Exopolysaccharides-Mediated ZnO Nanoparticles for the Treatment of Aquatic Diseases in Freshwater Fish Oreochromis mossambicus.

Bacterial fish disease outbreaks are a key concern for aquaculture. Complementary feed additives such as immunostimulants can serve as an ideal solution for disease prevention. Herein, we scrutinized the efficacy of exopolysaccharides (EPSs) from probiotic Bacillus licheniformis and EPS-mediated zinc oxide nanoparticles (EPS-ZnO NPs) for a diet to evaluate growth parameters, antioxidant enzyme activities, and immune stimulation together with disease resistance against Aeromonas hydrophila and Vibrio parahaemolyticus in Mozambique tilapia Oreochromis mossambicus. Fish were separated into seven groups, with six experimental groups fed with EPS and EPS-ZnO NPs at 2, 5, and 10 mg/g and a control fed a basal diet. The fish ingesting feed supplemented with EPS and EPS-ZnO NPs at 10 mg/g showed improved growth performance. Cellular and humoral-immunological parameters were tested in serum and mucus after 15 and 30 days of feeding. These parameters were substantially enriched with a 10 mg/g diet (p < 0.05) of EPS and EPS-ZnO NPs in comparison with the control. Furthermore, the EPS and EPS-ZnO NP supplemental diet actively enhanced the antioxidant response (glutathione peroxidase, superoxide dismutase, and catalase). In addition, the supplemental diet of EPS and EPS-ZnO NPs lowered the death rate and improved the disease resistance of O. mossambicus following assessment with A. hydrophila and V. parahaemolyticus at 50 µL. Hence, the overall results suggest that the supplemental diet of EPS and EPS-ZnO NPs might be used to ensure aquaculture feed additives.

Facile synthesis of haemocyanin-capped zinc oxide nanoparticles: Effect on growth performance, digestive-enzyme activity, and immune responses of Penaeus semisulcatus.

ZnO nanoparticles (NPs) synthesized using haemocyanin (Hc-ZnONPs) purified from haemolymph of Penaeus semisulcatus were characterized using various techniques. HR-TEM and SEM microscopy indicated Hc-ZnONPs had a typical size of 20-50 nm and were spherical. The objective of current investigation was to assess the effects of dietary supplementation of Hc-ZnONPs on the development and activity of digestive and metabolic enzymes, as well as the antioxidant levels in P. semisulcatus. Trial basal diets were supplemented with Hc-ZnONPs at rates of 0, 10, 20, 40, 60, and 80 mg kg-1 (dry feed weight) and were fed to P. semisulcatus for 30 d. For 60 mg kg-1 Hc-ZnONPs-supplemented feed, significantly (P < 0.05) enhanced endurance, development, and activity of the digestive enzyme were observed. The enzymatic antioxidants and metabolic enzymes activities in the muscle exhibited no significant changes when 10-60 mg kg-1 Hc-ZnONPs-supplemented feed was fed to P. semisulcatus. Conversely, feeding the P. semisulcatus with 80 mg kg-1 Hc-ZnONPs produced a harmful outcome, with significant increase in the enzymatic antioxidants and metabolic enzymes. Consequently, 80 mg kg-1 Hc-ZnONPs was identified as lethal to P. semisulcatus. Hence, it is proposed that the diet of P. semisulcatus can be supplemented with up to 60 mg kg-1 Hc-ZnONPs for improving the endurance, development and immunity.

Microbial exopolymer-capped selenium nanowires - Towards new antibacterial, antibiofilm and arbovirus vector larvicides?

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, LC50 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.

Crustin-capped selenium nanowires against microbial pathogens and Japanese encephalitis mosquito vectors - Insights on their toxicity and internalization.

Herein, we reported a method to synthesize selenium nanowires (Cr-SeNWs) relying to purified cysteine-rich antimicrobial peptide crustin in presence of ascorbic acid. Cr-SeNWs were characterized by UV-vis, XRD, FTIR and Raman spectroscopy, as well as SEM, HR-TEM and EDAX. The UV-vis spectroscopy peak was noted at 350 nm. XRD showed the crystalline nature of Cr-SeNWs through diffraction peaks observed 2θ at 12° and 28° corresponding to (020), and (241) lattice planes, respectively. HR-TEM results shed light on the size of Cr-SeNWs, ranging from 17 to 47 nm. Raman spectroscopy and EDAX analysis of Cr-SeNWs showed presence of 57% selenium element. Furthermore, Cr-SeNWs showed higher antimicrobial activity on Gram-positive bacteria (Staphylococcus aureus, Enterococcus faecalis) over Gram-negative ones (Pseudomonas aeruginosa, Escherichia coli). The zone of inhibition was larger on S. aureus (50 µg/ml = 4.0 mm, 75 µg/ml = 7.2 mm) and E. faecalis (50 µg/ml = 3.1 mm, 75 µg/ml = 5.1 mm), over P. aeruginosa (50 µg/ml = 2.1 mm, 75 µg/ml = 4.8 mm), E. coli (50 µg/ml = 1.3 mm, 75 µg/ml = 4.3 mm) bacteria. The antibiofilm activity of Cr-SeNWs was also investigated and biofilm reduction was observed at 75 µg/ml. In addition, Cr-SeNWs were highly effective as larvicides against Zika virus and Japanese encephalitis mosquito vectors, i.e., Culex quinquefasciatus and Culex tritaeniorhynchus, with LC50 values of 4.15 and 4.85 mg/l, respectively. The nanowire toxicity and internalization was investigated through confocal laser scanning microscopy and histological studies. To investigate the potential of Cr-SeNWs for real-world applications, we also evaluated Cr-SeNWs in hemolytic assays, showing no cytotoxicity till 5 mg/ml. Besides, higher antioxidant activity at the concentration at 100 µg/ml was noted, if compared with purified crustin. The strong antioxidant potential of this nanomaterial can be helpful to boost the shelf-life potential of Cr-SeNWs-based pesticides and antimicrobials.