GC-MS based metabolomic profiling of Aporosa cardiosperma (Gaertn.) Merr. leaf extracts and evaluating its therapeutic potential.

Aporosa cardiosperma is a plant species majorly found in the Indian Western Ghats that belongs to the phyllanthaceae family with ethnobotanical importance. Using a Fourier Transform-Infrared Spectrometer (FT-IR) and Gas Chromatography-Mass Spectrometry (GC-MS) for evaluating leaf extracts of A. cardiosperma, significant functional groups and metabolite constituents were determined, and its total flavonoid, phenol, and tannin content were quantified. Further, its antibacterial efficacy was investigated against microorganisms that cause fish and human disease and are resistant to common antibiotics, including Staphylococcus aureus, Bacillus subtilis, Mycobacterium tuberculosis, Klebsiella pneumoniae, Aeromonas hydrophila, and Pseudomonas aeruginosa. Regarding the outcomes of GC-MS analysis, the primary metabolites in the A. cardiosperma leaf extracts were heneicosane (57.06%), silane (13.60%), 1-heptadecene (10.09%), 3-hexadecene (9.99%), and pentadecane (9.54%). In comparison to other solvents, methanolic extract of A. cardiosperma leaves had increased phenolic, flavonoid, and tannin content; these findings are consistent with in vitro antioxidant potential and obtained that the methanolic extract (100 µg/mL) exhibited the higher percentage of inhibition in DPPH (82.35%), FRAP (86.20%), metal chelating (72.32%), and ABTS (86.06%) antioxidant assays respectively. Similar findings were found regarding the antibacterial efficacy against pathogenic bacteria. Comparatively, to other extracts, methanolic extracts showed more significant antibacterial activity at a lower minimum inhibitory concentration (MIC) value (250 µg/mL), whilst ethyl acetate and hexane solvent extracts of A. cardiosperma leaves had higher MIC values 500 µg/mL and 1000 µg/mL respectively. The antimicrobial potential was validated by investigating bacterial growth through the extracts acquired MICs and sub-MICs range. Bacterial growth was completely inhibited at the determined MIC range. In conclusion, A. cardiosperma leaf extract's phytochemical fingerprint has been determined, and its potent antibacterial and antioxidant activities were discovered. These findings of the current study will pave the way for developing herbal treatments from A. cardiosperma for various fish and human diseases.

Syrupy herbal formulation of green bean pod extract of Phaseolus vulgaris L.: Formulation optimization by central composite design, and evaluation for anti-urolithiatic activity.

The green bean pods of Phaseolus vulgaris L. are traditionally used as a folk remedy for treating calcium oxalate kidney stones. The current research aimed to develop a syrup formulation containing green bean pod extract for anti-urolithiatic activity. The syrup was prepared using a simple blending method and optimized through a central composite design (CCD) with two independent variables: the ratio of pod juice (PJ) to sugar solution (SS) ranging from 1:0.5 to 1:1.5, and the percentage of CMC from 0.2% to 0.4% w/v. These variables were analyzed for their impact on viscosity (CP) and sedimentation percentage, helping to identify the best formulation out of 13 variants. The finalized formulation (F-opt) underwent assessment for physicochemical characteristics such as organoleptic properties, viscosity, density, sedimentation rate, and stability. Additionally, a microbiological assessment was performed utilizing the spread plate method. Further, it was evaluated for in vitro, ex vivo, and in vivo anti-urolithiatic activity in rat models for 28 days and compared with that of the reference standard (Cystone syrup). Additionally, acute toxicity was assessed in albino Swiss mice. Histopathological evaluations were then conducted on the kidneys of the Wistar rats that had been used for the in vivo studies, providing insight into the treatment effects on kidney tissue structure. The optimized formulation (F-opt) was a green, viscous, clear syrup with a pH of 5.8, a viscosity of 256.38 CP, a density of 1.31 g/ml, and a sedimentation rate of 0.69%. The optimized formulation was found to be stable, showing no significant changes in physicochemical and microbiological properties. The results of the in vitro, ex vivo, and in vivo anti-urolithiatic studies indicated that the optimized formulation effectively inhibited the aggregation of calcium oxalate. The acute toxicity studies revealed no mortality or adverse effects for both the optimized formulation and pure bean pod juice at a dose of 2000 mg/kg body weight. Histopathological examination revealed that rats treated with the optimized formulation exhibited a significant reduction in both the number and size of calcium oxalate deposits within various parts of the renal tubules. It can be concluded that the syrupy formulation of Phaseolus vulgaris L. green bean pod extract demonstrated significant anti-urolithiatic activity. This activity could be due to its diuretic properties and its ability to inhibit the formation of calcium oxalate crystals. However, limitations of the study included a lack of elucidation of the mechanism and limited generalizability of the findings.

