Synthesis and characterization of mesoporous zinc oxide nanoparticles and evaluation of their biocompatibility in L929 fibroblasts.

OBJECTIVES: This study aimed to synthesize and characterize mesoporous zinc oxide nanoparticles (ZnO NPs) and also to evaluate the cytotoxicity of mesoporous ZnO NPs on L929 mouse fibroblast cell lines using 3-(4,5- dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay. MATERIALS AND METHODS: The synthesized mesoporous ZnO NPs were extensively characterized using X-ray diffraction analysis (XRD), transmission electron microscopy (TEM), Brunauer-Emmett-Teller (BET) analysis, field emission scanning electron microscopy (FESEM), energy dispersive X-ray spectra (EDAX), Fourier-transform infrared spectroscopy (FTIR), and dynamic light scattering (DLS). The cytotoxicity of mesoporous ZnO NPs was assessed by MTT assay. The study groups for cytotoxicity assay were normal saline, 0.1% calcined mesoporous ZnO NP solution, 1% calcined mesoporous ZnO NP solution, 0.1% noncalcined mesoporous ZnO NP solution, 1% noncalcined mesoporous ZnO NP solution, 0.1% ZnO NP solution, 1% ZnO NP solution, 2% chlorhexidine, and phosphate-buffered saline (PBS). The percentages of mean ± standard deviation of viable cells were analyzed. RESULTS: Characterization of mesoporous ZnO NPs revealed that all the particles were in a more or less spherical shape with a wide particle size distribution of 70-100 nm. TEM image showed the uniformed and aggregated ZnO NPs with a typical size of 10-15 nm. BET analysis showed a mesoporous structure for the prepared mesoporous ZnO NPs. According to the MTT assay, chlorhexidine had the lowest cell viability percentage. Cell viability percentages of 0.1% mesoporous ZnO NP solutions (calcined and noncalcined) were statistically, significantly higher than 0.1% ZnO NP solution (p < .05). Cell viability percentages of 0.1% calcined and noncalcined mesoporous ZnO NP solutions and 0.1% ZnO NP solution were statistically, significantly higher than the 1% solutions (p < .05). CONCLUSION: Mesoporous ZnO NPs exhibited less cytotoxicity against L929 mouse fibroblast cell lines compared to CHX and ZnO NPs, hence are safe to use.

Biosynthesis of ZnO, Bi2O3 and ZnO-Bi2O3 bimetallic nanoparticles and their cytotoxic and antibacterial effects.

This work introduces an easy method for producing Bi2O3, ZnO, ZnO-Bi2O3 nanoparticles (NPs) by Biebersteinia Multifida extract. Our products have been characterized through the outcomes which recorded with using powder X-ray diffractometry (PXRD), Raman, energy dispersive X-ray (EDX), field emission-scanning electron microscopy (FE-SEM), and Fourier-transform infrared (FT-IR) techniques. The finding of SEM presented porous structure and spherical morphology for Bi2O3 and ZnO NPs, respectively. While FE-SEM image of bimetallic nanoparticles showed both porous and spherical morphologies for them; so that spherical particles of ZnO have sat on the porous structure of Bi2O3 NPs. According to the PXRD results, the crystallite sizes of Bi2O3, ZnO and ZnO-Bi2O3 NPs have been obtained 57.69, 21.93, and 43.42 nm, respectively. Antibacterial performance of NPs has been studied on Staphylococcus epidermidis and Pseudomonas aeruginosa bacteria, to distinguish the minimum microbial inhibitory concentration (MIC). Antimicrobial outcomes have showed a better effect for ZnO-Bi2O3 NPs. Besides, wondering about the cytotoxic action against cancer cell lines, the MTT results have verified the intense cytotoxic function versus breast cancer cells (MCF-7). According to these observations, obtained products can prosper medical and biological applications.

Green synthesis of zinc oxide nanoparticles using aqueous extract of shilajit and their anticancer activity against HeLa cells.

