Effect of cassic acid on immunity and immune-reproductive genes transcription in Clarias gariepinus against Edwardsiella tarda.

The present study was investigated the dietary administration of cassic acid (CA) on growth, innate immunity, transcription profiles of estrogen and follicle-stimulating hormones as well as lysozyme enzyme determined in Clarias gariepinus against Edwardsiella tarda. The weight gain (WG), protein efficacy ratio (PER), and feed conversion ratio (FCR) were significantly improved in infected fish fed dietary administration with CA at 5 and 10 mg kg-1 diets. The survival is higher (96.7% and 98.3%) in the infected groups fed at 5 and 10 mg kg-1 CA diets. The red (RBC) and white (WBC) blood cells, hemoglobin (Hb), and packed cell volume (PCV) was found significantly high in the infected fish feeding at 5 and 10 mg kg-1 CA diets. Total protein and albumin were significantly increased with 5 and 10 mg kg-1 CA diets among weeks 1-4 while the globulin and albumin: globulin ratio increased of these diet only after week 2. The phagocytic and respiratory burst activities were enhanced statistically the infected fish fed at 10 mg kg-1 CA diet group whereas the production of superoxide anion (SOA) and nitric oxide (NO) were significantly increased at 5 and 10 mg kg-1 CA diets. The lymphocyte proliferation and myeloperoxidase (MPO) activity were significantly high the infected fish when fed at 5 and 10 mg kg-1 CA diets after 2nd week whereas the alternate complement activity (ACP), generation of reactive oxygen species (ROS), and lysozyme activity (Lyz) were observed at 5 and 10 mg kg-1 CA diets among weeks 1-4. The accumulative mortality was 10% in infected fish fed at 1 and 5 mg kg-1 CA diets whereas 15% mortality found with 10 mg kg-1 CA diet. The highest levels of estrogen receptor alpha (ERα) mRNA expression found in gonad while the highest levels of follicle stimulating hormone-beta subunit (FSH-ß) mRNA expression found in testis of the infected fish given at 5 and 10 mg kg-1 CA diets. The up-regulation of chick-type lysozyme (c-Lyz) mRNA was observed at 5 and 10 mg kg-1 CA diets after 2nd week while goose-type lysozyme (g-Lyz) mRNA was up-regulation amongst weeks 1-4 of these diets. The present study suggested that E. tarda challenged fish after feeding with 5 and 10 mg kg-1 CA diets did not affect growth and hemato-biochemical parameter, but it enhanced nonspecific immune system and improving ERα, FSH-ß, c-Lyz, and g-Lyz mRNA expression in C. gariepinus against E. tarda.

Effect of Rhododendron arboreum Leaf Extract on the Antioxidant Defense System against Chromium (VI) Stress in Vigna radiata Plants.

In the current investigation, we studied role of Rhododendron leaf extract in Vigna radiata grown under chromium metal stress. We observed that seed treatment with Rhododendron leaf extract resulted in the recuperation of seedling growth under chromium toxicity. Seed treatment with Rhododendron leaf extract significantly improved the contents of anthocyanin and xanthophyll pigments under stress. The antioxidative defense system triggered after Rhododendron extract treatment, resulting in the increased actions of antioxidant enzymes. Oxidative stress induced by the assembly of reactive oxygen species was reduced after Rhododendron extract treatment under chromium toxicity as indicated by the enhanced contents of non-enzymatic antioxidants, namely ascorbic acid, tocopherol, and glutathione. Furthermore, Rhododendron leaf extract treatment under chromium metal stress also encouraged the biosynthesis of organic acids, polyphenols, as well as amino acids in Vigna radiata. Statistical analysis of the data with multiple linear regression also supported that Rhododendron leaf extract can effectively ease chromium metal-induced phytotoxicity in Vigna radiata.

Crocus sativus L. Extract Containing Polyphenols Modulates Oxidative Stress and Inflammatory Response against Anti-Tuberculosis Drugs-Induced Liver Injury.

The purpose of this study is to analyze the polyphenolic rich extract of Crocus sativus L. petals (CSP) in modulating liver oxidative stress and inflammatory response status against rifampicin isoniazid (INH-RIF) drug-induced liver injury. The INH-RIF was administered for 14 days with varying doses in Wistar rats, while silymarin was administered as standard dose. We report the defensive impacts of CSP against INH-RIF induced liver oxidative stress and proinflammatory cytokine. The CSP treatment at both doses significantly controlled all modulating biochemical hepatic injury indicators and resulted in the attenuation of arbitral INH-RIF damage. The components present in CSP identified by LC-ESI-Q-TOF-MS were found to be flavonoids and fatty acids. It can be inferred that CSP possesses a hepatoprotective capacity against INH-RIF-mediated hepatic injury, which may prove to be a medically beneficial natural product for the management of drug-induced liver injury.

