Microbial Composition in Soil and Water column of the Everglades Stormwater Treatment Areas

Hanh Nguyen, R.Bhomia, J. Meyer, M. Fujimoto, and K. R. ReddyDepartment of Soil and Water SciencesUniversity of Floridahtn25@ufl.edu

Why study microbiome in the Everglades Stormwater Treatment Areas (STAs)?

“Bacteria have sculpted the world we live in”- Dianne Newman

STAs are designed to remove nutrients

Well controlled system

Lots of abiotic, biotic, and hydrology data

available

Lack of research on microbiome

Research goals

What is the structure of microbial communities in STAs?Alpha-diversity Beta-diversity

Why do these microbes have their particular

distributions?

What are the functions of these microbes?

Methods

Sampling: 2 cells x 3 locations x 5 depths x 3 replicatesMethods:

DNA extractions16S survey: V4 primersIllumina sequencingBioinformaticapproaches: QIIME, R

waterEAV-FW1SAV-FW3

Inflow Inflow

midflow midflow

outflow outflow

Concepts

• Alpha-diversity: how many species are there (measure of the

diversity within sample)

• Beta-diversity: How one sample is different from others

(measure of the difference between samples)

• Phyla: The highest classification level

• OTU: lowest classification level (unique sequences)

Higher Alpha-diversity in Floc~RAS > pre-STA1 > pre-STA-2 > water; EAV > SAV

EAV SAV

Alpha-diversity is highly correlated with TP

Four main phyla ~80% total relative abundance in water samples

Proteobacteria

Cyanobacteria

Actinobacteria

Bacteroidetes

EAV

In-fl

owEA

V m

id-fl

owEA

V ou

t-flo

wSA

V In

-flow

SAV

mid

-flow

SAVo

ut-fl

ow

Euryacheaeota was enriched in RAS

others others

Acidobacteria, Nitrospirae, Gemmatimonadete and NC10 were enriched in pre-STA2 soils

Euryarchaeota

Microbial compositions vary most in water samples> pre-STAs but more conservative Floc and RAS

others

Water samples have distinct microbial structures at OTU level

Water

Floc & RAS

Pre-STA soil

Microbial structures of Floc and RAS are more similar, pre-STA1 is more similar to pre-STA2

PC2 (11%)

PC1 (37%)

PC3 (10%)

Floc & RAS

Pre-STA soil

N/P & C/P ratios best explain the patchiness showed in PC1

PC1 (37%)

PC2 (11%)

PC3 (10%) White ->green (150:2300)

C/P ratioPC2 (11%)

PC3 (10%)

PC1 (37%)

White ->green (9 – 150)

N/P ratio

PC1 (37%)

PC2 (11%)

PC3 (10%) White ->green (10-22 g/kg)

C/N ratio

C/N ratios does not explain the patchiness showed in PC1

TP, TC and TN does not explain the patchiness or the clustering of the samples

PC2 (11%)

PC3 (10%)

PC1 (37%)

White ->green (0.2 – 2.5 g/kg)

Total PhosphorusPC2 (11%)

PC3 (10%)

PC1 (37%)

Total carbonPC2 (11%)

PC3 (10%)

PC1 (37%)White ->green (167-500 g/kg)

The samples of same vegetation treatment are grouped together

PC2 (11%)

PC3 (10%)PC1 (37%)

Emergent Aquatic Vegetation

Submerged Aquatic Vegetation

Take home messages• Alpha-diversity: Floc and RAS > pre-STA1> pre-STA2 >

water; EAV> SAV• Beta-diversity: Microbial composition in both cells

clustered best by by N/P & C/P ratios, vegetation types, depths

• Alpha-diversity affected by TP, beta-diversity affected by N/P or C/P ratio.

• Future studies need to link microbial compositions to microbial activities

Acknowledgements

•South Florida Water Management District•UF-Wetland Biogeochemistry Laboratory•UF-SWS-Microbial Ecology Laboratory

• A. Ryan Blaustein• M. Nicole Miller

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