Toxicity at the Edge of Life: A Review on Cyanobacterial Toxins from Extreme Environments.

Cyanotoxins are secondary metabolites produced by cyanobacteria, of varied chemical nature and toxic effects. Although cyanobacteria thrive in all kinds of ecosystems on Earth even under very harsh conditions, current knowledge on cyanotoxin distribution is almost restricted to freshwaters from temperate latitudes. In this review, we bring to the forefront the presence of cyanotoxins in extreme environments. Cyanotoxins have been reported especially in polar deserts (both from the Arctic and Antarctica) and alkaline lakes, but also in hot deserts, hypersaline environments, and hot springs. Cyanotoxins detected in these ecosystems include neurotoxins-anatoxin-a, anatoxin-a (S), paralytic shellfish toxins, ß-methylaminopropionic acid, N-(2-aminoethyl) glycine and 2,4-diaminobutyric acid- and hepatotoxins -cylindrospermopsins, microcystins and nodularins-with microcystins being the most frequently reported. Toxin production there has been linked to at least eleven cyanobacterial genera yet only three of these (Arthrospira, Synechococcus and Oscillatoria) have been confirmed as producers in culture. Beyond a comprehensive analysis of cyanotoxin presence in each of the extreme environments, this review also identifies the main knowledge gaps to overcome (e.g., scarcity of isolates and -omics data, among others) toward an initial assessment of ecological and human health risks in these amazing ecosystems developing at the very edge of life.

Phylogeography of cylindrospermopsin and paralytic shellfish toxin-producing nostocales cyanobacteria from mediterranean europe (Spain).

Planktonic Nostocales cyanobacteria represent a challenge for microbiological research because of the wide range of cyanotoxins that they synthesize and their invasive behavior, which is presumably enhanced by global warming. To gain insight into the phylogeography of potentially toxic Nostocales from Mediterranean Europe, 31 strains of Anabaena (Anabaena crassa, A. lemmermannii, A. mendotae, and A. planctonica), Aphanizomenon (Aphanizomenon gracile, A. ovalisporum), and Cylindrospermopsis raciborskii were isolated from 14 freshwater bodies in Spain and polyphasically analyzed for their phylogeography, cyanotoxin production, and the presence of cyanotoxin biosynthesis genes. The potent cytotoxin cylindrospermopsin (CYN) was produced by all 6 Aphanizomenon ovalisporum strains at high levels (5.7 to 9.1 µg CYN mg(-1) [dry weight]) with low variation between strains (1.5 to 3.9-fold) and a marked extracellular release (19 to 41% dissolved CYN) during exponential growth. Paralytic shellfish poisoning (PSP) neurotoxins (saxitoxin, neosaxitoxin, and decarbamoylsaxitoxin) were detected in 2 Aphanizomenon gracile strains, both containing the sxtA gene. This gene was also amplified in non-PSP toxin-producing Aphanizomenon gracile and Aphanizomenon ovalisporum. Phylogenetic analyses supported the species identification and confirmed the high similarity of Spanish Anabaena and Aphanizomenon strains with other European strains. In contrast, Cylindrospermopsis raciborskii from Spain grouped together with American strains and was clearly separate from the rest of the European strains, raising questions about the current assumptions of the phylogeography and spreading routes of C. raciborskii. The present study confirms that the nostocalean genus Aphanizomenon is a major source of CYN and PSP toxins in Europe and demonstrates the presence of the sxtA gene in CYN-producing Aphanizomenon ovalisporum.

Effect of salinity on scytonemin yield in endolithic cyanobacteria from the Atacama Desert.

Cyanobacteria inhabiting extreme environments constitute a promising source for natural products with biotechnological applications. However, they have not been studied in-depth for this purpose due to the difficulties in their isolation and mass culturing. The Atacama Desert suffers one of the highest solar irradiances that limits the presence of life on its hyperarid core to endolithic microbial communities supported by cyanobacteria as primary producers. Some of these cyanobacteria are known to produce scytonemin, a UV-screening liposoluble pigment with varied biotechnological applications in cosmetics and other industries. In this work we carried out a strain selection based on growth performance among 8 endolithic cyanobacteria of the genera Chroococcidiopsis, Gloeocapsa and Gloeocapsopsis isolated from non-saline rocks of the Atacama Desert. Then we investigated the influence of NaCl exposure on scytonemin production yield. Results in the selected strain (Chroococcidiopsis sp. UAM571) showed that rising concentrations of NaCl lead to a growth decrease while triggering a remarkable increase in the scytonemin content, reaching maximum values at 20 g L-1 of NaCl over 50-fold higher scytonemin contents than those obtained without NaCl. Altogether, these findings point out to cyanobacteria from the Atacama Desert as potentially suitable candidates for pilot-scale cultivation with biotechnological purposes, particularly to obtain scytonemin.

