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The prevalence of infectious diseases in the aquaculture industry and a limited number of safe and effective oral vaccines has imposed a challenge not only for fish immunity but also a threat to human health. The availability of fish oral vaccines has expanded recently, but little is known about how well they work and how they affect the immune system. The unsatisfactory efficacy of existing oral vaccinations is partly attributable to the antigen degradation in the adverse gastrointestinal environment of fishes, the highly tolerogenic gut environment, and inferior vaccine formulation. To overcome such challenges in designing: an easier, cost-efficient, and effective vaccination method, several encapsulation methods are being adopted to safeguard antigens from the intestinal atmosphere for their immunogenic functions. Oral vaccination is easily degraded by gastric acids and enzymes before reaching the immunological site; however, this issue can be solved by encapsulating antigens in poly-biodegradable nanoparticles, transgenic designed bacteria, plant systems, and live feeds. To enhance the immunological impact, each antigen delivery method operates at a different level. Utilizing nanotechnology, it has been possible to regulate vaccination parameters, target particular cells, and lower the antigen dosage with potent nanomaterials such as chitosan, poly D,L-lactic-co-glycolic acid (PLGA) as vaccine carriers. Live feeds such as Artemia salina can be utilized as bio-carrier, owing to their appropriate size and non-filter feed system, through a process called bio-encapsulation. It ensures the protection of antigens over the fish intestine and ensures complete uptake by immune cells in the hindgut for increased immune response. This review comprises recent advances in oral vaccination in aquaculture in terms of an encapsulation approach that can aid in future research. [ABSTRACT FROM AUTHOR]

The role of mosquito vectors in spreading disastrous diseases to living organisms, especially to humans is inevitable and undeniable. The impacts of the available chemical and synthetic insecticides on non-specific organisms as well as on nature are being the reason behind the search for target-specific, biocompatible and eco-friendly alternatives. The Madhuca longifolia seed extract and cry proteins from Bacillus thuringiensis -based nanocomposites (Cp- Ml -ZnO NCs) were produced to conquer the above-mentioned issues. The Cp- Ml -ZnO NCs (100 μg/mL) expressed better scavenging potentiality on 2,2-diphenyl-1-picryl-hydrazyl-hydrate (DPPH) radicals than Ml seed extract and Ml -ZnO NPs. The susceptibility of tested vector larvae to the Cp- Ml -ZnO NCs was Ae. aegypti ˃An. stephensi ˃ Cx. quinquefasciatus along with LC 50 -27.73, 34.81, and 42.54 μg/mL concentration. The target specificity and biocompatibility of Cp- Ml -ZnO NCs were authenticated by the results obtained by evaluating the efficacy on D. similis , A. salina , P. reticulata , G. affinis , and RBCs of goat blood. Thus the Cp- Ml -ZnO NCs could be adopted for the control of vector larvae. • Nanocomposites (Cp- Ml -ZnO NCs) are successfully prepared using core shell technique. • Cp- Ml -ZnO NCs are having better antioxidant activity than Ml -ZnO NPs and extract. • Mosquito larval population was drastically controlled by the synthesized NCs. • Fortunately, they are biocompatible with non-specific Daphnia , Artemia and etc. [ABSTRACT FROM AUTHOR]

Lycopersicon esculentum leaf-extract was utilized in the synthesis of zinc-oxide nanoparticles (Le-ZnO-NPs) using zinc acetate as the precursor. The synthesized ZnO-NPs were attested by the detection of a notable absorption peak at 343 nm, appearing in UV–Vis spectra due to surface-plasmon-resonance. The XRD-spectrum recorded at 2θ values coincides with that of lattice planes. HR-TEM results revealed that Le-ZnO-NPs exhibited a crystalline structure with hexagonal wurtzite shape (10–50 nm size). The effectiveness of inhibition of the microbial biofilms of Proteus vulgaris, Enterococcus faecalis and Candida albicans was analyzed at a concentration of 100 µg ml−1. Moreover, Le-ZnO-NPs showed a significant range of anti-coagulant activity, in contrast to commercial anti-coagulant (heparin). Cytotoxicity studies showed that Le-ZnO-NPs (100 µg ml−1) are non-toxic to murine macrophage cells, but toxic to human cervical cancer (HeLa) cells. Indeed, a reduction in the viability of HeLa-cells and a notable variation in the apoptotic cell structure were apparent. [ABSTRACT FROM AUTHOR]

Abstract Arboviral diseases and microbial pathogens resistant to commercially available drugs are on the rise. Herein, a facile microbial-based approach was developed to synthesize selenium nanowires (Se NWs) using microbial exopolymer (MEP) extracted from the Bacillus licheniformis (probiotic bacteria). MEP-Se NWs were characterized using UV–Visible, XRD, FTIR, HR-TEM, FE-SEM and EDX. An UV–Visible peak was detected at 330 nm while XRD spectrum data pointed out the crystalline nature of MEP-Se NWs. FTIR spectrum revealed functional groups with strong absorption peaks in the range 3898.52–477.97 cm−1. FE-SEM and HR-TEM revealed that the obtained structures were nanowires of 10–30 nm diameter. Se presence was confirmed by EDX analysis. MEP-Se NWs at 100 μg/ml highly suppressed the growth of both Gram (−) and Gram (+) bacteria. Further, microscopic analysis evidenced that 75 μg/ml MEP-Se NWs suppressed biofilm formation. Hemolytic assays showed that MEP-Se NWs were moderately cytotoxic. In addition, LC 50 values lower than 10 μg/ml were estimated testing MEP-Se NWs on both Aedes aegypti and Culex quinquefasciatus 3rd instar larvae. Morphological and histological techniques were used to elucidate on the damages triggered in mosquito tissues, with special reference to midgut, post-exposure to MEP-Se NWs. Therefore, based on our findings, MEP-Se NWs can be considered for entomological and biomedical applications, with special reference to the management of biofilm forming microbial pathogens and arbovirus mosquito vectors. Graphical Abstract Unlabelled Image Highlights • Se nanowires (Se NWs) were synthesized using microbial exopolymers (MEP) from Bacillus licheniformis Dahb1. • MEP-Se NWs were characterized by UV–Vis, XRD, FTIR, FE-SEM, HR-TEM and EDX. • MEP-Se NWs revealed great antioxidant and antibiofilm activity. • Low hemolytic activity of MEP-Se NWs pointed out their biocompatible nature. • MEP-Se NWs were highly toxic to Aedes aegypti and Culex quinquefasicatus larvae. [ABSTRACT FROM AUTHOR]

Abstract Murraya koenigii berry extract based zinc oxide nanoparticles (Mk -ZnO NPs) were synthesized by simple co-precipitation method and examined for bacteriostatic and larvicidal efficiency. Synthesized Mk -ZnO NPs were characterized by UV–Vis spectroscopy at 336 nm. X-Ray diffraction (XRD) showed crystalline nature as hexagonal. Fourier transform infrared spectroscopy (FTIR) spectrum exhibited strong peak at 3442.80 cm−1. Field emission scanning electron microscopy (FE-SEM) showed hexagonal shape of the particle. Transmission electron microscopy (TEM) measured 10–15 nm sized Mk -ZnO NPs. EDX peaks confirm 71.99% of zinc and 11.42% of oxide in Mk -ZnO NPs. Minimum inhibitory concentration (MIC) analysis reveals Mk -ZnO NPs inhibit growth of Gram positive (Staphylococcus aureus , Lysinibacillus fusiformis) and Gram negative (Proteus vulgaris , Providencia vermicola) bacteria at 40 and 50 μg mL−1 respectively. Live & dead assay confirms that Mk -ZnO NPs inhibits bacterial growth at 50 μg mL−1. Bacterial biofilm thickness significantly reduced by Mk -ZnO NPs at 50 μg mL−1. In vitro toxicity of Mk -ZnO NPs on RAW 264.7 macrophages determines 90–50% cell viability at concentrations of 10–100 μg mL−1. In vivo toxicity assay results indicate the lethal concentration of Artemia nauplii were LC 50− 78.73 μg mL−1 and LC 90− 130.03 μg mL−1. Larvicidal activity of Mk -ZnO NPs towards mosquito larvae of Culex quinquefasciatus were observed at LC 50− 2.1 μg mL−1 and LC 90− 12.1 μg mL−1. Finally the study discloses, potential bacteriostatic effect and mosquito larvae controlling capacity of Mk -ZnO NPs. Graphical Abstract Unlabelled Image Highlights • Inexpensive and ecofriendly ZnO NPs synthesized by using M. koenigii berry extract. • Quantified bacteriostatic upshots of Mk -ZnO NPs were analyzed and determined. • In vitro toxicity of Mk -ZnO NPs tested against RAW 264.7 macrophages. • In vivo toxicity effect of Mk -ZnO NPs on Artemia salina. • Larvicidal activity of Mk -ZnO NPs examined against larvae of C. quinquefasciatus. [ABSTRACT FROM AUTHOR]

