Incidence of mud crab reovirus (MCRV) outbreak in polyculture ponds of Andhra Pradesh, south east coast of India.

Reovirus designated as Mud crab reovirus (MCRV) is associated with the mass mortalities of mud crabs resulting in significant economic loss to crab and shrimp-mud crab polyculture farmers in the Nagayalanka, Krishna district, Andhra Pradesh. The 100 % chronic mass mortalities have been attributed to the outbreak of Mud crab reovirus (MCRV) in the polyculture farms. The moribund crabs showed autotomy, discoloration of carapace, loss of appetite, slow movement and loose gills. Histopathological observations of the infected mud crabs showed an atrophied hepatopancreas, complete degeneration of tissues along with viral inclusions in hepatopancreas, gills and muscles. Further analysis using Transmission electron microscopy (TEM), showed that the viral particles had a diameter of 70 nm and exhibited a non-enveloped, icosahedral shape arranged in a crystalline manner. The virus mainly infects the connective tissue of hepatopancreas, gills, muscle and develops in the cytoplasm. RT-PCR reconfirmed the presence of reovirus in the hepatopancreas of spontaneously infected mud crab Scylla serrata. The current study shows the importance of monitoring the MCRV prevalence in polyculture farms to minimize its spread and precautionary measures can be taken by screening the brooders from the crab hatchery and stocking of wild crabs without screening should be avoided in order to prevent MCRV outbreak.

Synergistic induction of apoptosis in lung cancer cells through co-delivery of PLGA phytol/α-bisabolol nanoparticles.

This study explored the potential of poly-(lactic-co-glycolic) acid (PLGA) nanoparticles to enhance the effectiveness of anticancer treatments through combination therapy with phytol and α-bisabolol. The encapsulation efficiency of the nanoparticles was investigated, highlighting the role of ionic interactions between the drugs and the polymer. Characterization of PLGA-Phy+Bis nanoparticles was carried out using DLS with zeta potential and HR-TEM for size determination. Spectrophotometric measurements evaluated the encapsulation efficiency, loading efficiency, and in vitro drug release. FTIR analysis assessed the chemical interactions between PLGA and the drug actives, ensuring nanoparticle stability. GC-MS was employed to analyze the chemical composition of drug-loaded PLGA nanocarriers. Cytotoxicity was evaluated via the MTT assay, while Annexin V-FITC/PI staining and western blot analysis confirmed apoptotic cell death. Additionally, toxicity tests were performed on L-132 cells and in vivo zebrafish embryos. The study demonstrates high encapsulation efficiency of PLGA-Phy+Bis nanoparticles, which exhibit monodispersity and sizes of 189.3±5nm (DLS) and 268±54 nm (HR-TEM). Spectrophotometric analysis confirmed efficient drug encapsulation and release control. FTIR analysis revealed nanoparticle structural stability without chemical interactions. MTT assay results demonstrated the promising anticancer potential of all the three nanoparticle types (PLGA-Phy, PLGA-Bis, and PLGA-Phy+Bis) against lung cancer cells. Apoptosis was confirmed through Annexin V-FITC/PI staining and western blot analysis, which also revealed changes in Bax and Bcl-2 protein expression. Furthermore, the nanoparticles exhibited non-toxicity in L-132 cells and zebrafish embryo toxicity tests. PLGA-Phy+Bis nanoparticles exhibited efficient encapsulation, controlled release, and low toxicity. Apoptosis induction in A549 cells and non-toxicity in healthy cells highlight their clinical potential.

An effective bio-inspired synthesis of platinum nanoparticles using Caulerpa sertularioides and investigating their antibacterial and antioxidant activities.

In this study, we report the synthesis of platinum nanoparticles (Cs-PtNPs) using an aqueous extract of Caulerpa sertularioides as a reducing agent. Cs-PtNPs were characterized by UV-Vis spectroscopy, fourier transform infrared (FT-IR) spectroscopy, X-ray diffraction (XRD), field emission electron microscopy (FE-SEM), energy-dispersive X-ray spectroscopy (EDAX), high-resolution transmission electron microscopy (HR-TEM) and dynamic light scattering (DLS) analysis. Cs-PtNPs are spherical with a particle size of 6-22 nm. Cs-PtNPs have been shown to have highly effective antioxidant activities with 74% for DPPH, 63% for reducing power, and 59% for total antioxidant at 1 mg/ml, and results were compared with standard L-ascorbic acid. Furthermore, the Cs-PtNPs demonstrated excellent antibacterial activity against the Gram-negative bacteria, Vibrio parahaemolyticus with the highest zone of inhibition (18 mm) at 50 µg/ml. Moreover, Artemia nauplii showed less toxicity when treated with Cs-PtNPs at 150 µg/ml, indicating that the Cs-PtNPs are less toxic and environment friendly.