Morphological, molecular, and biochemical study of cyanobacteria from a eutrophic Algerian reservoir (Cheffia).

The cyanobacteria management in water bodies requires a deep knowledge of the community composition. Considering the reliable and thorough information provided by the polyphasic approach in cyanobacteria taxonomy, here we assess the cyanobacterial community structure of the Cheffia reservoir from Algeria. Cyanobacteria were identified on the basis of morphological traits and next-generation sequencing (NGS); toxins-related genes were localized in addition to the identification of toxins; temperature and nutrient level of water samples were also determined. The polyphasic approach was essential for cyanobacteria investigation; 28 genera were identified through 16S rRNA metabarcoding with the dominance of taxa from Microcystis (34.2%), Aphanizomenon (20.1%), and Planktothrix (20.0%), and morphological analysis revealed the association in this water body of five species within the genus Microcystis: M. aeruginosa, M. novacekii, M. panniformis, M. ichthyoblabe, and M. flos-aquae. The presence of mcyE genotypes was detected; moreover, HPLC-PDA and LC-ESI-MS/MS revealed the production of microcystin-LR. Results obtained in our study are very important since this ecosystem is used for water supply and irrigation; as a consequence, a good water management plan is essential.

Microbial colonizers of microplastics in an Arctic freshwater lake.

Microplastics (MPs) have been found everywhere as they are easily transported between environmental compartments. Through their transport, MPs are quickly colonized by microorganisms; this microbial community is known as the plastisphere. Here, we characterized the plastisphere of three MPs, one biodegradable (PHB) and two non-biodegradables (HDPE and LDPE), deployed in an Arctic freshwater lake for eleven days. The plastisphere was found to be complex, confirming that about a third of microbial colonizers were viable. Plastisphere was compared to microbial communities on the surrounding water and microbial mats on rocks at the bottom of the lake. Microbial mats followed by MPs showed the highest diversity regarding both prokaryotes and eukaryotes as compared to water samples; however, for fungi, MPs showed the highest diversity of the tested substrates. Significant differences on microbial assemblages on the three tested substrates were found; regarding microbial assemblages on MPs, bacterial genera found in polar environments such as Mycoplana, Erythromicrobium and Rhodoferax with species able to metabolize recalcitrant chemicals were abundant. Eukaryotic communities on MPs were characterized by the presence of ciliates of the genera Stentor, Vorticella and Uroleptus and the algae Cryptomonas, Chlamydomonas, Tetraselmis and Epipyxis. These ciliates normally feed on algae so that the complexity of these assemblages may serve to unravel trophic relationships between co-existing taxa. Regarding fungal communities on MPs, the most abundant genera were Betamyces, Cryptococcus, Arrhenia and Paranamyces. MPs, particularly HDPE, were enriched in the sulI and ermB antibiotic resistance genes (ARGs) which may raise concerns about human health-related issues as ARGs may be transferred horizontally between bacteria. This study highlights the importance of proper waste management and clean-up protocols to protect the environmental health of pristine environments such as polar regions in a context of global dissemination of MPs which may co-transport microorganisms, some of them including ARGs.

Response of Endolithic Chroococcidiopsis Strains From the Polyextreme Atacama Desert to Light Radiation.

