Year Established: 2020 Start Date: 2020-03-01 End Date: 2021-02-28
Total Federal Funds: $22,030 Total Non-Federal Funds: $45,372
Principal Investigators: Dr. Babur Mirza
Abstract: Cyanobacteria-dominated harmful algal blooms (cyanoHABs) are increasing in occurrence worldwide. The wide distribution of cyanobacteria in aquatic environments leads to the risk of water contamination by cyanotoxins, which generate environmental and public health issues. Measurements of cyanobacterial cell densities or pigment contents can be used for early detection of cellular growth and bloom monitoring, but this approach is not accurate enough to identify or predict actual cyanobacterial risk. Because not all aquatic blooms are composed of cyanobacteria, eukaryotic phytoplankton such as diatoms and green algae can also cause algal blooms which are generally non-toxigenic. Traditionally, cyanobacterial species within cyanoHABs have been identified using various morphological or cellular characteristics such as pigmentation, cellular arrangements, presence of specialized cells and gas vacuoles. However, many of these differential cell characters can vary under different growth conditions and even some characteristics can be lost during cultivation. Furthermore, the correct microscopic identification of cyanobacteria heavily relies on the expertise of the individuals which can easily vary from one person to another. Considering these limitations in the microscopic based identification of toxigenic cyanobacteria, we propose to use Next Generation, or Next-Gen DNA sequencing to assess the diversity of cyanobacteria and three key cyanotoxin; microcystins, saxitoxins, and cylindrospermopsin, associated genes within a microbial impaired water ecosystem, the Little Sac Watershed. Direct measurement of cyanotoxins using various analytical methods (enzyme-linked immunosorbent assay, LC/MS, GC/MS) can be a useful approach to assess the toxigenic potential of a cyanoHABs. However, these analytical methods are expensive, time-consuming, and can be only detected high concentrations of cyanotoxins. This approach becomes costprohibitive when it comes to testing a water sample for the presence of many different cyanotoxins. In the proposed study, we will use a combination of Next-Gen DNA sequencing along with quantitative PCR of three specific cyanotoxins genes associated with the presence of three most commonly reported cyanotoxins (microcystins, saxitoxins, and cylindrospermopsin toxins). The goal of this study is to assess the diversity and relative abundance of different cyanobacteria species, and whether these species are capable of producing commonly reported cyanotoxin within the Little Sac Watershed ecosystem. This project will also provide research experience and training opportunities for three students. These students will learn the DNA extraction, DNA library preparation, qPCR, and Next-Gen DNA sequencing and data analysis.