ABSTRACT
BACKGROUND: In addition to its role in the digestive system, the peritrophic membrane (PM) provides a physical barrier protecting the intestine from abrasion and against pathogens. Because of its sensitivity to RNA interference (RNAi), the notorious pest insect, the Colorado potato beetle (CPB, Leptinotarsa decemlineata), has become a model insect for functional studies. Previously, RNAi-mediated silencing of Mannosidase-Ia (ManIa), a key enzyme in the transition from high-mannose glycan moieties to paucimannose N-glycans, was shown to disrupt the transition from larva to pupa and the metamorphosis into adult beetles. While these effects at the organismal level were interesting in a pest control context, the effects at the organ or tissue level and also immune effects have not been investigated yet. To fill this knowledge gap, we performed an analysis of the midgut and PM in ManIa-silenced insects. RESULTS: As marked phenotype, the ManIaRNAi insects, the PM pore size was found to be decreased when compared to the control GFPRNAi insects. These smaller pores are related to the observation of thinner microvilli (Mv) on the epithelial cells of the midgut of ManIaRNAi insects. A midgut and PM proteome study and reverse transcription quantitative polymerase chain reaction (RT-qPCR) analysis with a selection of marker genes was performed to characterize the midgut cells and understand their response to the silencing of ManIa. In agreement with the loss of ManIa activity, an accumulation of high-mannose N-glycans was observed in the ManIa-silenced insects. As a pathogen-associated molecular pattern (PAMP), the presence of these glycan structures could trigger the activation of the immune pathways. CONCLUSION: The observed decrease in PM pore size could be a response to prevent potential pathogens to access the midgut epithelium. This hypothesis is supported by the strong increase in transcription levels of the anti-fungal peptide drosomycin-like in ManIaRNAi insects, although further research is required to elucidate this possibility. The potential immune response in the midgut and the smaller pore size in the PM shed a light on the function of the PM as a physical barrier and provide evidence for the relation between the Mv and PM. © 2022 Society of Chemical Industry.
Subject(s)
Coleoptera , Solanum tuberosum , Animals , RNA Interference , Solanum tuberosum/metabolism , Mannosidases/genetics , Mannosidases/metabolism , Mannosidases/pharmacology , Mannose/metabolism , Mania , Digestive System/metabolism , Larva/genetics , Insecta/metabolism , Polysaccharides/metabolism , Polysaccharides/pharmacologyABSTRACT
Carotenoids are essential micronutrients for animals, and they can only be obtained from the diet for mollusk as well as other animals. In the body, carotenoids undergo processes including absorption, transport, deposition, and metabolic conversion; however, knowledge of the involved genes is still limited. To elucidate the molecular mechanisms of carotenoid processing and identify the related genes in Pacific oyster (Crassostrea gigas), we performed a comparative transcriptome analysis using digestive gland tissues of oysters on a beta-carotene supplemented diet or a normal diet. A total of 718 differentially expressed genes were obtained, including 505 upregulated and 213 downregulated genes in the beta-carotene supplemented group. Function Annotation and enrichment analyses revealed enrichment in genes possibly involved in carotenoid transport and storage (e.g., LOC105342035), carotenoid cleavage (e.g., LOC105341121), retinoid homeostasis (e.g., LOC105339597) and PPAR signaling pathway (e.g., LOC105323212). Notably, down-regulation of mRNA expressions of two apolipoprotein genes (LOC105342035 and LOC105342186) by RNA interference significantly decreased the carotenoid level in the digestive gland, supporting their role in carotenoid transport and storage. Based on these differentially expressed genes, we propose that there may be a negative feedback mechanism regulated by nuclear receptor transcription factors controlling carotenoid oxygenases. Our findings provide useful hints for elucidating the molecular basis of carotenoid metabolism and functions of carotenoid-related genes in the oyster.
