Chloroform Fraction from Methanol Extract of Starfish Acanthaster planci Stimulates Catfish (Clarias sp.) Macrophage Immunomodulatory Activity

Catfish (Clarias sp.) is Indonesia's most important and popular freshwater commodity, widely cultured and intensively grown. This study is aimed to find out the effects of the chloroform fraction from a methanol extract of Acanthaster planci on non-specific immunity in Clarias sp. Thin Layer Chromatography analysis of the methanol extract's chloroform fraction yielded three fractions: FrKl3.1, FrKl3.2, and FrKl3.3. The FrKl3.3 fraction of Acanthaster planci had a significant influence on Phagocytic Activity at doses of 0.1 (54.09 ± 8.99 %), 0.3 (48.16 ± 3.34 %), 0.5 (50.39 ± 5.71 %), and 0.7 mg/kg Body Weight (46.58 ± 0.37 %). At 0.5 mg/kg Body Weight (4.03 ± 1.40 %), the Phagocytic Index of FrKl3.2 was significantly higher than the control. At 0.5 mg/kg Body Weight (38.22 ±1.43 cell 106/ml) and 0.7 mg/kg BW (40.41 ± 1.92 cell 106/ml), the number of leukocytes/white blood cells was significantly higher than the control. Nitroblue Tetrazolium and Total Plasma Protein were not significantly different from the control. Based on the results,  the FrKl3.3 was leading on phagocytic capacity and number of leukocytes

Carrageenan Extraction of Kappaphycus alvarezii Seaweed from Nusa Lembongan Waters Using Different Alkaline Treatments

Kappaphycus alvarezii is a carrageenan-producing red seaweed that is widely cultivated in Nusa Lembongan waters, Bali, Indonesia. Carrageenan is generally extracted using an alkaline base,. iIn this study three different types of alkali were used to extract carrageenan originating from Nusa Lembongan Waters. This study aims to determine the quality of the extraction. The three alkalis used were KOH, NaOH, Ca(OH)2, and distilled water as a control. The 20 g dried seaweed was weighed, washed with the tap running water, and immersed in 0.15% alkaline solution (KOH, NaOH, Ca(OH)2) as well as distilled water for 24 hrs. Followed by rinsing with running water until pH was neutral. The solution was soaked and heated at 100ºC for 2 hours with a ratio of seaweed and water 1 kg in 20 L, and filtered. The extract was mixed with 1.25% KCI. The carrageenan precipitate was put in the oven at 60-80ºC until dry for 48 hours. The yield test results showed that the use of alkaline Ca(OH)2 resulted in a yield of 29.28% better than alkaline KOH (28.18%) and NaOH (27.7%). Based on the analysis of Fourier Transform Infrared Spectroscopy (FTIR), extraction using alkaline Ca(OH)2 showed functional groups characteristic of iota-carrageenan, whereas using alkaline KOH and NaOH showed functional groups characteristic of kappa-carrageenan

Sensitivity of Vibrio parahaemolyticus, V. vulnificus and V. harveyi Against Chloroxylenol (4-Chloro-3,5-dimethylphenol , C8H9ClO) Antiseptic and Pine Oil Disinfectant

Vibrio spp. genus is known as a marine indigeneous bacteria. Vibrio parahaemolyticus, V. vulnificus and  V. harveyi are pathogenic Vibrio. This study aims to assess the sensitivity of three Vibrio species (V parahaemolyticus, V. vulnificus and V. harveyi) isolated from shrimp pond against two type of disinfectant with different active compound namely Chloroxylenol (4-Chloro-3,5-dimethylphenol, C8H9ClO) and pine oil. The assessment was done by Kirby-Bauer disk diffusion methods in Zobell agar media with two different concentration (10 and 100 ppm) and replicated in three times. Sensitivity of Vibrio spp. was analized based on the inhibition zone activity produced by disinfectant. Results showed that sensitivity of Vibrio spp. against disinfectant Chloroxylenol 4.8% at 100 ppm were higher than 10 ppm. The increment of V parahaemolyticus was 182 %, V. vulnificus was 47 % and V. harveyi was 43 %, respectively.  Susceptibility of antiseptic with Chloroxylenol 4.8% at 100 ppm was arised to 152 % (V. parahaemolyticus), 43 % (V. vulnificus) and 31 % (V. harveyi) when compared to 2.5% pine oil disinfectant. It can be concluded that Chloroxylenol  4,8 % active compound and pine oil were able to inhibit the Vibrio spp. growth.