Abstract In the present study, immunoenhancing effect of β-1, 3 glucan binding protein based selenium nanowire (Ph β-GBP-SeNWs) in common carp, Cyprinus carpio was assessed. Biological based selenium nanoform was synthesized, using crustacean immune molecule β-GBP purified from the haemolymph of Paratelphusa hydrodromus. The morphological property of Ph β-GBP-SeNWs was analyzed through TEM which reveals, the synthesized nanowire exhibits approximately 30–50 nm width with smooth surface. For this current study, fish were fed with experimental diet includes Ph β-GBP, sodium selenite, selenomethionine and Ph β-GBP-SeNWs supplemented diet at different concentrations (0.5 mg, 1 mg and 2 mg) for 30 days. The growth performance, cellular and humoral immune responses (myeloperoxidase, reactive oxygen species, alkaline phosphatase and lysozyme activity) and antioxidant enzymes (glutathione peroxidase and catalase activity) in the fish fed with Ph β-GBP-SeNWs supplemented diet were significantly increased in dose-dependent manner, which was observed at two different interval period (15th and 30th day). Also, Ph β-GBP-SeNWs supplemented diet fed fish gain resistant after challenged with aquatic pathogen Aeromonas hydrophila and the relative survival percentage was increased. Agar disc diffusion and BacLight assay clearly demonstrated the antibacterial property of plasma of fish fed with Ph β-GBP-SeNWs supplemented diet against aquatic pathogen A. hydrophila , Vibrio parahaemolyticus and Vibrio alginolyticus. Moreover, confocal laser scanning microscopic analysis clearly showed that, Ph β-GBP-SeNWs supplemented diet fed fish plasma was more efficient in disrupting the architecture of bacterial colonies and thereby reduced the thickness of biofilm. Thus, the present study indicates that, incorporation of Ph β-GBP-SeNWs in the diet enhances the fish immune responses and disease resistance against aquatic pathogens. Graphical abstract Image 1 Highlights • β-GBP was purified from the Paratelphusa hydrodromus haemolymph. • Ph β-GBP based selenium nanowires (Ph β-GBP SeNWs) were synthesized and characterized. • Ph β-GBP SeNWs supplemented diet and its impact on immune parameters were studied. [ABSTRACT FROM AUTHOR]

The use of natural biopolymers in the synthesis of nanomaterials can have a low cost and eco-friendly approach. ZnO nanoparticles synthesized through biological method has been reported to have biomedical applications to control pathogenic microbes as it is cost effective compared to commonly used physical and chemical methods. In this work we would like to report the “bioinspired” synthesis of ZnO nanopowders (ZnO-NPs) using type 1 collagen. Collagen based ZnO NPs (Cl-ZnO NPs) were bio-physically characterized by UV–vis Spectroscopy, XRD, FTIR, HR-TEM, EDX and Zeta potential analysis. HR-TEM recorded the presence of hexagonal wurtzite structure of Cl-ZnO NPs with particle size ranged between 20–50 nm. Further, Cl-ZnO NPs exhibited antibacterial and antibiofilm activity against Gram positive Streptococcus mutans, Gram negative Proteus vulgaris and fungi Candida albicans at 75 μg/ml. Moreover, the cytotoxicity assay demonstrated that the Cl-ZnO NPs was not toxic to murine (RAW 264.7) macrophage cells up to 75 μg/ml. However, it exhibited cytotoxicity against human liver cancer (HepG2) cells at 75 μg/ml. The HepG2 cell viability was significantly reduced at 75 μg/ml. In addition, the ecotoxicity of Cl-ZnO NPs on the freshwater micro crustacean Daphnia longicephala showed no mortality up to 250 μg/ml. The current study clearly demonstrated that the Cl-ZnO NPs had greater potential for antimicrobial and anticancer activities. [ABSTRACT FROM AUTHOR]

Graphical abstract Highlights • Purification of crustin (Pp -Cru) from Portunus pelagicus was characterized by HPLC, CD, XRD, and FTIR analyses. • We reported an enhancement of the immune system by Pp -Cru through in vivo and in vitro analysis. • Synergic antibacterial and antibiofilm properties of Pp -Cru were observed against key bacterial pathogens. • Live and dead assay showed ultra-low bacterial cell viability post-treatment with Pp -Cru. • In vivo study of crustin was assessed with Artemia salina as a model crustacean. Abstract Marine organisms represent a huge source of novel compounds for the development of effective antimicrobial drugs. The present study focus on the purification of the antimicrobial peptide crustin from the haemolymph of the blue swimmer crab, Portunus pelagicus , by blue Sepharose CL-6B matrix assisted affinity column chromatography. Crustin showed a single band with a molecular mass of 17 kDa in SDS-PAGE analysis. The XRD analysis exhibited peaks at 32° and 45° while a distinct peak with a retention time of 1.8 min resulted in high performance liquid chromatography (HPLC) pointing out the crystalline nature and purity of crustin, respectively. Crustin purified from P. pelagicus (Pp -Cru) showed immunological activities, triggering encapsulation, phagocytosis on Sepharose beads and yeast (Saccharomyces cerevisiae) respectively. Furthermore, encapsulation of GFP tagged V. parahaemolyticus in Artemia salina and challenging study were assessed under CLSM and the potential of Pp -Cru was examined in vivo. In addition, the growth reduction and biofilm inhibition potential of Pp -Cru on S taphylococcus aureus, Enterococcus faecalis (Gram- positive bacteria) and Pseudomonas aeruginosa, Escherichia coli (Gram-negative bacteria) was evidenced by inverted and confocal laser scanning microscopic analysis, revealing that 100 μg/ml of Pp -Cru can disrupt the biofilm matrix thereby the thickness of biofilm was significantly reduced. Overall, the present investigation might provide a sensitive platform to realize the significant function of Pp -Cru in crustacean immune mechanism as well as its potential to bacterial growth inhibitor. The functional properties of purified Pp -Cru antimicrobial peptide may lead to a superior understanding of innate immune response in P. pelagicus species, which suggest the promising application for drug development in aquaculture. [ABSTRACT FROM AUTHOR]

• AI methods Accelerated Drug Screening and Design with structure and Ligand-based methods. • AI Learning models for categorizing data-driven drug discovery. • Applications and Limitations of AI models for drug discovery in pharmaceutical industries. • Need for balance between AI-driven predictions and experimental validations. Artificial intelligence (AI), particularly deep learning as a subcategory of AI, provides opportunities to accelerate and improve the process of discovering and developing new drugs. The use of AI in drug discovery is still in its early stages, but it has the potential to revolutionize the way new drugs are discovered and developed. As AI technology continues to evolve, it is likely that AI will play an even greater role in the future of drug discovery. AI is used to identify new drug targets, design new molecules, and predict the efficacy and safety of potential drugs. The inclusion of AI in drug discovery can screen millions of compounds in a matter of hours, identifying potential drug candidates that would have taken years to find using traditional methods. AI is highly utilized in the pharmaceutical industry by optimizing processes, reducing waste, and ensuring quality control. This review covers much-needed topics, including the different types of machine-learning techniques, their applications in drug discovery, and the challenges and limitations of using machine learning in this field. The state-of-the-art of AI-assisted pharmaceutical discovery is described, covering applications in structure and ligand-based virtual screening, de novo drug creation, prediction of physicochemical and pharmacokinetic properties, drug repurposing, and related topics. Finally, many obstacles and limits of present approaches are outlined, with an eye on potential future avenues for AI-assisted drug discovery and design. [ABSTRACT FROM AUTHOR]

