The present study was carried out with the aim to isolate an antibacterial pigment from seaweed-associated bacterium. The bacterium was identified as Halolactibacillus alkaliphilus MSRD1 by 16S rRNA sequencing. The isolated bacterium was cultured in 50% Luria-Bertani seawater broth (LB-SWB) with 1% glycerol. The pigment was extracted with 99% ethanol and analyzed by UV-Vis spectroscopy at 490 nm. The candidate bacterium was optimized with various NaCl concentrations from 5 to 20%. The results inferred that the bacterium produce maximum pigment at 5% NaCl level. The candidate bacterium H. alkaliphilus MSRD1 was found to be producing the maximum pigment during the 120-h incubation. The protein content of the pigment was found to be maximum of 72% at the end of the 120-h incubation. The extracted pigment was stable up to 80 °C, pink at acidic pH (1 to 5) and orange at basic pH (8 to 12). The isolated pigment was fractionated by silica gel column chromatography. Fractionated pigment was characterized by TLC, FT-IR, and SDS-PAGE. In the antibacterial context, the pigment was highly inhibited Staphylococcus aureus and Salmonella typhi with the zone of inhibition 16 and 14 mm, respectively. According to SDS-PAGE, the size of the pigment was approximately 80 kDa. The H. alkaliphilus MSRD1 has high capacity to produce the pigment with antibacterial properties. This could be effectively used in the future.
Somatic antigens of F. gigantica, G. explanatum, S. spindale and hydatid cyst ingredients were analysed to identify the cross-reactive antigens among them using Western blot technique. When probed with F. gigantica infected cattle sera, the immunodominant 156 kDa and 28 kDa proteins of F. gigantica was found common amongst the antigens prepared from hydatid cysts ingredients like germinal layer, fertile and sterile, hydatid fluid, fertile and sterile, while another protein of 34 kDa was shared between F. gigantica and antigen prepared from protoscolices. In F. gigantica-buffalo system the proteins of 34 kDa and 28 kDa were found reactive with most of the antigens tested. Immunoaffinity chromatography using, F. gigantica infected rabbit immunoglobulins as legands isolated the immunodominant 34 kDa and 28 kDa proteins in dimer form and the same were found immunodominant in F. gigantica-cattle, F. gigantica-buffalo and F. gigantica-sheep system. No cross-reaction was noted with the sera of goats experimentally infected with Paramphistomum epiclitum. ELISA with the immunodominant proteins of 34 kDa and 28 kDa could be a feasible diagnostic tool for the early detection of bovine fasciolosis.
An attempt has been made to identify the potential seaweed for antifouling property due to the growing need for environmentally safe antifouling systems. The antibacterial, antimicroalgal, and antimussel foot adherence potentials of methanol, dichloromethane, and hexane extracts of the chosen seaweeds such as Padina tetrastromatica, Caulerpa taxifolia, and Amphiroa fragilissima have been compared against copper sulfate. Among the extracts, the maximum antibacterial activities were exhibited by the methanol extract of P. tetrastromatica. The minimum inhibitory concentration (MIC) of the methanolic extract of P. tetrastromatica was found to be 10 and 1 μg/ml against test biofilm bacteria and diatoms, respectively. The antimussel foot adherence assay indicated that the extract had inhibited the foot adherence of the green mussels Perna viridis with the effective concentration (EC50) of 25.51 ± 0.03 μg/ml, and lethal concentration for 50 % mortality (LC50) was recorded at 280.22 ± 0.12 μg/ml. Based on the prolific results, the crude methanolic extract of P. tetrastromatica was subjected to purification using silica gel column and thin-layer chromatography (TLC). Then, the active compounds of the bioassay-guided fraction (F13) were identified using gas chromatography coupled with mass spectroscopy (GC-MS), and it was observed that fatty acids were the major components, which may be responsible for the antifouling properties.
The antibacterial potential of centric marine diatoms has been compared against the clinical pathogens and identified the potential diatom by rbcL gene sequencing. Totally, five diatoms namely Odontella aurita, Thalassiosira subtilis, Chaetoceros curvisetus, Skeletonema costatum and Coscinodiscus centralis were isolated from Cuddalore coastal waters. The diatoms were morphologically identified and isolated using micro capillary-pipette and serial dilution method. The isolated diatoms were cultured in Guillard's f/2 medium to get biomass for the antibacterial study. The dried biomass of the cultured diatoms was individually extracted with methanol, ethanol and hexane. All the obtained extracts were tested against Staphylococcus haemolyticus, Proteus vulgarius and Vibrio alginolyticus. The crude ethanol extract of O. aurita was exhibited highest zone of inhibition against all the test pathogens. The MIC of O. aurita was recorded as 50 μg/ml against both Staphylococcus haemolyticus and Proteus vulgarius whilst 75 μg/ml against Vibrio alginolyticus. This study indicates that O. aurita possesses antibacterial activities but the release of antibiotics depends on physical or chemical rupture of algal cells and extractive solvents. Based on the maximum antibacterial activity, O. aurita was further identified by rbcL gene sequencing. The rbcL gene could be an identical region for the species level identification of diatoms.
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