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.

Potentials of Aloe barbadensis inclusion in fish feeds on resilience to Aeromonas hydrophila infection in freshwater fish Labeo rohita.

Aquatic bacterial pathogens can cause severe economic loss in aquaculture industry. An opportunistic pathogen, Aeromonas hydrophila is responsible for Motile Aeromonas Septicemia, leading to high mortality rates in fish. The present study was focused on the efficacy of Aloe barbadensis replacing fishmeal diets on hematological, serum biochemical, antioxidant, histopathological parameters, and disease resistance against A. hydrophila infection in Labeo rohita. Isonitrogenous fishmeal replaced diets (FMR) were prepared with varying levels of A. barbadensis at D1 (0%) (control), D2 (25%), D3 (50%), D4 (75%) and D5 (100%) then fed to L. rohita. After 60 days of post-feeding, the experimental fish were challenged with A. hydrophila. Blood and organs were collected and examined at 1- and 15-days post infection (dpi). The results demonstrated that on 1 dpi, white blood cells (WBC), total protein, cholesterol and low-density lipoprotein (LDL) levels were significantly increased in D3 diet fed groups. The D2 and D3 diet fed group showed decreasing trends of serum glutamic pyruvic transaminase (SGPT) and antioxidant enzymes activity on 15 dpi. The histopathological architecture results clearly illustrated that the D3 diet fed group had given a higher protective effect by reducing the pathological changes associated with A. hydrophila infection in liver, intestine and muscle. Higher percentage of survival rate was also observed in D3 diet fed group. Therefore, the present study suggested that the dietary administration of A. barbadensis up to 50% fishmeal replacement (D3 diet) can elicit earlier antioxidant activity, innate immune response and improve survival rate in L. rohita against A. hydrophila infection.

Indole-3-acetic acid impacts biofilm formation and virulence production of Pseudomonas aeruginosa.

Bacterial pathogenesis involves complex mechanisms contributing to virulence and persistence of infections. Understanding the multifactorial nature of bacterial infections is crucial for developing effective interventions. The present study investigated the efficacy of indole-3-acetic acid (IAA) against Pseudomonas aeruginosa with various end points including antibacterial activity, minimum inhibitory concentration (MIC), virulence factor production, biofilm inhibition, bacterial cell detachment, and viability assays. Results showed significant biofilm inhibition, bacterial cell detachment, and modest effects on bacterial viability. Microscopic analysis confirmed the disintegrated biofilm matrix, supporting the inhibitory effect of IAA. Additionally, molecular docking studies revealed potential mechanisms of action through active bond interactions between IAA and virulence proteins. These findings highlight IAA as an effective antibiofilm agent against P. aeruginosa.

Evaluation of Phytochemical Screening, Pigment Content, In Vitro Antioxidant, Antibacterial Potential and GC-MS Metabolite Profiling of Green Seaweed Caulerpa racemosa.

Exploration of seaweeds to unravel their bioactive metabolites from the perspective of wider applications gained substantial importance. The present study was performed to investigate the total phenolic, flavonoid, tannin content, antioxidant activity and antibacterial potential of various solvent extracts of green seaweed Caulerpa racemosa. The methanolic extract showed higher phenolic (11.99 ± 0.48 mg gallic acid equivalents/g), tannin (18.59 ± 0.54 mg tannic acid equivalents/g) and flavonoid (33.17 ± 0.76 mg quercetin equivalents/g) content than other extracts. Antioxidant activity was determined by using 2,2-diphenylpicrylhydrazyl (DPPH) and 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) assay with different concentrations of C. racemosa extracts. The methanolic extract showed higher scavenging potential in both the DPPH and ABTS activity with the inhibition value of 54.21 ± 1.39% and 76.62 ± 1.08%, respectively. Bioactive profiling was also identified by using Gas chromatography-mass spectrometry (GC-MS) and Fourier transform infrared (FT-IR) techniques. These studies revealed the presence of valuable bioactive compounds in C. racemosa extracts and these compounds might be responsible for antimicrobial, antioxidant, anticancer and anti-mutagenic properties. Major compounds identified in GC-MS were 3,7,11,15-Tetramethyl-2-hexadecen-1-ol, 3-hexadecene and Phthalic acid. In terms of antibacterial activity, C. racemosa has promising antibacterial potential against aquatic pathogens Aeromonas hydrophila, Aeromonas veronii and Aeromonas salmonicida. Further evaluation studies focusing aquatic related aspects would reveal the novel bioproperties and applications of C. racemosa.

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.

