muramidase and Starvation

Aquatic animals are frequently suffered from starvation due to restricted food availability or deprivation. It is currently known that gut microbiota assists host in nutrient acquisition. Thus, exploring the gut microbiota responses would improve our understanding on physiological adaptation to starvation. To achieve this, we investigated how the gut microbiota and shrimp digestion and immune activities were affected under starvation stress. The results showed that the measured digestion activities in starved shrimp were significantly lower than in normal cohorts; while the measured immune activities exhibited an opposite trend. A structural equation modeling (SEM) revealed that changes in the gut bacterial community were directly related to digestive and immune enzyme activities, which in turn markedly affected shrimp growth traits. Notably, several gut bacterial indicators that characterized the shrimp nutrient status were identified, with more abundant opportunistic pathogens in starved shrimp, although there were no statistical differences in the overall diversity and the structures of gut bacterial communities between starved and normal shrimp. Starved shrimp exhibited less connected and cooperative interspecies interaction as compared with normal cohorts. Additionally, the functional pathways involved in carbohydrate and protein digestion, glycan biosynthesis, lipid and enzyme metabolism remarkably decreased in starved shrimp. These attenuations could increase the susceptibility of starved shrimp to pathogens infection. In summary, this study provides novel insights into the interplay among shrimp digestion, immune activities and gut microbiota in response to starvation stress.

Topics: Acid Phosphatase; Amylases; Animals; Bacteria; Digestion; Gastrointestinal Microbiome; Hepatopancreas; Lipase; Muramidase; Penaeidae; Pepsin A; RNA, Ribosomal, 16S; Starvation; Stomach; Stress, Physiological; Superoxide Dismutase

The present study aimed to test starvation-induced oxidative stress in the cinnamon clownfish Amphiprion melanopus illuminated by light-emitting diodes (LEDs): red (peak at 630 nm), green (peak at 530 nm), and blue (peak at 450 nm) within a visible light. We investigated the oxidative stress induced by starvation for 12 days during illumination with 3 LED light spectra through measuring antioxidant enzyme (superoxide dismutase [SOD] and catalase [CAT]) mRNA expression and activity; CAT western blotting; and measuring lipid peroxidation [LPO]), plasma H(2)O(2), lysozyme, glucose, alanine aminotransferase (AlaAT), aspartate aminotransferase (AspAT), and melatonin levels. In green and blue lights, expression and activity of antioxidant enzyme mRNA were significantly lower than those of other light spectra, results that are in agreement with CAT protein expression level by western blot analysis. Also, in green and blue lights, plasma H(2)O(2), lysozyme, glucose, AlaAT, AspAT, and melatonin levels were significantly lower than those in other light spectra. These results indicate that green and blue LEDs inhibit oxidative stress and enhance immune function in starved cinnamon clownfish.

Topics: Alanine Transaminase; Animals; Aspartate Aminotransferases; Blood Glucose; Catalase; Fish Proteins; Gene Expression; Glutathione Peroxidase; Hydrogen Peroxide; Light; Lipid Peroxidation; Liver; Malondialdehyde; Melatonin; Muramidase; Oxidative Stress; Perciformes; Starvation; Superoxide Dismutase

Lack of enteral feeding, with or without parenteral nutritional support, is associated with increased intestinal permeability and translocation of bacteria. Such translocation is thought to be important in the high morbidity and mortality rates of patients who receive nothing by mouth. Recently, Paneth cells, important constituents of innate intestinal immunity, were found to be crucial in host protection against invasion of both commensal and pathogenic bacteria. This study investigates the influence of food deprivation on Paneth cell function in a mouse starvation model. Quantitative PCR showed significant decreases in mRNA expression of typical Paneth cell antimicrobials, lysozyme, cryptdin, and RegIIIγ, in ileal tissue after 48 hours of food deprivation. Protein expression levels of lysozyme and RegIIIγ precursor were also significantly diminished, as shown by Western blot analysis and IHC. Late degenerative autophagolysosomes and aberrant Paneth cell granules in starved mice were evident by electron microscopy, Western blot analysis, and quantitative PCR. Furthermore, increased bacterial translocation to mesenteric lymph nodes coincided with Paneth cell abnormalities. The current study demonstrates the occurrence of Paneth cell abnormalities during enteral starvation. Such changes may contribute to loss of epithelial barrier function, causing the apparent bacterial translocation in enteral starvation.