Subject(s)
Crassostrea/genetics , Crassostrea/metabolism , Dietary Supplements , Gene Expression Profiling , beta Carotene/metabolism , Amino Acid Sequence , Animals , Apolipoproteins/chemistry , Apolipoproteins/genetics , Apolipoproteins/metabolism , Base Sequence , Digestive System/metabolism , Gene Expression Regulation , Molecular Sequence Annotation , Phylogeny , RNA Interference , RNA, Messenger/genetics , RNA, Messenger/metabolism , RNA-Seq , Reproducibility of Results , Vitamin A/metabolismABSTRACT
RATIONALE: Lipid correction models use elemental carbon-to-nitrogen ratios to estimate the effect of lipids on δ13 C values and provide a fast and inexpensive alternative to chemically removing lipids. However, the performance of these models varies, especially in whole-body invertebrate samples. The generation of tissue-specific lipid correction models for American lobsters, both an ecologically and an economically important species in eastern North America, will aid ecological research of this species and our understanding of the function of these models in invertebrates. METHOD: We determined the δ13 C and δ15 N values before and after lipid extraction in muscle and digestive glands of juvenile and adult lobster. We assessed the performance of four commonly used models (nonlinear, linear, natural logarithm (LN) and generalized linear model (GLM)) at estimating lipid-free δ13 C values based on the non-lipid-extracted δ13 C values and elemental C:N ratios. The accuracy of model predictions was tested using paired t-tests, and the performance of the different models was compared using the Akaike information criterion score. RESULTS: Lipid correction models accurately estimated post-lipid-extraction δ13 C values in both tissues. The nonlinear model was the least accurate for both tissues. In muscle, the three other models performed well, and in digestive glands, the LN model provided the most accurate estimates throughout the range of C:N values. In both tissues, the GLM estimates were not independent of the post-lipid-extraction δ13 C values, thus reducing their transferability to other datasets. CONCLUSIONS: Whereas previous work found that whole-body models poorly estimated the effect of lipids in invertebrates, we show that tissue-specific lipid correction models can generate accurate and precise estimates of lipid-free δ13 C values in lobster. We suggest that the tissue-specific logarithmic models presented here are the preferred models for accounting for the effect of lipid on lobster isotope ratios.
Subject(s)
Carbon Isotopes/chemistry , Lipids/chemistry , Nephropidae/chemistry , Animals , Carbon Isotopes/metabolism , Digestive System/chemistry , Digestive System/metabolism , Lipid Metabolism , Mass Spectrometry , Muscles/chemistry , Muscles/metabolism , Nephropidae/metabolism , Nitrogen Isotopes/chemistry , Nitrogen Isotopes/metabolism , Shellfish/analysisABSTRACT
Early-life gut microbial colonisation is known to influence host physiology and development, shaping its phenotype. The developing gastro-intestinal tract of neonatal piglets provides a "window of opportunity" for programming their intestinal microbiota composition and corresponding intestinal development. Here, we investigated the impact of early feeding on jejunum and colon microbiota composition, and intestinal maturation in suckling piglets. From two days of age, early-fed (EF; n = 6 litters) piglets had access to solid feed containing a mixture of fibres till weaning (day29) in addition to sow's milk, whereas the control (CON; n = 6 litters) piglets exclusively fed on sow's milk. Early feeding elicited a significant impact on the colon microbiota, whereas no such effect was seen in the jejunal and ileal microbiota. Quantified eating behavioural scores could significantly explain the variation in microbiota composition of EF piglets and support their classification into good, moderate, and bad eaters. Members of the Lachnospiraceae family, and the genera Eubacterium, Prevotella, and Ruminococcus were quantitatively associated with eating scores. EF piglets were found to have a decreased pH in caecum and colon, which coincided with increased short-chain fatty acid (SCFA) concentrations. Moreover, they also had increased weights and lengths of several intestinal tract segments, as well as a decreased villus-crypt ratio in jejunal mucosa and an increased abundance of proliferative cells in colon mucosa. The approaches in this study indicate that early feeding of a mixed-fibre (pre-weaning) diet changes the microbiota composition, pH, and fermentation products in the distal gut of piglets, while it also alters both macroscopic and microscopic intestinal measurements. These results exemplify the potential of early feeding to modulate intestinal development in young piglets.
Subject(s)
Digestive System/physiopathology , Gastrointestinal Microbiome/physiology , Intestinal Mucosa/microbiology , Animal Feed , Animals , Animals, Newborn , Cell Proliferation/physiology , Diet , Dietary Fiber/metabolism , Dietary Supplements , Digestive System/metabolism , Digestive System/microbiology , Fatty Acids, Volatile/metabolism , Fermentation/physiology , Hydrogen-Ion Concentration , Intestinal Mucosa/metabolism , Milk/metabolism , SwineABSTRACT
PURPOSE: Accumulation of Gamma aluminium oxide nanoparticles γ-Al2O3 NPs significant impact on aquatic ecosystems. However, the toxicity of γ-Al2O3 NPs in aquatic organisms has been limited investigated. This study investigated histopathological changes and antioxidant responses induced by different concentrations of γ-Al2O3 NPs in Mytilus galloprovincialis. MATERIAL AND METHODS: In this study, mussels were exposed to different concentrations of 5 nm γ-Al2O3 NPs (0, 5, 20 and 40 mg/L) for 96 h under controlled laboratory conditions. Gill and digestive gland from mussels were assessed to histopathological (light microscopy, histopathological condition indices, digestive gland tubule types), SOD, CAT, GPx activities. RESULTS: Histopathological indices calculated higher, and significantly different in all exposure groups compared to the control group in gill and digestive gland (p < 0.05). Atrophic phase tubules proportion very high in 20 and 40 mg/L γ-Al2O3 NPs exposure groups. No significant changes in CAT activities in the gill and digestive gland (p > 0.05). Superoxide dismutase (SOD) activities significantly different (p ≤ 0.05) in the digestive gland from 20 mg/L γ-Al2O3 NPs exposures, and GPx activities significantly different (p < 0.05) in gill from 40 mg/L γ-Al2O3 NPs exposures. CONCLUSION: These results indicate that contamination of γ-Al2O3 NPs negatively affects the aquatic organism.
