Nanocomposite of Half-Fin Anchovy Hydrolysates/Zinc Oxide Nanoparticles Exhibits Actual Non-Toxicity and Regulates Intestinal Microbiota, Short-Chain Fatty Acids Production and Oxidative Status in Mice

The nanocomposite of half-fin anchovy hydrolysates (HAHp) and zinc oxide nanoparticles (ZnO NPs) (named as HAHp(3.0)/ZnO NPs) demonstrated increased antibacterial activity compared to either HAHp(3.0) or ZnO NPs as per our previous studies. Also, reactive oxygen species (ROS) formation was detected in Escherichia coli cells after treatment with HAHp(3.0)/ZnO NPs. The aim of the present study was to evaluate the acute toxicity of this nanocomposite and to investigate its effect on intestinal microbiota composition, short-chain fatty acids (SCFAs) production, and oxidative status in healthy mice. The limit test studies show that this nanoparticle is non-toxic at the doses tested. The administration of HAHp(3.0)/ZnO NPs, daily dose of 1.0 g/kg body weight for 14 days, increased the number of goblet cells in jejunum. High-throughput 16S ribosomal RNA gene sequencing of fecal samples revealed that HAHp(3.0)/ZnO NPs increased Firmicutes and reduced Bacteriodetes abundances in female mice. Furthermore, the microbiota for probiotic-type bacteria, including Lactobacillus and Bifidobacterium, and SCFAs-producing bacteria in the Clostridia class, e.g., Lachnospiraceae_unclassified and Lachnospiraceae_UCG-001, were enriched in the feces of female mice. Increases of SCFAs, especially statistically increased propionic and butyric acids, indicated the up-regulated anti-inflammatory activity of HAHp(3.0)/ZnO NPs. Additionally, some positive responses in liver, like markedly increased glutathione and decreased malonaldehyde contents, indicated the improved oxidative status. Therefore, our results suggest that HAHp(3.0)/ZnO NPs could have potential applications as a safe regulator of intestinal microbiota or also can be used as an antioxidant used in food products.