In the present study, ZnO nanoparticles have been synthesized using an aqueous extract of shilajit. The nanoparticles were characterized using different techniques such as UV (ultraviolet-visible spectrophotometer), FTIR (Fourier transform infrared), XRD (X-ray diffraction), particle size analysis, SEM (scanning electron microscope) and EDAX (Energy-dispersive X-ray) analysis. The UV absorption peak at 422.40 nm was observed for ZnO nanoparticles. SEM analysis showed the shape of nanoparticles to be spherical, FTIR spectrum confirmed the presence of zinc atoms, particle size analysis showed the nanoparticle size, EDAX confirmed the purity of ZnO nanoparticles whereas XRD pattern similar to that of JCPDS card for ZnO confirmed the presence of pure ZnO nanoparticles. The in vitro anticancer activity of ZnO nanoparticles against the HeLa cell line showed the IC50 value of 38.60 µg/mL compared to reference standard cisplatin. This finding confirms that ZnO nanoparticles from shilajit extract have potent cytotoxic effect on human cervical cancer cell lines.

Effects of zinc supplementation from organic and inorganic sources on growth, blood biochemical indices, and intestinal microarchitecture in broilers.

In poultry nutrition, zinc supplementation is typically achieved through the addition of zinc oxide or zinc sulfate to the feed. The alternative approach of organic sources utilizes an organic ligand to bind zinc (Zn), resulting in higher bioavailability. Thus, a study was conducted to assess and compare the impact of a methionine-complexed Zn versus an inorganic Zn on growth, blood biochemical profile, gut histomorphology, and fecal excretion of Zn in broilers. The experimental design included two treatments: the addition of a zinc amino acid complex or zinc oxide to the basal diet. The zinc amino acid complex was supplemented at a dose equivalent to the inorganic zinc (Zn-80), while the organic zinc was provided at levels of 20, 40, and 80 mg/kg to a total of 400 broilers. There were five treatments in total, and each treatment was replicated four times. Broilers supplemented with an organic form of Zn at the level of 80 mg/kg had significantly (p < 0.05) higher body weight gain and lower feed conversion ratio (F/G). Significantly (p < 0.05) higher Zn excretion was recorded in broilers supplemented with inorganic Zn supplementation. Significantly (p < 0.05) higher villus length and width, their ratio, and lower (p < 0.05) crypt depth were observed in birds supplemented with 80 mg/kg organic Zn. From the results of the present study, it was concluded that Zn from an organic source at the rate of 80 mg/kg was superior in terms of growth performance, intestinal histomorphology and less excretion of Zn to the environment in broilers.

Mesenchymal stem cells osteogenic differentiation by ZnO nanoparticles and polyurethane bimodal foam nanocomposites.

Mesenchymal stem cells with tissue repair capacity involve in regenerative medicine. MSCs can promote bone repair when employed with nano scaffolds/particles. Here, the MTT and Acridine Orange assay enabled the cytotoxic concentration of Zinc oxide nanoparticles and Polyurethane evaluation. Following culturing adipose tissue-derived MSCs, ADSCs' proliferation, growth, and osteogenic differentiation in the presence of PU with and without ZnO NPs is tracked by a series of biological assays, including Alkaline Phosphatase activity, Calcium deposition, alizarin red staining, RT-PCR, scanning electron microscope, and immunohistochemistry. The results showed boosted osteogenic differentiation of ADSCs in the presence of 1% PU scaffold and ZnO NPS and can thus apply as a new bone tissue engineering matrix. The expression level of Osteonectin, Osteocalcin, and Col1 increased in PU-ZnO 1% on the 7th and 14th days. There was an increase in the Runx2 gene expression on the 7th day of differentiation in PU-ZnO 1%, while it decreased on day 14th. In conclusion, Polyurethane nano scaffolds supported the MSCs' growth and rapid osteogenic differentiation. The PU-ZnO helps not only with cellular adhesion and proliferation but also with osteogenic differentiation.

Construction of photocatalytic coating for alleviating the shriveling of postharvest fruit cucumber after simulated transportation.

The shriveling of fruit cucumber was commonly occurred during supply chain, photocatalyst exposed to UV light can endow the coatings with ethylene removal capacity to reduce the respiration of fruit and water loss. The study developed a novel photodynamic technology responsive photocatalytic coating with exceptional ultraviolet (UV) photocatalytic degradation of ethylene ability to decay the shriveling of postharvest fruit cucumber during supply chain. This coating involved the integration of Carbon dots (CDs)-loaded nano ZnO and the skillful selection of pullulan (Pul) and apple pectin (AP) matrix. The CDs/ZnO coatings boasted an impressive array of photocatalytic degradation of ethylene and adhesion properties, including high ethylene removal rates of 32.04 % in 60 min UV light stimulation. The decrease of cell-wall strength, degradation of the cell wall polysaccharides and water loss resulted in cucumber shriveling. Compared with CK sample, after UV-CDs/ZnO coating treatment, the higher firmness and cell wall polysaccharides were found in cucumbers with lower cell wall degrading enzymes activities, weight loss and water movement, which was associated with the decrease of respiration and ethylene accumulation. The UV-CDs/ZnO coatings possessed promising potential for alleviating the shriveling of postharvest fruit cucumber and applications in fruits preservation in the future.