Subject(s)
Aluminum Oxide/toxicity , Antioxidants/metabolism , Digestive System/drug effects , Gills/drug effects , Metal Nanoparticles/toxicity , Mytilus/drug effects , Oxidative Stress/drug effects , Water Pollutants, Chemical/toxicity , Animals , Biomarkers/metabolism , Digestive System/metabolism , Digestive System/pathology , Ecosystem , Gills/metabolism , Gills/pathology , Mytilus/metabolismABSTRACT
Rat models of human type 1 diabetes have been shown to be of great importance for the elucidation of the mechanisms underlying the development of autoimmune diabetes. The three major well-established spontaneous rat models are the BioBreeding (BB) diabetes-prone rat, the Komeda diabetes-prone (KDP) rat, and the IDDM (LEW.1AR1-iddm) rat. Their distinctive features are described with special reference to their pathology, immunology, and genetics and compared with the situation in patients with type 1 diabetes mellitus. For all three established rat models, a distinctive genetic mutation has been identified that is responsible for the manifestation of the diabetic syndrome in these rat strains.
Subject(s)
Diabetes Mellitus, Experimental/genetics , Diabetes Mellitus, Experimental/immunology , Diabetes Mellitus, Type 1/genetics , Diabetes Mellitus, Type 1/immunology , Age of Onset , Animals , Cytokines/immunology , Diabetes Mellitus, Experimental/drug therapy , Diabetes Mellitus, Experimental/metabolism , Diabetes Mellitus, Type 1/drug therapy , Diabetes Mellitus, Type 1/metabolism , Digestive System/immunology , Digestive System/metabolism , Drug Evaluation, Preclinical , Female , Humans , Islets of Langerhans/immunology , Islets of Langerhans/metabolism , Male , Rats , Rats, Inbred Strains , Selective Breeding/genetics , T-Lymphocytes/immunology , T-Lymphocytes/metabolismABSTRACT
Rhubarb is commonly used to treat constipation in China for its function of promoting intestinal movement and optimum water content in feces. However, its mechanism of mucus secretion is vague. The aim of the study is to investigate the role of mast cells and enteric neurons in rhubarb extract (RE)-induced mucus secretion in the rat colon. Immunofluorescence was used to detect histamine receptors. Western blotting and 3,3'-diaminobenzidine (DAB) were applied to explore the content changes of mast cells activation. The changes in colonic goblet cells (GCs) were determined by means of PAS/AB staining. An intestinal perfusion system with a Bradford protein assay kit was directly to estimate in vitro secretion. And the cytokines were investigated with ELISA. The longitudinal aspect of this study indicate that the number and water content of faecal pellets were enhanced after the administration of different doses of RE accompanied by mast cells accumulated and increased the content of interferon (IFN) -γ or decreased the levels of interleukin (IL) -10 at doses of 3 and 6 g/kg. Pretreatment with ketotifen, mast cell stabilizer, had partially inhibited on RE-induced mucus secretion. Furthermore, RE induced the release of acetylcholine and mucin-2 in the colonic tissue and the histamine levels from the faeces. The results suggest that RE induced colonic mucus secretion involves mast cell activation and some cytokine.
Subject(s)
Colon/metabolism , Digestive System/drug effects , Drugs, Chinese Herbal/pharmacology , Mast Cells/metabolism , Rheum/chemistry , Animals , Cell Aggregation/drug effects , Cell Degranulation/drug effects , Colon/drug effects , Colon/pathology , Cytokines/genetics , Digestive System/metabolism , Digestive System/pathology , Disease Models, Animal , Drugs, Chinese Herbal/chemistry , Exocytosis/drug effects , Humans , Mast Cells/drug effects , Mast Cells/pathology , Mucus/drug effects , Mucus/metabolism , RatsABSTRACT
Essential oils are a promising alternative to insecticides. We investigated the LD50 of oils extracted from Piper corcovadensis, P. marginatum, and P. arboreum after 48 h topical contact with Spodoptera frugiperda larvae using morphometry, histochemistry and immunohistochemistry of the midgut and fat body. Chromatography revealed that E-caryophyllene was the principal compound common to the Piper species. The essential oils of P. corcovadensis, P. marginatum and P. arboreum caused deleterious changes in the midgut of S. frugiperda larvae. P. corcovadensis oil produced the lowest LD50 and significant histopathological alterations including elongation of the columnar cells, formation of cytoplasmic protrusions, reduction in carbohydrate, increased apoptotic index and decreased cell proliferation. P. arboreum oil caused histopathological alterations similar to P. corcovadensis, but caused the highest rate of cell proliferation and increased regenerative cells, which indicated rapid regeneration of the epithelium. Our findings demonstrated the insecticidal potential of P. corcovadensis for control of S. frugiperda owing to the significant damage it inflicted on S. frugiperda midgut.