Antioxidant molecular mechanism of adenosyl homocysteinase from cyanobacteria and its wound healing process in fibroblast cells.

An antioxidant molecule namely, adenosyl homocysteinase (AHc) was identified from the earlier constructed transcriptome database of Spirulina, where it was cultured in a sulphur deprived condition. From the AHc protein, a small peptide NL13 was identified using bioinformatics tools and was predicted to have antioxidant property. Further, the peptide was synthesised and its antioxidant mechanism was addressed at molecular level. NL13 was subjected to various antioxidant assays including DPPH assay, HARS assay, SARS Assay, NO assay and ABTS assay, where NL13 exhibited significant (P < 0.05) potential antioxidant activity compared to its antioxidant control, Trolox. Cytotoxicity was performed on Human whole blood and the cell viability was performed on VERO fibroblast cells. In both assays, it was found that NL13 did not exhibit any cytotoxic effect towards the cells. Further, the intracellular ROS was performed on Multimode reader followed by imaging on fluorescence microscope which showed scavenging activity even at lower concentration of NL13 (31.2 µM). An effective wound healing property of NL13 on VERO cells was confirmed by analysing the cell migration rate at two different time intervals (24 and 48 h). Overall, the study shows that NL13 peptide scavenges the intracellular oxidative stress.

Study the immunomodulation of anthracenedione in striped dwarf catfish, Mystus vittatus against pathogenic bacteria, Aeromonas hydrophila.

Anthracenedione is a derivative of anthraquinone aromatic organic natural pigments found in senna, aloe latex, rhubarb, cascara, lichens, and fungi having broad range of bioactivity, including anti-cancer, anti-inflammatory, anti-microbial, anti-fungal, anti-oxidant, anti-viral activities suggesting potential for clinical purpose of many diseases. The effect of anthracenedione enriched diet on growth, hematology, innate and adaptive immune parameters as well as protection from Aeromonas hydrophila in Mystus vittatus was reported. The weight gain, feed intake, specific growth rate (SGR), and feed conversion ratio (FCR) were significantly increased in uninfected groups fed with 5 mg kg-1 diet. The red blood cells (RBC) and white blood cells (WBC) count and the percentage of lymphocytes were significantly augmented in both infected and uninfected groups feeding with any diet. The percentage of monocytes, eosinophils, neutrophils and the biochemical profile such as total protein, albumin, and globulin also were significantly increased in the infected and uninfected groups fed with 5 mg kg-1 enriched diet. The innate and adaptive immune parameters such as phagocytic activity, immunoglobulin M (IgM), respiratory burst activity, complement activity, and lysozyme activity were significantly increased in uninfected and infected groups fed with 5 or 10 mg kg-1 diets but not with 1 mg kg-1 diet. The serum superoxide dismutase (SOD) activity is significantly increased in the uninfected and infected fish fed with 5 mg kg-1 diet but the increase was not significantly observed in 1 or 10 mg kg-1 diets. The nitric oxide (NO) production is significantly elevated in both uninfected and infected groups fed with 5 mg kg-1 diet. On the other hand, the lymphocyte proliferation and myeloperoxidase (MPO) activity were significantly increased the infected and uninfected groups fed with 5 and 10 mg kg-1 diets. The cumulative mortality was found 5% with 1 and 5 mg kg-1 diet groups while it was observed 10% mortality with 10 mg kg-1 diet group. Based on the results, it is observed that feeding the uninfected and infected groups with 5 mg kg-1 anthracenedione diet resulted in better improvement of growth, hematological, biochemical, and innate as well as adaptive immune parameters in M. vittatus against A. hydrophila.

Effect of symbiotic supplemented diet on innate-adaptive immune response, cytokine gene regulation and antioxidant property in Labeo rohita against Aeromonas hydrophila.