A scaffold containing zinc oxide for Schwann cell-mediated axon growth.

Objective.Schwann cells (SCs) transplanted in damaged nervous tissue promote axon growth, which may support the recovery of function lost after injury. However, SC transplant-mediated axon growth is often limited and lacks direction.Approach.We have developed a zinc oxide (ZnO) containing fibrous scaffold consisting of aligned fibers of polycaprolactone (PCL) with embedded ZnO nanoparticles as a biodegradable, bifunctional scaffold for promoting and guiding axon growth. This scaffold has bifunctional properties wherein zinc is released providing bioactivity and ZnO has well-known piezoelectric properties where piezoelectric materials generate electrical activity in response to minute deformations. In this study, SC growth, SC-mediated axon extension, and the presence of myelin basic protein (MBP), as an indicator of myelination, were evaluated on the scaffolds containing varying concentrations of ZnOin vitro. SCs and dorsal root ganglion (DRG) neurons were cultured, either alone or in co-culture, on the scaffolds.Main results.Findings demonstrated that scaffolds with 1 wt.% ZnO promoted the greatest SC growth and SC-mediated axon extension. The presence of brain-derived neurotrophic factor (BDNF) was also determined. BDNF increased in co-cultures for all scaffolds as compared to SCs or DRGs cultured alone on all scaffolds. For co-cultures, cells on scaffolds with low levels of ZnO (0.5 wt.% ZnO) had the highest amount of BDNF as compared to cells on higher ZnO-containing scaffolds (1 and 2 wt.%). MBP immunostaining was only detected in co-cultures on PCL control scaffolds (without ZnO).Significance.The results of this study demonstrate the potential of the ZnO-containing scaffolds for SC-mediated axon growth and its potential for use in nervous tissue repair.

ZnO nanoparticles mediated by Azadirachta indica as nano fertilizer: Improvement in physiological and biochemical indices of Zea mays grown in Cr-contaminated soil.

The current investigation aimed at evaluating the impact of Azadirachta indica-mediated zinc oxide nanoparticles (Ai-ZnONPs) on the growth and biochemical characteristics of maize (sweet glutinous 3000) under exposure to 50 mg kg-1Ai-ZnONPs with Cr (VI) concentrations of 50 and 100 mg kg-1. The results indicate that plants exposed to Cr (VI) only experienced a decline in growth parameters. Conversely, the inclusion of Ai-ZnONPs caused a noteworthy increase in physiological traits. Specifically, shoot and root fresh weight increased by 28.02% and 16.51%, and 63.11% and 97.91%, respectively, when compared to Cr-50 and 100 treatments. Additionally, the SPAD chlorophyll of the shoot increased by 91.08% and 15.38% compared to Cr-50 and 100 treatments, respectively. Moreover, the antioxidant enzyme traits of plant shoot and root, such as superoxide dismutase (SOD 7.44% and 2.70%, and 4.45% and 3.53%), catalase (CAT 1.18% and 3.20%, and 5.03% and 5.78%), and peroxidase (POD 0.31% and 5.55%, and 4.72% and 3.61%), exhibited significant increases in Cr 50 and 100 treatments, respectively. The addition of Ai-ZnONPs to the soil also enhanced soil nutrient status and reduced Cr (VI) concentrations by 40.69% and 19.82% compared to Cr-50 and 100 treated soils. These findings suggest that Ai-ZnONPs can trigger the activation of biochemical pathways that enable biomass accumulation in meristematic cells. Further investigations are required to elucidate the mechanisms involved in growth promotion.

Antimicrobial mechanisms of ZnO nanoparticles to phytopathogen Pseudomonas syringae: Damage of cell envelope, suppression of metabolism, biofilm and motility, and stimulation of stomatal immunity on host plant.