Subject(s)
Fat Body/drug effects , Fat Body/pathology , Oils, Volatile/pharmacology , Piper/metabolism , Animals , Digestive System/metabolism , Digestive System/pathology , Fat Body/metabolism , Insecticides/metabolism , Insecticides/pharmacology , Larva/drug effects , Oils, Volatile/chemistry , Piper/chemistry , Plant Oils/metabolism , Plant Oils/pharmacology , SpodopteraABSTRACT
Enteroendocrine cells in the insect midgut are thought to secrete peptide hormones in response to the nutritional state. However, the role of dietary compounds in inducing peptide hormone secretion from enteroendocrine cells in insects remains unknown. In the present study, we demonstrated that several dietary compounds from mulberry leaves, including glucose, amino acids, and the secondary metabolite chlorogenic acid, induced significant secretion of tachykinin-related peptides from isolated silkworm midguts at the luminal concentrations measured in fed larvae. This study provides evidence that the insect midgut senses a non-nutritious secondary metabolite in addition to nutrient metabolites to monitor luminal food status and secretes a feeding regulatory hormone, suggesting that a unique dietary sensory system modulates insect feeding via enteroendocrine control.
Subject(s)
Amino Acids/pharmacology , Bombyx/metabolism , Chlorogenic Acid/pharmacology , Glucose/pharmacology , Tachykinins/metabolism , Animals , Bombyx/growth & development , DNA, Complementary/genetics , Digestive System/metabolism , Enteroendocrine Cells/metabolism , Larva/growth & development , Larva/metabolism , Morus/metabolism , Plant Leaves/metabolism , Secondary Metabolism , Tachykinins/geneticsABSTRACT
BACKGROUND: Measures to improve bird performance have been sought due to the imminent phase out of in-feed antibiotics in poultry and continued demand for higher poultry feeding efficiency. Increasing grain particle size and dietary fibre may improve gizzard function, digestive efficiency and nutrient absorption. This study was conducted to evaluate the effect increased particle size of corn and inclusion of sugarcane bagasse (SB) on mRNA expression of genes encoding digestive enzymes and nutrient transporters in broilers. RESULTS: A total of 336 day-old Ross 308 males were assigned in a 2 × 2 factorial arrangement of treatments with corn particle size - coarse 3576 µm or fine 1113 µm geometric mean diameter, and SB - 0 or 2% inclusion. Feed conversion ratio (FCR), weight gain and feed intake were measured from d 0-10 and d 10-24. The relative gizzard weight and mRNA expression of genes encoding digestive enzymes and intestinal nutrient transporters were measured on d 24. During d 10-24, a particle size × SB interaction was observed for FCR (P < 0.01), where birds fed coarsely ground corn (CC) with 2% SB had lower FCR than those fed CC without SB. A particle size × SB interaction was observed for both expression of pepsinogen A and C (P < 0.01) which were negatively correlated with FCR on d 24. Addition of 2% SB upregulated pepsinogen A and C only in CC fed birds. Further, 2% SB also upregulated pancreatic amylase (AMY2A) and intestinal cationic amino acid transporter-1 (CAT1). Inclusion of dietary CC upregulated duodenal amino peptidase N (APN), jejunal alanine, serine, cysteine and threonine transporter-1 (ASCT1), and ileal peptide transporter-2 (PepT2). CONCLUSION: These results suggest that both SB and coarse particle size modulate expression of genes encoding important digestive enzymes and nutrient transporters and thus are directly related to bird performance. These findings provide insights into the combination effects of dietary fiber and particle size in the future management of broiler feeding.
