Comparative biotoxicity study for identifying better alternative insecticide especially green nano-emulsion which used as mosquitocides.

This research work was planned to test biosafety of different nanomaterials on the different animals models. These nanoparticles were previously used as potential insecticides of mosquito larvae. The biosafety of these nanoproducts were evaluated on certain organs of non target animals that associated with mosquito breeding sites in Egypt. Animal organs such as the kidneys of rats, toads, and the fish's spleen were used as models to study the biological toxicity of these nanomaterials. After 30 days of the animals receiving the nanomaterials in their water supply, different cell mediated immune cells were assessed in these tissues. Both TNF-α and BAX immuno-expression were also used as immunohistochemical markers. Histopathology was conducted to detect the effect of the tested nanoproducts at the tissue level of the liver and kidneys of both the rats and toads. Green nanoemulsion of the lavender essential oil was relatively more effective, safe, and biodegradable to be used as insecticides against mosquito larvae than the metal-based nanomaterials.

Candida albicans Infection Model in Zebrafish (Danio rerio) for Screening Anticandidal Drugs.

BACKGROUND: Candida albicans is an opportunistic fungal pathogen which causes systemic infections in human. In this study, C. albicans infection model was developed in zebrafish to understand the host-pathogen interactions for straightforward anticandidal drug screening. METHODS: To develop the infection, 1 × 106 cells of C. albicans suspended in phosphate-buffered saline were deposited in zebrafish dorsal muscle by manually operated syringe. The infection progression was externally assessed by a scale of wound-healing events, based on visible changes of yeast deposited in the muscle tissues. Chemotherapy was carried out with known antifungal drugs (fluconazole, nystatin, and amphotericin B) and a potential antifungal agent, chitosan silver nanocomposites (CAgNC), after the infection as direct exposure in the water. Histopathological analysis was performed to identify the pathogen virulence and the host-pathogen interaction during the infection. RESULTS: The light microscopic observations and histopathological analysis revealed the yeast-hyphae transition at the site of infection (at 72 hpi) and progression of the infection in the host tissues. The larval survival rate under fluconazole (up to 80 µg mL-1) and nystatin (up to 20 µg mL-1) was > 90% and for CAgNC it was 40% at 36 h post-exposure (hpe). The infection progression was suppressed with the fungicidal treatments. Among inflammatory genes, il-1ß has been highly upregulated (14.68-fold) at 24 h post infection (hpi). Both il-1ß and tnf-α were moderately upregulated in infected fish gills at 72 hpi. Among the C. albicans antioxidant genes, cat1 and sod2 have been upregulated during the infection, and relative expression folds were increased from low to moderate levels with the time. DISCUSSION: We demonstrate the approach for the development of artificial infection model of zebrafish with C. albicans. By this mini vertebrate zebrafish model, researchers will be able to study novel anticandidal compounds in vivo with respect to the host, pathogen, and their interactions.

Metagenomics analysis of gut microbiota and immune modulation in zebrafish (Danio rerio) fed chitosan silver nanocomposites.

In this study, we evaluated the effects of chitosan silver nanocomposites (CAgNCs) supplemented diet on gut microbial community, goblet cell density, gut morphometry and mRNA expression of immune related and mucin encoding genes in zebrafish. Zebrafish gut microbiota analysis results clearly showed the reduction of phylum Proteobacteria. However, they remained as the major bacterial group in gut with CAgNCs supplemented diet, while the abundance of phylum Fusobacteria and phylum Bacteroidetes were increased notably compared to the control diet fed fish. Total goblet cell density was significantly increased at 30 and 60 days in CAgNCs supplemented group (1.6-fold and 2.0-fold, respectively) compared to the control group indicating enhanced immune function in the gut. CAgNCs supplementation has also increased villi height significantly in the zebrafish mid gut at the end of 30 (95.5 ± 3.7 µm) and 60 days (144.40 ± 4.8 µm) compared to control diet fed fish at 30 (86.90 ± 3.7 µm) and 60 days (96.2 ± 4.8 µm). Furthermore, mRNA expression of immune related genes such as TNF-α (6.2-fold), IL-10 (5.0-fold), IL-12 (9.2-fold), IRF-1 (5.2-fold), Defbl1 (3-fold), Lyz (5.1-fold) and mucin encoding genes were significantly upregulated (above 2-fold) compared to that of control group. The current study revealed that CAgNCs supplemented diet engenders promising effects on fish gut immunity by enhancing beneficial microbial populations, goblet cell density, villi length, and transcriptional regulation of immune related and mucin encoding genes.

Bactericidal, anti-biofilm, anti-oxidant potency and catalytic property of silver nanoparticles embedded into functionalised chitosan gel.

Chitosan is a versatile biocompatible polysaccharide which has attracted great attention for gel synthesis. Its reducing character is specifically exploited for nanoparticle synthesis via green approach. A silver nanocomposite synthesized using this gel, with a novel gelling agent 2,4,6-trihydroxy benzaldehyde, was found to be a promising candidate for several applications including anti-bacterial, anti-biofilm and anti-oxidant activity as well as catalysis. The nanocomposite was well characterized using various spectroscopic and microscopic techniques such as IR, TGA, XRD, XPS, SEM and TEM. The nanocomposite exhibited high bactericidal activity against both S. aureus and E. coli. Further, it was evaluated for anti-biofilm forming property and its potency as antioxidant agent. The nanocomposite served as a catalyst for degradation of Methyl Orange and Rhodamine B at high concentrations (in the range of mM) with a catalytic efficiency of 98.58 % and 99.56 % within 3 min and 5 min respectively.