Nanoparticles have recently been employed as a new strategy to act as bactericides in agricultural applications. However, the effects and mechanisms of foliar deposition of nanoparticles on bacterial pathogens, plant physiology and particularly plant immunity have not been sufficiently understood. Here, we investigated the effects and mechanisms of ZnO NPs in controlling of tobacco wildfire caused by Pseudomonas syringae pv. tabaci, through the comprehensive analysis of biological changes of both bacteria and plants. The global gene expression changes of Pseudomonas syringae pv. tabaci supported that the functions of "protein secretion", "membrane part", "signal transducer activity", "locomotion", "chemotaxis" and "taxis" in bacteria, as well as the metabolic pathways of "bacterial chemotaxis", "two-component system", "biofilm formation", "ABC transporters" and "valine, leucine and isoleucine degradation" were significantly down-regulated by ZnO NPs. Correspondingly, we reconfirmed that the cell envelope structure, biofilm and motility of Pseudomonas syringae pv. tabaci were directly disrupted or suppressed by ZnO NPs. Different from completely killing Pseudomonas syringae pv. tabaci, ZnO NPs (0.5 mg/mL) potentially improved plant growth and immunity through enzymatic activity and global molecular response analysis. Furthermore, the changes of gene expression in ABA signaling pathway, ABA concentration and stomatal aperture all supported that ZnO NPs can specifically stimulate stomatal immunity, which is important to defend bacterial infection. Taken together, we proposed that both the inhibition or damage of motility, biofilm, metabolisms, virulence and cell envelope on P. syringae pv. tabaci, and the activation of the stomatal immunity formed two-layered antibacterial mechanisms of ZnO NPs on phytopathogenic bacteria.

Compensatory response of Hong Kong oysters to co-occurring stressors: Zinc oxide nanoparticles and low salinity exposure.

Zinc Oxide nanoparticles (ZnO NPs), due to their ubiquitous use in industrial and consumer applications, present potential risks to marine ecosystems and biota, especially oysters. The physiological and immunological health of marine species is highly dependent on salinity levels. However, the combined impact of lowered salinity and exposure to ZnO NPs, particularly on key marine species like oysters, is an area that requires more research. Our study aimed to examine these concurrent stressors' impacts on phenotypic markers, gill and hepatopancreas physiological indices, and hemocyte immune parameters of Crassostrea hongkongensis. We subjected six oyster cohorts to varied ZnO NPs concentrations and salinity levels over 21 days. Our findings reveal that individual exposure to ZnO NPs or diminished salinity disrupts oyster physiology, impacting metabolism, antioxidant capacity, immune response, and energy distribution through distinct mechanisms. Remarkably, low salinity constituted a more significant threat than isolated ZnO NPs. However, when confronted with combined stressors, oysters exhibited a compensatory response, attenuating individual stressors' detrimental effects. This adaptation was characterised by reduced apoptosis rates, increased calcium ion concentration in mature hemocytes, and a restoration of conditioned indices, hepatopancreas alkaline phosphatase, and gill catalase activity to baseline levels. Principal Component Analysis and Integrated Biomarker Responses validated this compensatory phenomenon. Partial Least Squares Pathway Model analysis underscored these stressors' profound implications on oyster health, primarily driven by stressor exposure rather than mere zinc concentrations, despite acknowledging zinc's immunosuppressive impact on oyster immunity. Our research emphasises the importance of assessing multiple stressors' cumulative effects on aquatic species' ecological resilience, accentuating the need for comprehensive analyses incorporating functional specificity among diverse organs and immune components, including gill, hepatopancreas, and the critical hemocytes.

Bee venom (Apis mellifera L.) rescues zinc oxide nanoparticles induced neurobehavioral and neurotoxic impact via controlling neurofilament and GAP-43 in rat brain.