Administration of probiotic, prebiotic or symbiotic supplemented diets boosts the antioxidant property, pro and/or anti-inflammatory cytokine gene transcription, innate-adaptive immunity, growth rate and feed digestibility with very low or no mortality in healthy and infected (both groups) in Labeo rohita against Aeromonas hydrophila is reported. The probiotic diet increased the white blood cell (WBC) count and globulin (GB) level significantly on or after 6th week whereas with the symbiotic diet the increase was noted two weeks earlier in both groups; the total protein (TP) level also increased significantly when fed with probiotic diet on weeks 6 and 8, whereas with symbiotic diet the significant increase manifested earlier at 4th week itself. The serum phagocytic activity (PA), respiratory burst activity (RBA), complement C3 (CC3) level, alternative complement pathway (ACP), lysozyme activity (LA), and immunoglobulin M (IgM) levels in head kidney (HK) leucocytes increased significantly (P < 0.05) in both groups fed with probiotic diet on weeks 6 and 8; with symbiotic diet from weeks 2-8; but with prebiotic diet only on 8th week. With probiotic diet the superoxide dismutase (SOD) and catalase (CAT) activities increased significantly (P < 0.05) on weeks 6 and 8; with symbiotic diet from weeks 4-8 but the prebiotics diet only on 8th week. However, glutathione peroxidase (GPx) activity increased significantly (P < 0.05) with probiotic diet on weeks 6 and 8 and with symbiotic diet from weeks 4-8. When healthy fish fed with any supplementation diet for a period of 30 days there was no mortality while 5%, 10%, and 10% mortality was observed in infected group fed with symbiotic, probiotic, and prebiotic supplementation diets. In head kidney (HK) leucocytes, the IL-1ß, IL-8, TNF-α, and NF-κB gene transcriptions were significantly up-regulation in both groups when fed with probiotic diet on weeks 6 and 8, symbiotic diet from weeks 4-8 while the prebiotic diet only on 8th week. The iNOS expression was up-regulation significantly in both groups fed with probiotic and symbiotic diets on weeks 6 and 8; however, with any diet, the relative IL-10 and TGF-ß gene expressions were down-regulated. The present study suggested that dietary administration of symbiotic diet elicited earlier antioxidant activity, innate-adaptive immune response, immune related cytokine gene modulation, and disease protection earlier i.e. on 4th week than with probiotic or prebiotic diets in L. rohita against A. hydrophila.

Resolving the pathogenicity factors of a novel opportunistic fungus Schizophyllum commune at molecular level.

Schizophyllum commune is a well-known mushroom forming fungi which is an edible one due to its nutritive value. It exhibits a special wood degrading mechanism to grow in decay matters by releasing a series of enzymes. These enzymes might make them an opportunistic pathogen which has been reported to infect various animals and human beings too. Although these fungi were identified as human and animal pathogens, their mechanisms of pathogenesis and the key virulence factors involved in disease establishment are not known. In this study, we reported this fungal infection in freshwater fish for the first time and its morphological features. Further, we employed RNA-seq technique to identify the major virulence factors involved in the pathogenesis in fish and the network of interaction between the identified virulence factors were analysed. Also, we confirmed the virulence roles of this fungus during infection by qRT-PCR analysis. This study emphasizes the virulence nature of the common mushroom forming food fungus and the involvement of enzymes such as phosphoinositide phospholipase C, hexosaminidase and few toxins such as pesticidal and insecticidal crystal proteins which opened a new avenue in the virulence nature of edible mushrooms.

Design and characterization of a novel Arthrospira platensis glutathione oxido-reductase-derived antioxidant peptide GM15 and its potent anti-cancer activity via caspase-9 mediated apoptosis in oral cancer cells.

Glutathione oxido-reductase (GR) is a primary antioxidant enzyme of most living forms which protects the cells from oxidative metabolism by reducing glutathione (GSH) from its oxidized form (GSSG). Although the antioxidant role of the enzyme is well characterized, the specific role of conserved N' peptide sequence in antioxidant mechanism remains unclear. In this study, we have identified an RNA sequence encoding GR enzyme from spirulina, Arthrospira platensis (Ap) and the changes in its gene expression profile was analysed during H2O2 stress. Results showed that H2O2 (10 mM) stimulated the expression of ApGR throughout the timeline of study (0, 5, 10, 15 and 20 days) with highest expression at 5th day post-exposure which confirmed the antioxidant role of ApGR in spirulina during H2O2 induced oxidative stress. A dithiol containing short antioxidant peptide, 39GGTCVIRGCVPKKLM53 (GM15) from ApGR was predicted and its radicals (superoxide and hydroxyl radical) scavenging potential was confirmed by in vitro cell-free assays. GM15 (12.5 µM) reduced the intracellular generalized oxidative stress level, as measured using DCFDA assay in H2O2 exposed leucocytes without affecting any of the cellular population. Further, the biomedical application of the radical scavenging property of GM15 was validated in oral carcinoma (KB) cells where GM15 exhibited significant cytotoxicity. Also, GM15 exhibited heterogenous effects on intracellular oxidative stress level in KB cells: at lower concentration (6.25 µM), the peptide reduced oxidative stress whereas, at higher concentration (25 µM) it increased the intensity of oxidative stress. GM15 (25 µM) induced caspase-9 mediated apoptosis in KB cells along with membrane disruption and DNA degradation which are confirmed by propidium iodide (PI) internalization and comet assays, respectively. Overall, the study shows that GM15 peptide i) scavenges superoxide, hydroxyl radicals, and influences intracellular oxidative stress, and ii) has anti-cancer effect in oral cancer cells.

