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The excessive proliferation of cyanobacteria in surface waters is a widespread problem worldwide, leading to the contamination of drinking water sources. Short- and long-term solutions for managing cyanobacterial blooms are needed for drinking water supplies. The goal of this research was to investigate the cyanobacteria community composition using shotgun metagenomics in a short term, in situ mesocosm experiment of two lakes following their coagulation with ferric sulfate (Fe 2 (SO 4 ) 3 ) as an option for source water treatment. Among the nutrient paramenters, dissolved nitrogen was related to Microcystis in both Missisquoi Bay and Petit Lac St. François, while the presence of Synechococcus was related to total nitrogen, dissolved nitrogen, dissolved organic carbon, and dissolved phosphorus. Results from the shotgun metagenomic sequencing showed that Dolichospermum and Microcystis were the dominant genera in all of the mesocosms in the beginning of the sampling period in Missisquoi Bay and Petit Lac St. François, respectively. Potentially toxigenic genera such as Microcystis were correlated with intracellular microcystin concentrations. A principal component analysis showed that there was a change of the cyanobacterial composition at the genus level in the mesocosms after two days, which varied across the studied sites and sampling time. The cyanobacterial community richness and diversity did not change significantly after its coagulation by Fe 2 (SO 4 ) 3 in all of the mesocosms at either site. The use of Fe 2 (SO 4 ) 3 for an onsite source water treatment should consider its impact on cyanobacterial community structure and the reduction of toxin concentrations.

The Lake in Central Park (LCP) and Prospect Park Lake (PPL) in New York City (NYC), USA, are lakes within two of the most visited parks in the USA. Five years of nearshore sampling of these systems revealed extremely elevated levels of cyanobacteria and the toxin, microcystin, with microcystin levels averaging 920 µg L−1 and chlorophyll a from cyanobacterial (cyano-chla) populations averaging 1.0 × 105 µg cyano-chla L−1. Both lakes displayed elevated levels of orthophosphate (DIP) relative to dissolved inorganic nitrogen (DIN) during summer months when DIN:DIP ratios were < 1. Nutrient addition and dilution experiments revealed that N consistently limited cyanobacterial populations but that green algae were rarely nutrient limited. Experimental additions of public drinking water that is rich in P and, to a lesser extent N, to lake water significantly enhanced cyanobacterial growth rates in experiments during which N additions also yielded growth enhancement. Collectively, this study demonstrates that the extreme microcystin levels during blooms in these highly trafficked lakes represent a potential human and animal health threat and that supplementation of these artificial lakes with public drinking water to maintain water levels during summer may promote the intensity and N limitation of blooms.

Gaining a better understanding of the interactive effect of projected atmospheric CO 2 level increase and the Earth's rising temperature on plant chemistry (nutritional and defensive characteristics) of transgenic crops is essential when attempting to forecast the responses of target and non-target insects to climate change. In this study, effects of carbon dioxide (CO 2 ; elevated versus ambient), temperature (T; high versus low), and their interactions on leaf nitrogen content (N%) and C:N ratio of transgenic Bt rice and its non- Bt isoline grown under low- and high-N fertilizer were systematically analyzed together with the resulting insect population dynamics of a non-target planthopper Nilaparvata lugens (Stâl) in open-top-chamber experiments. The results indicated that under low-N treatment, elevated CO 2 at low T (i.e., eCO 2 ) (compared to ambient CO 2 at low T, i.e., CK) significantly decreased N% and Bt -toxin content and significantly increased C:N ratio in leaf sheath and leaf of Bt rice, especially during the tillering stage, whereas inverse effects of high T were shown on the plant chemistry of Bt rice, especially during heading stage. The combination of elevated CO 2 and high T (i.e., Combined) (in contrast to CK) significantly increased N% and decreased C:N ratio in leaf sheath of Bt rice during the heading stage under low-N fertilizer, while significantly decreased N% and increased C:N ratio in leaf of Bt rice during the tillering stage, regardless of fertilizer-N level, and significantly increased Bt -toxin content in leaf sheath and leaf during the tillering stage under both low- and high-N. Moreover, no discernable relationships between Bt -toxin content and N% or leaf C:N ratio were observed at any CO 2 or N levels evaluated. Furthermore, transgenic treatment, temperature and fertilizer-N level interactions, and CO 2 and fertilizer-N level interactions all significantly affected the population dynamics of N. lugens. Specifically, high-N significantly enhanced the population dynamics of N. lugens fed on non-Bt rice grown under eTemp and Bt cultivar significantly reduced the population dynamics of N. lugens under eCO 2 regardless of N fertilizer levels. The study demonstrates that the planting of transgenic Bt rice would not increase the risk of increased N. lugens severity under the combined condition of elevated CO 2 and increased temperature, particularly under moderate level of N fertility., Competing Interests: The authors declare no conflict of interest.

