Enhanced bactericidal, antibiofilm and antioxidative response of Lawsonia inermis leaf extract synthesized ZnO NPs loaded with commercial antibiotic.

Globally, antibiotic resistance is a challenging issue in healthcare sector. The emergence of multiple drug-resistant bacteria has forced us to modify existing medicines and or formulate newer medicines that are effective and inexpensive. In this perspective, this study involves the formation of zinc oxide nanoparticles (ZnO NPs) by utilizing the Lawsonia inermis (Li) leaf extract. The prepared L. inermis leaf extract mediated ZnO NPs (Li-ZnO NPs) were bio-physically characterized. The antibacterial and radical scavenging effects of Li-ZnO NPs were evaluated. In addition, ZnO NPs were conjugated with standard antibiotic (ciprofloxacin) and its drug loading efficiency, drug release and antibacterial efficacy were tested and compared with non-drug loaded ZnO NPs. An absorbance peak at 340 nm was noted for Li-ZnO NPs. After conjugation with the drug, two absorbance peaks- one at 242 nm characteristic of ciprofloxacin and the other at 350 nm characteristics of ZnO NPs were observed. The crystallite size was 18.7 nm as determined by XRD. The antibacterial effect was higher on Gram-positive (S. aureus and S. pyogenes) than the Gram-negative pathogens (E. coli and K. pneumoniae). Inhibition of S. aureus and S. pyogenes biofilm at 100 µg mL-1were, respectively, 97.5 and 92.6%. H2O2 free radicals was inhibited to 90% compared to the standard ascorbic acid at 100 µg mL-1. After drug loading, the FTIR spectrum confirmed the existence of ciprofloxacin peaks at 965 cm-1 and Zn-O bond at 492 cm-1. The drug loading capacity of 15 nm sized ZnO NPs was higher (58, 75, 90 and 95% at 1, 2.5, 5 and 10% drug concentrations, respectively) compared to 20 nm. Similarly, the percentage of drug (ciprofloxacin) released from 15 nm ZnO NPs were increased to 90% at 10% drug-loaded samples, respectively. Also, the antibiotic loaded ZnO NPs had significant antibacterial effects against tested bacteria compared to Li-ZnO NPs and ciprofloxacin alone. This revealed that the antibiotic loaded ZnO NPs offer a sustainable route to treat multi-drug-resistant bacterial infections.

Exopolysaccharide from Lactococcus hircilactis CH4 and Lactobacillus delbrueckii GRIPUMSK as new therapeutics to treat biofilm pathogens, oxidative stress and human colon adenocarcinoma.

Naturally occurring biopolymers like exopolysaccharides (EPS) secreted by lactic acid bacteria (LAB) has gained significant attention as they are cost effective, renewable and safe. In order to prevent the rapid increase in antibiotic resistant bacteria, the EPS of LAB offers novel approach of targeting the antibiotic resistant pathogens by limiting their effects on environment. Accordingly, in this study, the production, purification, characterization and biological properties of exopolysaccharides from Lactococcus hircilactis strain CH4 and Lactobacillus delbrueckii strain GRIPUMSK were performed. The optimization of lactic acid bacterial strains for exopolysaccharide production was done by response surface methodology and changing the carbon sources in the growth media. The carbohydrate and protein of exopolysaccharide 1 were 79.7 % and 8.7 % respectively and exopolysaccharide 2 were 75.2 % and 9.3 % respectively. When compared with the commercial emulsifier sodium dodecyl sulfate, both the exopolysaccharides have shown good emulsifying activity. Both the exopolysaccharides were linear homo-polysaccharide as determined by Fourier transform infrared spectroscopy and Nuclear magnetic resonance spectra. Scanning electron microscopy showed that the exopolysaccharides were porous and capable of holding water. The exopolysaccharides were partially crystalline as confirmed by X-ray diffraction spectra. Exopolysaccharides from L. hircilactis and L. delbrueckii exhibited significant antimicrobial activity against H. pylori, S. flexneri, S. pyogenes, E. faecalis and C. albicans. Both the exopolysaccharides revealed significant 2,2-diphenyl-1-picrylhydrazyl and hydrogen peroxide scavenging ability with the IC50 value of 100 µg/ml and 80 µg/ml respectively. Exopolysaccharides from L. hircilactis and L. delbrueckii at 100 µg/ml showed significant anticancer activity on HT-29 cells with 58.4 % and 58.7 % respectively. These findings proved that exopolysaccharides from the two selected lactic acid bacterial strains could be explored as natural bioactive carbohydrate polymer for biomedical applications.