Insecticidal efficacy of nanomaterials used to control mosquito, Culex quinquefasciatus Say, 1823 with special reference to their hepatotoxicity in rats.

The present study aimed to develop a novel methodology for controlling the mosquito larvae using different nanoparticles, with special reference to their effect on rats (a non-target mammalian model). The mosquito species of Culex quinquefasciatus was reared in the laboratory. Chitosan, silver nanoparticles and their combination as well as lavender (Lavandula officinalis) nanoemulsion with different concentrations were tested as biological insecticides against the mosquito larvae. Mammalian toxicity of the used nanoparticles were evaluated using 27 adult male rats, experimental rats were divided into 9 equal groups (n=3). The nanoparticles were added to the drinking water for 30 days. At the end of the study, blood and tissue samples were collected to assess the levels of the serum alanine aminotransferase and aspartate aminotransferase, different genes expression as interleukin 6 (IL-6) and IL-1ß activity. Histopathological and immunohistochemical studies using two markers (TNF-α and BAX expression) were also applied. The LC50 and LC90 were recorded for each tested nanoparticles, and also the changes of the treated mosquito larvae cuticle were assessed using the scanning electron microscopy. Green nanoemulsion (Lavandula officinalis) was more effective than metal (silver) or even biodegradable (chitosan) nanoparticles in controlling of Culex quiquefasciatus mosquito larvae, and also it proved its safety by evaluation of the mammalian hepatotoxicity of the tested nanoparticles.

Evaluation of the antiparasitic activity of the chitosan-silver nanocomposites in the treatment of experimentally infested pigeons with Pseudolynchia canariensis.

This study aimed to evaluate the efficacy of chitosan-silver nanocomposites in the treatment of experimentally infested pigeons with Pseudolynchia canariensis (P. canariensis) with evaluation of different immunological parameters before and after treatment. Therefore, fourteen birds were divided into 2 groups; group1(infested group including 12 birds) which subdivided into 6 sub-groups experimentally infested pigeons 2 pigeons each, and five group of them were treated with chitosan-silver nanocomposites and sub-group number 6 was treated with deltamethrin while, group 2 including two pigeons were kept as control negative ones. P. canariensis flies distributed under the wing and /or under the tail in infested group and these pigeons showed significantly lower RBCs and higher WBCs than that in non-infested pigeons. The cell mediated immune response against experimentally infested pigeons with P. canariensis was studied. P. canariensis infestation in pigeons have a negative impact on pigeon's blood parameters, increase TNF-α and IL-1ß cytokines levels. This study cleared out the role of P. canariensis in the induction of a case of oxidative stress indicated by high level of nitric oxide and malondialdehyde (MDA) with low antioxidant capacity in shape of reduced zinc concentration in the sera of experimentally infested pigeon. Chitosan-silver nanocomposite has a promising effect in the elimination of P. canariensis infestation in pigeons.

Anti-ESBL investigation of chitosan/silver nanocomposites against carbapenem resistant Pseudomonas aeruginosa.

In the present investigation functional chitosan/silver nanocomposites (CS/Ag NCs) were successfully synthesized and found to possess favorable antibacterial activity against extended spectrum beta-lactasame (ESBL) producing Pseudomonas aeruginosa. Powder X-ray diffraction showed that the obtained CS/Ag NCs are constituted of highly crystalline Ag nanoparticles (NPs) embedded in an amorphous CS matrix material. Transmission electron microscopy (TEM) analysis provided structural information about CS/Ag NCs, revealing the formation of spherical cluster structures constituted of Ag NPs with size ranging from 6 to 18 nm embedded in the amorphous CS matrix. The minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) of Ag NPs and CS/Ag NCs were found to inhibit the ESBL producing P. aeruginosa at 80 µg/mL (76%) and 50 µg/mL (92%), respectively. Confocal laser scanning microscope (CLSM) and scanning electron microscopy (SEM) images revealed that P. aeruginosa experienced reduced cell viability and morphological cell membrane damage at desired MIC. The in-vivo toxicity effect of Ag NPs and CS/Ag NCs suggested an increased mortality rate when Artemia franciscana were exposed for 24 h to increasing concentrations of Ag NPs and CS/Ag NCs. Anti-ESBL activity and toxicity effect of CS/Ag NCs revealed that these NCs possess promising antibacterial properties to overcome numerous communicable bacterial strains.

Combined antibacterial/tissue regeneration response in thermal burns promoted by functional chitosan/silver nanocomposites.

We report the combined antibacterial/tissue regeneration responses to thermal burns promoted by functional chitosan/silver nanocomposites (CS/nAg) with ultralow silver content (0.018wt.%, 7-30nm). Our approach allows one to produce CS/nAg nanocomposites without silver nanoparticles (nAg) agglomeration, with bactericide potency higher than 1wt.% of nAg (ca. 10nm) content and, promoting the healing process in controlled thermal burns. CS/nAg films exhibit high antibacterial activity against S. aureus and P. aeruginosa after 1.5h of incubation, demonstrating the bacterial penetration into hydrated films and their interaction with nAg. Additionally, exceptional healing of induced thermal burns was obtained by increasing myofibroblasts, collagen remodeling, and blood vessel neoformation. These factors are associated with epiderma regeneration after 7days of treatment with no nAg release. Our results corroborate the controlled synthesis of nAg embedded in CS matrix with combined antibacterial/biocompatibility properties aiming to produce functional nanocomposites with potential use in wound dressing and health care applications.