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The aim of the present study was to elucidate the anti-diabetic effects of the crude polysaccharide and rhamnose-enriched polysaccharide derived from G. lithophila on streptozotocin (STZ)-induced diabetic Wistar rats. Treatment with crude polysaccharide and rhamnose-enriched polysaccharide showed increases in body weight and pancreatic insulin levels and a decrease in blood glucose levels compared with control diabetic rats. The blood concentrations of total cholesterol (TC), triglycerides (TGs), low-density lipoprotein (LDL) and very-low-density lipoprotein (VLDL) decreased, and high-density lipoprotein (HDL) increased both in the crude polysaccharide- and rhamnose-enriched polysaccharide-treated rats. Superoxide dismutase (SOD) and glutathione peroxidase (GPx) levels increased, and malondialdehyde (MDA) levels decreased in the livers, kidneys and pancreases of crude polysaccharide- and rhamnose-enriched polysaccharide-treated rats. Immunohistological examination further confirmed that restoration of the normal cellular size of the islets of Langerhans and the rebirth of β-cells were found to be greater in the body region than in the head and tail regions of the pancreas. The crude polysaccharide- and rhamnose-enriched polysaccharide-treated diabetic rats showed normal blood glucose levels and insulin production, and reversed cholesterol levels and enzymatic actions. Therefore, rhamnose-enriched polysaccharide from G. lithophila acts as a potent anti-diabetic agent to treat diabetes and can lead to the development of an alternative medicine for diabetes in the future.

The present study was performed to utilize the microalgae biomass as a potential source for fatty acids and to evaluate its bioactivity through an antioxidant assays for the microalga Nannochloropsis sp. The alga comprises maximum saturated fatty acids (52.23 ± 0.481%), followed by polyunsaturated fatty acids (25.49 ± 0.043%) and monounsaturated fatty acids (22.28 ± 0.643%), respectively. Proximate composition and the active functional group Nannochloropsis sp. were analyzed by standard biochemical methods and Fourier transform infrared (FTIR) analysis. Antioxidant potential of the methanolic extract was evaluated by total antioxidant activity, 1,1-diphenyl-2-picrylhydrazyl (DPPH), superoxide radical scavenging, and hydroxyl radical scavenging assays. In all these assays, activity was expressed in concentration dependent manner and maximum activity was recorded at higher concentration of 250 µg/ml. This study concludes that Nannochloropsis sp. possessed the potential antioxidant properties and fatty acid profile. So, this alga can be used as a natural antioxidant to prevent the oxidative stress and various industrial applications like aqua feed formulation, biofuel production, and biomedicine in the future.

This study aimed to investigate the phytochemical profile, the antibacterial and antioxidant activities of mass cultured Spirulina platensis and the structural characterization of the methanolic extracts. S. platensis was isolated from estuary and mass cultured in the laboratory, methanol was used for extraction, and the yield was calculated as 16%. The seven phytochemicals were quantified by standard procedure, and the phenol was found maximum 96.7 µg/mg followed by alkaloid 89.5 µg/mg. The antibacterial activity was performed by disc diffusion method, and the maximum activity was noted against Salmonella typhi (19 mm) followed by Escherichia coli (12 mm). The antioxidant effect was recorded as concentration-dependent, and in all, the antioxidant effect was recorded in higher concentration level including as standards. In FT-IR spectral analysis, the active functional groups alkyl halide and amine were recorded simultaneously in 1H NMR analysis, 11 chemical shifts were recorded, and it revealed the stretching of alkane, allylic, benzylic, ketone, alkyne and esters. From the results, the present study concluded that S. platensis is an alternate source of pharmaceutical industry due to its phytochemical profile and bioactive potential.