Topics: Animals; Autophagy; Bacterial Translocation; Ileum; Immunity, Innate; Immunologic Techniques; Male; Mice; Mice, Inbred C57BL; Microscopy, Electron; Muramidase; Pancreatitis-Associated Proteins; Paneth Cells; Permeability; Protein Precursors; Proteins; Real-Time Polymerase Chain Reaction; RNA, Messenger; Starvation

Lysozymes can hydrolyze bacteria and play an important role in animal digestion and innate immunity. The cDNA of a chicken-type lysozyme gene (Mdlys) was cloned from housefly (Musca domestica). The 484 bp full-length cDNA contains a 426 bp open reading frame (ORF) that encodes MdLys of 141 amino acids. Phylogenetic analysis indicated that the MdLys was similar to chicken-type lysozymes. Spatio-temporal expression of Mdlys was analyzed by RT-PCR. The Mdlys transcript can be detected in both midgut and fat body and was expressed at a relatively lower level at the embryo stage. Mdlys mRNA was upregulated 2 h post bacterial challenge, maintained for 2 to 6 h, and slightly declined from 12 to 24 h post-injection. Western blot analysis showed that MdLys was highly expressed in midgut and was also detected in the hemolymph and fat body. MdLys expression was slightly increased in midgut after challenging with Escherichia coli or Staphylococcus aureus. Its expression was also slightly increased in the fat body after challenging with S. aureus, but no obvious change occurred after E. coli challenge. MdLys expression in the hemolymph was not affected by bacterial challenge. In the developmental stages, MdLys expression levels had no obvious change from the first instar to the pupae stage. There was also no variation under 24 h starvation stress. Recombinant MdLys displayed inhibitory activity against Gram-negative and Gram-positive bacteria. Together, these results suggest that MdLys may play an important role in the innate immunity of houseflies.

Topics: Animals; Anti-Bacterial Agents; Bacteria; Chickens; Cloning, Molecular; Gene Expression Regulation; Gene Expression Regulation, Developmental; Houseflies; Humans; Immunity, Innate; Muramidase; Phylogeny; Recombinant Proteins; Starvation; Time Factors

Topics: Animals; Drug Synergism; Emetine; Guinea Pigs; In Vitro Techniques; Mitochondria, Muscle; Muramidase; Myocardium; Proteins; Sodium Dodecyl Sulfate; Spectrophotometry, Ultraviolet; Starvation; Succinate Dehydrogenase

Postnuclear supernates from homogenates of skeletal muscle from rats subjected to starvation, injections of Triton WR-1339, dextran-500, and dextran + corticosterone were fractionated by means of rate and isopycnic zonal centrifugation in sucrose-0.02 M KCl gradients. Zonal fractions were analyzed for protein, RNA, cytochrome oxidase, and up to six acid hydrolases. The results indicate the presence of two groups of lysosome-like particles. One group contributes approximately 95% of the cathepsin D and acid phosphatase activity and 75% of the acid ribonuclease, beta-glucuronidase, and arylsulfatase activity in muscle. It is characterized by a modal equilibrium density of 1.18 that is decreased by starvation, but is not shifted by dextran-500 or Triton WR-1339. The second group has a higher proportion of acid ribonuclease, beta-glucuronidase, and arylsulftase; the equilibrium density can be shifted by dextran-500 and Triton WR-1339. It is suggested that this group of lysosomes is derived from macrophages and other connective tissue cells, whereas the former group represents lysosome-like particles from muscle cells.

Topics: Acid Phosphatase; Animals; Cathepsins; Cell Fractionation; Centrifugation, Zonal; Corticosterone; Detergents; Dextrans; Dialysis; Electron Transport Complex IV; Glucuronidase; Lysosomes; Methods; Muramidase; Muscle Proteins; Muscles; Phagocytosis; Proteins; Rats; Ribonucleases; RNA; Starvation; Time Factors; Tissue Extracts