Subject(s)
Animal Feed/analysis , Chickens/genetics , Diet/veterinary , Dietary Fiber/administration & dosage , Dietary Supplements , Zea mays/chemistry , Animal Nutritional Physiological Phenomena , Animals , Biological Transport , Chickens/growth & development , Digestive System/enzymology , Digestive System/metabolism , High-Throughput Nucleotide Sequencing , MaleABSTRACT
Senegalese sole is one of the most promising fish species cultivated in the Southern European countries. This study was aimed at assessing the effects of microalgae biomass added to diets for Senegalese sole juveniles on fish growing and condition status. Three isoproteic (52%) and isolipidic (10%) were formulated containing 15% Tisochrysis lutea (TISO), Nannochloropsis gaditana (NAN), or Scenedesmus almeriensis (SCE) biomass, respectively. An experimental microalgae-free diet (CT) and a commercial diet (COM) were used as controls. Fish were fed at 3% of their body weight for 85 days. Final body weight of fish fed microalgae-supplemented diets did not differ from group fed CT diet. Fish-fed CT, TISO, NAN, and SCE showed higher growth performance and nutrient utilization figures than specimen-fed COM diet. The highest carcass lipid content was found in COM group (141 g kg-1), and no differences were observed in body protein content. Ash was significantly higher in TISO, NAN, and SCE groups compared to fish-fed CT. Muscle EPA and DHA contents were not modified owing to the different dietary treatments. The n3/n6 and EPA/DHA ratios in muscle were similar in all the experimental groups. The quantification of digestive proteolytic activities did not differ among experimental groups, although differences in the protease pattern in digestive extracts by zymography were revealed in those fish fed on COM diet. Both α-amylase activity in the intestinal lumen and leucine aminopeptidase in the intestinal tissue were significantly lower in COM fish. Specimens fed on SCE diet showed a higher leucine aminopeptidase activity associated to the intestinal tissue compared to NAN-fed fish (0.40 and 0.25 U g tissue-1, respectively). The ultrastructural study revealed that the dietary inclusion of algal biomass, especially T. lutea and N. gaditana, had a positive impact on the absorptive capacity of the intestinal mucosa. The highest values for the parameters microvilli length and microvilli absorption surface were observed in fish fed on NAN diet (1.99 µm and 45.93 µm2, respectively). Even though further studies aimed at optimizing commercial formulas for Senegalese sole are required prior to any large-scale practical utilization, the results obtained clearly suggest the potential of microalgae as dietary ingredients for this fish species.
Subject(s)
Animal Nutritional Physiological Phenomena , Biomass , Body Composition , Diet/veterinary , Flatfishes/physiology , Microalgae/chemistry , Animal Feed , Animals , Digestive System/metabolism , Fatty Acids/metabolism , Flatfishes/growth & development , Freeze DryingABSTRACT
How FACs-producing generalist and specialist herbivores regulate their FACs-hydrolyzing enzyme L-ACY-1 to balance FACs' beneficial vs. detrimental effects remains unknown. To address this question, we compared L-ACY-1 expression in Helicoverpa armigera and Helicoverpa assulta, a pair of closely related sibling species differing mainly in their host range, by the same sets of hostplants, protein to digestible carbohydrate (P:C) ratios, or allelochemical. L-ACY-1 expression remained low/unchanged in H. armigera, but was induced by hot pepper fruits and repressed by cotton bolls in H. assulta. The representative allelochemicals of the tested hostplants significantly (capsaicin) or insignificantly (gossypol and nicotine) induced L-ACY-1 expression in H. armigera, but insignificantly inhibited (capsaicin and gossypol) or induced (nicotine) it in H. assulta. L-ACY-1 expression remained low/unaltered on balanced (P50:C50 and P53:C47) or protein-biased diets and induced on carbohydrate-biased diets in H. armigera, but was at the highest level on balanced diets and reduced on either protein- or carbohydrate-biased diets in H. assulta. Furthermore, L-ACY-1 expression was significantly higher in H. assulta than in H. armigera for most of feeding treatments. Such expressional divergences suggest that FACs are utilized mainly for removal of excessive nitrogen in generalists but for nitrogen assimilation in specialists.
Subject(s)
Amidohydrolases/metabolism , Moths/enzymology , Amidohydrolases/chemistry , Amidohydrolases/genetics , Amino Acid Sequence , Amino Acids , Animals , Base Sequence , Carbohydrates/analysis , DNA, Complementary/genetics , Digestive System/metabolism , Fatty Acids , Larva/enzymology , Moths/genetics , Pheromones/pharmacology , Phylogeny , Transcription, GeneticABSTRACT
The aim of this study was to investigate the effects of dietary vitamin E deficiency on systematic pathological changes and oxidative stress in fish. A total of 320 healthy common carp (Cyprinus carpio) were randomized into four groups; the control group was fed a basal diet supplemented with 100 IUkg-1 of vitamin E, while the three experimental groups were fed the same basal diet with reduced vitamin E content (0, 25, or 50 IUkg-1). Findings showed that fish in the experimental groups mainly presented with sekoke disease, exophthalmia, leprnorthsis, and ascites. Histopathological and ultrastructural changes comprised nutritional myopathy with muscle fiber denaturation and necrosis, and multi-tissue organ swelling, degeneration, and necrosis. Compared with the control group, RBC count, hemoglobin content, vitamin E concentration, and superoxide dismutase activity were significantly lower in all three experimental groups. However, malondialdehyde content was considerably higher in experimental groups than in the control group. However, there was no difference in glutathione peroxidase activity among groups. In conclusion, dietary vitamin E deficiency (<100 IUkg-1) can cause severe injury and, in particular, oxidative damage in common carp. The oxidative damage might be a main influence caused by vitamin E deficiency in fish. These findings reveal the complete systematic pathological effect of vitamin E deficiency in common carp, which may be applicable to other fish and animals.