Polysaccharide extracted from Taraxacum platycarpum root exerts immunomodulatory activity via MAPK and NF-κB pathways in RAW264.7 cells.

ETHNOPHARMACOLOGICAL RELEVANCE: Taraxacum platycarpum Dahlst. (Korean dandelion) is a medicinal herb used in traditional medicine in Korea to treat various disease such as furuncles, mammitis, hepatitis, jaundice. Moreover, a decoction prepared from T. platycarpum leaves and stems is an effective treatment for cancer, glycosuria, liver disease, pleurodynia, and stomach problems. AIM OF THE STUDY: The main objective of this work was to study the composition and structural properties of polysaccharides (TPP) from Taraxacum platycarpum Dahlst. root and investigate the immunostimulatory activity on RAW264.7 cells. MATERIALS AND METHODS: TPP was extracted from T. platycarpum using hot water extraction, ethanol precipitation method and its fractionated using DEAE-Sepharose fast flow column. The composition, molecular weight, and structural characterization of TPP and its fractions were evaluated by various techniques. Further, the immunostimulatory activity of polysaccharides was tested on murine macrophage cell line RAW264.7 by various in vitro assays. The structure effect of TPP on RAW264.7 cells was studied by the removal of sulfate (desulfation) and protein (deproteinization) contents from TPP. RESULTS: We obtained three fractions namely TPP-1, TPP-2, and TPP-3 which mainly consisted of carbohydrates (75.55, 52.71, and 48.41%), sulfate (8.42, 15.19, and 27.67%), uronic acid (1.27, 6.56, and 4.39%), and protein (8.15, 24.85, and 9.73%). The average molecular weight of the fractions was 56.7, 108.2, and 132.3 × 103 g/mol, respectively. The polysaccharides activate the RAW264.7 cell to produce a significant amount of NO and upregulate the various mRNA expression by the activation of MAPK and NF-κB pathways via TLR4, TLR2, and CR3 receptors. The structurally modified deproteinated derivative (DP-TPP-2) more effectively decreases the NO production which means the protein content of TPP-2 mainly contributes to the RAW264.7 cells activation. The structure of DP-TPP-2 primarily consists of 1 â†’ 2)-Galp, 1 â†’ 6)-Glup, 1 â†’ 2) - Rhap, and 1 â†’ 5) - Arap glycosidic linkages. CONCLUSIONS: The present study demonstrated that the polysaccharide isolated from T. platycarpum shows admirable immunostimulatory by the activation of MAPK and NF-κB pathways through TLR4, TLR2, and CR3 receptors. The protein content of polysaccharides mainly contributes to the RAW264.7 cells activation. Our study results could be useful for developing a new immunostimulant agent.

Surveillance of disease incidence in shrimp farms located in the east coastal region of India and in vitro antibacterial efficacy of probiotics against Vibrio parahaemolyticus.

We surveyed 130 shrimp farms located on the eastern coast of India to determine the prevalence of emerging diseases in Litopenaeus vannamei and Penaeus monodon. Live shrimps were collected from the farms based on external symptoms. The biochemical, molecular, and histopathology results confirmed infection with Enterocytozoon hepatopenaei (32.4%), Vibrio parahaemolyticus (27.7%), White Spot Syndrome Virus (25.4%), Vibrio alginolyticus (16.1%), Vibrio harveyi (13.1%), Monodon-type baculovirus (4.61%), and infectious Hematopoietic Necrosis Virus (2.3%) in the collected shrimps. Enterocytozoon hepatopenaei (EHP) occurred more frequently in L. vannamei than P. monodon, with the microsporidian spores in the hepatopancreas. In P. monodon, Monodon-type Baculovirus infection (33.3%) was dominant and small percentages of WSSV, IHHNV, V. alginolyticus, and V. harveyi were observed. A few ponds were observed with co-infection of EHP and WSSV (7.6%), V. parahaemolyticus and WSSV (4.6%) and also V. parahaemolyticus and EHP (6.1%). Among the Vibrio spp, V. parahaemolyticus showed the highest percentage of infection in L. vannamei. Overall, we found that shrimp were chiefly infected with EHP and V. parahaemolyticus. The impact of water quality parameters on shrimp diseases was not addressed in this study. In an antibiotic susceptibility study, V. parahaemolyticus isolated from L. vannamei ponds was susceptible to nitrofurantoin, chloramphenicol, oxytetracycline and tetracycline, but resistant to erythromycin and nalidixic acid. In a preliminary in vitro antibacterial activity assay, probiotics against V. parahaemolyticus showed high inhibitory activity and the results encourage further in-depth studies on the efficacy of probiotics for disease control and prevention in shrimp farms.