Bioactive Analysis of Antibacterial Efficacy and Antioxidant Potential of Aloe barbadensis Miller Leaf Extracts and Exploration of Secondary Metabolites Using GC-MS Profiling.

Aloe barbadensis Miller (ABM) is a traditional medicinal plant all over the world. Numerous studies were conducted to exhibit its medicinal properties and most of them were concentrated on its metabolites against human pathogens. The current research work evaluates the attributes of different polar-based extracts (ethanol, methanol, ethyl acetate, acetone, hexane, and petroleum ether) of dried Aloe barbadensis leaf (ABL) to investigate its phytochemical constituents, antioxidant potential (DPPH, ABTS), phenolic, tannin, flavonoid contents, identification of bioactive compounds, and functional groups by gas chromatography-mass spectrometry (GC-MS) and fourier transform infrared spectroscopy (FT-IR) respectively, and comparing antibacterial efficacy against human pathogens, aquatic bacterial pathogens, and zoonotic bacteria associated with fish and human. The present results showed that the methanolic extract of ABL showed higher antioxidant activity (DPPH-59.73 ± 2.01%; ABTS-74.1 ± 1.29%), total phenolic (10.660 ± 1.242 mg GAE/g), tannin (7.158 ± 0.668 mg TAE/g), and flavonoid content (49.545 ± 1.928 µg QE/g) than that of other solvent extracts. Non-polar solvents hexane and petroleum ether exhibited lesser activity among the extracts. In the case of antibacterial activity, higher inhibition zone was recorded in methanol extract of ABL (25.00 ± 0.70 mm) against Aeromonas salmonicida. Variations in antibacterial activity were observed depending on solvents and extracts. In the current study, polar solvents revealed higher antibacterial activity when compared to the non-polar and the mid-polar solvents. Diverse crucial bioactive compounds were detected in GC-MS analysis. The vital compounds were hexadecanoic acid (30.69%) and 2-pentanone, 4-hydroxy-4-methyl (23.77%) which are responsible for higher antioxidant and antibacterial activity. Similar functional groups were identified in all the solvent extracts of ABL with slight variations in the FT-IR analysis. Polar-based solvent extraction influenced the elution of phytocompounds more than that of the other solvents used in this study. The obtained results suggested that the ABM could be an excellent source for antioxidant and antibacterial activities and can also serve as a potential source of effective bioactive compounds to combat human as well as aquatic pathogens.

Evaluating Biofilm Inhibitory Potential in Fish Pathogen, Aeromonas hydrophila by Agricultural Waste Extracts and Assessment of Aerolysin Inhibitors Using In Silico Approach.

