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\begin{document}

\title{DNA metabarcoding of mock communities highlights potential biases when
assessing Neotropical fish diversity}



\author[1]{Heron Hilário}%
\author[2]{Izabela Mendes}%
\author[3]{Naiara Sales}%
\author[1]{Daniel Carvalho}%
\affil[1]{Pontifícia Universidade Católica de Minas Gerais}%
\affil[2]{UFMG}%
\affil[3]{Universidade de Lisboa}%


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  \date{May 18, 2024}


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\begin{abstract}
Despite the increasing popularity of DNA metabarcoding in the assessment
of aquatic ecosystems using fish eDNA or ichthyoplankton, challenges
have hampered its broader application in the Neotropical freshwaters.
Using five mock communities composed of fish species from two
Neotropical river basins, we evaluated the influence of DNA
concentration and choice of mitochondrial 12S molecular markers (MiFish,
NeoFish and Teleo) on species detection and Relative Read Abundance
(RRA) using DNA metabarcoding. Of the three 12S markers analysed, only
MiFish detected all species from all mock communities. The performance
of a taxonomy-free approach using ASV/MOTUs was not as precise as
assigning DNA reads to species using a curated 12S library that includes
approximately 100 fish species, since more than one ASV/MOTU was
observed for the same specimen. Thus, here we showcase the importance of
a custom reference database to allow precise assignment of Neotropical
fish species in metabarcoding studies and that the RRA is dependent on
community composition, marker and DNA concentration. We highlight the
importance of controlled experiments using known species communities
before large investments are made in assessing biodiversity using
non-invasive methods that apply DNA metabarcoding.%
\end{abstract}%



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