Banca de QUALIFICAÇÃO: PEDRO HENRIQUE SOUSA FERRO

Uma banca de QUALIFICAÇÃO de MESTRADO foi cadastrada pelo programa.
STUDENT : PEDRO HENRIQUE SOUSA FERRO
DATE: 03/12/2021
TIME: 10:00
LOCAL: Web Conferência - Google Meet - meet.google.com/xzj-fecs-daj.
TITLE:

Determination of environmental genomic material (eDNA) viability time for detection via PCR of invasive species in fish farms.

KEY WORDS:

aquaculture, primer, invasive species, metabarcoding.

PAGES: 45
BIG AREA: Ciências Agrárias
AREA: Medicina Veterinária
SUMMARY:

One of the challenges in fish production is the presence of accompanying fauna, which can develop together with the target species throughout the cultivation cycle, increasing the biomass of the pounds, the dissemination of pathogens, reducing water quality and the gain of target species weight. Environmental DNA is a complex mixture of genomic DNA from whole organisms or parts of them, present in environmental samples. The use of environmental DNA to assess distributions of aquatic communities is promising, but sampling and analysis schemes need to be adapted according to specific objectives. In order for this technique to help producers of aquatic organisms and be viable on a commercial scale, it is necessary to evaluate the specificity of the primers, to maximize the effectiveness and minimize the risk of false diagnoses, and to determine the viability time of the genomic material environment in the water. The sensitivity of a commercial primer and the viability time of environmental DNA from fish in a lentic environment (pounds) were evaluated. For the analysis of sensitivity and viability time, water from three 300 L culture tanks with a known number of individuals of Oreochromis niloticus was used. The water from the growing tanks was transferred to three other 60 L polyethylene tanks, previously cleaned and treated with nucleic acid denaturing agents. For the analysis of trace DNA, a water sample was collected from each tank approximately 10 cm below the surface and in three stages: collection on the day of water transfer (T0), 15 days after transfer (T15) and 30 days after transfer (T30). Water samples were concentrated and DNA was extracted by the PureLink™ kit. Libraries were quantified by qPCR and sequenced on the Miseq sequencer (Illumina). The results were collated with the MitoFish fish mitochondrial genome database. It was possible to identify DNA sequences in two samples from the T0 collection that corresponded to the species Oreochromis niloticus, one with 114861 and the other with 2019 sequences. In one sample, only one DNA sequence was identified, and it was not possible to identify the species. Samples from T15 and T30 collections are in laboratory processing. Positive PCR amplification of the environmental DNA present in the water samples confirmed the efficiency of the primer and this highly reproducible, fast and technically easy methodology. Traces of Oreochromis niloticus could be detected from the environmental DNA, allowing for field use to have management and tracking responses to invasive species present in the reservoir.

BANKING MEMBERS:
Presidente - 2613308 - DELANO DIAS SCHLEDER
Interna - 1046884 - ELIZABETH SCHWEGLER
Interno - 2017813 - RICARDO EVANDRO MENDES
Interna - 2408296 - SORAYA REGINA SACCO