Subject(s)
Animal Nutritional Physiological Phenomena , Carps/metabolism , Nutritional Status , Oxidative Stress , Vitamin E Deficiency/metabolism , Animals , Biomarkers/blood , Carps/blood , Digestive System/drug effects , Digestive System/metabolism , Digestive System/ultrastructure , Kidney/drug effects , Kidney/metabolism , Kidney/ultrastructure , Muscle, Skeletal/drug effects , Muscle, Skeletal/metabolism , Muscle, Skeletal/ultrastructure , Myocardium/metabolism , Myocardium/ultrastructure , Time Factors , Vitamin E Deficiency/pathologyABSTRACT
The hypothesis of this work was that oxidative and nitrosative metabolism in the digestive gland (DG) of two limpet populations (intertidal and subtidal) of the Antarctic species Nacella concinna show different behavior when they were exposed to either intermittent (intertidal) or constant (subtidal) natural Fe. Total Fe content and labile Fe pool were higher in the DG of the subtidal compared to the intertidal population. However, no significant differences between populations were seen on the Fe atoms content of the isolated ferritin. Ascorbyl radical content was 2.0±0.4 and 6.5±0.8pmol/mg FW in the DG of the intertidal and subtidal animals, respectively. Lipid damage, assessed as content of thiobarbituric reactive substances, was different between the tissues of intertidal and subtidal samples, 491±102 and 1242±367pmol/mg FW, respectively. Catalase and superoxide dismutase activities showed no differences between the limpets. Nitric oxide (NO) content was 25±3 and 22±2pmol/mg FW in DG from intertidal and subtidal animals, respectively. NO synthase-like (NOS-like) activity was evaluated supplementing the samples with the enzyme co-factors, and the inhibitory effect of Nω-nitro-L-arginine methyl ester hydrochloride was tested. NO generation rate was 3.4±0.3 and 4.7±0.6pmol/minmg FW in DG from the intertidal and subtidal population, respectively. These results showed that the oxidative condition of the limpet population constantly covered by the Fe enriched water is more affected than the intertidal population. However, the nitrosative metabolism seems to be independent of the environmental high Fe content since similar NO steady state concentration and NOS-like activity were measured in both populations.
Subject(s)
Digestive System/metabolism , Gastropoda/metabolism , Iron/metabolism , Nitric Oxide/metabolism , Animals , Antarctic Regions , Catalase/metabolism , Digestive System/drug effects , Electron Spin Resonance Spectroscopy , Gastropoda/drug effects , Iron/toxicity , Nitric Oxide Synthase/metabolism , Oxidation-Reduction/drug effects , Oxidative Stress/drug effects , Seawater/chemistry , Superoxide Dismutase/metabolism , Thiobarbituric Acid Reactive Substances/metabolismABSTRACT
This study was conducted to establish the multicomponent sequential metabolism (MSM) method based on comparative analysis along the digestive system following oral administration of licorice (Glycyrrhiza uralensis Fisch., leguminosae), a traditional Chinese medicine widely used for harmonizing other ingredients in a formulae. The licorice water extract (LWE) dissolved in Krebs-Ringer buffer solution (1 g/mL) was used to carry out the experiments and the comparative analysis was performed using HPLC and LC-MS/MS methods. In vitro incubation, in situ closed-loop and in vivo blood sampling were used to measure the LWE metabolic profile along the digestive system. The incubation experiment showed that the LWE was basically stable in digestive juice. A comparative analysis presented the metabolic profile of each prototype and its corresponding metabolites then. Liver was the major metabolic organ for LWE, and the metabolism by the intestinal flora and gut wall was also an important part of the process. The MSM method was practical and could be a potential method to describe the metabolic routes of multiple components before absorption into the systemic blood stream. Copyright © 2015 John Wiley & Sons, Ltd.
Subject(s)
Chromatography, High Pressure Liquid/methods , Glucosides/metabolism , Glycyrrhiza , Mass Spectrometry/methods , Animals , Digestive System/metabolism , Glucosides/administration & dosage , Male , Rats , Rats, Sprague-DawleyABSTRACT
Barley malt, a product of controlled germination, has been shown to produce high levels of butyric acid in the cecum and portal serum of rats and may therefore have anti-inflammatory effects. The aim of the study was to investigate how four barley malts, caramelized and colored malts, 50-malt and 350-malt, differing in functional characteristics concerning beta-glucan content and color, affect short-chain fatty acids (SCFA), barrier function and inflammation in the hindgut of rats fed high-fat diets. Male Wistar rats were given malt-supplemented high-fat diets for four weeks. Low and high-fat diets containing microcrystalline cellulose were incorporated as controls. All diets contained 70 g kg(-1) dietary fiber. The malt-fed groups were found to have had induced higher amounts of butyric and propionic acids in the hindgut and portal serum compared with controls, while cecal succinic acid only increased to a small extent. Fat increased the mRNA expression of tight junction proteins and Toll-like receptors (TLR) in the small intestine and distal colon of the rats, as well as the concentration of some amino acids in the portal plasma, but malt seemed to counteract these adverse effects to some extent. However, the high content of advanced glycation end-products (AGE) in caramelized malt tended to prohibit the positive effects on occludin in the small intestine and plasma amino acids seen with the other malt products. In conclusion, malting seems to be an interesting process for producing foods with positive health effects, but part of these effects may be destroyed if the malt contains a high content of AGE.