Nanomedicine for drug resistant pathogens and COVID-19 using mushroom nanocomposite inspired with bacteriocin - A review.

Multidrug resistant (MDR) pathogens have become a major global health challenge and have severely threatened the health of society. Current conditions have become worse as a result of the COVID-19 pandemic, and infection rates in the future will rise. It is necessary to design, respond effectively, and take action to address these challenges by investigating new avenues. In this regard, the fabrication of metal NPs utilized by various methods, including green synthesis using mushroom, is highly versatile, cost-effective, eco-compatible, and superior. In contrast, biofabrication of metal NPs can be employed as a powerful weapon against MDR pathogens and have immense biomedical applications. In addition, the advancement in nanotechnology has made possible to modify the nanomaterials and enhance their activities. Metal NPs with biomolecules composite prevent the microbial adhesion and kills the microbial pathogens through biofilm formation. Bacteriocin is an excellent antimicrobial peptide that works well as an augmentation substance to boost the antimicrobial effects. As a result, we concentrate on the creation of new, eco-compatible mycosynthesized metal NPs with bacteriocin nanocomposite via electrostatic, covalent, or non-covalent bindings. The synergistic benefits of metal NPs with bacteriocin to combat MDR pathogens and COVID-19, as well as other biomedical applications, are discussed in this review. Moreover, the importance of the adverse outcome pathway (AOP) in risk analysis of manufactured metal nanocomposite nanomaterial and their future possibilities were also discussed.

Antimicrobial, anti-biofilm, antioxidant and cytotoxic effects of bacteriocin by Lactococcus lactis strain CH3 isolated from fermented dairy products-An in vitro and in silico approach.

The current study aimed to screen bacteriocin producing LAB from different dairy products and evaluation of their biological properties. Initially, 12 (4-chess, 4-curd, and 4-yohurt) LAB species were isolated and only 4 isolates alone were selected based on their clear yellow halo zone around the colonies in the selective medium. The selected 4 isolates were identified based on their morphological and biochemical characteristics. Among them, the strain CH3 have showed better antimicrobial effects on selected human pathogens. The isolated strain CH3 were further identified as Lactococcus lactis strain CH3 (MZ636710) by SEM imaging and 16 s rRNA molecular sequencing. Bacteriocin was extracted from L. lactis strain CH3 and partially purified using 60 % ammonium sulphate and then completely purified by G-50 column chromatography. The purified bacteriocin showed a specific activity of 5859.37 AU/mg in 24.7 % of recovery and 10.9-fold purification. The molecular weight of bacteriocin was 3.5 kDa as observed in SDS-PAGE. The bacteriocin showed sensitivity to proteolytic enzymes and resistance to high temperature, wide range of pH, organic solvents and detergents. FT-IR spectral studies of bacteriocin detected the existence of OH/NH-stretching, CH, and COC and CO bonds. NMR spectrum showed one doublet and 4 various singlet peaks at different ppm, indicating the occurrence of six amino acids in the structure of purified bacteriocin. The purified bacteriocin have shown stronger antimicrobial and anti-biofilm activity against selected human pathogens at 100 µg/mL. SEM showed the evidence of structural deformation and loss of membrane integrity of bacterial cells treated with bacteriocin. Bacteriocin exhibited greater DPPH radical scavenging potential with an EC50 value of 12.5 µg/mL. Bacteriocin have not shown significant toxicity on normal human dermal fibroblast (NHDF) cells (83.2 % at 100 µg/ mL). Furthermore, in silico studies using molecular modeling and docking were performed to know the proteins involved in antimicrobial action. The results suggests that bacteriocin could be an alternative to combat AMR pathogens and more suitable for food and dairy industries to preserve food without contamination.

High synergistic antibacterial, antibiofilm, antidiabetic and antimetabolic activity of Withania somnifera leaf extract-assisted zinc oxide nanoparticle.

