Comparitive Metagenomics of Toxic Freshwater Cyanobacteria Bloom Communities on Two Continents

Abstract Toxic cyanobacterial blooms have persisted in freshwater systems around the world for centuries and appear to be globally increasing in frequency and severity. Toxins produced by bloom-associated cyanobacteria can have drastic impacts on the ecosystem and surrounding communities, and bloom biomass can disrupt aquatic food webs and act as a driver for hypoxia. Little is currently known regarding the genomic content of the Microcystis strains that form blooms or the companion heterotrophic community associated with bloom events. To address these issues, we examined the bloom-associated microbial communities in single samples from Lake Erie (North America), Lake Tai (Taihu, China), and Grand Lakes St. Marys (OH, USA) using comparative metagenomics. Together the Cyanobacteria and Proteobacteriacomprised \u3e90% of each bloom bacterial community sample, although the dominant phylum varied between systems. Relative to the existing Microcystis aeruginosa NIES 843 genome, sequences from Lake Erie and Taihu revealed a number of metagenomic islands that were absent in the environmental samples. Moreover, despite variation in the phylogenetic assignments of bloom-associated organisms, the functional potential of bloom members remained relatively constant between systems. This pattern was particularly noticeable in the genomic contribution of nitrogen assimilation genes. In Taihu, the genetic elements associated with the assimilation and metabolism of nitrogen were predominantly associated withProteobacteria, while these functions in the North American lakes were primarily contributed to by the Cyanobacteria. Our observations build on an emerging body of metagenomic surveys describing the functional potential of microbial communities as more highly conserved than that of their phylogenetic makeup within natural systems

Insight Into the Molecular Mechanisms for Microcystin Biodegradation in Lake Erie and Lake Taihu

Microcystins are potent hepatotoxins that are frequently detected in fresh water lakes plagued by toxic cyanobacteria. Microbial biodegradation has been referred to as the most important avenue for removal of microcystin from aquatic environments. The biochemical pathway most commonly associated with the degradation of microcystin is encoded by the mlrABCD (mlr) cassette. The ecological significance of this pathway remains unclear as no studies have examined the expression of these genes in natural environments. Six metatranscriptomes were generated from microcystin-producing Microcystis blooms and analyzed to assess the activity of this pathway in environmental samples. Seventy-eight samples were collected from Lake Erie, United States/Canada and Lake Tai (Taihu), China, and screened for the presence of mlr gene transcripts. Read mapping to the mlrcassette indicated transcripts for these genes were absent, with only 77 of the collective 3.7 billion reads mapping to any part of the mlr cassette. Analysis of the assembled metatranscriptomes supported this, with only distantly related sequences identified as mlrABC-like. These observations were made despite the presence of microcystin and over 500,000 reads mapping to the mcy cassette for microcystin production. Glutathione S-transferases and alkaline proteases have been previously hypothesized to be alternative pathways for microcystin biodegradation, and expression of these genes was detected across space and time in both lakes. While the activity of these alternative pathways needs to be experimentally confirmed, they may be individually or collectively more important than mlr genes in the natural environment. Importantly, the lack of mlr expression could indicate microcystin biodegradation was not occurring in the analyzed samples. This study raises interesting questions about the ubiquity, specificity and locality of microcystin biodegradation, and highlights the need for the characterization of relevant mechanisms in natural communities to understand the fate of microcystin in the environment and risk to public health

Molecular prediction of lytic vs lysogenic states for Microcystis phage: Metatranscriptomic evidence of lysogeny during large bloom events