Accumulation, Chronicity, and Induction of Oxidative Stress Regulating Genes Through Allium cepa L. Functionalized Silver Nanoparticles in Freshwater Common Carp (Cyprinus carpio).

Green evolutionary products such as biologically fabricated nanoparticles (NPs) pose a hazard to aquatic creatures. Herein, biogenic silver nanoparticles (AgNPs) were synthesized by the reaction between ionic silver (AgNO3) and aqueous onion peel extract (Allium cepa L). The synthesized biogenic AgNPs were characterized with UV-Visible spectrophotometer, XRD, FT-IR, and TEM with EDS analysis; then, their toxicity was assessed on common carp fish (Cyprinus carpio) using biomarkers of haematological alterations, oxidative stress, histological changes, differential gene expression patterns, and bioaccumulation. The 96 h lethal toxicity was analysed with various concentrations (2, 4, 6, 8, and 10 mg/l) of biogenic AgNPs. Based on 96 h LC50, sublethal concentrations (1/15th, 1/10th, and 1/5th) were given to C. carpio for 28 days. At the end of experiment, the bioaccumulations of Ag content were accumulated mainly in the gills, followed by the liver and muscle. At an interval of 7 days, the haematological alterations showed significance (p < 0.05) and elevation of antioxidant defence mechanism reveals the toxicity of biogenic synthesized AgNPs. Adverse effects on oxidative stress were probably related to the histopathological damage of its vital organs like gill, liver, and muscle. Finally, the fish treated with biogenic synthesized AgNPs were significantly (p < 0.05) downregulates the oxidative stress genes such as Cu-Zn SOD, CAT, GPx1a, GST-α, CYP1A, and Nrf-2 expression patterns. The present study provides evidence of biogenic synthesized AgNPs influence on the aquatic life through induction of oxidative stress.

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.

Synthesis, morphological analysis, antibacterial activity of iron oxide nanoparticles and the cytotoxic effect on lung cancer cell line.

Focusing on the huge importance associated in developing functional materials, this research study describes the synthesis, characterization of morphology, bactericidal activity and cytotoxic effect of iron oxide nanoparticles (IONPs). IONPs have been successfully fabricated through thermal decomposition of a diiron(III) complex precursor. The morphology of the nanoparticle has been delineated with different spectroscopic and analytic methods. Scanning and transmission electron microscopy (FE-SEM and HR-TEM) analyses estimate the cross linked porous structure of IONPs with an average size ~97 nm. Dynamic light scattering (DLS) study of IONPs determines the hydrodynamic diameter as 104 nm. The cytotoxic behavior of IONPs has been examined against human lung cancer cell line (A549) through different fluorescence staining studies which ensure the mode of apoptosis for cell death of A549. Furthermore, measurement of reactive oxygen species suggests the destruction of mitochondrial membrane of Staphylococcus aureus, leading to effective bactericidal propensity which holds a good promise for IONPs to become a clinically approved antibacterial agent.

GST M1-T1 null allele frequency patterns in geographically assorted human populations: a phylogenetic approach.

Genetic diversity in drug metabolism and disposition is mainly considered as the outcome of the inter-individual genetic variation in polymorphism of drug-xenobiotic metabolizing enzyme (XME). Among the XMEs, glutathione-S-transferases (GST) gene loci are an important candidate for the investigation of diversity in allele frequency, as the deletion mutations in GST M1 and T1 genotypes are associated with various cancers and genetic disorders of all major Population Affiliations (PAs). Therefore, the present population based phylogenetic study was focused to uncover the frequency distribution pattern in GST M1 and T1 null genotypes among 45 Geographically Assorted Human Populations (GAHPs). The frequency distribution pattern for GST M1 and T1 null alleles have been detected in this study using the data derived from literatures representing 44 populations affiliated to Africa, Asia, Europe, South America and the genome of PA from Gujarat, a region in western India. Allele frequency counting for Gujarat PA and scattered plot analysis for geographical distribution among the PAs were performed in SPSS-21. The GST M1 and GST T1 null allele frequencies patterns of the PAs were computed in Seqboot, Gendist program of Phylip software package (3.69 versions) and Unweighted Pair Group method with Arithmetic Mean in Mega-6 software. Allele frequencies from South African Xhosa tribe, East African Zimbabwe, East African Ethiopia, North African Egypt, Caucasian, South Asian Afghanistan and South Indian Andhra Pradesh have been identified as the probable seven patterns among the 45 GAHPs investigated in this study for GST M1-T1 null genotypes. The patternized null allele frequencies demonstrated in this study for the first time addresses the missing link in GST M1-T1 null allele frequencies among GAHPs.