Determination of environmental genomic material (eDNA) viability time for detection via PCR of invasive species in fish farms and quantification of marine shrimp in clear water.
aquaculture, primer, metabarcoding.
One of the challenges of the aquaculture sector is to seek more precise methodologies for identification and quantification of animals in pounds throughout the production cycle, as greater precision in these estimates is vital for achieving a more productive and sustainable creation. In order for this technique to help fish farmers and be viable on a commercial scale, it is necessary to evaluate the specificity of the primers, in order to maximize the effectiveness and minimize the risks of false diagnoses, and the determination of the time of viability of the eDNA in the water. The sensitivity of commercial primers and the viability time of eDNA from fish in a lentic environment and the quantification of shrimp in clear water ponds were evaluated. The water samples were concentrated and the DNA was extracted by the PureLink™ kit. Libraries were quantified by qPCR and sequenced on the Miseq sequencer (Illumina). The results were compared with the mitochondrial genome database. In the experiment with tilapia, DNA sequences were identified that corresponded to the species Oreochromis niloticus. In the experiment with shrimp, it was not possible to identify DNA sequences in the samples collected. The positive PCR amplification of the environmental DNA present in the water samples confirmed the efficiency of the primer and of this highly reproducible, fast and technically easy methodology.