In this study, we purified β-GBP from hemolymph of Scylla serrata crabs using affinity chromatography. The purified S . serrata β-GBP ( Ss -β-GBP) had 100 kDa molecular mass in the SDS-PAGE. MALDI-TOF/TOF analysis was conducted, revealing that the purified 100 kDa protein had 96% similarity with β-GBP of Astacus leptodactylus . Ss -β-GBP was characterized using high-performance liquid chromatography (HPLC), X-ray diffraction (XRD) analysis, circular dichroism (CD) and Fourier transform infrared (FTIR) spectroscopy, which confirmed the structure of the Ss -β-GBP. The purified Ss -β-GBP was functionally analyzed by yeast agglutination and phagocytic reaction assays. Moreover, the PO enhancing ability of Ss -β-GBP was evidenced through PO activity. Specifically, the antibacterial activity of the Ss -β-GBP against Gram-positive ( Enterococcus faecalis and Staphylococcus aureus ) and Gram-negative ( Escherichia coli and Pseudomonas aeruginosa ) bacteria was evaluated by determining its minimum inhibitory concentration (MIC) < 60 μg/ml for all tested species. Furthermore, the antibiofilm efficacy of Ss -β-GBP at 50 and 100 μg/ml was outlined using light microscopy and confocal laser scanning microscopy (CLSM). Bacterial viability assays also outlined the dose-dependent activity of Ss -β-GBP based on the ratio of live/dead bacterial cells. The results of this study revealed that crab-borne Ss -β-GBP might be widely used to suppress the growth of pathogenic bacteria. [ABSTRACT FROM AUTHOR]

Prophenoloxidase is a conserved Cu-containing enzyme acting as a major defense molecule in the immune response of crustaceans. In the present research, we purified prophenoloxidase from the haemolymph of Portunus pelagicus ( Pp -proPO) by Blue Sepharose CL-6B chromatography. Pp -proPO exhibited only one band with molecular weight of 75 kDa on SDS-PAGE. The purified Pp -proPO was characterized through X-ray diffraction (XRD) and high-performance liquid chromatography (HPLC). Pp -proPO showed phagocytic activity on the yeast Saccharomyces cerevisiae as well as encapsulation on sepharose CL-6B beads associated with CM sepharose and beads of sodium alginate. Pp -proPO also led to strong agglutination on human erythrocytes. Furthermore, Pp -proPO showed magnified PO activity when altered with activated particles acting as pathogen combined molecular patterns (PAMPs), metal ions or other chemicals. Pp -proPO showed relevant antibiofilm activity on Gram negative bacteria Pseudomonas aeruginosa and Escherichia coli . Overall, the above results allowed us to claim that Pp -proPO play a key role in immune defense mechanisms of P. pelagicus crabs, in particular towards microbial pathogens; notably we added basic information to the functional characterization of Pp -proPO, as well as to understand its immunological role in crustaceans defense systems. [ABSTRACT FROM AUTHOR]

The green synthesis of metal nanoparticles using phytochemical from marine seaweeds is a fast-growing research field in nanotechnology. Here, the biosynthesis of zinc oxide nanoparticles was achieved using the hot water extract of Sargassum wightii . The hot water extract prepared from S. wightii (H Sw ) and ZnO NPs were studied by UV–visible and FTIR spectroscopy, SEM and XRD. Then, both products were evaluated for antibiofilm activity towards aquatic pathogens. The nanoparticles' immunostimulating potential on green tiger prawns, Penaeus semisulcatus was studied through immersion and dietary administration. Shrimp immune parameters (i.e., total hemocytes count (THC), respiratory bursts (RBs), phenoloxidase (PO) and superoxide dismutase (SOD) activity) were significantly affected by exposure or ingestion of ZnO nanoparticles. In addition, the hot water extract and ZnO nanoparticles had high antibiofilm activity against Gram-positive ( B. subtilis, S. aureus ) and Gram-negative ( S. sonnei, P. aeruginosa ) microbial pathogens. It was accomplished that the ZnO nanoparticles can be used as the bacteriostatic and immunostimulant agents through immersion and dietary administration enhancing immunity of green tiger shrimp. Furthermore, the toxicity effects of ZnO nanoparticles were 100% at 24 h on Aedes aegypti 3 rd instar larvae at the concentration of 100 μg/mL and the greatest efficacy was accomplished by H Sw ZnO NPs against the Ae. aegypti after 24 h (LC 50 49.22; LC 90 86.96 mg/mL), if compared to the seaweed extract alone. Morphological and histological damages triggered by nanoexposure were investigated. [ABSTRACT FROM AUTHOR]

Recently, several immunostimulants such as β-glucan, microbial and plant products have been used as dietary supplements to combat disease outbreaks in aquaculture. The present study investigates the potential of Portunus pelagicus β-1, 3 glucan binding protein based zinc oxide nanoparticles ( Pp β-GBP-ZnO NPs) supplemented diet on growth, immune response and disease resistance in Mozambique tilapia, Oreochromis mossambicus . The immune-related protein β-GBP was purified from the haemolymph of P. pelagicus using Sephadex G-100 affinity column chromatography. Pp β-GBP-ZnO NPs was physico- chemically characterized and experimental feed was formulated. Fish were separately fed with commercial diet (control-group I) and Pp β-GBP (group II, III, IV), Pp β-GBP-ZnO NPs (group V, VI, VII), chem-ZnO NPs (VIII, IX, X) mixed diet at the concentration of 0.001%, 0.002% and 0.004% respectively. Triplicate groups of O. mossambicus were fed with experimental diets twice a day for 30 days. Fish receiving Pp β-GBP-ZnO NPs supplemented diet showed a significant increase (P < 0.05) in growth performance. Cellular immune responses (myeloperoxidase activity, lysozyme activity and reactive oxygen species activity) and humoral immune responses (complement activity, antiprotease activity and alkaline phosphatase activity) were evaluated at an interval of 15 days during the feeding trial. Results demonstrate that both cellular and humoral immune responses were substantially increased (P < 0.05) in fish fed with 0.004% of Pp β-GBP-ZnO NPs supplemented diet than others. Antibiofilm potential of Pp β-GBP-ZnO NPs against Aeromonas hydrophila was visualized through confocal laser scanning microscopy (CLSM), which reveals reduction in the preformed biofilm thickness to 10 μm  at the concentration of 50 μg/ml. Furthermore, after 30 days of feeding trial, fish were challenged with aquatic fish pathogen A. hydrophila (1 × 10 7  cells ml −1 ) through intraperitoneal injection. Challenge study displayed a reduced mortality rate in fish fed with diet containing Pp β-GBP-ZnO NPs. Thus our study suggests that dietary supplementation of Pp β-GBP-ZnO NPs at 0.004% may have a potential effect to enhance the immune system and survival of O. mossambicus . [ABSTRACT FROM AUTHOR]

Marine organisms are an outstanding source of molecules with wide biological activities. In this research, we investigated the antimicrobial protein haemocyanin isolated from the haemolymph of flower crab ( Portunus pelagicus ) using gel filtration chromatography. Purified P . pelagicus haemocyanin ( Pp -Hc) was subjected to SDS-PAGE analysis and the fraction containing Pp -Hc ran as a distinct band, its molecular weight was 78 kDa. The Pp -Hc was characterized through HPLC, XRD, CD and FTIR analyses. The antibacterial potential of Pp -Hc was evaluated against five Gram-positive and ten Gram-negative bacteria through the agar well diffusion method, which clearly revealed the high bactericidal activity of Pp -Hc. Furthermore, Pp -Hc was studied for its antibiofilm activity against five biofilm-forming Gram-negative bacteria, i.e., Vibrio alginolyticus , Vibrio harveyi , Vibrio parahaemolyticus , Pseudomonas aeruginosa , and Proteus vulgaris . Confocal laser scanning microscopy (CLSM) and light microscopy evidenced significant antibiofilm properties of Pp -Hc at 100 μg/ml concentration. Based on exopolysaccharide (EPS) quantification and cell surface hydrophobicity (CSH) index, the antibiofilm action of Pp -Hc was linked to its ability to reduce EPS production and increase cell surface hydrophobicity. The pathogen recognition mechanism of purified Pp -Hc showed a wide spectrum of binding activities, leading to high toxicity against both Gram-positive and Gram-negative bacterial species. Furthermore, the purified Pp -Hc showed high antifungal activity against the pathogen Candida albicans . Haemocyanin is not only involved in transporting oxygen, it has a relevant function in the flower crab immune system. This work adds useful knowledge for aquaculture concerns dealing with microbial pathogen control. Overall, our results outlined the high potential of purified Pp -Hc as a starting material to develop new and effective antimicrobial drugs for aquaculture purposes. [ABSTRACT FROM AUTHOR]

