Partitional Clustering and Differential Abundance Analysis Reveal the Community Structure of eDNA in the Los Angeles River

In this study we sought to investigate the impact of urbanization, presence of concrete river bottom, and nutrient pollution on microbial communities along the L.A. River. Six molecular markers were evaluated for identification of bacteria, plants, fungi, fish, and invertebrates in 90 samples. PCA (principal components analysis) was used with PAM (partitioning around medoids) clustering to reveal community structure and an NB (Negative binomial) model in DESeq2 was used for differential abundance analysis. PCA and factor analysis exposed the main axes of variation but were sensitive to outliers. Differential abundance of Proteobacteria was associated with soft bottom sites, and there was an apparent balance in the abundance of organisms responsible for nitrogen cycling. Nitrogen cycling was explained by differential abundance of ammonia oxidizing archaea, the complete ammonia oxidizers Nitrospira sp., nitrate reducing bacteria Marmoricola sp., and nitrogen fixing bacteria Devosia sp. which were differentially abundant at soft-bottom sites (p adj < 0.002). In contrast, differential abundance of several Cyanobacteria and other anoxygenic phototrophs was associated with the concrete bottom sites, which suggested the accumulation of excess nitrogen. The soft bottom sites tended to be represented by differential abundance of aerobes, whereas the concrete-associated species tended to be alkaliphilic, saliniphilic, calciphilic, sulfate dependent, and anaerobic. In Glendale Narrows, downstream from multiple water reclamation plants, there were differential abundance of cyanobacteria and algae, however indicator species for low nutrient environments and ammonia-abundance were also present. There was differential abundance of ascomycetes associated with Arroyo Seco and a differential abundance of Scenedesmaceae green algae and cyanobacteria in Maywood, in the analysis which compared suburban with urban river communities. The proportion of Ascomycota to Basidiomycota within the LA River differed from the expected proportion based on published worldwide freshwater and river 18S data; the shift in community structure was most likely associated with the extremes of urbanization. This study indicates that extreme urbanization can result in overrepresentation of cyanobacterial species that could cause reductions in water quality and safety.