Nanotechnology is currently gaining immense attention to combat food borne bacteria, and biofilm. Diabetes is a common metabolic disease affecting majority of people. A better therapy relies on phytomediated nanoparticle synthesis. In this study, W. somnifera leaf extract-assisted ZnO NPs (Ws-ZnO NPs) was synthesized and characterized. From HR-TEM analysis, it has been found that the hexagonal wurtzite particle is 15.6 nm in size and - 12.14 mV of zeta potential. A greater antibacterial effect of Ws-ZnO NPs was noticed against E. faecalis and S. aureus at 100 µg mL-1. Also, the biofilm of E. faecalis and S. aureus was greatly inhibited at 100 µg mL-1 compared to E. coli and P. aeruginosa. The activity of α-amylase and α-glucosidase enzyme was inhibited at 100 µg mL-1 demonstrating its antidiabetic potential. The larval and pupal development was delayed at 25 µg mL-1 of Ws-ZnO NPs. A complete mortality (100%) was recorded at 25 µg mL-1. Ws-ZnO NPs showed least LC50 value (9.65 µg mL-1) compared to the uncoated ZnO NPs (38.8 µg mL-1) and leaf extract (13.06 µg mL-1). Therefore, it is concluded that Ws-ZnO NPs are promising to be used as effective antimicrobials, antidiabetic and insecticides to combat storage pests.

Biopolymer K-carrageenan wrapped ZnO nanoparticles as drug delivery vehicles for anti MRSA therapy.

Kappa-Carrageenan wrapped ZnO nanoparticles (KC-ZnO NPs) was synthesized, physico-chemically characterized and evaluated their biocompatibility and antimicrobial therapy against MRSA. XRD showed the highly crystalline and hexagonal phase structure of ZnO NPs. FETEM confirmed the spherical and hexagonal shaped particle with the mean size of 97.03 ± 9.05 nm. The synthesized KC-ZnO NPs exhibited significant antibacterial activity against MRSA. The biofilm growth of MRSA was greatly inhibited at 100 µg/ml as observed through live and dead cell assay. KC-ZnO NPs have shown invitro anti-inflammatory activity (82%) at 500 µg/ml. KC-ZnO NPs was non-toxic to NIH3T3 mouse embryonic fibroblasts cell lines. Further, no apoptotic and necrotic mediated death in NIH3T3 mouse embryonic fibroblasts cells were noticed by flow cytometric analysis. KC-ZnO NPs have good biocompatibility as recorded by the least hemolysis percentage (<3%) up to 100 µg/ml, which is much lesser than the acceptable limit. In addition, ecosafety analysis has shown that KC-ZnO NPs and kappa karrageenan (0-500 µg/ml) caused no mortality of A. salina after 48 h. However, bare zinc acetate has shown 35% mortality of A. salina after 48 h. The results conclude that KC-ZnO NPs could be a novel antibacterial therapy for the treatment of MRSA associated infectious.

Anti-cancer, anti-biofilm, and anti-inflammatory properties of hen's albumen: A photodynamic approach.

The albumen plays a major role in the protection of eggs against microorganisms. It contains an arsenal of natural antimicrobial molecules and antibacterial proteins, including the well-known ovotransferrin and lysozyme, which exert their activities against a range of bacteria. In the present study, the hen's albumen extract treated with the dried insect body of blister beetle M. pustulata was assessed for antibacterial, antibiofilm, anti-inflammatory and anti-proliferative activity. The zone of inhibition against Gram positive E. faecalis and S. aureus was 10.8 mm and 12.1 mm respectively at 100 µg mL-1. However, it was 13.6 mm and 15.3 mm for Gram negative P. aeruginosa and P. vulgaris respectively. The biofilm of tested bacteria was significantly inhibited at 100 µg mL-1. The hydrophobicity of bacterial biofilms was considerably condensed after treatment with the hen's albumen extracts at 100 µg mL-1. The anti-inflammatory activity of hen's albumen extracts was confirmed by the inhibition of cyclooxygenase (COX) enzyme to 84.91% at 100 µg mL-1 with the relative IC50 of 8.26 µg mL-1. The albumen extract effectively inhibited the viability (23.61%) of HepG2 hepatic cancer cells at 100 µg mL-1. The anti-proliferative activity of the albumen extracts was further revealed by the induction of HepG2 apoptotic cell morphology. This study concludes that the hen's albumen extract treated with M. pustulata is a natural therapeutic agent to treat biofilm associated clinical bacteria, inflammations and human hepatic cancer cells.

Garlic clove extract assisted silver nanoparticle - Antibacterial, antibiofilm, antihelminthic, anti-inflammatory, anticancer and ecotoxicity assessment.