The physical characteristics such as specific gravity, solidification point, saponification value, iodine, and acid value of liver oil were analyzed, and it has showed good characters similar to fish oil. The heavy metal was found in permissible limit, ash, and moisture was recorded in moderate level, protein, and carbohydrate were found as trace amount; the bad cholesterol was noticed lower and good cholesterol was high than fish oil. Ten fatty acids were recorded, the oleic acid, ω-3, EPA, DHA, and α- linolenic acid were recorded maximum level. The essential and non-essential amino acids were estimated and the fat soluble vitamins (A and E) were noticed higher level. The minerals Zn and Fe were found moderate level. The liver oil has showed antibacterial effect against 7 bacterial strains. The six antioxidant activities of liver oil have showed concentration depended. The Sepioteuthis lessoniana liver oil has showed against A549 cell lines.

The chemical composition, structural features and bioactivities of chitosan derivatives were studied and reported here. Chitin and chitosan was extracted by chemical methods and yield was recorded 35% and 41%, moisture and ash has showed 7.4, 0.73, 1.2 and 0.79% and Ca was recorded high level (590 ppm). The IR spectrum has showed NH primary and secondary amines, OH group and several sugar stretching. N-acetyl glucosamine (GlcNAc), H-2 proton of glucosamine (GlcN), Anomeric protons, H-1 [GlcN (H-1D), GlcNAc (H-1A)] were noticed in 1H NMR and anomeric, methyl carbon atoms in 13C NMR. Rough surface, irregular block, crystalline with cluster and porosity structure was noticed by SEM observation. Antioxidant effect has showed good and concentration depended in four antioxidant assays. The α-amylase and β-glucosidase enzyme inhibition effect of chitosan has showed promising and dose depend also anticoagulant potential. Chitosan could be used in pharmaceutical industry and tissue engineering.

The present study was aimed to investigate the nutritional composition, chemical components and anti-inflammatory activity of C. vulgaris. The isolated microalga was mass cultured in laboratory by selective media with optimum conditions. Protein content was found to be higher for 45.23% followed by carbohydrate (23.43%) and total lipid (18.12%). Minerals components was recorded by the following order: Magnesium > Calcium > Iron > Manganese > Zinc > Copper. Seven vitamins were estimated, vitamin B3 was recorded higher (13.3 mg/100 g) and B12 has shown lesser amount (0.21 mg/100 g). The pigments chlorophyll a, b and carotenoids were found to be 4.7 mg/g, 4.2 mg/g and 6.11 mg/g respectively. Secondary metabolite was extracted by methanol and chloroform and further screened for the in vitro anti-inflammatory effect through the inhibition of albumin denaturation, antiproteinase, hypotonicity-induced haemolysis and anti-lipoxygenase assays and results have been recorded by concentration dependent. From the results, the anti-inflammatory activities of the methanolic extract were found higher than the chloroform extract at 500 µg/ml. The functional groups of the potent methanolic extract were studied by FT-IR analysis which revealed the presence of alkane, nitro and carboxyl groups.

The present study was aimed to evaluate the nutritional fact and anticancer effect of liver oil extracted from the squid Loligo duvauceli. The ash and moisture content was noted as 3.21 and 2.13% whereas the total protein, lipid and carbohydrate, recorded 23.44, 21.23 and 2.13% respectively. The water and fat soluble vitamins and minerals were found in good level. The lipid profile was noted similarly to cod liver oil at the same time the good cholesterol was found in high level. Ten fatty acids were found especially DHA, EPA, omega-3 in high level. The essential amino acids were recorded at high level whereas Non-essential amino acids, moderate level. The anticancer ability of oil against lung cancer cell line (A549) showed better activity, up to 70% inhibition was noted at 960 µg/ml with the CTC50 of 260 µg/ml. In addition to this, there cytotoxicity effect was seen in Vero (normal) cell lines. The present study concludes that the L. duvauceli liver oil is a good source of nutrition and provides good health benefits to human.