This study investigated the possible beneficial role of the bee venom (BV, Apis mellifera L.) against zinc oxide nanoparticles (ZNPs)-induced neurobehavioral and neurotoxic impacts in rats. Fifty male Sprague Dawley rats were alienated into five groups. Three groups were intraperitoneally injected distilled water (C 28D group), ZNPs (100 mg/kg b.wt) (ZNPs group), or ZNPs (100 mg/kg.wt) and BV (1 mg/ kg.bwt) (ZNPs + BV group) for 28 days. One group was intraperitoneally injected with 1 mL of distilled water for 56 days (C 56D group). The last group was intraperitoneally injected with ZNPs for 28 days, then BV for another 28 days at the same earlier doses and duration (ZNPs/BV group). Depression, anxiety, locomotor activity, spatial learning, and memory were evaluated using the forced swimming test, elevated plus maze, open field test, and Morris water maze test, respectively. The brain contents of dopamine, serotonin, total antioxidant capacity (TAC), malondialdehyde (MDA), and Zn were estimated. The histopathological changes and immunoexpressions of neurofilament and GAP-43 protein in the brain tissues were followed. The results displayed that BV significantly decreased the ZNPs-induced depression, anxiety, memory impairment, and spatial learning disorders. Moreover, the ZNPs-induced increment in serotonin and dopamine levels and Zn content was significantly suppressed by BV. Besides, BV significantly restored the depleted TAC but minimized the augmented MDA brain content associated with ZNPs exposure. Likewise, the neurodegenerative changes induced by ZNPs were significantly abolished by BV. Also, the increased neurofilament and GAP-43 immunoexpression due to ZNPs exposure were alleviated with BV. Of note, BV achieved better results in the ZNPs + BV group than in the ZNPs/BV group. Conclusively, these results demonstrated that BV could be employed as a biologically effective therapy to mitigate the neurotoxic and neurobehavioral effects of ZNPs, particularly when used during ZNPs exposure.

The inclusion of engineered ZnO nanoparticles and bulk ZnSO4 in the growth medium distinctively modulate the root and leaf metabolome in bean plants.

Together with toxicity, beneficial effects on plant growth have been ascribed to nanoparticles (NPs). This study aimed to survey the growth performance and metabolome adjustment of beans grown in a growth medium containing ZnONPs at different concentrations and compared with bulk ZnSO4 as a positive control. Growth parameters showed a reduction in shoot height starting from the lowest (25 mg L-1 ) concentration of ZnONPs. In comparison, growth was inhibited from 50 mg L-1 ZnSO4 , suggesting more toxic effects of nano forms of Zn. Untargeted metabolomics allowed us to unravel the biochemical processes involved in both promising and detrimental aspects. Multivariate statistics indicated that the tested Zn species substantially and distinctively altered the metabolic profile of both roots and leaves, with more metabolites altered in the former (435) compared with leaves (381). Despite having Zn forms in the growth medium, also leaf metabolome underwent a significant and extensive modulation. In general, the elicitation of secondary metabolism (N-containing compounds, phenylpropanoids, and phytoalexins) and the down-accumulation of fatty acid biosynthesis compounds were common responses to different Zn forms. However, an opposite trend could be observed for amino acids, fatty acids, carbohydrates, and cofactors being down-accumulated in ZnONPs treatment. Osmolytes, especially in ZnSO4 treatment, contributed to mitigating the effect of Zn toxicity and maintaining plant growth. Overall, the results indicated a complexity of tissue-specific and Zn-dependent response differences, resulting in distinctive metabolic perturbations.

Combined cerium and zinc oxide nanoparticles induced hepato-renal damage in rats through oxidative stress mediated inflammation.

The toxicity profiles of nanoparticles (NPs) used in appliances nowadays remains unknown. In this study, we investigated the toxicological consequences of exposure to cerium oxide (CeO2) and zinc oxide (ZnO) nanoparticles given singly or in combination on the integrity of liver and kidney of male Wistar rats. Twenty (20) rats were allotted into four groups and treated as: Control (normal saline), CeO2NPs (50 µg/kg), ZnONPs (80 µg/kg) and [CeO2NPs (50 µg/kg) + ZnONPs (80 µg/kg)]. The nanoparticles were given to the animals through the intraperitoneal route, three times per week for four repeated weeks. Results revealed that CeO2 and ZnO NPs (singly) increased serum AST and ALT by 29% & 57%; 41% & 18%, and co-administration by 53% and 23%, respectively. CeO2 and ZnO NPs increased hepatic and renal malondialdehyde (MDA) by 33% and 30%; 38% and 67%, respectively, while co-administration increased hepatic and renal MDA by 43% and 40%, respectively. The combined NPs increased hepatic NO by 28%. Also, CeO2 and ZnO NPs, and combined increased BAX, interleukin-1ß and TNF-α by 45, 38, 52%; 47, 23, 82% and 41, 83, 70%, respectively. Histology revealed hepatic necrosis and renal haemorrhagic parenchymal in NPs-treated rats. Summarily, CeO2 and ZnO NPs produced oxidative injury and induced inflammatory process in the liver and kidney of experimental animals.