Facile and low cost garlic clove extract based silver nanoparticles was synthesized and its broad spectrum of therapeutic activity including antibiofilm, antiparasitic and anti-breast cancer activity was evaluated. The synthesized garlic­silver nanoparticles (G-AgNPs) were characterized by various physico-chemical techniques. G-AgNPs showed good optical property, highly crystalline nature, spherical shape and uniformly dispersed with size measuring between 10 and 50 nm. G-AgNPs have shown greater anti-bacterial and antibiofilm activity on clinically important pathogens methicillin-resistant S. aureus and P. aerigunosa at 100 µg ml-1. The efficacy of G-AgNPs against earthworm evidenced its effectiveness as anti-helminthic agent in treating intestinal parasites. The significant inhibition of BSA protein denaturation proves its anti-inflammatory property. In addition, G-AgNPs have shown remarkable anticancer effect and significantly inhibited the human breast cancer cell (MCF-7) viability at 100 µg ml-1 after 24 h. A noticeable change in the morphology of MCF-7 cells was also noticed. G-AgNPs were non-toxic to human HEK293 embryonic cells. Also, the non-toxic nature of G-AgNPs to C. cornuta and no morphological, physiological changes proved its safety to the environment. It is concluded that G-AgNPs have a broad range of biological applications and it can be used as an eco-friendly material without having negative effects in the environment.

In vitro antagonistic activity and the protective effect of probiotic Bacillus licheniformis Dahb1 in zebrafish challenged with GFP tagged Vibrio parahaemolyticus Dahv2.

In vitro antagonistic activity and the protective effect of probiotic Bacillus licheniformis Dahb1 in zebrafish (Danio rerio) challenged with GFP tagged Vibrio parahaemolyticus Dahv2 was studied. The cell free extract of probiotic B. licheniformis Dahb1 at 100 µg mL-1 showed growth inhibition of V. parahaemolyticus Dahv2 in vitro. B. licheniformis Dahb1 also inhibited the biofilm growth of GFP tagged V. parahaemolyticus Dahv2 at 100 µg mL-1 in vitro. The growth and survival of zebrafish was tested using probiotic B. licheniformis Dahb1. Weight (1.28 g) of zebrafish that received the cell free extract was much higher than in control (1.04 g). The mortality of zebrafish infected with GFP tagged V. parahaemolyticus Dahv2 at 107 Cfu mL-1 (Group IV) was 100%, whereas a complete survival of zebrafish that received the cell free extract of B. licheniformis Dahb1 at 107 Cfu mL-1 (Group VII) was observed after 30 days. The number of GFP tagged V. parahaemolyticus Dahv2 colonies in the intestine and gills significantly reduced after treatment with the cell free extract of B. licheniformis Dahb1. Furthermore, a significant decrease in the fluorescent colonies of GFP tagged V. parahaemolyticus Dahv2 was observed after treatment with the cell free extract of B. licheniformis Dahb1 under confocal laser scanning microscopy (CLSM). In conclusion, the cell free extract of B. licheniformis Dahb1 could prevent Vibrio infection by enhancing the growth and survival of zebrafish.

Biological control of the Pulse beetle, Callosobruchus maculatus in stored grains using the entomopathogenic bacteria, Bacillus thuringiensis.

In the present study, we reported the biological control of stored product insect pest, Callosobruchus maculatus using the entomopathogenic bacteria, Bacillus thuringiensis. A significant delay in the larval, pupal and total development period of C. maculatus was observed after treatment with B. thuringiensis at 4 × 108 cells/mL. Furthermore, B. thuringiensis are highly effective in the control of C. maculatus and produced 100% mortality at 4 × 108 cells/mL. The LC50 value was estimated to be 3 × 107 cells/mL. In addition, a significant decrease in the activity of mid-gut α-amylase, cysteine protease, α & ß-glucosidases, lipase, glutathione S-transferase (GST) and lactate dehydrogenase (LDH) was observed after treatment with B. thuringiensis at 4 × 108 cells/mL. This study concludes that B. thuringiensis are more effective against C. maculatus and could be used as a potential biological control agent in the management of stored product insect pests in the future.

Two potential uses for silver nanoparticles coated with Solanum nigrum unripe fruit extract: Biofilm inhibition and photodegradation of dye effluent.

