Isolation, identification, cloning and phylogenetic analysis of the micro algae Prorocentrum minimum (Pavillard) J. Schiller 1933 Isolated from the Bay of Callao - Peru

Abstract

Harmful Algal Blooms (HAB) or Red Tides are water discolorations visible to the naked
eye due to the proliferation of one or more planktonic microorganisms such as microalgae. These
can reach high levels and produce adverse effects on human health, as well as cause damage to
other marine organisms near the coast. For this reason, there was interest in isolating, identifying
and making a phylogenetic study of the species Prorocentrum minimum found in the Bay of
Callao, Peru. Several samples of seawater were taken for their subsequent morphological
identification, and the technique of purification of the microalgae was carried out. At the same
time, a standard of Prorocentrum minimum was acquired from the Provasoli- Guillard National
Center for Culture of Marine Phytoplankton (CCMP), USA. It was carried out the growth curve
for the concentration of microalgal density, later it was carried out the DNA extraction and the
molecular phylogeny from the sequences of the ribosome LSU rRNA subunits of the species
Prorocentrum minimum. P. minimum and P gracile were identified, which were close epibenthic
species because they were more recent evolutionarily. Likewise, to confirm the presence of some
marine biotoxin absence in Prorocentrum minimum culture, it was established a quantitative
analysis of the animal's dose response in the mouse bioassay. It was possible to obtain the
purification of Prorocentrum minimum culture and to standardize the work protocol; it was
confirmed the phylogeny of microalgae species. It was not possible to obtain DSP toxin from
microalgae culture.