Detection and identification of potentially toxic cyanobacteria in Polish water bodies.

Abstract

The main goal of this study was to determine the distribution of potentially toxic cyanobacteria in 39 selected Polish water bodies. From the water bodies with blooms and also from those in which blooms were not visible 87 samples were investigated. For the first time samples from ponds localized in villages with high agricultural activities were included. Lakes for which microcystin concentrations had been determined before were included as a reference for the research. The detection of cyanobacteria was conducted by microscopic observation as well as by PCR amplification of the rpoC1 gene fragment. Cyanobacteria were present in 75 out of 87 samples. The presence of potentially toxic cyanobacteria was detected by amplification of the mcyB and mcyE genes, which are involved in the biosynthesis of microcystins. Both genes were detected in 7 out of 9 blooms investigated. In the case of samples collected from water bodies in which blooms were not observed, the mcyB and mcyE genes were detected in 20 out of 36. In order to identify the cyanobacteria occurring in selected reservoirs, 16S plus ITS clone libraries were constructed. The method allowed distinguishing 18 different genotypes. After sequence analysis, cyanobacteria belonging to genera Microcystis, Planktothrix, Anabaena, Pseudanabaena, Synechocystis, Synechococcus and Woronichinia were identified. Results confirmed the usefulness of the rpoC1 and mcy genes for monitoring water bodies and detection of potentially toxic cyanobacteria. Application of molecular markers allowed detecting potentially toxic cyanobacteria before the bloom was visible. This is the first comprehensive study concerning cyanobacteria present in different types of Polish water bodies performed using molecular markers.

Polyphasic approach using multilocus analyses supports the establishment of the new aerophytic cyanobacterial genus Pycnacronema (Coleofasciculaceae, Oscillatoriales)

“…) were used for sequencing of rbc L gene and the primers rpo C3 and rpo C4 (Glowacka et al. ), for rpo C1 gene. DNA fragments were assembled into contigs using the Phred/Phrap/Consed software (Ewing and Green , Ewing et al.…”

Cyanobacteria Characteristics and Methods for Isolation and Accurate Identification of Cyanotoxins: A Review Article

“…However, depending on only toxin-encoding gene amplification may lead to unreliable results regarding the presence of toxins, as the gene cluster may not be expressed by the cell. For instance, up to 21% of P. rubescens strains with mcyS genes tested negative for microcystin production (7), while some strains only had fragments of toxin gene cluster or mutations had occurred within these genes, which made them unable to express toxin genes (25). Therefore, in addition to identifying gene-encoding toxins, detection of gene expression and toxins is equally important for accurate surveillance of toxic cyanobacteria.…”

“…However, bacteria can regulate gene transcription and toxin production, based on their environment (see section 4) through several gene expression modifications. In addition, mutations may occur within these genes or fragments, or all toxin gene clusters may be lost; consequently, toxin gene clusters cannot express proteins (25). Confirmation of toxic cyanobacteria based on only amplification of toxin-encoding genes may lead to false positive results, as PCR does not confirm cyanotoxin gene expression.…”

Influence of cyanobacteria, mixotrophic flagellates, and virioplankton size fraction on transcription of microcystin synthesis genes in the toxic cyanobacterium Microcystis aeruginosa

“…In aquatic environments, several strains and species of cyanobacteria generally co‐occur (Al‐Tebrineh et al., ; Glowacka, Szefel‐Markowska, Waleron, Lojkowska, & Waleron, ; Via‐Ordorika et al., ; Zwart et al., ), and studies suggest that microcystin producers may have an advantage in the competition for micronutrients such as iron (Lukač & Aegerter, ; Utkilen & Gjølme, ). Interaction of cyanobacteria with other members of the microbial community by means of secondary metabolites has been described and is an ongoing and expanding field of study (Kaplan, Weiss, & Sukenik, ; Kaplan et al., ).…”

“…In addition, colony formation as a response to grazing danger has been observed but is not restricted to toxic strains (Fulton & Paerl, 1987a;Yang, Kong, Shi, & Cao, 2006). In aquatic environments, several strains and species of cyanobacteria generally co-occur (Al-Tebrineh et al, 2012;Glowacka, Szefel-Markowska, Waleron, Lojkowska, & Waleron, 2011;Via-Ordorika et al, 2004;Zwart et al, 2005), and studies suggest that microcystin producers may have an advantage in the competition for micronutrients such as iron (Lukač & Aegerter, 1993;Utkilen & Gjølme, 1995).…”