The present study evaluated the dietary supplementation of probiotic Bacillus licheniformis Dahb1 on the growth performance, immune parameters and antioxidant enzymes activities in serum and mucus as well as resistance against Aeromonas hydrophila in Mozambique tilapia Oreochromis mossambicus . Fish (24 ± 2.5 g) were fed separately with three diets, 1) commercial diet (control), 2) diet containing probiotic at 10 5  cfu g −1 (D1) and 3) diet containing probiotic at 10 7  cfu g −1 (D2) for 4 weeks. Growth performance in term of final weight (FW) specific growth rate (SGR) and feed conversion ratio (FCR), immune parameters of total protein (TP), alkaline phosphatase (ALP), myeloperoxidase (MPO), lysozyme (LYZ), reactive oxygen species (ROS), reactive nitrogen species (RNS) and antioxidant parameters of superoxide dismutase (SOD) and glutathione peroxidase (GPx) in serum and mucus were evaluated after 2 nd and 4 th weeks. The FW, SGR, and FCR of fish fed with D1 and D2 significantly improved ( p  < 0.05). The activities of ALP, LYZ and MPO in the mucus were significantly higher ( p  < 0.05) in fish that fed D1 and D2. The TP, ROS, RNS, SOD and GPx in the serum were significantly higher ( p  < 0.05) in fish that fed D1 and D2. In addition, the challenge test showed that fish fed D1 and D2 enhanced significantly ( p  < 0.05) the resistance against A. hydrophila (1 × 10 7  cells ml −1 ). In conclusion, probiotic B. licheniformis Dahb1 can be applied in diet at 10 7  cfu g −1 to improve healthy status and resistance against A. hydrophila in tilapia farming. [ABSTRACT FROM AUTHOR]

The present study reports the biological synthesis of silver nanoparticles using crustacean immune molecule β-1, 3 glucan binding protein (β-GBP) purified from the haemolymph of blue swimmer crab Portunus pelagicus . The characterization of synthesized β-GBP based silver nanoparticles ( Pp β-GBP-AgNPs) was made by UV-Vis spectroscopy, XRD, FTIR and TEM analysis. UV-Vis spectra recorded the strong absorbance peak at 420 nm due to its surface plasmon resonance. The XRD analysis revealed the crystalline nature of synthesized nanoparticles with Bragg's reflection peaks at (111), (200), (220), (311) planes. FTIR analysis showed the possible functional groups at 3422, 2926, 2847, 1648, 1556, 1407, 1016 and 669 cm −1 . The mean particle size of Pp β-GBP-AgNPs was 33–47 nm revealed by TEM analysis. Pp β-GBP-AgNPs exhibit appreciable antibacterial activity against Enterococcus faecalis and Pseudomonas aeruginosa when compared to chemical based AgNPs (Chem-AgNPs). The antibiofilm property of Pp β-GBP-AgNPs was assessed through light microscopy and confocal laser scanning microscopy analysis (CLSM), which clearly demonstrates, thickness of E. faecalis and P. aeruginosa preformed biofilm was reduced to 11 μm & 8 μm from 47 μm & 45 μm respectively. Moreover, exopolysaccharide (EPS) quantification and cell surface hydrophobicity (CSH) index exhibited that, Pp β-GBP-AgNPs had the potential to disturb structural integrity of biofilm by upset EPS matrix and bacterial adhesion to hydrocarbons. In addition, the cytotoxic effect of Pp β-GBP-AgNPs was evaluated against human cervical cancer cells (HeLa). Pp β-GBP-AgNPs effectively inhibit the viability of HeLa cells at 50 μg/ml concentration and the morphological changes in Pp β-GBP-AgNPs treated HeLa cells were observed under phase contrast microscopy. [ABSTRACT FROM AUTHOR]

The successful treatment of multi-drug resistant microbial pathogens represents a major challenge for public health management. Here, chitosan-alginate (CS/ALG) microspheres with narrow size distribution were fabricated by ionically cross linking method using Ca 2+ ions as agents for polymer solidification. The physicochemical properties of CS/ALG microspheres, such as surface morphology and size, were studied by SEM. The functional group interactions were confirmed by Fourier transform infrared (FTIR) spectroscopy. SEM revealed that the CS/ALG microspheres were spherical in shape with smooth surfaces, size was 50–100 μm. The synthesized CS/ALG microspheres showed antibacterial and antibiofilm activity on bacteria of public health relevance. CS/ALG microspheres exhibited antibacterial activity at the concentration of 5–20 μg, with significant inhibitory zones on multiple antibiotic resistant pathogens, including Gram positive Staphylococcus aureus, Enterococcus faecalis, and Gram negative Pseudomonas aeruginosa and Proteus vulgaris . Furthermore, in situ light microscopy and confocal laser scanning microscopy (CLSM) showed that CS/ALG microspheres inhibited the bacterial biofilm formation in S. aureus , E. faecalis P. aeruginosa and P. vulgaris after a single treatment with 40 μg. Overall, our findings underlined that chemically synthesized CS/ALG biomaterial has high antibacterial and antibiofilm activity against a number of microbial pathogens of interest for human health, thus this synthesis route can be further exploited for drug development in current biomedical science. [ABSTRACT FROM AUTHOR]

Dipteran flies are responsible of myiasis, a common parasitic infestation leading to the invasion of living, necrotic or dead tissues. They also transmit mycobacterial infections to livestock. In the present investigation, seven plant species employed in Southern India for medical and ethno-veterinary purposes were extracted using ethanol or water and tested against second instar larvae of Lucilia sericata. 100% larval mortality was observed testing Lobelia leschenaultiana ethanol extract at 60 mg/L (LC 50 = 3.4 mg/L). Based on the highest percentage of larval mortality, we selected L. leschenaultiana for fabrication of ZnO nanoparticles to control L. sericata . Zinc acetate showed 82.2% mortality at 60 mg/L (LC 50 = 38.2 mg/L) whereas L. leschenaultiana- coated ZnO nanoparticles (Ll-ZnO NPs) showed 100% mortality at 6 mg/L, the LC 50 was 0.78 mg/L. Overall, our research represents an attempt to integrate current ethno-veterinary knowledge from native tribes of Southern India, in order to identify effective botanicals for the development of nano-biopesticides against livestock pests and parasites. [ABSTRACT FROM AUTHOR]

The bioactivity of semiconductor nanocomplexes has been poorly studied in the field of pesticide science. In this research, the synthesis of zinc nanoparticles was accomplished through new effortless green chemistry process, using the Ulva lactuca seaweed extract as a reducing and capping agent. The production of U. lactuca -fabricated ZnO nanoparticles ( Ul -ZnO Nps) was characterized by powder X-ray diffraction (XRD), UV–visible, Fourier transform infrared (FTIR) spectroscopy, selected area electron diffraction (SAED) analysis and transmission electron microscopy (TEM). The U. lactuca -fabricated ZnO NPs were tested for their photodegradative action against organic dyes, as well as for antibiofilm and larvicidal activities. The UV visible absorbance spectrum of Ul -ZnO NPs exhibited the absorbance band at 325 nm and TEM highlighted average crystallite sizes of nanoparticles of 10–50 nm. Methylene blue (MB) dye was efficiently corrupted under sunlight in presence of Ul -ZnO NPs. Excellent bactericidal activity was shown by the Ul -ZnO Nps on Gram positive ( Bacillus licheniformis and Bacillus pumilis ) and Gram negative ( Escherichia coli and Proteus vulgaris ) bacteria. High antibiofilm potential was noted under both dark and sunlight conditions. The impact of a single treatment with Ul -ZnO NPs on biofilm architecture was also analyzed by confocal laser scanning microscopy (CLSM) on both Gram positive and Gram negative bacteria. Moreover, Ul -ZnO NPs led to 100% mortality of Aedes aegypti fourth instar larvae at the concentration of 50 μg/ml within 24 h. The effects of ZnO nanoparticle-based treatment on mosquito larval morphology and histology were monitored. Overall, based on our results, we believe that the synthesis of multifunctional Ul -ZnO Nps using widely available seaweed products can be promoted as a potential eco-friendly option to chemical methods currently used for nanosynthesis of antimicrobials and insecticides. [ABSTRACT FROM AUTHOR]

The use of natural polymers in drug design plays an important role in biomedical applications. Combinations of nanoparticles (NPs) and biopolymers have been shown to be useful for many purposes. This study focused on gelatin-coated zinc oxide NPs synthesized by co-precipitation. The particles were characterized by UV–Vis spectrum, showing a main peak at 375 nm. The stability and crystalline nature of the particles was evaluated by Zeta potential and X-ray diffraction analysis. Fourier transform infrared spectroscopy (FTIR) revealed the possible functional groups of Ge-ZnO NPs, with strong bands at 3851, 3447, and 2923 cm − 1 . Moreover, transmission electron microscopy (TEM) highlighted the presence of spherically shaped Ge-ZnO NPs that were 20 nm in size. Energy dispersive analysis X-ray (EDX) analysis showed that the zinc elemental content of Ge-ZnO NPs was 59.10%. The results of antibacterial activity assays revealed higher inhibition of Ge-ZnO NPs against Gram-negative Pseudomonas aeruginosa at 100 μg/ml over that against Gram-positive Enterococcus faecalis . Greater inhibition of biofilm formation was observed for Gram-negative bacteria compared to Gram-positive bacteria. In addition, Ge-ZnO NPs effectively inhibited the biofilm growth of the fungus Candida albicans at 50 μg/ml. Ge-ZnO NPs reduced the viability of hepatocarcinoma cancer cell lines at 100 μg/ml. Moreover, in chick embryos, notable anti-angiogenesis effects were observed for Ge-ZnO NPs and zinc acetate at 50 μg/ml compared to that observed testing gelatin. Overall, based on the results, Ge-ZnO NPs may be used as a novel agent for the control of biofilm-forming microbial pathogens. [ABSTRACT FROM AUTHOR]