Aeromonas hydrophila, an opportunistic bacteria, causes several devastating diseases in humans and animals, particularly aquatic species. Antibiotics have been constrained by the rise of antibiotic resistance caused by drug overuse. Therefore, new strategies are required to prevent appropriate antibiotic inability from antibiotic-resistant strains. Aerolysin is essential for A. hydrophila pathogenesis and has been proposed as a potential target for inventing drugs with anti-virulence properties. It is a unique method of disease prevention in fish to block the quorum-sensing mechanism of A. hydrophila. In SEM analysis, the crude solvent extracts of both groundnut shells and black gram pods exhibited a reduction of aerolysin formation and biofilm matrix formation by blocking the QS in A. hydrophila. Morphological changes were identified in the extracts treated bacterial cells. Furthermore, in previous studies, 34 ligands were identified with potential antibacterial metabolites from agricultural wastes, groundnut shells, and black gram pods using a literature survey. Twelve potent metabolites showed interactions between aerolysin and metabolites during molecular docking analysis, in that H-Pyran-4-one-2,3 dihydro-3,5 dihydroxy-6-methyl (-5.3 kcal/mol) and 2-Hexyldecanoic acid (-5.2 kcal/mol) showed promising results with potential hydrogen bond interactions with aerolysin. These metabolites showed a better binding affinity with aerolysin for 100 ns in molecular simulation dynamics. These findings point to a novel strategy for developing drugs using metabolites from agricultural wastes that may be feasible pharmacological solutions for treating A. hydrophila infections for the betterment of aquaculture.

Sustainable fabrication of hybrid silver-copper nanocomposites (Ag-CuO NCs) using Ocimum americanum L. as an effective regime against antibacterial, anticancer, photocatalytic dye degradation and microalgae toxicity.

In this study, a sustainable fabrication of hybrid silver-copper oxide nanocomposites (Ag-CuO NCs) was accomplished utilizing Ocimum americanum L. by one pot green chemistry method. The multifarious biological and environmental applications of the green fabricated Ag-CuO NCs were evaluated through their antibacterial, anticancer, dye degradation, and microalgae growth inhibition activities. The morphological features of the surface functionalized hybrid Ag-CuO NCs were confirmed by FE-SEM and HR-TEM techniques. The surface plasmon resonance λmax peak appeared at 441.56 nm. The average hydrodynamic size distribution of synthesized nanocomposite was 69.80 nm. Zeta potential analysis of Ag-CuO NCs confirmed its remarkable stability at -21.5 mV. XRD and XPS techniques validated the crystalline structure and electron binding affinity of NCs, respectively. The Ag-CuO NCs demonstrated excellent inhibitory activity against Vibrio cholerae (19.93 ± 0.29 mm) at 100 µg/mL. Anticancer efficacy of Ag-CuO NCs was investigated against the A549 lung cancer cell line, and Ag-CuO NCs exhibited outstanding antiproliferative activity with a low IC50 of 2.8 ± 0.05 µg/mL. Furthermore, staining and comet assays substantiated that the Ag-CuO NCs hindered the progression of the A549 cells and induced apoptosis as a result of cell cycle arrest at the G0/G1 phase. Concerning the environmental applications, the Ag-CuO NCs displayed efficient photocatalytic activity against eosin yellow degradation up to 80.94% under sunlight irradiation. Microalgae can be used as an early bio-indicator/prediction of environmental contaminants and toxic substances. The treatment of the Ag-CuO NCs on the growth of marine microalgae Tetraselmis suecica demonstrated the dose and time-dependent growth reduction and variations in the chlorophyll content. Therefore, the efficient multifunctional properties of hybrid Ag-CuO NCs could be exploited as a regime against infective diseases and cancer. Further, the findings of our investigation witness the remarkable scope and potency of Ag-CuO NCs for environmental applications.

Exploration of marine red seaweed as a dietary fish meal replacement and its potentiality on growth, hematological, biochemical, and enzyme activity in freshwater fish Labeo rohita.