Subject(s)
Butyric Acid/metabolism , Digestive System/metabolism , Glycation End Products, Advanced/analysis , Hordeum/chemistry , Hordeum/metabolism , Seeds/growth & development , Tight Junction Proteins/genetics , Toll-Like Receptors/genetics , Animals , Cecum/metabolism , Diet, High-Fat/adverse effects , Food Handling , Germination , Glycation End Products, Advanced/metabolism , Hordeum/growth & development , Male , Rats , Rats, Wistar , Seeds/chemistry , Seeds/metabolism , Tight Junction Proteins/metabolism , Toll-Like Receptors/metabolism , beta-Glucans/analysis , beta-Glucans/metabolismABSTRACT
Chitinolytic enzymes have an important physiological significance in immune and digestive systems in plants and animals, but chitinase has not been identified as having a role in the digestive system in molluscan. In our study, a novel chitinase homologue, named Ca-Chit, has been cloned and characterized as the oyster Crassostrea angulate. The 3998bp full-length cDNA of Ca-Chit consisted of 23bp 5-UTR, 3288 ORF and 688bp 3-UTR. The deduced amino acids sequence shares homologue with the chitinase of family 18. The molecular weight of the protein was predicted to be 119.389 kDa, with a pI of 6.74. The Ca-Chit protein was a modular enzyme composed of a glycosyl hydrolase family 18 domain, threonine-rich region profile and a putative membrane anchor domain. Gene expression profiles monitored by quantitative RT-PCR in different adult tissues showed that the mRNA of Ca-Chit expressed markedly higher visceral mass than any other tissues. The results of the whole mount in-situ hybridization displayed that Ca-Chit starts to express the visceral mass of D-veliger larvae and then the digestive gland forms a crystalline structure during larval development. Furthermore, the adult oysters challenged by starvation indicated that the Ca-Chit expression would be regulated by feed. All the observations made suggest that Ca-Chit plays an important role in the digestive system of the oyster, Crassostrea angulate.
Subject(s)
Chitinases/metabolism , Crassostrea/enzymology , Digestive System/enzymology , Amino Acid Sequence , Animals , Base Sequence , Chitinases/classification , Chitinases/genetics , Cloning, Molecular , Crassostrea/genetics , Crassostrea/growth & development , DNA, Complementary/chemistry , DNA, Complementary/genetics , Digestive System/metabolism , Eating , Gene Expression Regulation, Developmental , In Situ Hybridization , Larva/enzymology , Larva/genetics , Larva/growth & development , Molecular Sequence Data , Phylogeny , Reverse Transcriptase Polymerase Chain Reaction , Sequence Analysis, DNA , Sequence Homology, Amino Acid , StarvationABSTRACT
Increasing evidence indicates that the gut microbiota can be altered to ameliorate or prevent disease states, and engineering the gut microbiota to therapeutically modulate host metabolism is an emerging goal of microbiome research. In the intestine, bacterial urease converts host-derived urea to ammonia and carbon dioxide, contributing to hyperammonemia-associated neurotoxicity and encephalopathy in patients with liver disease. Here, we engineered murine gut microbiota to reduce urease activity. Animals were depleted of their preexisting gut microbiota and then inoculated with altered Schaedler flora (ASF), a defined consortium of 8 bacteria with minimal urease gene content. This protocol resulted in establishment of a persistent new community that promoted a long-term reduction in fecal urease activity and ammonia production. Moreover, in a murine model of hepatic injury, ASF transplantation was associated with decreased morbidity and mortality. These results provide proof of concept that inoculation of a prepared host with a defined gut microbiota can lead to durable metabolic changes with therapeutic utility.
