Presentation Title

Detection of Cyanobacteria and Their Toxins Using PCR for Safe Algae-based Feed Operations

Faculty Mentor

Shelton Murinda

Start Date

18-11-2017 2:15 PM

End Date

18-11-2017 3:15 PM

Location

BSC-Ursa Minor 85

Session

Poster 3

Type of Presentation

Poster

Subject Area

biological_agricultural_sciences

Abstract

Toxic cyanobacteria are photosynthetic algae that are commonly observed in nitrogen rich water supplies, as a contaminant. Algae have the ability to convert nitrogen into animal feed by removing dissolved organic matter from nutrient rich sources such as manure and waste water. Our goal is to develop molecular techniques to determine which classes of cyanobacteria are toxic, and to detect and control their production before the growth becomes toxic. This experiment is part of a larger sustainability project focused on converting livestock manure into safe algae-based feed. The algae are produced in paddle-wheeled model ponds that can potentially become contaminated by toxic strains. Multiple species of pure cyanobacteria were cultured and their DNA was isolated and purified. The 16S rRNA and rpoC1 gene sequences were targeted for their universal detection of cyanobacteria using PCR. To determine which cyanobacteria species are capable of producing toxins, five gene clusters in the toxin synthesis pathways were targeted: mcy (microcystin), nda (nodularin), cyr (cylindrospermopsin), ana (anatoxin-a) and sxt (saxitoxin). The PCR-amplified fragments were separated using gel electrophoresis, and sequenced. These results enabled us to identify the quality control strains with high sensitivity and specificity, and distinguish cyanobacteria from non-cyanobacteria. Analysis of the quality control strains determined that some have the potential to produce mcy, and ana. The toxin production was detected using ELISA kits. The correlation between the presence of toxin genes and toxin production, as well as their quantitation with qPCR is an area of interest. This protocol will regularly detect and identify the presence of cyanobacteria species that can produce toxins in algae producing ponds, and allow for their control in algae-based livestock feeds.

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Nov 18th, 2:15 PM Nov 18th, 3:15 PM

Detection of Cyanobacteria and Their Toxins Using PCR for Safe Algae-based Feed Operations

BSC-Ursa Minor 85

Toxic cyanobacteria are photosynthetic algae that are commonly observed in nitrogen rich water supplies, as a contaminant. Algae have the ability to convert nitrogen into animal feed by removing dissolved organic matter from nutrient rich sources such as manure and waste water. Our goal is to develop molecular techniques to determine which classes of cyanobacteria are toxic, and to detect and control their production before the growth becomes toxic. This experiment is part of a larger sustainability project focused on converting livestock manure into safe algae-based feed. The algae are produced in paddle-wheeled model ponds that can potentially become contaminated by toxic strains. Multiple species of pure cyanobacteria were cultured and their DNA was isolated and purified. The 16S rRNA and rpoC1 gene sequences were targeted for their universal detection of cyanobacteria using PCR. To determine which cyanobacteria species are capable of producing toxins, five gene clusters in the toxin synthesis pathways were targeted: mcy (microcystin), nda (nodularin), cyr (cylindrospermopsin), ana (anatoxin-a) and sxt (saxitoxin). The PCR-amplified fragments were separated using gel electrophoresis, and sequenced. These results enabled us to identify the quality control strains with high sensitivity and specificity, and distinguish cyanobacteria from non-cyanobacteria. Analysis of the quality control strains determined that some have the potential to produce mcy, and ana. The toxin production was detected using ELISA kits. The correlation between the presence of toxin genes and toxin production, as well as their quantitation with qPCR is an area of interest. This protocol will regularly detect and identify the presence of cyanobacteria species that can produce toxins in algae producing ponds, and allow for their control in algae-based livestock feeds.