels of intestinal inflammatory factor (interleukin 8, transcription factor p65, tumor necrosis factor alpha) relative expression displayed an opposing trend. Additionally, dietary MDA exerted a linear influence on muscle color and texture characteristics. In conclusion, high doses of MDA (5-80 mg/kg) reduced the growth performance of striped catfish, attributed to linear damage to the gastrointestinal tract, a linear decrease in antioxidant function, and the occurrence of an inflammatory response. High doses of MDA (>40 mg/kg) were observed to significantly increase dorsal muscle b-value and induce muscle yellowing.

Effect of Dietary Astragalus Fermentation Products on Growth, Intestinal Microflora and Disease Resistance in Largemouth Bass Micropterus salmoides.

Fermentation of Astragalus by Lactobacillus plantarum and Bacillus coagulans can increase the release of active components and degrade its macromolecular substances. This study investigated the effect of fermentation products (Astragalus + L. plantarum + B. coagulans, ALB) on largemouth bass. We specifically focused on growth performance, serum biochemical indices, intestinal microbial diversity, intestinal enzyme activity, immune gene expression and resistance to infections by Aeromonas hydrophila and largemouth bass ranavirus (LMBRaV). The largemouth bass were divided into five groups based on the amount of ALB added to the feed as following, (1) ALB0 (no ALB, ALB0.5 [0.5% addition of ALB], ALB1 [1% addition of ALB], ALB3 [3% addition of ALB], ALB5 [5% addition of ALB]). The feeding trial spanned 28 days. Comprehensively comparing the feeding results of different ALB concentration, the ALB0.5 group showed the best effect. The ALB0.5 group had significantly increased weight gain rate, alkaline phosphatase, total protein, albumin, digestive enzymes activities of lipase, trypsin and increased intestinal villi and thickness of muscularis propria. And it decreased feed conversion ratio, aspartate aminotransferase and alanine aminotransferase of largemouth bass. Furthermore, the ALB0.5 group improved the richness and diversity of the intestinal microbiota. Increased abundance of dominant phylum and genus in the intestine of largemouth bass included Fusobacteria and Cetobacterium, which promoted the growth and immune performance of largemouth bass. After infection with A. hydrophila and LMBRaV, the survival rates were higher in ALB addition experimental groups than in the ALB0 group, respectively. And the survival rate of ALB0.5 group was higher than other groups. Meanwhile, the ALB added to the feed could regulated the immune gene expression (Mx, IRF-3, TNF-α, IL-1β and IL-10), which also promoted the largemouth bass resistance to disease. In summary, adding 0.5% ALB to the diet of largemouth bass can boost its growth performance, immune genes expression, intestinal health and disease resistance.

Comparative analysis of intestinal structure, enzyme activity, intestinal microbiota and gene expression in different segments of pufferfish (Takifugu Obscurus)

The structure of fish intestines does not have a clear regional division, while the function of the intestines may be related to their structure. Therefore, in this study, the delimitation of intestinal segments in pufferfish (Takifugu obscurus) was achieved by morphological analysis. Subsequently, enzyme activity, intestinal microbiota, and gene expression were examined to compare the differences among the pufferfish various segments. According to four morphological parameters: height of mucosa folds (HF), width of mucosa folds (WF), thickness of muscularis (TM), and cross-sectional area (CSA), the pufferfish's intestine was divided into anterior intestine (AI), middle intestine (MI), and posterior intestine (PI). The activity levels of amylase, lipase, and trypsin in the AI and MI were significantly higher than these in the PI. According to the analysis of 16S rDNA, the dominant microbiota at the phylum level in the different segments were Epsilonbacteraeota, Spirochaetes, and Proteobacteria. At the genus level, there were variations observed in the relative abundance of Brevinema, Mycobacterium, Bradyrhizobium, and Microvirga. α diversity analysis revealed that the richness indexes (Ace and Chao1) were the lowest in the MI, while β diversity analysis revealed significant difference in intestinal microbial community composition among the three intestinal segments. Furthermore, RNA-Seq was used to identify differential expression genes (DEGs) and biological pathways among the different intestinal segments. The DEGs between the AI and MI were enriched in pancreatic secretion and protein digestion and absorption, those between AI and PI were involved in ascorbate and aldarate metabolism and glutathione metabolism, and those between MI and PI were involved in steroid biosynthesis, fat digestion and absorption, vitamin digestion and absorption, and glycine, serine and threonine metabolism. In conclusion, the presented results compare and analyze the differences in various intestinal segments of pufferfish, which will be conductive to future exploration of the functions of these different segments.