Silver nanoparticle was green synthesized involving the unripe fruit extracts of Solanum nigrum (Sn-AgNPs). The synthesized Sn-AgNPs was bio-physically characterized by UV-Vis spectroscopy, X-ray diffraction (XRD), Fourier transform infrared spectroscopy (FTIR) and Transmission electron microscopy (TEM). UV-Vis recorded the absorbance spectra at 443 nm. XRD analysis clearly demonstrated the crystalline nature of Sn-AgNPs with Bragg's reflection peaks at 111, 200, 220 and 311 lattice planes. The FTIR spectrum of Sn-AgNPs showed strong bands at 3432, 1555, 1455, 862 and 406 cm-1 which corresponds at O-H, C-H, C-C, C-OH and C-N groups respectively. TEM exhibited the spherical shape of Sn-AgNPs with particle size between 20 and 30 nm. The antibacterial effects of Sn-AgNPs were tested on clinically important biofilm forming Gram positive (Bacillus pumulis and Enterococcus faecalis) and Gram negative (Proteus vulgaris and Vibrio parahaemolyticus) bacteria. The greater inhibition of B. pumulis and E. faecalis was observed at 100 µg mL-1 of Sn-AgNPs compared to P. vulgaris and V. parahaemolyticus. The biofilm inhibition potential of Sn-AgNPs was greater against Gram positive bacteria than that of Gram negative bacteria. Furthermore, Sn-AgNPs effectively degraded the industrial effluent methyl orange dye by photocatalysis. It is concluded that Sn-AgNPs could be used as an effective therapeutics against the biofilm of clinically important bacteria. The green synthesized Sn-AgNPs can be employed to degrade dye effluents and prevent environmental pollution as well.

Bacillus thuringiensis coated zinc oxide nanoparticle and its biopesticidal effects on the pulse beetle, Callosobruchus maculatus.

Insect pests belonging to the genus Callosobruchus are the major cause of damage to stored pulse crops. Recently, nanotechnology has emerged as a promising tool for pest control. In the present study, we report for the first time the synthesis and biological evaluation of Bacillus thuringiensis coated zinc oxide nanoparticles (Bt-ZnO NPs) on the pulse beetle, Callosobruchus maculatus. The biologically synthesized Bt-ZnO NPs were extensively characterized using UV-Vis spectroscopy, X-ray diffraction (XRD), Fourier transform infrared spectroscopy (FTIR), Transmission electron microscopy (TEM) and Zeta potential. The bio-physical characterization revealed that the Bt-ZnO NPs has a hexagonal wurtzite structures with an average particle size of 20nm. In addition, zeta potential measurement demonstrated that the Bt-ZnO NPs are negatively charged (-12.7mV) and are moderately stable. The biopesticidal effect of Bt-ZnO NPs was tested against the pulse beetle, C. maculatus. Treatment with Bt-ZnO NPs reduced the fecundity (eggs laid) and hatchability of C. maculatus in a dose-dependent manner. A significant delay in the larval, pupal and total development period of C. maculatus was observed after treatment with Bt-ZnO NPs at 25µg/mL. Furthermore, Bt-ZnO NPs are highly effective in the control of C. maculatus and caused 100% mortality at 25µg/mL. The LC50 value was estimated to be 10.71µg/mL. In addition, treatment with Bt-ZnO NPs decreased the mid-gut α-amylase, cysteine protease, α-glucosidase and glutathione S-transferase (GST) activity in C. maculatus. Our results suggest that Bt-ZnO NPs are effective against C. maculatus and could be used as nanobiopesticides in the control of stored grain insect pests in the future.

A novel antimicrobial therapy for the control of Aeromonas hydrophila infection in aquaculture using marine polysaccharide coated gold nanoparticle.

In the present study, we prepared fucoidan coated Au-NPs (Fu-AuNPs), and examined its antimicrobial activity against Aeromonas hydrophila. The green synthesized Fu-AuNPs were bio-physically characterized by Ultraviolet-visible (UV-Vis) spectroscopy, X-ray Diffraction (XRD), Fourier Transform Infrared spectroscopy (FTIR), Higher Transmission Electron Microscopy (HR-TEM), Zeta potential analysis and Energy Dispersive X-ray spectroscopy (EDX). Fu-AuNPs were crystalline in nature, spherical to triangular in shape, with particle size ranging within 10-100 nm. The synthesized Fu-AuNPs at 100 µg mL-1 showed inhibition zone against A. hydrophila (23.2 mm) which is much higher than that of commercial antibiotic chloramphenicol (17.3 mm). The biofilm inhibitory activity of Fu-AuNPs against Gram negative (Aeromonas hydrophila) was higher. Light and confocal laser scanning microscopic observations showed that the Fu-AuNPs at 100 µg mL-1 inhibited the biofilm of A. hydrophila. The cytotoxicity study indicated that Fu-AuNPs were effective in inhibiting the viability of human cervical cancer cells (HeLa) at 100 µg mL-1. In another experiment, the antibacterial effect of Fu-AuNPs on tilapia, Oreochromis mossambicus were evaluated in vivo. The mortality rate of O. mossambicus infected by A. hydrophila was much higher (90%), whereas, the mortality of O. mossambicus that received Fu-AuNPs followed by challenge with A. hydrophia was reduced to 30%. This study concludes that Fu-AUNPs are effective in the control of A. hydrophila infections in O. mossambicus.