The present study reports the purification of novel immune molecule β-1, 3 glucan binding protein from the heamolymph of the Indian white shrimp, Fenneropenaeus indicus ( Fi β-GBP). The purified Fi β-GBP had 95 kDa molecular weight in SDS-PAGE analysis. MALDI-TOF/TOF analysis revealed that the purified Fi β-GBP showed similarity to various crustacean proteins; 48 and 46% similarity was observed for β-1, 3 glucan binding protein of Chinese white shrimp Fenneropenaeus chinensis and banana shrimp Fenneropenaeus merguiensis, with MOWSE score of 3.11e + 12 and 2.05e + 8, respectively. The phenoloxidase activity (PO) of Fi β-GBP was evaluated and, in the presence of laminarin, PO activity increased significantly. Substrate specificity assay demonstrated that Fi β-GBP had the specific binding site for soluble or insoluble β-glucan (laminarin), since the PO activity increased in the presence of laminarin when compared to other sugars. Enzymatic activities revealed that the optimum temperature and pH for Fi β-GBP activating PO were 40 °C and pH 7–8. Moreover, even at 100 °C Fi β-GBP enhanced PO activity highlighting that Fi β-GBP was thermostable and thermophilic in nature. Among various divalent metallic ions, Fi β-GBP significantly promoted the PO activity in presence of Mg 2+ and Ca 2+ . The breakdown of para nitroanilide from N α −Benzoyl- l -Arginine 4-Nitroanilide hydrochloride showed that serine protease activity was induced by Fi β-GBP and also increased concentration of Fi β-GBP evoked the activity. Furthermore, hemolytic activity tests revealed that PO reaction product induced RBC membrane damage and cell shrinkage. Lastly, Baclight bacterial viability assays showed maximum killing effect of PO reaction product on both Gram positive and Gram negative bacteria. [ABSTRACT FROM AUTHOR]

The effective treatment of cancer and bacterial pathogens are two key challenges in modern nanomedicine. Here, zinc oxide nanoparticles (ZnO NPs) were fabricated using the crustacean immune molecule β-1, 3- glucan binding protein ( Ph β-GBP, 100 kDa) purified from the heamolymph of Paratelphusa hydrodromus . β-GBP coated zinc oxide nanoparticles ( Ph β-GBP-ZnO NPs) were characterized by UV–vis spectroscopy, X-ray diffraction (XRD), Fourier transform infrared spectroscopy (FTIR), and high resolution-transmission electron microscopy (HR-TEM) analyses. Ph β-GBP-ZnO NPs inhibited the growth of Staphylococcus aureus and Proteus vulgaris . Protein and nucleic acid leakage assays showed that Ph β-GBP-ZnO NPs facilitate membrane permeability leading to cell death. The antibacterial activity of Ph β-GBP-ZnO NPs was due to the high level of reactive oxygen species (ROS) release from bacterial cells post-treatment with 75 μg/mL of Ph β-GBP-ZnO NPs. Confocal laser scanning microscopy pointed out that biofilm thickness was highly reduced post-treatment with nanoparticles. Cytotoxicity on human liver carcinoma (HepG2) cells highlighted that 75 μg/mL of Ph β-GBP-ZnO NPs inhibited viability of HepG2 cells. Phase contrast microscopy showed key morphological changes of HepG2 cells post-treatment with Ph β-GBP-ZnO NPs. Overall, Ph β-GBP-ZnO NPs can be further considered for the development of novel drugs against microbial pathogens and HepG2 cells. [ABSTRACT FROM AUTHOR]

The tick Rhipicephalus (Boophilus) microplus is a key vector of bacterial and protozoan diseases causing heavy economic losses directly and indirectly in animal husbandry. In the past decades, the control of ticks faced some major issues, such as the rapid development of resistance in targeted vectors and non-target effects on human health and the environment, due to the employ of synthetic acaricides and repellents. Eco-friendly pesticides for treating and controlling animal parasites such as ticks were mainly from medicinal plants and thus they form the richest entity for manufacturing resources for drugs. Even though there are efforts made to discover reliable plant-based acaricides to control ectoparasites in animal husbandry, the effective control of R. (B.) microplus ticks still represent a major challenge in current veterinary entomology. Recently, a wide number of promising attempts have been conducted to use herbal preparations and green-fabricated nanoparticles for the control of R. (B.) microplus . The aim of this review is to critically summarize and discuss the use of herbal preparations used in ethno-veterinary as well as green-fabricated nanoparticles as novel acaricides for the control of the cattle tick R. (B.) microplus. [ABSTRACT FROM AUTHOR]

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 . [ABSTRACT FROM AUTHOR]

The green synthesis of pesticides and antibiotics is gaining increasing importance nowadays. We fabricated Euphorbia rothiana-capped Ag nanoparticles (Er-AgNPs), testing their antibiofilm and growth inhibition potential on various microbial pathogens. We evaluated the toxicity of E. rothiana-capped Ag nanoparticles on larvae of the Zika virus vector Aedes aegypti, shedding light on histological changes post-exposure to the plant leaf extract and Er-AgNPs. Nanoparticles were characterized by UV-Vis, and FTIR spectroscopy, XRD and TEM. Light microscopy showed the greatest inhibition of biofilm formation in Gram positive and Gram-negative bacteria post-treatment with E. rothiana-capped Ag nanoparticles at 75 μg/ml. Confocal microscopy confirmed the biofilm interruption and disintegration post-treatment with nanoparticles at 75 μg/ml. E. rothiana extract and Er-AgNPs showed 100% larvicidal activity on A.aegypti at 50 and 8 mg/L, respectively. AgNO led to 100% mortality of A. aegypti larvae at 40 mg/L after 48 h. Stereomicroscopic and histopathological analysis reported tissue damages and loss of cuticular parts in mosquito larvae. [ABSTRACT FROM AUTHOR]

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 20 nm. In addition, zeta potential measurement demonstrated that the Bt -ZnO NPs are negatively charged (− 12.7 mV) 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 25 μg/mL. Furthermore, Bt -ZnO NPs are highly effective in the control of C. maculatus and caused 100% mortality at 25 μg/mL. The LC 50 value was estimated to be 10.71 μg/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. [ABSTRACT FROM AUTHOR]

The control of Zika virus mosquito vectors and well as the development of drugs in the fight against biofilm-forming microbial pathogens, are timely and important challenges in current bionanoscience. Here we focused on the eco-friendly fabrication of Ag nanostructures using the seed extract of Pedalium murex , an ancient Indian medicinal plant. Initial confirmation of Ag nanoparticles (AgNPs) production was showed by a color change from transparent to dark brown. The UV–Visible spectrum (476 nm), X-ray diffraction peaks (101, 200, 220 and 311) and Fourier transform infrared spectroscopy shed light on the production of green-capped AgNPs. Morphological structure analysis using HR-TEM showed that the AgNPs were mostly hexagonal in shape with rough edges, and a size of 20–30 nm. The larvicidal potential of P . murex seed extract and AgNPs fabricated using the P . murex seed extract ( Pm -AgNPs) was tested on fourth instar mosquito larvae of the Zika virus vector Aedes aegypti . Maximum efficacy was achieved by Pm –AgNPs against Ae . aegypti after 24 h (LC 50 34.88; LC 90 64.56 mg/ml), if compared to the P . murex seed extract. Histopathological analyses showed severe damages to the hindgut and larval muscles in NPs-treated Ae . aegypti larvae. The sub-MIC concentrations of Pm -AgNPs exhibited significant anti-biofilm activity against Gram positive ( Enterococcus faecalis , Staphylococcus aureus ) and Gram negative ( Shigella sonnei , Pseudomonas aeruginosa ) bacterial pathogens, as showed by EPS and MTP assays. Light and CLSM microscopic studies highlighted a significant impact of P . murex seed extract and Pm –synthesized AgNPs on the surface topography and architecture of bacterial biofilm, both in Gram positive and Gram negative species. Overall, results reported here contribute to the development of reliable large-scale protocols for the green fabrication of effective mosquito larvicides and biofilm inhibitors. [ABSTRACT FROM AUTHOR]