The present study investigated the dietary fishmeal replacement by marine red seaweed (Halymenia dilatata) meal (RSM) on growth performance, feed utilization, chemical body composition, hematological constituents, digestive, antioxidant, and metabolic enzymes in freshwater fish Labeo rohita (Rohu) fingerlings. The fish were fed with RSM-free control diet (RSM0) and four experimental diets, which replaced fish meal (FM) with varying levels of RSM (25%, 50%, 75%, and 100%, represented as RSM25, RSM50, RSM75, and RSM100 respectively). After a 60-day feeding trial, the survival rate (SR), growth performance (length gain, weight gain, and specific growth rate), protein efficiency ratio, chemical body composition (protein, lipid, and ash), and digestive enzymes (amylase and protease) were significantly increased (P < 0.05) in the fish fed with RSM50 diet containing 39% protein level. The sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) revealed a higher staining intensity of muscle proteins in fish fed with the RSM50 diet. However, the hematological constituents (hemoglobin, hematocrit, red blood cell, white blood cell, mean corpuscular volume, mean corpuscular hemoglobin, and mean corpuscular hemoglobin concentration), antioxidant enzyme - superoxide dismutase, and metabolic enzymes (glutamic oxaloacetic transaminase and glutamic pyruvic transaminase) were not significantly altered in RSM50 diet when compared to control. In contrast, hematological constituents were decreased (P < 0.05), and antioxidant and metabolic enzymes were increased in rohu fed with RSM75 and RSM100 (P < 0.05). Furthermore, these findings suggest that RSM might be adopted at a pace of 37% (estimated polynomial second-order regression) and is found to be beneficial for freshwater fish L. rohita diets that enhance growth and immune responses. The current study recommended substituting (50%) of marine red seaweed (Halymenia dilatata) for fish meal significantly improves the growth performance, chemical body composition, and digestive enzymes of L. rohita and this could be a valuable natural replacement for fishmeal to reduce the production cost of aquatic feed.

Biofabrication of ecofriendly copper oxide nanoparticles using Ocimum americanum aqueous leaf extract: analysis of in vitro antibacterial, anticancer, and photocatalytic activities.

Nanotechnology tends to be a swiftly growing field of research that actively influences and inhibits the growth of bacteria/cancer. Noble metal nanoparticles (NPs) such as silver, copper, and gold have been used to damage bacterial and cancer growth over recent years; however, the toxicity of higher NPs concentrations remains a major issue. The copper oxide nanoparticles (CuONPs) were therefore fabricated using a simple green chemistry approach. Biofabricated CuONPs were characterized using UV-visible, FE-SEM with EDS, HR-TEM, FT-IR, XRD, Raman spectroscopy, and XPS analysis. Formations of CuONPs have been observed by UV-visible absorbance peak at 360.74 nm. The surface morphology of the CuONPs showed the spherical structure and size (~ 68 nm). The EDS spectrum of CuONPs has proved to be the key signals of copper (Cu) and oxygen (O) components. FT-IR analysis, to validate the important functional biomolecules (O-H, C=C, C-H, C-O) are responsible for reduction and stabilization of CuONPs. The monoclinic end-centered crystalline structures of CuONPs were confirmed with XRD planes. The electrochemical oxygen states of the CuONPs have been studied using spectroscopy of the Raman and X-ray photoelectron. After successful preparation, CuONPs examined their antibacterial, anticancer, and photocatalytic activities. Green-fabricated CuONPs were promising antibacterial candidate against human pathogenic gram-negative bacteria Escherichia coli, Vibrio cholerae, Salmonella typhimurium, Klebsiella pneumoniae, Aeromonas hydrophila, and Pseudomonas aeruginosa. CuONPs were demonstrated the excellent anticancer activity against A549 human lung adenocarcinoma cell line. Furthermore, CuONPs exhibited photocatalytic degradation of azo dyes such as eosin yellow (EY), rhodamine 123 (Rh 123), and methylene blue (MB). Biofabricated CuONPs may therefore be an important biomedical research for the aid of bacterial/cancer diseases and photocatalytic degradation of azo dyes.