Control of biofilm forming clinically important bacteria by green synthesized ZnO nanoparticles and its ecotoxicity on Ceriodaphnia cornuta.

Zinc oxide nanoparticles were synthesized using the aqueous leaf extracts of Plectranthus barbatus (Plb-ZnO NPs) and characterized by UV-visible spectroscopy, XRD, FTIR, SEM and EDS. UV-Visible spectra recorded the absorbance peak of Plb-ZnO NPs at 343 nm. SEM analyses showed the spherical shape of Plb-ZnO NPs with the particle size between 30 and 60 nm. Plb-ZnO NPs exhibited antibacterial and antibiofilm activity against Gram positive Bacillus subtilis at all tested concentrations. In contrast, Plb-ZnO NPs showed antibacterial and antibiofilm activity against Gram negative Vibrio parahaemolyticus and Proteus vulgaris only at 100 µg/ml. The Atomic absorption spectrometry (AAS) revealed that Zn2+ dissolution was 1.87 and 8.8 µg/L at 10 and 160 µg/L of Plb-ZnO NPs respectively. The body accumulation of Zn2+ was increased from 0.8 µg/g body weight to 3.5 µg/g body weight when C. cornuta exposed to 10 µg/L and 160 µg/L respectively. Plb-ZnO NPs were toxic to Ceriodaphnia cornuta neonates (LC50: 28 µg/L). Plb-ZnO NPs caused 100% mortality of C. cornuta at 160 µg/L after 24 h. However, zinc acetate does not cause any mortality of C. cornuta upto 350 µg/L. The light and confocal laser scanning microscopic images evidenced the uptake and accumulation of Plb-ZnO NPs on the internal gut regions of C. cornuta at 160 µg/L after 5, 10, 15, 20 and 24 h. Abnormalities in the swimming behaviour such as erratic swimming (ERR), migration to bottom (BOT) and migration to water surface (SUR) of C. cornuta were noticed after treatment with different concentrations of Plb-ZnO NPs.

Biological therapeutics of Pongamia pinnata coated zinc oxide nanoparticles against clinically important pathogenic bacteria, fungi and MCF-7 breast cancer cells.

The overuse of antimicrobics and drugs has led to the development of resistance in a number of pathogens and parasites, which leads to great concerns for human health and the environment. Furthermore, breast cancer is the second most common cause of cancer death in women. MCF-7 is a widely used epithelial cancer cell line, derived from breast adenocarcinoma for in vitro breast cancer studies, since the cell line has retained several ideal characteristics particular to the mammary epithelium. In this scenario, the development of novel and eco-friendly drugs are of timely importance. Green synthesis of nanoparticles is cost effective, environmental friendly and does not involve the use of toxic chemicals or elevate energy inputs. This research focused on the anticancer activity of Pongamia pinnata seed extract-fabricated zinc oxide nanoparticles (Pp-ZnO NPs) on human MCF-7 breast cancer cells, antibiofilm activity against bacteria and fungi was also investigated. Nanoparticles were characterized by UV-Vis spectroscopy, X-ray diffraction (XRD), Fourier transform infrared spectroscopy (FTIR), Scanning electron microscopy (SEM) and Energy dispersive X-ray spectroscopy (EDX). Pp-ZnO NPs effectively inhibited the growth of Gram positive Bacillus licheniformis (zone of inhibition: 17.3 mm) at 25 µg ml-1 followed by Gram negative Pseudomonas aeruginosa (14.2 mm) and Vibrio parahaemolyticus (12.2 mm). Pp-ZnO NPs also effectively inhibited the biofilm formation of C. albicans at 50 µg ml-1. Cytotoxicity studies revealed that a single treatment with Pp-ZnO NPs significantly reduced the cell viability of breast cancer MCF-7 cells at doses higher than 50 µg ml-1. Morphological changes in the Pp-ZnO NPs treated MCF-7 breast cancer cells were observed using phase contrast microscopy. This study concludes that the green synthesized Pp-ZnO NPs may be used as an effective antimicrobial and antibreast cancer agents.