Effects of aloe-emodin on innate immunity, antioxidant and immune cytokines mechanisms in the head kidney leucocytes of Labeo rohita against Aphanomyces invadans.

The effect of aloe-emodin incorporated diets on innate immune response, disease resistance, pro and/or anti-inflammatory cytokine gene transcription in Labeo rohita against Aphanomyces invadans is reported for the first time. In healthy and infected groups fed with 5 mg aloe-emodin enriched diet the white blood cell (WBC) count increased significantly (p > 0.05) after 6th week. In both groups fed with any enriched diet the biochemical parameters such as albumin, globulin, and albumin/globulin ratio did not vary significantly; however with 5 mg aloe-emodin diet the albumin and globulin levels increased significantly (p > 0.05) after 6th week. The serum phagocytic activity (PA), respiratory burst activity (RBA), serum complement C3 (CC3), and lysozyme activity (LA) did not increase with any diet between weeks 2 and 4, whereas with 5 mg aloe-emodin diet increased significantly in both groups after 6th week. The pro-inflammatory cytokines such as IL-1ß, IL-8, TNF-α, and iNOS significantly modulated the expression in both groups on being fed with 5 mg aloe-emodin incorporation diet on 8th week. Healthy fish fed with any aloe-emodin diet did not suffer mortality. However, the infected fish fed with 1, 5, and 10 mg kg-1 aloe-emodin diets registered 5%, 10%, and 15% mortality. The present study indicates that healthy and infected L. rohita exhibited enhanced innate immune response, disease resistance, pro and/or anti-inflamatory cytokine gene transcription levels against A. invadans.

Ameliorative efficacy of bioencapsulated Chironomous larvae with Shilajit on Zebrafish (Danio rerio) exposed to Ionizing radiation.

Using Zebrafish (Danio rerio) as a model organism, we evaluated the radioprotective and antioxidant effects of the Indian traditional medicine Shilajit exposed to X-Ray. The Zebrafish were divided into three experimental groups and control group, each group containing ten fish. The three experimental fish groups, group I, group II and group III were fed with 3, 5 and 7ppm shilajit encapsulated Chironomous larvae and group IV served as a control fed with non- encapsulated larvae. After 60 days of feeding trial, fish were irradiated with X-Ray at a single acute dose of 1Gy. 72h of post-irradiation, each experimental fish were observed for its morphological, behavioral, clinical symptoms, antioxidant levels and DNA damage were evaluated. Among the experimental groups 5ppm shilajit encapsulated Chironomous larvae fed fish group shows the most significant radioprotective effects compared with control and other experimental fish groups. The present study indicates that shilajit have significant radioprotective and antioxidant enhancing capability. The humus substance of shilajit may be the factor responsible to react with radiation-derived or radiation related reactive species on zebrafish.

Effect of Shilajit enriched diet on immunity, antioxidants, and disease resistance in Macrobrachium rosenbergii (de Man) against Aeromonas hydrophila.

The effect of diet supplemented with Shilajit, a multi-component natural mineral substance on the antioxidant activity, immune response, and disease resistance in freshwater prawn, Macrobrachium rosenbergii (de Man) against Aeromonas hydrophila is reported. The total hemocyte count (THC) and phagocytic activity significantly increased with 2 g kg(-1) supplemented diet on first week and with other enriched diets on weeks 2 and 4. The respiratory burst (RB) activity and glutathione peroxidase (GPx) activity were significantly increased with 2 g kg(-1) supplemented diet on weeks 1 and 2 whereas 2 and 4 g kg(-1) diets on week 4. The phenoloxidase (PO) activity increased significantly with 2 g kg(-1) diet only on second week and with other enriched diets only on fourth week. The superoxide dismutase (SOD) activity increased significantly with any enriched diet during the experimental period except with 6 g kg(-1) diets on first week. However, the glutathione reductase (GR) activity was enhanced significantly only with 2 g kg(-1) enriched diets on weeks 2 and 4. The cumulative mortality of the prawn fed with 2 and 4 g kg(-1) enriched diets was 10% and 15% whereas with 6 g kg(-1) diet the mortality was 20%. The results suggest that diet enriched with Shilajit at 2 g kg(-1) or 4 g kg(-1) positively enhances the antioxidant activity, immunity, and disease resistance in M. rosenbergii against A. hydrophila.