The ash, moisture, protein and carbohydrate content of the extracted oil were estimated as 4, 7, 4 and 1% respectively. The Vitamins A and E was found to be 59.14 and 17.4 mg/100 g. Nine fatty acids were found and the Linoleic acid, DHA and EPA were recorded as higher. The specific gravity, solidification point, refractive index, saponification value, iodine value and acid value were also calculated and found as 1.056, 1.2 °C, 1.465 µ, 50.5, 20.4 mg and 0.075 mg KOH/g. The antioxidant activity that includes total antioxidant activity, DPPH & Hydroxyl radical scavenging assay, reducing power and ferrous ion chelating activity of liver oil was studied by following standard methods and results of all the assays was found increasing with increase in concentration. Further purification of liver oil and studying its antioxidant property in animal model shall form the basis for its use as a potent alternate natural antioxidant and nutrition supplements.

The chitin and chitosan of S. prashadi was prepared through demineralization, deproteinzation, deacetylation process and sulfation were carried by chlorosulfonic acid in N,N-dimethylformamide. The sulfate content in chitosan was found to be 18.9%. The carbon, hydrogen and nitrogen composition of the sulfated chitosan were recorded 39.09%, 6.95% and 6.58% respectively. The structural analysis was done by using FT-IR and NMR spectroscopy technique. The DSC curves of sulfated chitosan showed a large endothermic peak resolved with To value of 54.57°C and TP value of 97.46°C. The morphology of sulfated chitin and sulfated chitosan were studied by SEM. The Further in vitro antioxidant activity of sulfated chitosan was screened by scavenging activity of superoxide radical assay, hydroxyl radical scavenging assay, metal-ion chelating effect and reducing power. Its anticoagulant activity was tested for human plasma with respect to Activated Partial Thromboplastin Time (APTT) and Prothrombin Time (PT). Results prove that sulfated chitosan has potent antioxidant and anticoagulant activity.

Exopolysaccharide (EPS) designated as HMEPS was isolated from Halolactibacillus miurensis and purified through gel filtration chromatography. EPS extracted from the supernatant yielded a 56.1% total carbohydrate content. The ash and moisture content were 12.1% and 1.5% respectively. Galactose and glucose were found as main monosaccharides of the HMEPS through HPLC analysis. FT-IR spectra of the HMEPS revealed its composition with hydroxyl, alkenes, amide and carboxyl as functional groups and aliphatic amine and alkynes at the fingerprint region. In vitro antioxidant activity was investigated against hydroxyl, DPPH, superoxide free radicals and the scavenging activity against all were found to be dose dependent proportionately. HMEPS showed higher reducing ability against superoxide radical and potency in chelating the ferrous ions. 10mg/ml of HMEPS was found equivalent to 2.7units of ascorbic acid through the total antioxidant assay. Phylogenetic relation of H. miurensis SEEN MKU3 (GenBank number KT803852) was plotted with MEGA 5.0.

The crude polysaccharide was extracted from Grateloupia lithophila through hot-water extraction and deproteinization. Further, fractionated by anion-exchange column using Q-Sepharose and purified by gel-permeation chromatography using Sepharose 4-LB column. The crude and purified polysaccharide contains high carbohydrate (75.7 and 89.7%), ash (18.2 and 3.2%) and moisture (14.8 and 1.3%); the protein and uronic acid were absent. The molecular weight of crude, fractionated and purified polysaccharide was found to be 37 kDa, 29 kDa and 24 kDa. The monosaccharide composition of the crude polysaccharide was found to be having rhamnose (79.82%), fructose (8.38%), galactose (3.95%), xylose (3.31%) and glucose (1.48%); whereas the purified polysaccharide reported higher amount of rhamnose (95.88%), 1.13% of xylose and 2.21% of fructose. The structural elucidation of the purified polysaccharide was conformed as α-l-rhamnose through polarimetry, FT-IR and 1H NMR spectroscopy.