Cyanobacteria exposed to high solar radiation make use of a series of defense mechanisms, including avoidance, antioxidant systems, and the production of photoprotective compounds such as scytonemin. Two cyanobacterial strains of the genus Chroococcidiopsis from the Atacama Desert - which has one of the highest solar radiation levels on Earth- were examined to determine their capacity to protect themselves from direct photosynthetically active (PAR) and ultraviolet radiation (UVR): the UAM813 strain, originally isolated from a cryptoendolithic microhabitat within halite (NaCl), and UAM816 strain originally isolated from a chasmoendolithic microhabitat within calcite (CaCO3). The oxidative stress induced by exposure to PAR or UVR + PAR was determined to observe their short-term response, as were the long-term scytonemin production, changes in metabolic activity and ultrastructural damage induced. Both strains showed oxidative stress to both types of light radiation. The UAM813 strain showed a lower acclimation capacity than the UAM816 strain, showing an ever-increasing accumulation of reactive oxygen species (ROS) and a smaller accumulation of scytonemin. This would appear to reflect differences in the adaptation strategies followed to meet the demands of their different microhabitats.

Unmasking the identity of toxigenic cyanobacteria driving a multi-toxin bloom by high-throughput sequencing of cyanotoxins genes and 16S rRNA metabarcoding.

Cyanobacterial harmful algal blooms (CyanoHABs) are complex communities that include coexisting toxic and non-toxic strains only distinguishable by genetic methods. This study shows a water-management oriented use of next generation sequencing (NGS) to specifically pinpoint toxigenic cyanobacteria within a bloom simultaneously containing three of the most widespread cyanotoxins (the hepatotoxins microcystins, MCs; and the neurotoxins anatoxin-a, ATX, and saxitoxins, STXs). The 2013 summer bloom in Rosarito reservoir (Spain) comprised 33 cyanobacterial OTUs based on 16S rRNA metabarcoding, 7 of which accounted for as much as 96.6% of the community. Cyanotoxins and their respective biosynthesis genes were concurrently present throughout the entire bloom event including: MCs and mcyE gene; ATX and anaF gene; and STXs and sxtI gene. NGS applied to amplicons of cyanotoxin-biosynthesis genes unveiled 6 toxigenic OTUs, comprising 3 involved in MCs production (Planktothrix agardhii and 2 Microcystis spp.), 2 in ATX production (Cuspidothrix issatschenkoi and Phormidium/Tychonema spp.) and 1 in STXs production (Aphanizomenon gracile). These toxigenic taxa were also present in 16S rRNA OTUs list and their relative abundance was positively correlated with the respective toxin concentrations. Our results point at MC-producing P. agardhii and ATX-producing C. issatschenkoi as the main contributors to the moderate toxin concentrations observed, and suggest that their distribution in Southern Europe is broader than previously thought. Our findings also stress the need for monitoring low-abundance cyanobacteria (<1% relative abundance) in cyanotoxicity studies, and provide novel data on the presence of picocyanobacteria and potentially ATX-producing benthic taxa (e.g., Phormidium) in deep thermally-stratified water bodies. This study showcases a straightforward use of amplicon metagenomics of cyanotoxin biosynthesis genes in a multi-toxin bloom thus illustrating the broad applicability of NGS for water management in risk-oriented monitoring of CyanoHABs.

Discovery of carotenoid red-shift in endolithic cyanobacteria from the Atacama Desert.

The biochemical responses of rock-inhabiting cyanobacteria towards native environmental stresses were observed in vivo in one of the Earth's most challenging extreme climatic environments. The cryptoendolithic cyanobacterial colonization, dominated by Chroococcidiopsis sp., was studied in an ignimbrite at a high altitude volcanic area in the Atacama Desert, Chile. Change in the carotenoid composition (red-shift) within a transect through the cyanobacteria dominant microbial community (average thickness ~1 mm) was unambiguously revealed in their natural endolithic microhabitat. The amount of red shifted carotenoid, observed for the first time in a natural microbial ecosystem, is depth dependent, and increased with increasing proximity to the rock surface, as proven by resonance Raman imaging and point resonance Raman profiling. It is attributed to a light-dependent change in carotenoid conjugation, associated with the light-adaptation strategy of cyanobacteria. A hypothesis is proposed for the possible role of an orange carotenoid protein (OCP) mediated non-photochemical quenching (NPQ) mechanism that influences the observed spectral behavior. Simultaneously, information about the distribution of scytonemin and phycobiliproteins was obtained. Scytonemin was detected in the uppermost cyanobacteria aggregates. A reverse signal intensity gradient of phycobiliproteins was registered, increasing with deeper positions as a response of the cyanobacterial light harvesting complex to low-light conditions.