IN VITRO ANTIOXIDANT ACTIVITY OF VARIOUS LEAF EXTRACTS OF CANTHIUM COROMANDELICUM(BURM.F.) ALSTON.

Objectives: The objective of this study was to estimate the total phenolic and flavonoid contents and to explore the antioxidant potential of various leaf extracts (chloroform, ethyl acetate, and ethanol) of Canthium coromandelicum, which is considered traditionally as an important medicinal plant.Methods: Antioxidant properties of the extracts were assessed using 1, 1- diphenyl -2- picrylhydrazyl and hydrogen peroxide radical scavenging assays for ascorbic acid equivalents. The total phenolic and flavonoid contents were also investigated to determine their correlation with the antioxidant activity of the leaf extracts and expressed in Gallic acid and quercetin equivalents, respectively.Results: The results showed that the content of total phenols and flavonoids was found to be high in ethyl acetate extract which was recorded as 61.02±1.30 mg Gallic acid equivalent (GAE/g) and 81.72±0.61 QE/g, respectively. Compared to other extracts, ethyl acetate leaf extract was found to possess high antioxidant activity at p<0.05 level, with high percentage of inhibition at 100 μg/ml concentration (82.70%) toward hydrogen peroxide radical scavenging with IC50 value 62.94 μg/ml. Statistically, two-tailed Pearson's correlation showed strong positive correlations between hydrogenperoxide radical scavenging activity and total phenolic contents (TPC) (r=1.000) at p<0.05 level.Conclusion: The results obtained in this study clearly signifies that the ethyl acetate leaf extract of C. coromandelicum has high content of total phenols which are correlated to its antioxidant activity and thus has the potential to use as a source of natural antioxidants and can be explored as a therapeutic agent in free radical induced diseases.Â

Bioencapsulation strategy and highly unsaturated fatty acids (HUFA) enrichment in Artemia franciscana nauplii by using marine trash fish Odonus niger liver oil

To investigate the maximum accumulation of highly unsaturated fatty acids (HUFA) in Artemia franciscana nauplii through bioencapsulation process, five different concentrations (0 - 4%) of emulsified Odonus niger liver oil were prepared. The prepared emulsions were used to enrich A. franciscana at different time intervals of 6, 12, 18 and 24 h. After the enrichment period, the fatty acid composition of the nauplii were analysed and estimated individually along with freshly hatched A.franciscana and O. niger liver oil. The HUFA content such as linoleic (18: 2n-6), linolenic (18: 3n-3), arachidonic (20: 4n-3 + n-6), eicosapentaenoic (20: 5-n3) and docosahexaenoic (22: 6n-3) acids were 12.87, 0.21, 2.66, 2.86 and 2.30% dry weight (DW), respectively, in O. niger liver oil, and 8.60, 17.20, 1.80, 2.40 and 0.1% DW, respectively, in freshly hatched A. franciscana. During 6 – 24 h of enrichment period, all the above HUFA increased considerably from 8.76 to 10.84, 17.24 to 23.84, 1.16 to 3.98, 2.45 to 5.88 and 0.30 to 2.69% DW, respectively. The increase in the level of individual HUFA of A. franciscana enriched with various concentrations of emulsified liver oil at different time durations showed a positive linear relationship and the correlation coefficient obtained were statistically significant (P< 0.05)

Screening and characterization of antimicrobial secondary metabolites from Halomonas salifodinae MPM-TC and its in vivo antiviral influence on Indian white shrimp Fenneropenaeus indicus against WSSV challenge