The sulfated mucopolysaccharide (GAG) was isolated from S. pharonis and the carbohydrate and protein content was found to be 62.4% and 3.9%. The disaccharide profile of sulfated GAG composed glucuronic acid, N-acetyl glucosamine and sulfate content by contributing 50.11%, 38.00% and 27.69% respectively. The carbon, hydrogen and nitrogen content of the sulfated GAG showed 14.80%, 1.68% and 2.99% respectively. The molecular weight of sulfated GAG was calculated as 27kDa and the structural characterization was done by Fourier Transform Infrared (FT-IR) and NMR Spectroscopy. The Activated Partial Thromboplastin Time (APTT) and Prothrombin Time (PT) of sulfated GAG were determined as 91 IU and 39.55 IU at 25μg/ml respectively. Further the sulfated GAG reported the cytotoxic effect (CC50) of 1100μg/ml concentration on Vero cell line. The sulfated GAG reported the anticancer activity against HeLa cell line with an inhibition rate of 18.65%-66.13% at 50-250μg/ml concentration. The sulfated GAG can be considered as a potent anticoagulant and anticancer drug in future.

To evaluate the antioxidant potential of phosphorylated chitosan from the Sepioteuthis lessoniana gladius that is thrown as waste in seafood processing industries. Chitin was extracted from the gladius of S. lessoniana by demineralization and deproteinization. Chitosan was obtained by deacetylation of chitin in strong alkali and phosphorylated by using ortho-phosphoric acid. The structure of phosphorylated chitosan was elucidated through Fourier Transform Infra Red (FT-IR) spectroscopy. The antioxidant efficacy of phosphorylated chitosan was assessed through scavenging activity against 2,2′-azinobis(3-ethylbenzothiazoline-6-sulfonic acid diammonium salt) (ABTS), 1, 1-diphenyl-2-picrylhydrazyl (DPPH), superoxide and hydroxyl radicals. The ability to chelate ferrous ion was also assessed. The yield and water solubility of phosphorylated chitosan was calculated as 81% and 78% respectively. The antioxidant activity of phosphorylated chitosan was found concentration dependent. The phosphorylated chitosan scavenges DPPH, ABTS, superoxide and hydroxyl radicals by 56.97%, 76.56%, 71.57% and 56.33% respectively and Inhibitory Concentration (IC50) values of scavenging those radicals were reported as 4.53, 0.46, 0.38 and 0.5 mg/ml, respectively. The chelating ability of phosphorylated chitosan was 59.56% with IC50 of 4.07 mg/ml. Moreover, the antioxidant activity of phosphorylated chitosan was compared with that of the standards such as Butylated Hydroxy Anisole (BHA), ascorbic acid, α-Tocopherol and Ethylene Diamine Tetra Acetic acid (EDTA). The results of the present investigation suggest that the phosphorylated chitosan with potent antioxidant property could be used as an alternative antioxidant that from a natural non-conventional source.

The Sulfated polysaccharide was purified through anion-exchange and gel permeation column chromatography. The isolated sulfated polysaccharide from C. corticata contains 84% of carbohydrate, 0% of protein, 19.7% of ash and 29.4% of moisture was found. The carbon, hydrogen, nitrogen and sulfur content as 33.19%, 5.91%, 7.21% and 3.75%. The molecular weight of sulfated polysaccharide was found to be 43kDa. The sugar was composed of (90.11%), glucose (5.47%), xylose (2.30%) and mannose (2.12%). The structural feature of sulfated polysachharide was studied through FT-IR and 1H NMR spectral analysis. Further the sulfated polysaccharide showed total antioxidant activity of 24.93%-75.21% at 50-250μg/ml, DPPH free radical scavenging activity of 23.12%-73.01% at 10-160μg/ml, ABTS scavenging activity of 15.8%-74.5% at 25-125μg/ml hydroxyl radical scavenging activity 12.87-69.19% at 25-125μg/ml and superoxide radical scavenging activity 28.10-78.11% at 50-250μg/ml respectively. The sulfated polysaccharide has shown good antibacterial activity against human pathogen.