Alginate from Sargassum sp. Improve the Hematology Performance and Oxygen Tolerance Exposure of Lates calcarifer

Indonesia's fish production is abundant, especially in aquaculture. Lates calcarifer is farmed fish species. The failure of L. calcarifer cultivation due to disease problems. We are utilizing a natural compound derived from tropical Sargassum sp. extracts, namely alginate. This study aims to determine and analyse the supplementation of alginate in the diet by oral administration to improve the fish's hematological performance and oxygen stress tolerance. There were one control and three treatments (2 g.kg-1, 4 g.kg-1, and 8 g.kg-1 alginate/feed). The experiment was a completely randomized design with three replications. Fish were reared in a 350 L fiber tank for 12 days at a density of 20 ind.tank-1. Stress oxygen tolerance was applied by rearing the 10 fish in 12 L fully plastic-wrapped containers. The parameters test analyzed were phagocytic activity and phagocytic index, red blood parameters which are hematocrit and hemoglobin. The survival rate after 5 hr anoxic exposure was also recorded. The best treatments were achieved at a dose of 6 g.kg-1 and 8 g.kg-1in all parameters, except the phagocytic index. The alginate addition at all oxygen stress treatments also performed a better survival rate compared to the control. Adding alginate to feed as feed supplementation by oral administration can boost hematological performance and higher tolerance from oxygen stress exposure

In vitro antibacterial activity of alkaloid extracts from green, red and brown macroalgae from western coast of Libya

Marine organisms and microorganisms are known to be a rich source of alkaloids with unique chemical feature and interesting biological activities. The current study presents the antibacterial effect of the alkaloid extracts of some green, red and brown algae were collected from western coast of Libya, against, Escherichia coli, Salmonella typhi, Klebsiella spp., and Pseudomonas aeruginosa, Staphylococcus aureus, Bacillus subtilis, Bacillus spp. and Staphylococcus epidermidis were investigated. Although alkaloid extracts of green algae inhibited all tested bacteria, maximum effect was exhibited by brown and red algae species. Thus, Cystoseira barbata alkaloid extract showed remarkable inhibition of human pathogen Klebsiella spp. Dictyopteris membranacea alkaloid extract also demonstrated similar considerable effect against S. typhi with MIC value 1.56 mg/ml. The pronounced antibacterial activity of C. barbata and D. membranacea can be attributed to their high alkaloid contents. These results suggest that red and brown algae secondary metabolites are important sources that could produce potential chemotherapeutic agents.Key words: Macroalgae, alkaloids, antibacterial activity

Effect of oxidized lipids on protein structure and cultured human cells.

Many foods, particularly fish, change in texture and become tough on frozen storage. In addition to ice crystal formation that causes protein denaturation, lipid oxidation products including aldehydes may induce protein cross-linking. In this study, Atlantic Mackerel was stored for up to 26 weeks at -80 and -10 °C. The formation of aldehydes was investigated by thiobarbituric (TEARS) test, HPLC and LC-MS. As expected, there was an increase in the production of aldehydes during storage at -10 °C compared with the control fillets at -80 °C. However, in addition to malonaldehyde (MDA) and hexanal (HEX), we report the formation of gluteraldehyde (GLA) and hydroxynonenal (4-HNE) in frozen fish for the first time. Frozen storage also decreased protein solubility, induced changes in protein conformation and increased formaldehyde and DMA formation. Changes in texture during storage, assessed by small deformation rheology showed increased elastic modulus (G') values. Aldehyde formation in fish and consequently the G' values were reduced in samples that were treated with antioxidant (green tea). A model study using BSA with various aldehydes showed a decrease in lysine availability, increased G' values and conformational changes including increased beta-sheet and tyrosine doublet ratio and decreased tryptophan and disulphide groups. Increased MW of BSA-aldehyde adducts as well as fragmentations were observed by LC-MS. Methyl linoleate (ML) and extracted mackerel oil were oxidised under ultraviolet (UV) radiation for 24, 48 and 72 hours and lipid peroxidation was assessed by measuring peroxide value (PV) and TEARS. To assess the cytotoxicity, oxidized lipids (0, 20, 40, 80 and 100 mug/ml) (ML or fish oil) were added to human colon cancer (Caco-2) cells (density 2 x 104 cells/well) and incubated for 24 h. Using the MTT assay, a decrease in cell viability was observed in all samples treated with UV oxidized ML or fish oil, at concentrations above 40 mug/ml. Caco-2 cells treated with 24 and 72 h oxidized mackerel oil were less damaged compared to cells treated with oxidized ML. Improved cell viability and decrease in lipid peroxidation were shown in cells pretreated with 50 muM EGCG. The mechanism of cell death either by necrosis or apoptosis was investigated. Oxidized 100 mug/ml ML or fish oil-treated caco-2 cell line showed characteristics of apoptosis. Morphological changes resulted from cellular membrane rupture, the formation of apoptotic bodies, DNA fragmentation and caspase-3 activation, detected by Western blotting; these features were visualized by bioimaging techniques such as light microscopy, fluorescence microscopy, Raman microspecfroscopy and atomic force microscopy

Effect of oxidized lipids on protein structure and cultured human cells

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