The positive effects of dietary inositol on juvenile hybrid grouper (♀Epinephelus fuscoguttatus × ♂ Epinephelus lanceolatu) fed high-lipid diets: Growth performance, intestinal digestive enzymes, tissue morphology, and intestinal microbiota

The aim of study was to explore the beneficial effects of adding inositol to high-lipid diets on intestinal digestive enzymes, tissue morphology, and intestinal microbiota in juvenile hybrid grouper (♀Epinephelus fuscoguttatus × ♂Epinephelus lanceolatu). Six isonitrogenous (CP 51 %) and isolipidic (CL 16 %) diets were formulated, each containing different levels of inositol: 0 % (J1, as the control group), 0.04 % (J2), 0.08 % (J3), 0.16 % (J4), 0.32 % (J5) and 0.64 % (J6) inositol. Grouper (initial body weight of 6.76 ± 0.34 g) were fed experimental diets continuously for 8 weeks. Results showed that inositol significantly increased the specific growth rate of J3, J4, J5, J6 groups compared to the control group (P > 0.05). Amylase, lipase, and trypsin activities generally increased with higher inositol levels. Amylase activities were significantly higher in J4, J5, and J6 groups compared to the control group (P < 0.05), while lipase and trypsin activities were significantly higher in all inositol-treated groups (P < 0.05). Inositol did not significantly affect intestinal plica height (P > 0.05), but increased plica width, muscularis thickness, and goblet cell number (P < 0.05). Analysis of intestinal microbiota revealed a higher number of OTUs in the J3 group compared to J1 and J6 groups (P < 0.05), with no significant differences in Ace and Chao1 indices (P > 0.05). The Simpson index was significantly higher in the J3 group compared to J1 group (P < 0.05), with no significant differences between J6 and J1, J3 groups (P > 0.05). The dominant intestinal phylum mainly consists of Proteobacteria and Cyanobacteria, while the dominant family mainly consists of Vibrionaceae and Burkholderiaceae. In summary, supplementing high-lipid diets with appropriate inositol can enhance digestive ability, improve the morphology of hybrid grouper intestines, increase the abundance of beneficial bacterial microbiota, maintain intestinal health, and promote growth.

Effects of dietary resveratrol supplementation on the growth performance, physiological health, transcriptomic response, and intestinal microflora of Litopenaeus vannamei

We evaluated the effects of resveratrol (RES) supplementation on the growth performance, physiological health, transcriptomic response, and intestinal health of Litopenaeus vannamei. L. vannamei (initial body weight: 0.40 ± 0.02 g) were fed using six types of experimental diets (R0, R50, R100, R200, R400, and R800, supplemented with 0, 50, 100, 200, 400 and 800 mg/kg of RES, respectively) for 56 days. The results showed that dietary RES had no significant effect on the growth of L. vannamei. The survival rate (SR) of shrimp in the R400 group was significantly higher than that in the R0 group after exposure to Vibrio parahaemolyticus. RES supplementation dramatically enhanced the immunity of L. vannamei, as indicated by the increased serum levels of important indicators of immune function. The activities of intestinal digestive enzymes increased dramatically with increasing RES supplementation. Transcriptome analysis revealed that adding RES had a considerable effect on various metabolic and immune-related pathways in L. vannamei, including beta-alanine metabolism, ascorbate and aldarate metabolism, and drug metabolism-other enzymes. 16S rDNA analysis indicated that RES supplementation improved the intestinal microflora composition of L. vannamei by enhancing the abundance of beneficial bacteria and reducing the abundance of harmful bacteria. In conclusion, this study indicated that RES supplementation at an appropriate dose (400 mg/kg) could improve the physiological health, regulate the response at the transcriptional level, and ameliorate the intestinal microflora structure of L. vannamei. These results offer novel ideas and theoretical support for using RES in L. vannamei cultures as an immune-boosting supplement.

Effects of different feeding rates on growth performance, intestinal digestive enzyme activity, and intestinal microbial composition of red swamp crayfish (Procambarus clarkii)

The study aimed to investigate the effects of different feeding rates on the growth, muscle composition, intestinal digestive enzyme activities, and intestinal microbiota composition of juvenile crayfish (Procambarus clarkii). A total of 240 crayfish with an initial body weight of 8.33±0.50 g were randomly distributed into five groups, and each group had three replicates. The five groups corresponded to five different feeding rates of 2%, 4%, 6%, 8%, and 10% body weight per day (BW/day). The feeding trial lasted for six weeks. The results showed that the final body weight (FBW), weight gain rate (WGR), and specific growth rate (SGR) first increased when the feed rate increased from 2% to 6 % (P&lt;0.05) and then showed a decreased trend with the further increase of the feeding rate, but no significant differences were observed. The feed conversion ratio (FCR) increased with a rise in feeding rate, which reached a peak at the feeding rate of 10% (P&lt;0.05). The survival rate (SR), hepatosomatic index (HSI), and condition factor (CF) were unaffected by the feeding rates (P&gt;0.05). The moisture, crude protein, crude fat, and ash contents of crayfish muscle remained unchanged among all feeding rate groups. Intestinal activities of lipase and amylase increase with the feeding rate (up to 6%) increasing (P&lt;0.05). The ACE and Chao indices in the groups fed at 2% of feeding rate were significantly lower than those at a feeding rate of 10% (P&lt;0.05). The relative abundance of Firmicutes in the groups fed at feeding rates of 4%, 6%, and 8% was higher than that in other groups (P&lt;0.05), Citrobacter in the group fed at feeding rate of 6% was lower than that in different groups (P&lt;0.05). The intestinal microbiota primarily functions in connection to amino acid, carbohydrate, and vitamin metabolism. Therefore, the optimum feeding rate could be 6.0% of the feeding rate of juvenile P. clarkii.