Nanobioremediation: a bacterial consortium-zinc oxide nanoparticle-based approach for the removal of methylene blue dye from wastewater.

Industrial effluents carrying dyes are considered a major environmental threat in the present era. Methylene blue (MB) dye is one of the key dyes of the thiazine group of dyes. It is broadly used in medical, textile, and various fields and is well known for its carcinogenicity and methemoglobin nature. Bacterial and other microbes-mediated bioremediation is becoming an emerging and significant section for the treatment of wastewater. Isolated bacteria were used for the bioremediation and nanobioremediation of methylene blue dye under varying conditions and parameters. A comparative study was conducted for the remediation of methylene blue dye using bacterial consortium, potential bacteria (isolated by scale-up method), and potential bacteria within zinc oxide nanoparticles. The decolorizing ability of bacteria was analyzed by UV visible spectrophotometer after stirring and static incubation in different time intervals of the isolates. Growth parameters and environmental parameters which include pH, initial dye concentration, and dose of nanoparticles were optimized with the minimal salt medium. An enzyme assay study was also done to check the effect of dye and nanoparticles on bacterial growth and the mode of action of degradation. The authors found that potential bacteria within ZnO nanoparticles showed enhanced decolorization efficiency (95.46% at pH 8) due to the properties of nanoparticles. On the other hand, the decolorization of MB dye by potential bacteria and the bacterial consortium was about 89.08 and 76.3%, respectively, for a 10-ppm dye concentration. During the enzyme assays study, the highest activity was observed for phenol oxidase, nicotinamide adenine dinucleotide (NADH), 2,6-Dichloroindophenol(DCIP), and laccase for nutrient broth having MB dye, MB dye, and ZnO NPs, while no such change was observed for manganese peroxidase enzyme activity. Nanobioremediation is a promising approach to removing such pollutants from the environment.

Biofertilizer effect of some zinc dissolving bacteria free and encapsulated on Zea mays growth.

Crop nutrition depends on zinc for enzymatic, oxidative, and metabolic processes. In the current study 20 different bacteria were isolated from five soil samples collected from different fields in Egypt. Bacterial isolates were screened for their ability to solubilize insoluble zinc oxide and zinc carbonate. The ability of selected isolates to tolerate soluble zinc was determined using different concentrations of (ZnSO4). Three bacterial isolates were selected with efficiency in solubilizing zinc oxide and zinc carbonate while tolerating high levels of soluble zinc. Molecular identification by 16S rRNA sequencing of the chosen isolates identified them as B3 (Acinetobacter calcoaceticus), B5 (Bacillus proteolyticus) and C6 (Stenotrophomonas pavanii). Sodium alginate beads formulated with the isolated bacteria were tested for stability under different storage conditions for 3 months. A pot experiment was conducted to study and compare the effect of using chosen isolates as an in vivo Zn solubilizer with amended ZnCO3 either alone or embedded in beads as carrier in the soil and its effect on growth parameters of Zea mays after 2 months. There was an increase in Zn uptake in all treatments compared to the control. However, plants grown in a pot treated with ZnCO3 and Acinetobacter calcoaceticus showed the highest zinc content and plant dry weight as compared to the control. Finally, selected isolates in both free and encapsulated forms showed improved plant growth parameters and higher zinc content and can be applied as biofertilizers to enhance soil fertility.

Multiple effects of ZnO nanoparticles on goldfish (Carassius auratus): Skin mucus, gut microbiota and stable isotope composition.

The skin and the gut are direct target tissues for nanoparticles, yet attention to effects of metal-based nanoparticles (MNPs) on these two and the discrepancy in these effects remain inadequate. Here, effects of ZnO nanoparticles (nZnO) on skin mucus and gut microbiota of goldfish (Carassius auratus) were investigated, as well as further elements turnover and metabolic variations. After 14 days of exposure, considerable variations in levels of biomarkers (protein, glucose, lysozyme and immunoglobulin M) in skin mucus demonstrated significant stress responses to nZnO. nZnO exposure significantly reduced the abundance of Cetobacterium in the gut while increased that of multiple pathogens, and further leading to down-regulation of pathways such as carbohydrate metabolism, translation, and replication and repair. Decreased δ15N values indicated declined N turnover in vivo, further demonstrating the negative effect of nZnO on metabolism in the organism. Integration analysis of each biomarker using the biomarker response index version 2 (IBRv2) revealed concentration-dependent effects of nZnO on skin mucus, while effects on physiology in vivo was not, demonstrating the discrepancy in the toxicity pathways and toxic effects of nZnO on different tissues. This work improved our understanding about the comprehensive toxicity of nZnO on aquatic organism.