Effects of graded levels of mineral mixtures (Aquamin® and Agrimin®) supplemented diets on growth, survival, proximate composition, and carcass mineralization of juvenile freshwater prawn, Macrobrachium malcolmsonii (H Milne-Edwards 1844).

The present study investigated the effects of Aquamin® mineral mixture (AQMM) and Agrimin® mineral mixture (AGMM) supplemented diets on growth and chemical composition of juvenile freshwater prawn Macrobrachium malcolmsonii (H. Milne-Edwards, 1844). Experimental diets containing 6 different AQMM levels (Trial 1: 0, 0.5, 1.0, 1.5, 2.0, and 2.5%) and 6 different AGMM levels (Trial 2: 0, 0.5, 1.0, 1.5, 2.0, and 2.5%) were formulated to feed juvenile prawn [initial body weight of 0.82 ± 0.05 g (Trial 1) and 1.24 ± 0.03 g (Trial 2)] for 60 days. Prawn fed diets containing 1.0% of AQMM and AGMM showed significantly increased (P < 0.05) survival rate, weight gain, specific growth rate, protein efficiency ratio, edible flesh weight, and proximate composition (moisture, crude protein, crude fat, and ash), while feed intake and feed conversion ratio were significantly decreased (P < 0.05). Whereas, 1.5-2.5% of AQMM- and AGMM-supplemented diets fed prawn showed a reverse trend when comparing other groups. Moreover, the minerals (macro elements: Ca, P, Mg, Na, and K; trace elements: Cu, Zn, and Fe) were significantly increased (P < 0.05) in the carcass of prawn when fed with 2.5% AQMM- and AGMM-supplemented diets. The present results suggest that the optimal dietary supplementation of AQMM and AGMM at a concentration of up to 1.06 and 1.02%, respectively (based on polynomial regression analysis), improved growth and enhanced the crude protein level of juvenile prawn.

Activity profile of innate immune-related enzymes and bactericidal of freshwater fish epidermal mucus extract at different pH.

The epidermal mucus of fish performs diverse functions from prevention of mechanical abrasion to limit pathogen invasions. The current experiment was designed to extract skin mucus proteins of three freshwater fish, i.e. common carp (Cyprinus carpio), mrigal (Cirrhinus mrigala) and rohu (Labeo rohita) with organic solvent (methanol) and dissolve in different pH of Tris-HCl buffers to examine the significance of pH in the solubilisation of skin mucus proteins. The protein profiles of different pH solubilised methanol fish skin mucus extracts were determined by SDS-PAGE. The non-specific immune enzymes, alkaline phosphatase, lysozyme and protease of fish skin mucus were compared and this present study demonstrated that these enzymes differed in their activity depending on pH buffers. The higher lysozyme and protease activity were observed at the pH of 8.0 and higher alkaline phosphatase activity in the pH 9.0 of C. mrigala fish skin mucus methanol extract. In addition, the bactericidal activity was evaluated against the pathogens Proteus vulgaris and Pseudomonas aeruginosa. The pH 8.0 of C. mrigala skin mucus extract revealed better bactericidal activity than other fish species mucus pH buffers against both P. vulgaris and P. aeruginosa. In the case of protein profile from SDS-PAGE, based on pH buffers and the solubilisation of proteins, differences in the resolution of bands were observed. The higher alkaline pH of 9.0 showed smeared gel bands in all the three fish skin mucus methanol extract. The present study suggests that methanol extracted C. mrigala fish skin mucus at pH 8.0 showed better innate immune enzymes and bactericidal activity. The additional examinations of C. mrigala skin mucus methanol extract in this pH aids in identifying novel bioactive molecules. This is the study of proteome of three fish species skin mucus in the effect of pH. Further analyses are required to evaluate proteins present in fish skin mucus extracted with methanol and the influence of pH on protein solubility. These findings could be helpful in exploring natural alternatives to antibiotics in aquaculture industry against infectious pathogens.