Bioengineered gold nanoparticles from marine seaweed Acanthophora spicifera for pharmaceutical uses: antioxidant, antibacterial, and anticancer activities.

In this study, we report a green synthesis of pharmaceutically active gold nanoparticles from marine red alga Acanthophora spicifera by the reduction of chloroauric acid. The formation of A. spicifera-mediated gold nanoparticles (As-AuNPs) was characterized by several analytical techniques. The crystalline and face-centered cubic (fcc) structure were confirmed by X-ray diffraction (XRD) analysis. Electron microscopy results confirmed that As-AuNPs were spherical and the average size of particles was < 20 nm. As-AuNPs hold a significant level of antioxidant activities than A. spicifera extract. As-AuNPs exhibited the highest antibacterial activity against Vibrio harveyi than Staphylococcus aureus at 100 µg/ml. Furthermore, As-AuNPs exhibited the utmost cytotoxicity against human colon adenocarcinoma (HT-29) cells and registered the half-maximal inhibitory concentration (IC50) at 21.86 µg/ml. These findings authenticated that the synthesized As-AuNPs possess a broad spectrum of biological activities, and it can be effectively applied in the field of aquaculture and biomedical application.

Inducing inflammatory response in RAW264.7 and NK-92 cells by an arabinogalactan isolated from Ferula gummosa via NF-κB and MAPK signaling pathways.

The polysaccharide isolated from F. gummosa (FGP) was found homogenous with a weight average molecular weight (Mw) of 50.0 × 103 g/mol and radius of gyration (Rg) of 105.3 nm. The FGP was an arabinogalactan with a backbone formed of →6)-ß-Galp-1→ residues having random branching points at C-3 extended with either ß-Galp-(1→3)-ß-Galp-(1→ or α-Araf-(1→ side chain residues. FGP exhibited proliferative effect on RAW264.7 cells and induced macrophages to exert proinflammatory response releasing NO and up-regulating the transcription of cytokines including TNF-α, IL-1ß, IL-6 and IL-12. The FGP induced NK-92 cells to up-regulate the expressions of TNF-α, IFN-γ, granzyme-B, perforin, NKG2D and FasL. The presence of p-NF- κB, p-ERK, p-JNK and p-p38 in RAW264.7 and NK-92 cells indicated their activation through NF-κB and MAPKs signaling pathways. These findings suggested that polysaccharides from F. gummosa are potent in boosting immune system and thus may be considered for further studies of biomedical applications.

Studies on isolation, characterization of fucoidan from brown algae Turbinaria decurrens and evaluation of it's in vivo and in vitro anti-inflammatory activities.

In the present study, the anti-nociception and anti-inflammatory activity of fucoidan isolated from T. decurrens on formalin induced paw-edema in mice model were investigated. The extracted fucoidan contain 54.86% of total sugar, 23.51% of sulfate and 3.4% of protein. The monosaccharide composition analysis revealed that fucoidan encompassed of fucose (59.3%), galactose (12.6%), mannose (9.6%), rhamnose (6.4%) and xylose (11.4%). Further, the structural characterization was done by UV-visible spectroscopy, X-ray diffraction, FT-IR and 1HNMR analysis. The fucoidan reduced the licking time thereby suggesting anti-nociceptive effect and decreased the size of paw swelling in the formalin induced inflammatory edema condition. The isolated fucoidan could significantly decreased the MDA and also increase the SOD, CAT, GPx, GST and GSH activity in paw edema tissue of formalin injected mice. Furthermore, fucoidan administration retained p65/NF-κB transcription factor in the cytosol thereby showing down regulation of the gene expression of pro-inflammatory mediators such as IL-1ß, COX-2 and MMP-9 in fucoidan treated mice. The anti-inflammatory effect of fucoidan was attributed to its capacity on modulating the levels of enzymatic antioxidants, master regulator NF-κB and pro-inflammatory cytokines. The fucoidan has reduced LPS induced cytotoxicity in IC-21 macrophage at a dose depended on manner.