Subject(s)
Biological Therapy/methods , Digestive System/microbiology , Hyperammonemia/microbiology , Hyperammonemia/therapy , Microbiota , Ammonia/metabolism , Animals , Bacteria/enzymology , Bacteria/genetics , Bacterial Proteins/genetics , Bacterial Proteins/metabolism , Bioengineering , Chemical and Drug Induced Liver Injury/therapy , Digestive System/metabolism , Disease Models, Animal , Feces/microbiology , Female , Genes, Bacterial , Hyperammonemia/metabolism , Male , Mice , Mice, Inbred C57BL , Mice, SCID , Microbiota/physiology , Time Factors , Urease/genetics , Urease/metabolismABSTRACT
UNLABELLED: The objective of this study was to investigate effects of encapsulated nitrate (EN) on enteric methane emission and N and energy utilization in beef heifers. Eight ruminally-cannulated beef heifers (451 ± 21 kg BW) were used in a replicated 4 × 4 Latin square design. Four experimental diets were prepared and fed once daily for ad libitum intake: control, 1%, 2%, and 3% EN (0.15, 0.9, 1.5, and 2.5% NO3(-) in dietary DM, respectively). The control diet (55% forage and 45% concentrate) included encapsulated urea, which was gradually replaced with EN for the EN diets (iso-nitrogenous; 12.5% CP). In each period, EN was increased stepwise by 1% every 4 d during adaptation. A 7-d washout period (control diet offered to all heifers) was provided between experimental periods. Dry matter intake tended to decrease (10.4 to 10.1 kg/d; linear, P = 0.06) with EN levels. Enteric methane yield was linearly decreased (21.3 to 17.4 g/kg DMI; P < 0.01) by EN, and methane production (g/d) recovered to the level from heifers fed the control diet on the first day when EN was withdrawn from the diet. Apparent total-tract digestibility of DM and OM increased (P = 0.03) or tended to increase (P = 0.06), respectively, with EN levels. Starch digestibility tended to be greater (P = 0.07) for EN vs. CONTROL: The concentrations of rumen ammonia-N and plasma urea-N decreased linearly (P < 0.01) with EN. Total urinary N and urea-N excretion as proportions of N intake were linearly decreased (46.3 to 41.4%, = 0.09 and 37.1 to 29.9%, P = 0.01, respectively) with EN addition. However, NO3(-)-N excretion in urine increased linearly (P < 0.01) with EN levels. Fecal N excretion was not affected (P = 0.47) by EN, although fecal NO-N excretion increased linearly (P < 0.01) with inclusion of EN (0.09 to 0.88% of total N, P < 0.01). Retained N tended to be increased (percentage of N intake; 16.6 to 21.4%, = 0.08) by the EN. Supplementary EN lowered (6.64 to 5.46% of GE intake [GEI], P < 0.01) energy losses by enteric methane mitigation, which increased ME supply (calculated; 56.5 to 58.8% of GEI, P = 0.01) without changes in calculated heat production (P = 0.24). As a result, retained energy tended to increase (P = 0.07) with EN levels. In conclusion, feeding EN to beef heifers lowered enteric methane production in a dose-response manner, which slightly increased energy supply. Total urinary N excretion was lowered for EN due to lower urinary urea-N excretion.
Subject(s)
Cattle/metabolism , Digestive System/drug effects , Energy Metabolism/drug effects , Methane/metabolism , Nitrates/pharmacology , Nitrogen/metabolism , Ammonia/metabolism , Animal Feed , Animal Nutritional Physiological Phenomena/drug effects , Animal Nutritional Physiological Phenomena/physiology , Animals , Body Weight/physiology , Capsules , Dietary Supplements , Digestive System/metabolism , Eating/physiology , Energy Metabolism/physiology , Female , Nitrates/administration & dosage , Rumen/metabolism , Urea/metabolismABSTRACT
Magnesium (Mg(2+)) is an essential ion to the human body, playing an instrumental role in supporting and sustaining health and life. As the second most abundant intracellular cation after potassium, it is involved in over 600 enzymatic reactions including energy metabolism and protein synthesis. Although Mg(2+) availability has been proven to be disturbed during several clinical situations, serum Mg(2+) values are not generally determined in patients. This review aims to provide an overview of the function of Mg(2+) in human health and disease. In short, Mg(2+) plays an important physiological role particularly in the brain, heart, and skeletal muscles. Moreover, Mg(2+) supplementation has been shown to be beneficial in treatment of, among others, preeclampsia, migraine, depression, coronary artery disease, and asthma. Over the last decade, several hereditary forms of hypomagnesemia have been deciphered, including mutations in transient receptor potential melastatin type 6 (TRPM6), claudin 16, and cyclin M2 (CNNM2). Recently, mutations in Mg(2+) transporter 1 (MagT1) were linked to T-cell deficiency underlining the important role of Mg(2+) in cell viability. Moreover, hypomagnesemia can be the consequence of the use of certain types of drugs, such as diuretics, epidermal growth factor receptor inhibitors, calcineurin inhibitors, and proton pump inhibitors. This review provides an extensive and comprehensive overview of Mg(2+) research over the last few decades, focusing on the regulation of Mg(2+) homeostasis in the intestine, kidney, and bone and disturbances which may result in hypomagnesemia.