Impact of Zinc Oxide Nanoparticles on the Composition of Gut Microbiota in Healthy and Autism Spectrum Disorder Children.

Rongrong Yu,Luqiong Lv,Temoor Ahmed,Muchen Zhang,Bin Li,Hubiao Jiang,Guoling Zhou

doi:10.3390/ma14195488

Rongrong Yu, Luqiong Lv + Show 5 more

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https://doi.org/10.3390/ma14195488

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Abstract

Autism spectrum disorder (ASD) seriously affects children’s health, while the gut microbiome has been widely hypothesized to be involved in the regulation of ASD behavior. This study investigated and compared the number, diversity, and population structure of gut microbiota between healthy and ASD children and their susceptibility to zinc oxide nanoparticles (ZnONPs) based on the measurement of live cell number, living/dead bacterial staining test, flow cytometry observation and bacterial community analysis using 16S rRNA gene amplicon sequencing. The result of this present study revealed that ASD children not only significantly reduced the live cell number and the community diversity of gut bacteria, but also changed the gut bacterial community composition compared to the healthy children. In addition, this result revealed that ZnONPs significantly reduced the number of live bacterial cells in the gut of healthy children, but not in that of ASD children. In contrast, ZnONPs generally increased the gut bacterial community diversity in both ASD and healthy children, while a greater increase was found in ASD children than that of healthy children. Furthermore, this study successfully isolated and identified some representative nanoparticle-resistant bacteria based on the color, shape, and edge of colony as well as the 16S rDNA sequence analysis. The community of nanoparticle-resistant bacteria differed in between healthy and ASD children. Indeed, the representative strains 6-1, 6-2, 6-3 and 6-4 from healthy children were identified as Bacillus anthracis, Escherichia coli, Bacillus cereus and Escherichia coli with sequence similarity of 97.86%, 99.86%, 99.03% and 99.65%, respectively, while the representative strains 8-1, 8-2 and 8-3 from ASD children were identified as Bacillus cereus, with sequence similarities of 99.58%, 99.72% and 99.72%, respectively. Overall, this study demonstrated that ZnONPs caused a change in number, diversity, and species composition of gut bacteria, but differed in healthy and ASD children.

Highlights

Autism spectrum disorder (ASD) is a severe neurodevelopmental disorder characterized by abnormal social interactions, impaired language, and stereotypical and repetitive behaviors [1,2,3,4]
This study investigated and compared the number, diversity, and community structure of gut bacteria from the fecal samples between healthy and ASD children using both the traditional plate counting method and 16S rRNA gene amplicon sequencing approaches
The effect of zinc oxide nanoparticles (ZnONPs) on gut bacteria were determined by a count of live cell number, living and dead bacterial cell staining and flow cytometry test, while the influence of ZnONPs in the composition of gut bacteria was determined by 16S rRNA gene sequence analysis of the obtained resistant strains in combination with 16S rRNA gene amplicon sequence analysis of fecal samples from healthy and ASD children

Summary

Introduction

Autism spectrum disorder (ASD) is a severe neurodevelopmental disorder characterized by abnormal social interactions, impaired language, and stereotypical and repetitive behaviors [1,2,3,4]. The number of ASD children has significantly increased in China in the last few years [2]. ASD is becoming a public health concern with an incidence rate of about 0.26% in Chinese children. Pharmacological treatments are able to reduce the behavioral problems, they are often found to be ineffective in treating the core symptoms of ASD (i.e., social deficits). Nogay and Nahikian-Nelms reported that most of the randomized clinical trials of ASD deficits treatments have shown inconclusive results because of clinical and genetic heterogeneity [6]

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