Sulfated galactan from Halymenia dilatata enhance the antioxidant properties and prevents Aeromonas hydrophila infection in tilapia fish: In vitro and in vivo study.

The structural characterization and pharmaceutical perspective of sulfated galactan from Halymenia dilatata (Hd-SG) were reported in this study. The Hd-SG consists of carbohydrate (58.5 ± 0.9%), sulfate (28.7 ± 0.9%) and protein (2.7 ± 0.2%). The existence of carbon (28.14%), hydrogen (5.50%), nitrogen (0.51%) and sulfur (8.26%) was confirmed in CHNS analysis. The Hd-SG was mainly comprising of galactose and mannose connected by (1 â†’ 4)-glycosidic linkages, and it shows the molecular weight of 900.9 × 103 g/mol in high-performance size exclusion chromatography (HPSEC). The Hd-SG exhibited the dose depended on antioxidant activities. The in vitro and in vivo studies proved the antibacterial efficacy of Hd-SG against Aeromonas hydrophila. The pre-treated Oreochromis fish with Hd-SG (2.0 g/0.1 kg of feed) showed the highest survival, antioxidant, and improved histological changes than the fish infected with A. hydrophila alone. These results concluded that the isolated Hd-SG has extensive therapeutic properties, and it can be used as preventive medicine.

Isolation, structural elucidation and immuno-stimulatory properties of polysaccharides from Cuminum cyminum.

The aim of the present study was to evaluate the effect of water-soluble polysaccharides from Cuminum cyminum to induce inflammatory response in immune cells and understand their underlying mechanisms. Weight average molecular weight (Mw) of polysaccharides varied between 191.4-512.2 × 103 g/mol. Polysaccharides induced RAW264.7 cells to release nitric oxide and express TNF-α, IL-1ß, IL-6 and IL-12 inflammatory cytokines. Polysaccharides activated NK-92 cells to produce TNF-α, IFN-γ, perforin, granzyme B, NKG2D and FasL. Activations of RAW264.7 and NK-92 cells were through NF-κB and MAPKs signal pathways indicated by the presence of phosphorylated NF-κB, ERK, JNK and p38 proteins. The polysaccharide structure was mainly constituted of →4)-Galp-(1→, →3)-Galp-(1→, →2)-Arap-(1→ and →2)-Arap-(1→ glycosidic linkages. Overall results suggested that polysaccharides from C. cyminum possessing lower MW and greater expanded conformation more effectively stimulate RAW264.7 and NK-92 cells and thus could be considered for further studies on their biomedical applications.

Isolation and structural characterization of sulfated polysaccharide from Spirulina platensis and its bioactive potential: In vitro antioxidant, antibacterial activity and Zebrafish growth and reproductive performance.

In this study, the sulfated polysaccharide (SPs) was isolated from Spirulina platensis. The isolated SPs contains carbohydrate, sulfate, protein and uronic acid at 38.7 ±â€¯0.30%, 21.3 ±â€¯0.87%, 7.1 ±â€¯0.15% and 7.9 ±â€¯0.4% respectively. The elemental analysis confirmed the presence of carbon (18.01 ±â€¯0.10%), hydrogen (1.83 ±â€¯0.02%) and nitrogen (3.43 ±â€¯0.01%). The monosaccharide composition and molecular weight of SPs were analyzed by high-performance liquid chromatography and size exclusion chromatography respectively. The monosaccharide composition analysis showed the existence of glucose, rhamnose, xylose, fucose, mannose, galactose and the molecular weight of SPs was 1016 kDa. Further, the characterization of SPs was done by UV-visible spectroscopy, X-ray diffraction, FT-IR, 1H NMR and 13C NMR analysis. The obtained SPs exhibited potent antioxidant activity in DPPH (76.45 ±â€¯0.49%), reducing power (absorbance: 1.3 ±â€¯0.02), hydrogen peroxide scavenging (66.3 ±â€¯1.16%), hydroxyl scavenging (68.6 ±â€¯3.2%), nitric oxide (81.36 ±â€¯1.85%) and total antioxidant (absorbance:1.66 ±â€¯0.02) activities at 5 mg/ml. In addition, SPs revealed the highest antibacterial efficacy against the pathogenic bacteria Vibrio vulnificus in disc diffusion, agar bioassay and protein leakage assays at 100 µg/ml. Furthermore, the supplementation of 2% SPs through a feed to the Danio rerio fish enhances the growth and reproductive performances. This finding confirmed that the isolated SPs from S. platensis possess pharmaceutical as well as nutritional properties.