Oreochromis mossambicus diet supplementation with Psidium guajava leaf extracts enhance growth, immune, antioxidant response and resistance to Aeromonas hydrophila.

In this research, we focused on the efficacy of aqueous and ethanol leaf extracts of Psidium guajava L. (guava) based experimental diets on the growth, immune, antioxidant and disease resistance of tilapia, Oreochromis mossambicus following challenge with Aeromonas hydrophila. The experimental diets were prepared by mixing powdered (1, 5 and 10 mg/g) aqueous and ethanol extract of guava leaf with commercial diet. The growth (FW, FCR and SGR), non-specific cellular immune (myeloperoxidase activity, reactive oxygen activity and reactive nitrogen activity) humoral immune (complement activity, antiprotease, alkaline phosphatase activity and lysozyme activity) and antioxidant enzyme responses (SOD, GPX, and CAT) were examined after 30 days of post-feeding. A significant enhancement in the biochemical and immunological parameters of fish were observed fed with experimental diets compared to control. The dietary supplementation of P. guajava leaf extract powder for 30 days significantly reduced the mortality and increased the disease resistance of O. mossambicus following challenge with A. hydrophila at 50 µl (1 × 107 cells ml-1) compared to control after post-infection. The results suggest that the guava leaf extract could be used as a promising feed additive in aquaculture.

Laurus nobilis leaf extract mediated green synthesis of ZnO nanoparticles: Characterization and biomedical applications.

The present study reports the green synthesis of zinc oxide nanoparticles using the aqueous leaf extract of Laurus nobilis (Ln-ZnO NPs) by co-precipitation method. The synthesized Ln-ZnO NPs were characterized by UV-Vis spectroscopy, FTIR, XRD, TEM, SEM and EDX. Ln-ZnO NPs were crystalline in nature, flower like and have hexagonal wurtzite structure with a mean particle size of 47.27nm. The antibacterial activity of Ln-ZnO NPs was greater against Gram positive (Staphylococcus aureus) bacteria than Gram negative (Pseudomonas aeruginosa) bacteria. The zone of inhibition against S. aureus was 11.4, 12.6 and 14.2mm at 25, 50 and 75µgmL-1. The zone of inhibition against P. aeruginosa was 9.8, 10.2 and 11.3mm at 25, 50 and 75µgmL-1. The light and confocal laser scanning microscopic images evidenced that Ln-ZnO NPs effectively inhibited the biofilm growth of S. aureus and P. aeruginosa at 75µgmL-1. The cytotoxicity studies revealed that Ln-ZnO NPs showed no effect on normal murine RAW264.7 macrophage cells. On the other hand, Ln-ZnO NPs were effective in inhibiting the viability of human A549 lung cancer cells at higher concentrations of 80µgmL-1. The morphological changes in the Ln-ZnO NPs treated A549 lung cancer cells were observed under phase contrast microscope.

Chitosan coated Ag/ZnO nanocomposite and their antibiofilm, antifungal and cytotoxic effects on murine macrophages.

In the present study, chitosan coated Ag/ZnO (CS/Ag/ZnO) nanocomposite was synthesized and characterized by UV-Vis spectroscopy (UV-Vis), X-ray diffraction (XRD), Fourier transform infrared spectroscopy (FTIR) and Scanning electron microscopy (SEM). The CS/Ag/ZnO nanocomposite exhibited antibacterial activity against Gram positive (B. licheniformis and B. cereus) bacteria at 8 µg mL-1 compared to Gram negative (V. parahaemolyticus. and P. vulgaris) bacteria. CS/Ag/ZnO nanocomposite effectively inhibited the biofilm growth of Gram positive bacteria compared to Gram negative bacteria at 30 µg mL-1. The hydrophobicity index and EPS (extracellular polysaccharide) production of both Gram positive and Gram negative bacteria was decreased after treatment with 30 µg mL-1 of CS/Ag/ZnO nanocomposite. CS/Ag/ZnO nanocomposite showed effective control of fungal C. albicans biofilm (92%) at 50 µg mL-1. The inhibition of bacterial and fungal biofilms was clearly visualized under light and confocal laser scanning microscopy (CLSM). CS/Ag/ZnO nanocomposite was observed to be non toxic to RAW264.7 murine macrophages and no changes in the morphology of macrophages was observed under phase contrast microscopy. The study concludes that CS/Ag/ZnO nanocomposite is the promising candidate to be used as biomaterial against bacterial and fungal infections without any toxicity risk.