Modulatory effect of thymol on the immune response and susceptibility to Aeromonas hydrophila infection in Nile tilapia fish exposed to zinc oxide nanoparticles.

Zinc oxide nanoparticles (ZnO-NPs) have many exciting properties that make their use in a continuous increase in various biomedical, industrial, and agricultural applications. This is associated with accumulation in the aquatic ecosystems and fish exposure with consequent deleterious effects. To determine the potential of thymol to counteract the immunotoxic effects of ZnO-NPs, Oreochromis niloticus was exposed to ZnO-NPs (⅕ LC50 =1.14 mg/L, for 28 days) with or without feeding a thymol-incorporated diet (1 or 2 g/kg diet). Our data demonstrated a reduction of aquaria water quality, leukopenia, and lymphopenia with a decrease in serum total protein, albumin, and globulin levels in exposed fish. At the same time, the stress indices (cortisol and glucose) were elevated in response to ZnO-NPs exposure. The exposed fish also revealed a decline in serum immunoglobulins, nitric oxide, and the activities of lysozyme and myeloperoxidase, in addition to reduced resistance to the Aeromonas hydrophila challenge. The RT-PCR analysis showed downregulation of antioxidant (SOD) superoxide dismutase and (CAT) catalase gene expression in the liver tissue with overexpression of the immune-related genes (TNF-α and IL-1ß). Importantly, we found that thymol markedly protected against ZnO-NPs-induced immunotoxicity in fish co-supplemented with thymol (1 or 2 g/kg diet) in a dose-dependent manner. Our data confirm the immunoprotective and antibacterial effects of thymol in ZnO-NPs exposed fish, supporting the potential utility of thymol as a possible immunostimulant agent.

ZnO-NPs alleviate aflatoxin B1-induced hepatoxicity in ducklings by promoting hepatic metallothionein expression.

Aflatoxin B1 (AFB1) is a mycotoxin widely present in animal feed and human food, posing a serious threat to animal and human health. This study was aim to illustrate the mechanism of the protective role of MT against AFB1-induced hepatotoxicity, as well as to explore the feasibility of enhancing the tolerance of poultry to AFB1 by upregulating the expression of hepatic MT. After being exposed to AFB1 (50 ng/kg) primary duckling hepatocytes, the cell viability, the antioxidant index (SOD and GPx) and the mRNA levels of MT downstream genes (PTGR, p53, TrxR, AR and Bcl-2) significantly (p < 0.05) decreased, while the intracellular formation of (AFBO)-DNA adduct content, apoptosis, and MDA content significantly (p < 0.05) increased. Interestingly, overexpression of MT in primary duckling hepatocytes markedly (p < 0.05) reversed the detrimental impact of AFB1 and increased the expression of MT downstream genes. HepG2 cells were applied to study the mechanism how MT works to relieve the hepatic toxicity of AFB1. The ZnO-NPs (20 µg/mL) + AFB1 (20 µg/mL) group significantly (p < 0.05) increased the cell viability, the expression of NRF2, NQO1 and SOD, and expression of MT and MTF-1, as well as significantly (p < 0.05) decreased LDH, ROS and apoptotic rate, comparing with the AFB1 group. While joint treatment with AFB1 and ZnO-NPs, the hepatic toxicity exerted by AFB1 alone was reversed, along with the translocation of MTF-1 from the cytoplasm to the nucleus and upregulated its expression. Duckling trails were further carried out. A total number of 96 1-day-old healthy Cherry Valley commercial ducklings were randomly allocated according to a 2 by 2 factorial arrangement of treatments with the main factors including oral administration of AFB1 (0 vs. 40 µg/kg) and dietary supplementation of ZnO-NPs (0 vs. 60 mg/kg) for 7 days. It showed that AFB1 exposure caused body weight loss (p < 0.05), impaired liver structure and failure in hepatic function (activity of ALT, AST and concentration of TP and GLU) (p < 0.05), and decreases in antioxidant capacity(activity of SOD, CAT and concentration of GSH) (p < 0.05), along with the decrease in hepatic concentration of Zn, increase in expression of apoptosis-related genes and protein CAS3 and mRNA Bcl-2 expression (p < 0.05), and suppressed mRNA levels of antioxidant-related genes MT, SOD1, NRF2, and NQO1 (p < 0.05). In accordance with the cell test, dietary supplementation with ZnO-NPs mitigated the toxicity exerted by AFB1. In conclusion, ZnO-NPs has the protective effects against AFB1-induced hepatocyte injury by activating the expression of MTF-1 and the ectopic induction of MT expression, providing detailed information on the detoxification ability of MT on AFB1.

