The presence of fungi in the indoor environment is associated with allergies and other respiratory symptoms. The aim of this study was to use sequencing and molecular methods, including next-generation sequencing (NGS) approaches, to explore the bacterial and fungal communities and their abundance in the indoor environment of houses (n = 20) with visible \"moldy\" (HVM) and nonvisible \"non-moldy\" (HNM) in Memphis, TN, USA. Dust samples were collected from air vents and ground surfaces, and the total DNA was analyzed for bacteria and fungi by amplifying 16S rRNA and ITS genes on the Illumina Miseq. Results indicated that Leptosphaerulina was the most abundant fungal genus present in the air vent and ground samples from HNM and HVM. At the same time, the most abundant bacterial genera in the air vent and ground samples were Propionibacterium and Streptococcus. The fungi community diversity was significantly different in the air vent samples. The abundance of fungal species known to be associated with respiratory diseases in indoor dust samples was similar, regardless of the visibility of fungi in the houses. The existence of fungi associated with respiratory symptoms was compared with several parameters like dust particulate matter (PM), CO2 level, temperature, and humidity. Most of these parameters are either positively or negatively correlated with the existence of fungi associated with respiratory diseases; however, none of these correlations were significant at p = 0.05. Our results indicate that implementing molecular methods for detecting indoor fungi may strengthen common exposure and risk assessment practices.

The application of graphene oxide (GO) has attracted increasing concerns in the past decade regarding its environmental impacts, except for the impact of GO on a metal-contaminated soil system, due to its special properties. In the present work, the effects of GO on the migration and transformation of heavy metals and soil bacterial communities in Cd-contaminant soil were systematically evaluated. Soil samples were exposed to different doses of GO (0, 1, and 2 g kg-1) over 60 days. The Community Bureau of Reference (BCR) sequential extraction procedure was used to reflect the interaction between GO and Cd. Several microbial parameters, including enzyme activities and bacterial community structure, were measured to determine the impacts of GO on polluted soil microbial communities. It was shown that Cd was immobilized by GO throughout the entire exposure period. Interestingly, the structure of the bacterial community changed. The relative abundance of the major bacterial phyla (e.g., Acidobacteria and Actinobacteria) increased, which was possibly attributed to the reduced toxicity of Cd in the presence of GO. However, GO exerted an adverse influence on the relative abundance of some phyla (e.g., WD272 and TM6). The diversity of bacterial communities was slightly restricted. The functional bacteria related to carbon and the nitrogen cycling were also affected, which, consequently, may influence the nutrient cycling in soil.

High-throughput sequencing of the 16S rRNA gene has considerably helped revealing the essential role of bacteria living on insect cuticles in the ecophysiology and behaviour of their hosts. However, our understanding of host-cuticular microbiota feedbacks remains hampered by the difficulties of working with low bacterial DNA quantities as with individual insect cuticle samples, which are more prone to molecular biases and contaminations. Herein, we conducted a methodological benchmark on the cuticular bacterial loads retrieved from two Neotropical ant species of different body size and ecology: Atta cephalotes (~15 mm) and Pseudomyrmex penetrator (~5 mm). We evaluated the richness and composition of the cuticular microbiota, as well as the amount of biases and contamination produced by four DNA extraction protocols. We also addressed how bacterial community characteristics would be affected by the number of individuals or individual body size used for DNA extraction. Most extraction methods yielded similar results in terms of bacterial diversity and composition for A. cephalotes (~15 mm). In contrast, greater amounts of artefactual sequences and contaminations, as well as noticeable differences in bacterial community characteristics were observed between extraction methods for P. penetrator (~5 mm). We also found that large (~15 mm) and small (~5 mm) A. cephalotes individuals harbour different bacterial communities. Our benchmark suggests that cuticular microbiota of single individual insects can be reliably retrieved provided that blank controls, appropriate data cleaning, and individual body size and functional role within insect society are considered in the experiment.