The control of multidrug-resistant bacteria as well as insect pests and vectors is timely and important now a days. The present study was designed to evaluate the in vitro antibacterial, antibiofilm and mosquito larvicidal effects of gold nanoparticles synthesized using the zein biopolymer (Ze-AuNPs) against Gram positive ( Bacillus pumilus and Bacillus subtilis ), Gram negative ( Shigella sonnei and Pseudomonas aeruginosa ) bacteria and third instar larvae of the dengue and Zika virus vector Aedes aegypti . The synthesized Ze-AuNPs were characterized by UV–vis spectroscopy, X-ray diffraction (XRD), Fourier transform infrared (FTIR) spectroscopy, scanning electron microscopy (SEM), and transmission electron microscopy (TEM). The antibacterial assays testing Ze-AuNPs at 100 μg/ml showed that the zones of inhibition against Gram positive species B. pumilus and B. subtilis were 13.9 and 14.2 mm, respectively, while for Gram negative S. sonnei and P. aeruginosa they were 18.1 and 18.4 mm, respectively. Light and confocal laser scanning microscopy (CLSM) confirmed the interruption and disintegration of bacterial biofilm post-treatment with Ze-AuNPs at 100 μg/ml. In larvicidal assays on A. aegypti , HAuCl 4 and Ze-AuNPs treated third instar larvae of A. aegypti showed LC 50 of 26.6 and 6.81 mg/L, respectively, and LC 90 of 81.1 and 13.6 mg/L respectively. The histopathological analysis of A. aegypti treated with Ze-AuNPs showed complete disintegration of abdominal region, particularly the midgut and caeca, with loss of lateral and caudal hairs. The stereomicroscopic visualization of A. aegypti treated with Ze-AuNPs showed the loss of upper head hair, lower head hair, antenna hair, lateral hair and caudal hair. Overall, the study concludes that Ze-AuNPs have excellent antibacterial, antibiofilm effects and has ability to control the larval populations of A. aegypti mosquitoes. [ABSTRACT FROM AUTHOR]

Our understanding of nanoparticle toxicity and fate in the aquatic environment is still patchy. In the present study, the toxicity of silver nanoparticles coated by Camellia sinensis ( Cs) leaf extract metabolites ( Cs-AgNPs) was investigated in comparison with C. sinensis leaf extract and AgNO on a micro-crustacean, Ceriodaphnia cornuta, and a fish Poecilia reticulata. 100% mortality of C. cornuta was observed post-exposure to AgNO (40 µg/ml) if compared to the Cs leaf extract and Cs-AgNPs, showing 30 and 56% mortality at the same concentration, respectively. In P. reticulata 100% mortality was observed testing AgNO and Cs-AgNPs post-exposure to 1 and 30 µg/ml, respectively. Light microscopy and CLSM images showed the accumulation of nanoparticles in the intestine of C. cornuta treated with Cs-AgNPs at 40 µg/ml. In addition, histological observations confirmed the abnormal tissue texture in nanoparticle-exposed P. reticulata, if compared to control fishes. Furthermore, C. cornuta and P. reticulata treated with Cs-AgNPs showed DNA damages compared to the control. Overall, these findings indicated relevant limits about the employ of silver-based pesticides in the environment, and also pointed out the Cs-AgNPs were less toxic to C. cornuta and P. reticulata if compared to silver ions. [ABSTRACT FROM AUTHOR]

Dengue and chikungunya are arboviral diseases mainly vectored by the mosquito Aedes aegypti. Presently, there is no treatment for these viral diseases and their prevention is still based on vector control measures. Nanopesticides fabricated using herbal extracts as reducing and capping agents currently represent an excellent platform for pest control. In this scenario, the present study assessed the acute toxicity of seven plants employed in ethno-veterinary medicine of southern India, as well as the green synthesis of zinc oxide nanoparticles, on third-instar larvae of A. aegypti. Larvae were exposed to extracts of the seven plants obtained with solvents of different polarity (acetone, ethanol, petroleum ether, and water) for 24 h. Maximum efficacy was observed for Lobelia leschenaultiana leaf extracts prepared using all the four solvent extracts (LC = 22.83, 28.12, 32.61, and 36.85 mg/L, respectively). Therefore, this plant species was used for the synthesis and stabilization of ZnO nanoparticles based on its maximum efficacy against third-instar larvae of A. aegypti. L. leschenaultiana-encapsulated ZnO nanoparticles showed 100% mortality when tested at 10 mg/L, the LC was extremely low, 1.57 mg/L. Zinc acetate achieved only 65.33% when tested at 60 mg/L, with a LC of 51.62 mg/L. Additionally, ZnO nanoparticles inhibited growth of Pseudomonas aeruginosa, Proteus vulgaris, Shigella sonnei, and Vibrio parahaemolyticus and also inhibited biofilm formation on selected microbila pathogens, showing impact on EPS production and hydrophobicity. Overall, our results suggest that L. leschenaultiana-fabricated ZnO nanoparticles have a significant potential to control A. aegypti mosquitoes and Gram-negative bacterial pathogens. [ABSTRACT FROM AUTHOR]

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. [ABSTRACT FROM AUTHOR]

Recently, the green synthesis of silver nanoparticles gained increasing attention due to interesting properties for optical, antimicrobial and pest control applications. However, their toxicity against micro-crustaceans and fishes has been scarcely explored, while most of the research efforts focused on mosquito control with the green-synthesized nanocomposites. In this study, we investigated the toxic effects of AgNO, Cissus quadrangularis (Cq)-synthesized AgNPs and Cq extract in two different study models, the larvae of Poecilia reticulata fishes and adults of the micro-crustacean Ceriodaphnia cornuta. In both species, AgNO and Cq-AgNPs showed high mortality rates even if tested at very low doses. Molecular analysis revealed high DNA damages induced by Cq-AgNPs on both aquatic organisms. Furthermore, light microscopy studies evidenced lesions in the gills and vacuolization in the gills and in the abdomen of P. reticulata larvae. Overall, our research pointed out that the exposure of aquatic organisms to AgNO or green-fabricated AgNPs can damage fishes and crustaceans, posing noteworthy risks to the aquatic ecosystems. [ABSTRACT FROM AUTHOR]

Haemocyanin (Hc) is an important non-specific immune macromolecule present in the haemolymph of both mollusks and crustaceans. In the present study, Hc was purified from the haemolymph of Indian white shrimp Fenneropenaeus indicus by gel filtration chromatography and it exhibits a single band with a molecular weight of 74 kDa on SDS-PAGE. The X-ray diffraction (XRD) and High performance liquid chromatography (HPLC) result of purified Hc express single peak at 31.5° be a sign of crystalline nature and appear as a single peak with a retention time of 5.6 min signify the homogeneity nature of the protein respectively. The purified Hc exhibited haemolytic activity against chicken erythrocytes. The haemolytic activity of purified Hc in optimum conditions observed to be pH 6.0, temperature 40 °C, in the presence of calcium. As well purified Hc exhibited the antibiofilm activity against both Gram positive and Gram negative bacteria. Moreover, the haemolysis can be inhibited to different degrees by osmoprotectants of diverse molecular masses, signifying that it follows a colloid-osmotic mechanism. This study conclude that purified Hc from F. indicus remarkably possess haemolytic and antibiofilm activity. [ABSTRACT FROM AUTHOR]

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 47.27 nm. 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 14.2 mm at 25, 50 and 75 μg mL −1 . The zone of inhibition against P. aeruginosa was 9.8, 10.2 and 11.3 mm at 25, 50 and 75 μg mL −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 75 μg mL −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 80 μg mL −1 . The morphological changes in the Ln -ZnO NPs treated A549 lung cancer cells were observed under phase contrast microscope. [ABSTRACT FROM AUTHOR]

In the present study, chitosan-ZnO composite was synthesized by stirring of chitosan purified from mud crab Scylla serrata shells with zinc chloride and sodium hydroxide. The physicochemical characteristics of chitosan-ZnO composite were studied using UV-Vis spec, Fourier transform infrared, X-ray diffraction and scanning electron microscopy. Chitosan-ZnO composite formation was confirmed by the functional groups stretching and bending vibrations in Fourier transform infrared. Scanning electron microscopy results showed the synthesized chitosan-ZnO composite was rod-like structure. X-ray diffraction results represent the hexagonal shape and crystalline size (30–60 nm) of zinc oxide in the chitosan-ZnO composite. Antibacterial activity of chitosan-ZnO composite demonstrated the effective growth control of a Gram-negative Vibrio parahaemolyticus and Gram-positive Bacillus lechiniformis bacteria isolated from aquatic environments. Light microscopy and confocal laser scanning microscopy also evidenced the antibiofilm activity of chitosan-ZnO composite against the V. parahaemolyticus and B. lechiniformis at the concentration of 40 and 60 µg/ml, respectively. [ABSTRACT FROM AUTHOR]