Potential ecotoxicity of substrate-enriched zinc oxide nanoparticles to Physalaemus cuvieri tadpoles.

Although the ecotoxicological effects of ZnO nanoparticles (ZnO NPs) have already been reported in different taxa, little is known about their impacts on amphibians. Thus, we aimed to evaluate the potential effects of exposure of Physalaemus cuvieri tadpoles to substrates enriched with ZnO NPs (and with its ionic counterpart, Zn+2, ZnCl2 - both at 100 mg/kg) previously used in the cultivation of Panicum maximum (Guinea grass). We showed that although exposure for 21 days did not impact the survival, growth, and development of tadpoles, we noted an increase in the frequency of erythrocyte nuclear abnormalities in the "ZnCl2" and "ZnONP" groups, which was associated with suppression of antioxidant activity in the animals (inferred by SOD and CAT activity and DPPH free radical scavenging capacity). In the tadpoles of the "ZnONP" group, we also noticed a reduction in creatinine and bilirubin levels, alpha-amylase activity, and an increase in alkaline phosphatase activity. But the treatments did not alter the activity of the enzymes lactate dehydrogenase and gamma-glutamyl-transferase and total protein and carbohydrate levels. On the other hand, we report a cholinesterase and hypotriglyceridemic effect in the "ZnCl2" and "ZnONP" groups. Zn bioaccumulation in animals, from ZnO NPs, from Zn+2 released from them, or both, has been associated with causing these changes. Finally, principal component analysis (PCA) and the values of the "Integrated Biomarker Response" index revealed that the exposure of animals to substrates enriched with ZnO NPs caused more pronounced effects than those attributed to its ionic counterpart. Therefore, our study reinforces the need to consider the environmental risks of using these nanomaterials for agricultural purposes for amphibians.

Susceptibility of Bacillus subtilis to Zinc Oxide Nanoparticles Treatment.

Aim: With the characteristics such as low toxicity, high total surface, ability to inhibit the growth of pathogenic microorganisms, zinc oxide nanoparticles (ZnO NPs), as one of the metallic nanoparticles, have been chosen as an antibacterial agent to treat various skin infections. The present study was aimed to determine the antibacterial potential of ZnO NPs on Bacillus subtilis, the Gram-positive bacterium that can cause skin and wound infections. Methods: B. subtilis was exposed to 5 to 150 µg/mL of ZnO NPs for 24 h. The parameters employed to evaluate the antimicrobial potential of ZnO NPs were the growth inhibitory effect on B. subtilis, the surface interaction of ZnO NPs on the bacterial cell wall, and also the morphological alterations in B. subtilis induced by ZnO NPs. Results: The results demonstrated a significant (p <0.05) inhibition of ZnO NPs on B. subtilis growth and it was in a dose-dependent manner for all the tested concentrations of ZnO NPs from 5 to 150 µg/mL at 24 h. Fourier transformed infrared (FTIR) spectrum confirmed the involvement of polysaccharides and polypeptides of bacterial cell wall in surface binding of ZnO NPs on bacteria. The scanning electron microscopy (SEM) was used to visualize the morphological changes, B. subtilis illustrated several surface alterations such as distortion of cell membrane, roughening of cell surface, aggregation and bending of cells, as well as, the cell rupture upon interacting with ZnO NPs for 24 h. Conclusion: The results indicated the potential of ZnO NPs to be used as an antibacterial agent against B. subtilis. The findings of the present study might bring insights to incorporate ZnO NPs as an antibacterial agent in the topical applications against the infections caused by B. subtilis.