Essential oils of Nigella sativa protects Artemia from the pathogenic effect of Vibrio parahaemolyticus Dahv2.

The anti-Vibrio activity of essential oils (EOs) of nine medicinal plants was tested against 28 Vibrio spp. isolated from diseased Fenneropenaeus indicus. EO of Nigella sativa exhibited anti-Vibrio activity against all Vibrio spp. and greater inhibition was noted for the isolate V2 which was identified as Vibrio parahaemolyticus Dahv2. Further, EO of N. sativa effectively inhibited V. parahaemolyticus Dahv2 with an inhibition zone of 23.9mm at 101.2µgml(-1). Moreover, EO of N. sativa revealed anti-biofilm activity at 101.2µgml(-1) against V. parahaemolyticus Dahv2 and inhibited the growth of V. parahaemolyticus Dahv2 at 100µgml(-1).In vivo experimental infection studies showed that the survival of Artemia spp. infected with V. parahaemolyticus Dahv2 at 1×10(3)cfuml(-1) was only 40%. However, the survival of Artemia spp. was significantly increased after challenge with 100µgml(-1) of EO of N. sativa. EO of N. sativa showed higher anti-oxidant potential and total phenol content than other EOs tested. The anti-oxidant activity of EO of N. sativa was highly correlated to their total phenolic contents (r=0.836, P<0.05). This observation suggests that EO of N. sativa protected the Artemia spp. after experimental infection of V. parahaemolyticus Dahv2.

GFP tagged Vibrio parahaemolyticus Dahv2 infection and the protective effects of the probiotic Bacillus licheniformis Dahb1 on the growth, immune and antioxidant responses in Pangasius hypophthalmus.

In this study, the pathogenicity of GFP tagged Vibrio parahaemolyticus Dahv2 and the protective effect of the probiotic strain, Bacillus licheniformis Dahb1 was studied on the Asian catfish, Pangasius hypophthalmus. The experiment was carried out for 24 days with three groups and one group served as the control (without treatment). In the first group, P. hypophthalmus was orally infected with 1 mL of GFP tagged V. parahaemolyticus Dahv2 at two different doses (10(5) and 10(7) cfu mL(-1)). In the second group, P. hypophthalmus was orally administrated with 1 ml of the probiotic B. licheniformis Dahb1 at two different doses (10(5) and 10(7) cfu mL(-1)). In the third group, P. hypophthalmus was orally infected first with 1 mL of GFP tagged V. parahaemolyticus Dahv2 followed by the administration of 1 mL of B. licheniformis Dahb1 (combined treatment) at two different doses (10(5) and 10(7) cfu mL(-1)). The growth, immune (myeloperoxidase, respiratory burst, natural complement haemolytic and lysozyme activity) and antioxidant (glutathione-S-transferase, reduced glutathione and total glutathione) responses of P. hypophthalmus were reduced after post infection of GFP tagged V. parahaemolyticus Dahv2 compared to control. However, after administration with the probiotic B. licheniformis Dahb1 at 10(5) cfu mL(-1), P. hypophthalmus showed significant increase in the growth, immune and antioxidant responses compared to 10(7) cfu mL(-1). On the otherhand, the growth, immune and antioxidant responses of P. hypophthalmus infected and administrated with combined GFP tagged Vibrio + Bacillus at 10(5) cfu mL(-1) were relatively higher than that of GFP tagged V. parahaemolyticus Dahv2 and control groups but lower than that of probiotic B. licheniformis Dahb1 groups. The results of the present study conclude that the probiotic B. licheniformis Dahb1 at 10(5) cfu mL(-1) has the potential to protect the P. hypophthalmus against V. parahaemolyticus Dahv2 infection by enhancing the growth, immune and antioxidant responses. The probiotic B. licheniformis Dahb1 would be effectively used in the treatment of aquatic diseases for improvement of aquaculture industry.