Sulfated polysaccharide was extracted from the internal shell (gladius) of Sepioteuthis lessoniana. The sulfated polysaccharide contained 61.3% of carbohydrate, 0.8% of protein, 28.2% of ash and 1.33% of moisture respectively. The elemental composition was analyzed using CHNS/O analyzer. The molecular weight of sulfated polysaccharide determined through PAGE was found to be as 66 kDa. Monosaccharides analysis revealed that sulfated polysaccharide was composed of rhamnose, galactose, xylose and glucose. The structural features of sulfated polysaccharide were analyzed by FT-IR and NMR spectroscopy. Further the sulfated polysaccharide was evaluated for its antibacterial activity against selected human clinical pathogens, namely Staphylococcus aureus, Klebsiella pneumoniae, Salmonella typhi, Vibrio cholerae, Klebsiella oxytoca, Escherichia coli, Salmonella paratyphi, Proteus mirabilis, Vibrio parahaemolyticus and Streptococcus pyogenes using agar well diffusion method. The polysaccharide has showed good antibacterial activity and MIC and MBC have also been evaluated. The anticancer activity was tested against HeLa cell line by MTT assay. The Cytotoxic Concentration (CC50) was observed as 700 μg/ml and the maximum anticancer activity of 62.89% was recorded at 200 μg/ml; whereas, the lowest of 9.87% was observed at 25 μg/ml. In conclusion, the sulfated polysaccharide is an alternate, non-toxic and cheap source of substance that showed good antibacterial and anticancer acitivity.

Sulfated polysaccharide was isolated from Monostroma oxyspermum through hot water extraction, anion-exchange and gel permeation column chromatography. The sulfated polysaccharide contained 92% of carbohydrate, 0% of protein, 7.8% of uronic acid, 22% of ash and 33% of moisture respectively. The elemental composition was analyzed using CHNS/O analyzer. The molecular weight of sulfated polysaccharide determined through PAGE was found to be as 55 kDa. Monosaccharides analysis revealed that sulfated polysaccharide was composed of rhamnose, fructose, galactose, xylose, and glucose. The structural features of sulfated polysaccharide were analyzed by NMR spectroscopy. Further the sulfated polysaccharide showed total antioxidant and DPPH free radical scavenging activity were as 66.29% at 250 μg/ml and 66.83% at 160 μg/ml respectively. The sulfated polysaccharide also showed ABTS scavenging ability and reducing power were as 83.88% at 125 μg/ml and 15.81% at 400 μg/ml respectively. The anticoagulant activity was determined for human plasma with respect to Activated Partial Thromboplastin Time (APTT) and Prothrombin Time (PT) was 20.09 IU and 1.79 IU at 25 μg/ml respectively. These results indicated that the sulfated polysaccharide from M. oxyspermum had potent antioxidant and anticoagulant activities.

The acid soluble collagen (ASC) and pepsin soluble collagen (PSC) were extracted from the outer skin of Sepiella inermis and further characterized partially. The yield of ASC was low (0.58% on dry weight basis); whereas the yield of PSC was comparatively more (16.23% on dry weight basis). The protein content in ASC and PSC was calculated as 20.24 and 69.56%, respectively (on dry weight basis). The sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) gel profile showed two bands for ASC and PSC with corresponding molecular weight of 86 and 67 kDa and 86, 63 and 58 kDa respectively. The differential scanning calorimetry (DSC) results showed that ASC withstand up to 75.93°C whereas the PSC withstand up to 75.05°C. The fourier transform infrared spectroscopy (FT-IR) spectrum of both ASC and PSC recorded 11 and 13 peaks, respectively. The fine structure of both ASC and PSC was also studied using scanning electron microscopy (SEM). Key words: Sepiella inermis, acid soluble collagen (ASC), pepsin soluble collagen (PSC), differential scanning calorimetry (DSC), fourier transform infrared (FT-IR) spectroscopy, scanning electron microscopy (SEM).