Lake Lesser Prespa and its adjacent pond, Vromolimni in Greece, is a shallow freshwater system and a highly protected area hosting an exceptional biodiversity. The occurrence of microcystins (MCs) producing cyanobacterial blooms in these waters during recent years can be harmful to the wildlife. We tested the hypothesis that both cyanobacterial biomass and MCs are strongly influenced by nutrients (eutrophication) and warming (climate change). Lake and pond water was collected from two sites in each water body in 2013 and incubated at three temperatures (20 °C, 25 °C, 30 °C) with or without additional nutrients (nitrogen +N, phosphorus +P and both +N and +P). Based on both biovolume and chlorophyll-a concentrations, cyanobacteria in water from Lesser Prespa were promoted primarily by combined N and P additions and to a lesser extent by N alone. Warming seemed to yield more cyanobacteria biomass in these treatments. In water from Vromolimni, both N alone and N+P additions increased cyanobacteria and a warming effect was hardly discernible. MC concentrations were strongly increased by N and N+P additions in water from all four sites, which also promoted the more toxic variant MC-LR. Hence, both water bodies seem particularly vulnerable to further N-loading enhancing MC related risks., Competing Interests: The authors declare no conflict of interest.

Scorpion stings are the main cause of human envenomation in Brazil and, for the treatment of victims, the World Health Organization (WHO) recommends the use of antivenoms. The first step to achieve effective antivenom is to use a good quality venom pool and to evaluate it, with LD 50 determination as the most accepted procedure. It is, however, time-consuming and requires advanced technical training. Further, there are significant ethical concerns regarding the number of animals required for testing. Hence, we investigated the correspondence between LD 50 results, in vitro assays, and a strong correlation with proteolytic activity levels was observed, showing, remarkably, that proteases are potential toxicity markers for Tityus serrulatus venom. The comparison of reversed-phase chromatographic profiles also has a potential application in venoms' quality control, as there were fewer neurotoxins detected in the venom with high LD 50 value. These results were confirmed by mass spectrometry analysis. Therefore, these methods could precede the LD 50 assay to evaluate the venom excellence by discriminating-and discarding-poor-quality batches, and, consequently, with a positive impact on the number of animals used. Notably, proposed assays are fast and inexpensive, being technically and economically feasible in Tityus serrulatus venom quality control to produce effective antivenoms., Competing Interests: The authors declare that they have no competing financial interests.