AbstractAntimicrobial secondary metabolites from extremophiles play a significant role in the pharmacological industry due to their stable and strong activity and it is used in the treatment of microbial infections. In the present work, Halomonas salifodinae MPM-TC (M. Peter Marian-T. Citarasu) was isolated from the solar salt works in India and identified by 16S rRNA sequencing. The secondary metabolites were extracted from H. salifodinae MPM-TC and tested for antibacterial activity against aquatic bacterial pathogens such as Vibrio harveyi, Vibrio parahaemolyticus, Pseudomonas aeruginosa and Aeromonas hydrophila isolated from infected fish/shrimp, and it effectively controlled them with more than 10mm of zone of inhibition. The metabolites were purified through silica column chromatography and in vitro antiviral activity was performed against White Spot Syndrome Virus (WSSV) using different fractions. Among the different tested fractions, fraction-III (F-III) was able to suppress WSSV replication. Shrimps challenged with a WSSV inoculum incubated with F-III and treated Fenneropenaeus indicus survived around twice as many as the controls. Gas chromatography–mass spectroscopic (GC–MS) analysis revealed that the antiviral active fraction contains around eight compounds including Perfluorotributylamine, Cyclopentane, 1-butyl-2-ethyl and 1,1′-Biphenyl]-3-amine. Further the active fraction F-III was incorporated in the artificial diets at the concentration of 200 (HS1), 400 (HS2) and 800 (HS3) μgkg−1 and fed to F. indicus for 30days. After 30days of culture, shrimps were challenged with virulent WSSV and studied for WSSV VP 28 gene expression, biochemical, haematological and immunological changes. Surprisingly, groups treated with lower concentrations of fraction F-III (HS1 or HS2) significantly (P<0.05) suppressed the viral replication. Different levels of protein and glucose, improved total haemocyte count (THC), coagulase activity and oxyhaemocyanin level all were comparable to controls. Also, immunological parameters such as prophenol oxidase and intracellular superoxide anion production were significantly increased (F=97.18; P⩽0.001 and F=5.70; P⩽0.05) in the groups treated with the three test concentrations. The presence of antiviral and immunostimulant active principles in the F-III fraction effectively suppressed the WSSV load and boosted F. indicus’s immune system. This research will help to develop novel antiviral drugs from plants against aquatic important pathogens

cDNA cloning and expression of vitamin-K Epoxide Reductase (VKOR) gene from orange spotted grouper Epinephelus coioides

The cDNA sequence encoding Vitamin-K Epoxide Reductase (VKOR) was constructed from hypothalamus and pituitary cDNA libraries of Orange spotted Grouper Epinephelus coioides. VKOR gene contains 610 nucleotides including the open reading frame (ORE) of length 399 bp. ORE was started with ATG codon at the position of 199 and terminated with a stop codon TAA of 585. ORE sequence had 128 amino acids and the predicted protein molecular weight is approximately 14.08 kDa (GenBank: FJ939281.1). Expression of VKOR gene in different tissues was further examined using one-step RT-PCR. RT PCR result revealed that, VKOR mRNA expression was detected only in liver, kidney, hypothalamus and pituitary. VKOR gene sequence of E. coioides aligned with related sequences by multiple sequence alignment by clustal X and phylogenetic tree was generated by Neighbor Joining method using Genious v 5.4. E. coioides VKOR had the highest identity and homology with the puffer fish, Tetraodon nigroviridis, Takifugu rubripes and Zebra fish Danio rerio. Based on the VKOR expression in the kidney, liver hypothalamus and pituitary of the seven year's old (sex reversal stage) E. coioides, it may be involved in sex reverse.The cDNA sequence encoding Vitamin-K Epoxide Reductase (VKOR) was constructed from hypothalamus and pituitary cDNA libraries of Orange spotted Grouper Epinephelus coioides. VKOR gene contains 610 nucleotides including the open reading frame (ORE) of length 399 bp. ORE was started with ATG codon at the position of 199 and terminated with a stop codon TAA of 585. ORE sequence had 128 amino acids and the predicted protein molecular weight is approximately 14.08 kDa (GenBank: FJ939281.1). Expression of VKOR gene in different tissues was further examined using one-step RT-PCR. RT PCR result revealed that, VKOR mRNA expression was detected only in liver, kidney, hypothalamus and pituitary. VKOR gene sequence of E. coioides aligned with related sequences by multiple sequence alignment by clustal X and phylogenetic tree was generated by Neighbor Joining method using Genious v 5.4. E. coioides VKOR had the highest identity and homology with the puffer fish, Tetraodon nigroviridis, Takifugu rubripes and Zebra fish Danio rerio. Based on the VKOR expression in the kidney, liver hypothalamus and pituitary of the seven year's old (sex reversal stage) E. coioides, it may be involved in sex reverse