Microcystis aeruginosa is a freshwater bloom-forming cyanobacterium capable of producing the potent hepatotoxin, microcystin. Despite increased interest in this organism, little is known about the viruses that infect it and drive nutrient mobilization and transfer of genetic material between organisms. The genomic complement of sequenced phage suggests these viruses are capable of integrating into the host genome, though this activity has not been observed in the laboratory. While analyzing RNA-sequence data obtained from Microcystis blooms in Lake Tai (Taihu, China), we observed that a series of lysogeny-associated genes were highly expressed when genes involved in lytic infection were down-regulated. This pattern was consistent, though not always statistically significant, across multiple spatial and temporally distinct samples. For example, samples from Lake Tai (2014) showed a predominance of lytic virus activity from late July through October, while genes associated with lysogeny were strongly expressed in the early months (June–July) and toward the end of bloom season (October). Analyses of whole phage genome expression shows that transcription patterns are shared across sampling locations and that genes consistently clustered by co-expression into lytic and lysogenic groups. Expression of lytic-cycle associated genes was positively correlated to total dissolved nitrogen, ammonium concentration, and salinity. Lysogeny-associated gene expression was positively correlated with pH and total dissolved phosphorous. Our results suggest that lysogeny may be prevalent in Microcystis blooms and support the hypothesis that environmental conditions drive switching between temperate and lytic life cycles during bloom proliferation

Circumstellar CO in metal-poor stellar winds: the highly irradiated globular cluster star 47 Tucanae V3

We report the first detection of circumstellar CO in a globular cluster. Observations with ALMA have detected the CO J=3-2 and SiO v=1 J=8-7 transitions at 345 and 344 GHz, respectively, around V3 in 47 Tucanae (NGC 104; [Fe/H] = -0.72 dex), a star on the asymptotic giant branch. The CO line is detected at 7 sigma at a rest velocity v_LSR = -40.6 km/s and expansion velocity of 3.2 +/- ~0.4 km/s. The brighter, asymmetric SiO line may indicate a circumstellar maser. The stellar wind is slow compared to similar Galactic stars, but the dust opacity remains similar to Galactic comparisons. We suggest that the mass-loss rate is set by the levitation of material into the circumstellar environment by pulsations, but that the terminal wind-expansion velocity is determined by radiation pressure on the dust: a pulsation-enhanced dust-driven wind. We suggest the metal-poor nature of the star decreases the grain size, slowing the wind and increasing its density and opacity. Metallic alloys at high altitudes above the photosphere could also provide an opacity increase. The CO line is weaker than expected from Galactic AGB stars, but its strength confirms a model that includes CO dissociation by the strong interstellar radiation field present inside globular clusters.Comment: 5 pages, accepted MNRAS Letter

Microcystin aids in cold temperature acclimation: Differences between a toxic Microcystis wildtype and non-toxic mutant

For Microcystis aeruginosa PCC 7806, temperature decreases from 26 °C to 19 °C double the microcystin quota per cell during growth in continuous culture. Here we tested whether this increase in microcystin provided M. aeruginosa PCC 7806 with a fitness advantage during colder-temperature growth by comparing cell concentration, cellular physiology, reactive oxygen species damage, and the transcriptomics-inferred metabolism to a non-toxigenic mutant strain M. aeruginosa PCC 7806 ΔmcyB. Photo-physiological data combined with transcriptomic data revealed metabolic changes in the mutant strain during growth at 19 °C, which included increased electron sinks and non-photochemical quenching. Increased gene expression was observed for a glutathione-dependent peroxiredoxin during cold treatment, suggesting compensatory mechanisms to defend against reactive oxygen species are employed in the absence of microcystin in the mutant. Our observations highlight the potential selective advantages of a longer-term defensive strategy in management of oxidative stress (i.e., making microcystin) vs the shorter-term proactive strategy of producing cellular components to actively dissipate or degrade oxidative stress agents