The pathways by which pathogens invade Fenneropenaeus indicus, the potential colonization in various tissues and the disease transmission mechanisms are unclear. The aims of the present study were to visualize the colonization and pathogenesis of GFP-tagged Vibrio parahaemolyticus, in various tissues of F. indicus to evaluate the pathogen interaction. Among the three strains isolated, a virulent strain VpDAHV2 was tagged with green fluorescent protein (GFP-VpDAHV2) and validated for both its growth characteristics and its virulence as a genuine model for F. indicus infection. VpDAHV2 was positive for toxR and tlh genes and negative for tdh genes. CLSM images revealed that maximum colonization was observed in the haemolymph of the F. indicus challenged with GFP-VpDAHV2. The haemolymph was the primary site for the colonization of GFP-VpDAHV2 in F. indicus. The enteric localization occurred independently of the flagellum or motility of GFP-VpDAHV2 through the intestinal route. The F. indicus infection model suggests that the haemolymph and the intestine represent the sites of infection by GFP-VpDAHV2, and hence are the active sites of pathogen interactions. GFP tagging of V. parahaemolyticus is a new and systemic approach to determine the presence of bacteria in vivo for the confirmation of host pathogen interactions in aquaculture studies. [ABSTRACT FROM AUTHOR]

Highlights: [•] β-GBP was purified from the haemolymph of E. tetragonum and physicochemically characterized. [•] Biological activity of E. tetragonum β-GBP substantiated by yeast agglutination and PO enhancing activity. [•] Crystalline “Y” shaped β-GBP of E. tetragonum helped out solving the β-GBP structure. [Copyright &y& Elsevier]

Abstract: The present study reports the purification and characterization of immune molecule lectin from the green tiger shrimp Penaeus semiculcatus, which involves the non-self-recognition mechanism to destroy the infectious pathogens that continuously threaten their survival. P. semisulcatus lectin (Ps-Lec) was purified by affinity chromatography with mannose coupled sepharose CL-4B column and showed the 37 and 118kDa subunits in SDS-PAGE. The surface morphology of purified Ps-Lec exhibits the crystalline and aggregated nature in Transmission electron microscopy (TEM) analysis. Functional analysis of purified Ps-Lec showed that the broad spectrum of bacterial agglutination activity against Gram-positive and Gram-negative bacteria. In addition, Ps-Lec had the ability to involve in the haemoagglutination activity, which agglutinates the several vertebrate erythrocytes tested, and the haemagglutination titres were observed under light microscopy. Interestingly, Ps-Lec also exhibits the fungal aggregation activity against Candida albicans, this is the first report regarding the β-glucan recognition ability of fungal aggregation of Ps-Lec. Moreover, phenoloxidase activity was triggered by the Ps-Lec when compared to control. The present study reveals that Ps-Lec is an important multifunctional humoral defence protein plays a significant role against the bacterial and fungal cells. [Copyright &y& Elsevier]

Aquaculture makes a significant contribution in the production of protein-rich food for human consumption. Aquaculture practices encounter many challenges, and one of the most devastating problems is disease outbreaks caused by microbial pathogens. To control disease outbreaks, several chemotherapeutics and antibiotics were used indiscriminately, which in turn leads to residual problems in the surrounding environment affecting higher animals and also humans. Immunostimulants are considered as an alternative for antibiotics, which will boost the immune system of the cultured organism, thus effectively countering the assault of pathogens. The use of plant materials as immunostimulant will be an ecofriendly approach for the control of pathogens. The botanicals present in the plants have a key role in enhancing the fish immunity. This review focuses on the importance of plant material as immunostimulant in the control of diseases in aquaculture. [ABSTRACT FROM AUTHOR]

Abstract: Alpha 2 macroglobulin (α2M) is a non-specific protease inhibitor involved in host defence mechanism in invertebrates. By c-DNA cloning and sequencing, α2M gene was identified from the haemocytes of Indian white shrimp Fenneropenaeus indicus with an open reading frame of 4599bp encoding 1532 amino acids with a signal sequence of 26 amino acids. The calculated molecular mass of the mature protein is 170.6kDa with an estimated pI of 6.13. The F. indicus α2M (Fi-α2M) sequence contains putative conserved functional domains including a GCGEQNM thioester region and a receptor-binding domain which are present in other invertebrate and vertebrate α2Ms. Sequence comparison and phylogenetic analysis revealed that Fi-α2M displays the highest similarity with Litopenaeus vannamei (96%). QRT-PCR analysis of challenge experiments by Vibrio harveyi, GFP tagged Vibrio parahaemolyticus (GFP-Vp) and WSSV infection of F. indicus significantly increased the α2M mRNA transcript up to 24h in the haemocytes. In addition, confocal laser scanning microscopy (CLSM) confirms that the GFP-Vp treated F. indicus have high proliferation of GFP-Vp in haemolymph and intestine at 24h. These results suggest that α2M is an important molecule in shrimp immune system. [Copyright &y& Elsevier]

This study focuses on the evaluation of the antimicrobial activity of essential oils ( EOs) from medicinal plants against Pseudomonas sp isolated from aquatic environments, and proves its controlling efficacy on Pseudomonas a eruginosa-Dahp1 using Confocal Laser scanning Microscopy. Twenty-seven Pseudomonas spp. ( Ps1- Ps27) were isolated using King's B medium and the selected P. aeruginosa-Dahp1 was confirmed using 16S r- DNA methods. Antimicrobial activity of 10 EOs was determined using agar well and disc diffusion methods. Ten EOs were extracted with six solvents and the crude extracts were tested against the Pseudomonas spp. through disc diffusion method. Among 10 EOs tested, maximum inhibitory activity was noted in EOs of Wringtia tinctoria against the P. aeruginosa-Dahp1 in the disc diffusion method. The MIC values (concentrations were expressed in Weight/Volume) of the EOs range from 0.5 to 9.054 mg mL−1. The EO of W. tinctoria showed the maximum activity at the concentration (w/v) of 2.060 mg mL−1. The EO of W. tinctoria with chloroform extract showed the maximum inhibition against P. aeruginosa-Dahp1. The CLSM proves the control and viability of 1 × 105 CFU mL−1 of P. aeruginosa-Dahp1 at a lower concentration (2.720 mg) of EOs with chloroform extracts of W. tinctoria. This study pivots for designing of new drugs using EOs of W. tinctoria against Pseudomonades in the aquaculture sites. [ABSTRACT FROM AUTHOR]

In aquaculture industries, there is an urgent need to develop microbial control strategies, to control disease outbreaks. In recent years, probiotics are considered as a valid alternative for the use of antibiotics in aquaculture to prevent high mortality and promote growth. In the present study, seven strains of bacteria such as Bacillus licheniformis (DAB1), Bacillus pumilus (DAB2), Bacillus sp. (DAB3), Pseudomonas aeruginosa (DAP1), Pseudomonas sp. (DAP2), Pseudomonas aeruginosa (DAP3), Pseudomonas aeruginosa (DAP4), and three pathogenic Vibrio parahaemolyticus (DAV1, DAV2, DAV3) were isolated from healthy and diseased Fenneropenaeus indicus collected from the east coast of Tamilnadu, India. The strains were identified by biochemical analysis and 16S rRNA sequence methods. Among the seven probiotic strains tested, the cell-free extract from DAB1 and DAP1 exhibited higher inhibitory activity of V. parahaemolyticus than other isolates under in vitro conditions. The LC of DAV1, DAV2, and DAV3 was found to be ∼10 CFU mL. Pathogenicity of three V. parahaemolyticus DAV1, DAV2, and DAV3 showed significant mortalities (40 %) in Artemia nauplii at inoculation densities of 10 CFU mL when compared to the controls (unchallenged nauplii). A significant reduction in mortality ( P < 0.001) was found by addition of 10 CFU mL of DAB1 and DAP1 strains in nauplii against the pathogens. In conclusion, the present study result reveals that DAB1 and DAP1 have potential applications for controlling pathogenic V. parahaemolyticus in Artemia culture systems and aquaculture practices. [ABSTRACT FROM AUTHOR]