Mineral composition and antioxidant activity of partially digested coffee “Kopi Luwak”

Background: Kopi Luwak is a rare type of coffee, that is partially digested in the Asian palm civet’s digestive tract before roasting. It therefore possesses a distinctive flavor and unique properties, shaped by the digestive enzymes in the civet's intestine. Objective: In this study, the differences of mineral composition, antioxidant potency and total polyphenol amount between commercially available Kopi Luwak and regular coffees (without digestive fermentation) were evaluated. Methods: Twenty-one regular type of coffee bean and 17 of Kopi Luwak bean were purchased from local markets or Kopi Luwak production company in Indonesia. The samples were then analyzed for the levels of 23 minerals (Al, As, Ba, Bi, Ca, Cd, Co, Cr, Cs, Cu, Fe, Li, Mg, Mn, Mo, Ni, Pb, Rb, Se, Sn, Sr, V and Zn) using ICP-MS system. The total polyphenol amount and antioxidant activity were evaluated using Folin-Ciocalteu assay and DPPH (1,1-Diphenyl-2-picrylhydrazyl) radical scavenging assay, respectively. Results: Of these 23 minerals, the concentrations of Mn and Ba were significantly lower in Kopi Luwak than in regular coffee. Otherwise, the concentrations of Cu and Rb in Kopi Luwak were significantly higher than those in regular coffee. The IC50 value on DPPH radical scavenging assay of Kopi Luwak (61.4±2.8 mg/dL) was significantly lower than that of regular coffee (93.6±1.9 mg/dL). Total polyphenol amount for Kopi Luwak was significantly higher than regular coffee. The correlation factor between these factors was r = -0.5910 and p &lt; 0.001. Conclusion: Kopi Luwak had lower concentrations of Mn and Ba, but higher levels of Cu and Rb compared to regular coffee. Additionally, Kopi Luwak exerted strong antioxidant activity and contained higher polyphenol amounts than regular coffee. Thus, our findings imply that Kopi Luwak is an important candidate for functional foods. Keywords: antioxidant potency, ICP-MS, Kopi Luwak, mineral composition, polyphenols

Intestinal DHA-PA-PG axis promotes digestive organ expansion by mediating usage of maternally deposited yolk lipids

Although the metabolism of yolk lipids such as docosahexaenoic acid (DHA) is pivotal for embryonic development, the underlying mechanism remains elusive. Here we find that the zebrafish hydroxysteroid (17-β) dehydrogenase 12a (hsd17b12a), which encodes an intestinal epithelial-specific enzyme, is essential for the biosynthesis of long-chain polyunsaturated fatty acids in primitive intestine of larval fish. The deficiency of hsd17b12a leads to severe developmental defects in the primitive intestine and exocrine pancreas. Mechanistically, hsd17b12a deficiency interrupts DHA synthesis from essential fatty acids derived from yolk-deposited triglycerides, and consequently disrupts the intestinal DHA-phosphatidic acid (PA)-phosphatidylglycerol (PG) axis. This ultimately results in developmental defects of digestive organs, primarily driven by ferroptosis. Our findings indicate that the DHA-PA-PG axis in the primitive intestine facilitates the uptake of yolk lipids and promotes the expansion of digestive organs, thereby uncovering a mechanism through which DHA regulates embryonic development.

Artemisinin supplementation improves growth, lipid metabolism, and intestinal microbiota of golden pompano (Trachinotus ovatus) fed high cottonseed meal diets

This study aimed to investigate the effect of artemisinin supplementation to high cottonseed meal (CSM) diet on the growth, lipid metabolism and intestinal microbiota of golden pompano (Trachinotus ovatus). Five diets were formulated, including the control (CM, with CSM replacing 16 % of fishmeal) and experimental groups (CA0, CA2, CA4, and CA8, with CSM replacing 36 % of fishmeal and supplemented with 0 %, 0.20 %, 0.40 %, and 0.80 % of artemisinin, respectively). 375 fishes (initial weight 22.45 ± 0.07 g) were randomized into five groups of triplicates each and fed in tanks for 8 weeks. Results indicated that 0.8 % artemisinin improved final weight, weight gain rate and specific growth rate, reduced the serum lipid levels under the condition of 36 % fishmeal replaced with CSM. In the liver, artemisinin supplementation alleviated hepatocyte vacuolization and lipid deposition induced by the high level CSM replacement fishmeal diet, reduced the activity of lipogenesis-related enzymes and increased lipolysis-related enzyme activity. In the intestine, supplementation with 0.8 % artemisinin significantly increased the activities of intestinal digestive enzymes. Supplementation with 0.8 % artemisinin significantly increased the diversity and stability of the intestinal microbiota of the golden pompano, while also reducing the abundance of potentially harmful Vibrio associated with high CSM levels. In conclusion, 0.8 % artemisinin supplementation mitigated the negative effects of high CSM diets on growth, lipid metabolism, and liver health in golden pompano by enhancing lipid metabolism, intestinal digestive enzyme activity, and modulating intestinal microbiota.