Future sustainability of freshwater resources is seriously threatened due to the presence  of  harmful  cyanobacterial  blooms,  and  yet,  the  number,  extent,  and  distribution  of  most  cyanobacterial toxins-including "emerging" toxins and other bioactive compounds-are poorly  understood.  We  measured  15  cyanobacterial  compounds-including  four  microcystins  (MC),  saxitoxin (SXT), cylindrospermopsin (CYL), anatoxin-a (ATX) and homo-anatoxin-a (hATX), two  anabaenopeptins (Apt), three cyanopeptolins (Cpt), microginin (Mgn), and nodularin (NOD)-in  six freshwater lakes that regularly experience noxious cHABs. MC, a human liver toxin, was present  in all six lakes and was detected in 80% of all samples. Similarly, Apt, Cpt, and Mgn were detected  in all lakes in roughly 86%, 50%, and 35% of all samples, respectively. Despite being a notable  brackish  water  toxin,  NOD  was  detected  in  the  two  shallowest  lakes-Wingra  (4.3  m)  and  Koshkonong (2.1 m). All compounds were highly variable temporally, and spatially. Metabolite  profiles were significantly different between lakes suggesting lake characteristics influenced the  cyanobacterial community and/or metabolite production. Understanding how cyanobacterial toxins  are  distributed  across  eutrophic  lakes  may  shed  light  onto  the  ecological  function  of  these  metabolites, provide valuable information for their remediation and removal, and aid in the  protection of public health.

The microcystin biodegradation potential of a natural bacterial community coexisting with a toxic cyanobacterial bloom was investigated in a water reservoir from central Spain. The biodegradation capacity was confirmed in all samples during the bloom and an increase of mlr A gene copies was found with increasing microcystin concentrations. Among the 24 microcystin degrading strains isolated from the bacterial community, only 28% showed presence of mlr A gene, strongly supporting the existence and abundance of alternative microcystin degradation pathways in nature. In vitro degradation assays with both mlr ⁺ and mlr - bacterial genotypes (with presence and absence of the complete mlr gene cluster, respectively) were performed with four isolated strains ( Sphingopyxis sp. IM-1, IM-2 and IM-3; Paucibacter toxinivorans IM-4) and two bacterial degraders from the culture collection ( Sphingosinicella microcystinivorans Y2; Paucibacter toxinivorans 2C20). Differences in microcystin degradation efficiencies between genotypes were found under different total organic carbon and total nitrogen concentrations. While mlr ⁺ strains significantly improved microcystin degradation rates when exposed to other carbon and nitrogen sources, mlr - strains showed lower degradation efficiencies. This suggests that the presence of alternative carbon and nitrogen sources possibly competes with microcystins and impairs putative non- mlr microcystin degradation pathways. Considering the abundance of the mlr - bacterial population and the increasing frequency of eutrophic conditions in aquatic systems, further research on the diversity of this population and the characterization and conditions affecting non- mlr degradation pathways deserves special attention., Competing Interests: The authors declare no conflict of interest.

As the biosynthesis of cylindrospermopsin (CYN) is assumed to depend on nitrogen availability, this study investigated the impact of nitrogen availability on intra- and extracellular CYN and deoxy-CYN (D-CYN) contents in three Aphanizomenon strains from temperate waters. Nitrogen deficient (-N) cultures showed a prolonged growth phase and intracellular toxin accumulation by a factor of 2-6. In contrast, cultures with additional nitrate supply (+N) did not accumulate CYN within the cells. Instead, the maximum conceivable CYN release estimated for dead cells (identified by SYTOX Green staining) was much lower than the concentrations of dissolved CYN actually observed, suggesting these cultures actively release CYN from intact cells. Furthermore, we found remarkably altered proportions of CYN to D-CYN: as batch cultures grew, the proportion of D-CYN increased by up to 40% in +N medium, whereas D-CYN remained constant or decreased slightly in -N medium. Since +N cultures showed similar toxin patterns as -P cultures with increased extracellular CYNs and higher proportion of D-CYN we conclude that nitrogen limitation may affect the way the cells economize resources, especially the yield from phosphorus pools, and that this has an impact on CYN production and release. For water management, these result imply that nutrient availability not only determines the abundance of potentially CYN-producing cyanobacteria, but also the amount of extracellular CYNs (challenging drinking-water treatment) as well as the ratio of D-CYN to CYN (affecting toxicity).