Abstract: A β-1, 3-Glucan binding protein (β-GBP) was isolated from green tiger shrimp Penaeus semisulcatus and purified using laminarin precipitation and affinity chromatography on laminarin-Sepharose 6B column respectively. P. semisulcatus β-GBP exhibits a single band with a molecular weight of 112 kDa on SDS-PAGE and pI of 5.9 in isoelectric focusing (IEF). Negative staining of P. semisulcatus β-GBP showed large aggregates with crystalline surface when viewed by Electron Microscopy. Circular dichroism spectra of P. semisulcatus β-GBP showed broad negative minimum wavelength extending from 200 to 250 nm can be attributed to the presence of β-sheets in its secondary structure. P. semisulcatus β-GBP comprises the specific binding affinity with the polysaccharide β-1, 3-glucans (laminarin), this recognition and binding leads to the activation of prophenoloxidase cascade. Interestingly, P. semisulcatus β-GBP also involved in the agglutination of baker's yeast, bacteria, erythrocytes (RBCs) and enhances the PO activity. Herein, we have investigated the importance of β-GBP in innate immune response of P. semisulcatus and they implicate the evolutionary link with similar proteins found in other invertebrates. [Copyright &y& Elsevier]

Penaeidae is a family of shrimp, and it contains many species of economic importance, such as the tiger prawn ( Penaeus monodon), white leg shrimp, Atlantic white shrimp and Indian prawn. Identification and population genotype structure of penaeid shrimp have been enhanced by molecular markers that can be classified into three types, namely allozyme, mitochondrial and nuclear markers. The widely used mitochondrial DNA markers are 12S rDNA, 16S rDNA, cytochrome b and control region. Random amplification of polymorphic DNA, amplified fragment length polymorphism, restriction fragment length polymorphism, single-stranded conformational polymorphism and microsatellites are the most commonly used nuclear markers for DNA fingerprinting. Molecular markers play a crucial role in penaeid shrimp to evaluate phenotypic and genetic variation, assess demographic bottleneck, study natural population structure, compare wild and hatchery populations, preserve genetic biodiversity, construct chromosome maps and detect whether genetic tag propagation-assisted rehabilitation programs are effective. Increase in the number of molecular markers, construction of high-density genetic maps and implementation of genomic resources (including genome sequencing) are expected to provide tools for the genetic improvement in these aquaculture species through marker-assisted selection. Molecular markers are versatile tools for the identification of populations with genetic crisis by comparing genetic diversities, which helps to establish management units within these threatened species. [ABSTRACT FROM AUTHOR]

Summary: A lipopolysaccharide (LPS) and β-1,3-glucan binding protein (LGBP) gene was cloned from hemocytes of kuruma shrimp Marsupenaeus japonicus by reverse-transcription polymerase chain reaction (RT-PCR), cloning and sequencing of overlapping PCR, and rapid amplification of cDNA ends (RACE) method. The open reading frame (ORF) of M. japonicus LGBP is 1062bp and encodes a 354 amino acid (aa) sequence with a 23aa signal peptide. The calculated molecular mass of the mature protein (331aa) is 40.15kDa with an estimated pI of 4.78. The M. japonicus LGBP sequence contains (1) two putative N-linked glycosylation sites, (2) two putative integrin-binding motifs, (3) a kinase C phosphorylation site (KCPS), (4) a glucanase motif (GM), and (5) two potential polysaccharide recognition motifs (polysaccharide binding motif (PsBM) and β-glucan recognition motif (GRM)), and with features of tryptophan-rich, slight homology to lysozyme, and slight homology to lectin. A sequence comparison showed that the deduced amino acids of M. japonicus LGBP has an overall high similarity to penaeid LGBP and βGBP (85.6–89.9%), lobster Homarus gammarus βGBP (77.0%), and crayfish Pacifastacius leniusculus LGBP (67.8%). The phylogenetic analysis revealed that M. japonicus LGBP grouped together with other crustacean LGBP and βGBP, and was close to termite GNBP, but was far way from moth βGBP, βGRP, fly GNBP, and mosquito βGRP. The LGBP of M. japonicus was strongly expressed in hemocytes. The LGBP mRNA transcript in hemocytes of M. japonicus was significantly upregulated 12–48h after a LPS injection, indicating activation of the innate immune system through the binding of the LGBP and LPS complex. [Copyright &y& Elsevier]

Abstract: The full-length sequence of α2-macroglobulin (α2-M) was cloned from the hemocytes of white shrimp Litopenaeus vannamei by reverse transcription polymerase chain reaction (RT-PCR), cloning and sequencing of overlapping PCR, and the rapid amplification of cDNA ends method. The α2-M cDNA consists of 4682bp with an open reading frame of 4479bp, a 52-bp 5′-untranslated region, and a 132-bp 3′-untranslated region. The predicted molecular mass of the mature protein (1475 amino acids) is 167.1kDa, with an estimated pI of 5.70. The L. vannamei α2-M sequence contains putative functional domains including a bait region, a GCGEQNM (985–991) thiol ester domain, and a receptor-binding domain. Sequence comparison and phylogenetic analyses show that α2-M deduced amino acid sequence of L. vannamei has a high level of similarity to that of tiger shrimp Penaeus monodon and kuruma shrimp Marsupenaeus japonicus. The α2-M transcript was mainly expressed in hemocytes, and was significantly higher in premoult stage than in intermoult and postmoult stages. [Copyright &y& Elsevier]

Abstract: An extracellular copper–zinc superoxide dismutase (ecCuZnSOD, or EC-SOD) gene was cloned from haemocytes of mud crab Scylla serrata. The ecCuZnSOD cDNA consists of 939bp with an open reading frame (ORF) of 585bp that encodes 195 amino acids (aa) with 31 residues of a signal peptide sequence. The predicted molecular mass of the mature protein (164aa) is 17.1kDa with an estimated pI of 6.89. Amino acids responsible for binding Cu (His84, 86, 101, and 163) and Zn (His101, 109, and 121, and Asp 124), two cysteines (95 and 189) that form a disulfide bond, two CuZnSOD signatures (GFHVHAEGDLS) from 82 to 92 and (GNAGGRAGCGLI) from 181 to 192, as well as a N-linked glycosylation site (NVSG) were conserved. The deduced amino acid sequence of S. serrata ecCuZnSOD showed similarity of 84% and 54% with the ecCuZnSOD of blue crab Callinectes sapidus, and crayfish Pacifastacus leniusculus, respectively. The phylogenetic analysis revealed that the ecCuZnSOD of S. serrata grouped together with ecCuZnSODs of crustaceans and insects, but was far way from the intracellular (ic)CuZnSODs of invertebrates and vertebrates. The ecCuZnSOD transcript in haemocytes significantly increased in 6–48h, and had returned to the original value by 72h after the β-glucan (βG) injection, whereas the ecCuZnSOD transcript significantly increased 6–72h after the peptidoglycan (PG) injection indicating induction of the immune system within a short time. [Copyright &y& Elsevier]

Abstract: An alpha 2-macroglobulin (α2-M) gene was cloned from the haemocytes of tiger shrimp Penaeus monodon by RT-PCR, cloning and sequencing of overlapping PCR and rapid amplification of cDNA ends (RACE) method. Analysis of the nucleotide sequence revealed that the α2-M cDNA consists of 4876bp with an open reading frame (ORF) of 4494bp, a 52bp 5′-untranslated region, and a 327bp 3′-untranslated region containing a poly A signal. The open reading frame encodes a protein of 1498 amino acids with 18 residues signal sequence. The predicted molecular mass of the mature protein (1480 amino acids) is 167.7kDa with an estimated pI of 5.30. The P. monodon α2-M sequence contains putative functional domains including a GCGEQNM thioester region, a bait region, and a receptor-binding domain which are present in other invertebrate and vertebrate α2-Ms. Sequence comparison showed that α2-M deduced amino acid sequence of P. monodon has an overall similarity of 85, 52 and 49% to that of kuruma shrimp Marsupenaeus japonicus, American horseshoe crab Limulus polyphemus and mud crab Scylla serrata, respectively. Alignment of the deduced amino acid sequence to other species α2-M showed that the overall structure is evolutionarily conserved and phylogentic analysis revealed that P. monodon α2-M is closely related to other arthropod α2-M, and displays the highest similarity to M. japonicus α2-M. The α2-M was mainly expressed in haemocytes, but not in eyestalk, gill, muscle, hepatopancreas, and intestine. Quantitative real-time RT-PCR analysis showed that α2-M mRNA transcript in haemocytes of P. monodon increased significantly in 12, 24 and 48h post-peptidoglycan (PG) injection, but returned to the original values in 72h post-PG injection. [Copyright &y& Elsevier]