Studies on structural properties and immune-enhancing activities of glycomannans from Schizophyllum commune.

Polysaccharides were extracted from Schizophyllum commune (a common mushroom) and their structural and immune-enhancing properties were investigated. Crude and fractions (F1 and F2) were composed of sugars (50.3-82.8%), proteins (1.46-20.1%), and sulfates (1.33-7.01%). Monosaccharide compositions of Cr and F1 were mainly composed of glucose (75.5% and 88.2%) with small amounts of mannose, galactose and xylose whereas the F2 was mainly composed of manose (55.2%) with minor amounts of galactose, glucose, and xylose. Their immune-enhancing activities were tested using RAW264.7 cells. Proliferation activity of RAW264.7 cells was over 100% after treatment with these polysaccharides. In addition, RAW264.7 cells produced large amounts of nitric oxide and various cytokines by up-regulating mRNA expression levels and the activation of nuclear factor-kappa (NF-κB) and mitogen-activated protein kinase (MAPK) after treatment with these polysaccharides. In addition, RAW264.7 cells were activated mainly through CR-3 and TLR-4 receptors. The backbone of F2 with excellent immune-enhancing activity was mainly linked by (1→3)-linked-mannopyranosyl and (1→2,3)-linked-mannopyranosyl residues.

Antibacterial efficacy of a fucoidan fraction (Fu-F2) extracted from Sargassum polycystum.

In this study therapeutic potential of fucoidan fraction-2 (Fu-F2) isolated from Sargassum polycystum was evaluated for the development of antibacterial agent against the human and animal pathogenic bacteria by in vitro and in vivo analysis. The Fu-F2 contained 51.12 ±â€¯0.86% of total sugar and 20.41 ±â€¯0.91% of sulfate. The structural characterization of Fu-F2 was performed by HPLC, FTIR and NMR analysis and reported in our earlier study. The in vitro antibacterial assays such as MIC, MBC, killing kinetics, disk diffusion, protein leakage, ROS and confocal laser scanning microscopy demonstrate that Fu-F2 possesses the highest antibacterial activity against the tested pathogens. Among the tested pathogens, the highest antibacterial activity (21 ±â€¯1.0 mm) was recorded at the concentration of 50 µg/ml against Pseudomonas aeruginosa and the lowest activity (16 ±â€¯0.53 mm) was registered against Staphylococcus aureus. In the in vivo analysis, the pre-treatment group with Fu-F2 at the concentration of 15 mg/0.1 kg through feed exhibited the highest survival (83.4%) and antioxidant activities (p < 0.05) than the fish infected with pathogen. Thus, the present findings suggest that the Fu-F2 of S. polycystum encompasses significant antibacterial properties and that can be used as a therapeutic agent for controlling the bacterial disease.

Investigation of antioxidant and anticancer potential of fucoidan from Sargassum polycystum.

The present study was aimed to evaluate the antioxidant and anticancer potential of fucoidan isolated from Sargassum polycystum. The isolated fucoidan was successfully purified by DEAE cellulose-ion exchange chromatography and dialysis. Totally four active fractions (F1-F4) were collected and explored its chemical constitution by calorimetric assays. Among them, fraction 2 (F2) showed the higher yield percentage, fucose and sulphate content. Further, monosaccharide composition, structural and functional properties of the F2 was analyzed by HPLC, FTIR and NMR. F2 shows highest DPPH radical scavenging activity (55.94 ±â€¯0.69%), reducing power (0.33 absorbance rate), hydrogen peroxide scavenging activity (71.76 ±â€¯2.14%) and nitric oxide radical scavenging activity (51.81 ±â€¯1.04%) at 1000 µg/ml. The cell viability of MCF-7 and HCT-15 cell lines was proportionate to the concentration of F2 with an estimated IC50 was 20 and 50 µg/ml respectively. The fluorescence and confocal laser scanning microscopic analysis demonstrated the apoptotic morphological changes and cell mediated death in F2 treated cancer cells. Higher amount of LDH release was found in the F2 treated cancer cells than the control group. Thus, the present finding proved that the isolated F2 encompasses significant antioxidant and anticancer property.