Bohm and Einstein-Sasaki Metrics, Black Holes and Cosmological Event Horizons

We study physical applications of the Bohm metrics, which are infinite sequences of inhomogeneous Einstein metrics on spheres and products of spheres of dimension 5 <= d <= 9. We prove that all the Bohm metrics on S^3 x S^2 and S^3 x S^3 have negative eigenvalue modes of the Lichnerowicz operator and by numerical methods we establish that Bohm metrics on S^5 have negative eigenvalues too. We argue that all the Bohm metrics will have negative modes. These results imply that higher-dimensional black-hole spacetimes where the Bohm metric replaces the usual round sphere metric are classically unstable. We also show that the stability criterion for Freund-Rubin solutions is the same as for black-hole stability, and hence such solutions using Bohm metrics will also be unstable. We consider possible endpoints of the instabilities, and show that all Einstein-Sasaki manifolds give stable solutions. We show how Wick rotation of Bohm metrics gives spacetimes that provide counterexamples to a strict form of the Cosmic Baldness conjecture, but they are still consistent with the intuition behind the cosmic No-Hair conjectures. We show how the Lorentzian metrics may be created ``from nothing'' in a no-boundary setting. We argue that Lorentzian Bohm metrics are unstable to decay to de Sitter spacetime. We also argue that noncompact versions of the Bohm metrics have infinitely many negative Lichernowicz modes, and we conjecture a general relation between Lichnerowicz eigenvalues and non-uniqueness of the Dirichlet problem for Einstein's equations.Comment: 53 pages, 11 figure

Joint analysis of stressors and ecosystem services to enhance restoration effectiveness

With increasing pressure placed on natural systems by growing human populations, both scientists and resource managers need a better understanding of the relationships between cumulative stress from human activities and valued ecosystem services. Societies often seek to mitigate threats to these services through large-scale, costly restoration projects, such as the over one billion dollar Great Lakes Restoration Initiative currently underway. To help inform these efforts, we merged high-resolution spatial analyses of environmental stressors with mapping of ecosystem services for all five Great Lakes. Cumulative ecosystem stress is highest in near-shore habitats, but also extends offshore in Lakes Erie, Ontario, and Michigan. Variation in cumulative stress is driven largely by spatial concordance among multiple stressors, indicating the importance of considering all stressors when planning restoration activities. In addition, highly stressed areas reflect numerous different combinations of stressors rather than a single suite of problems, suggesting that a detailed understanding of the stressors needing alleviation could improve restoration planning. We also find that many important areas for fisheries and recreation are subject to high stress, indicating that ecosystem degradation could be threatening key services. Current restoration efforts have targeted high-stress sites almost exclusively, but generally without knowledge of the full range of stressors affecting these locations or differences among sites in service provisioning. Our results demonstrate that joint spatial analysis of stressors and ecosystem services can provide a critical foundation for maximizing social and ecological benefits from restoration investments. www.pnas.org/lookup/suppl/doi:10.1073/pnas.1213841110/-/DCSupplementa

Physical drivers facilitating a toxigenic cyanobacterial bloom in a major Great Lakes tributary

The Maumee River is the primary source for nutrients fueling seasonal Microcystis-dominated blooms in western Lake Erie\u27s open waters though such blooms in the river are infrequent. The river also serves as source water for multiple public water systems and a large food services facility in northwest Ohio. On 20 September 2017, an unprecedented bloom was reported in the Maumee River estuary within the Toledo metropolitan area, which triggered a recreational water advisory. Here we (1) explore physical drivers likely contributing to the bloom\u27s occurrence, and (2) describe the toxin concentration and bacterioplankton taxonomic composition. A historical analysis using 10-years of seasonal river discharge, water level, and local wind data identified two instances when high-retention conditions occurred over ≥ 10 d in the Maumee River estuary: in 2016 and during the 2017 bloom. Observation by remote sensing imagery supported the advection of cyanobacterial cells into the estuary from the lake during 2017 and the lack of an estuary bloom in 2016 due to a weak cyanobacterial bloom in the lake. A rapid-response survey during the 2017 bloom determined levels of the cyanotoxins, specifically microcystins, in excess of recreational contact limits at sites within the lower 20 km of the river while amplicon sequencing found these sites were dominated by Microcystis. These results highlight the need to broaden our understanding of physical drivers of cyanobacterial blooms within the interface between riverine and lacustrine systems, particularly as such blooms are expected to become more prominent in response to a changing climate

Deep and abyssal ocean warming from 35 years of repeat hydrography

Global and regional ocean warming deeper than 2000 m is investigated using 35 years of sustained repeat hydrographic survey data starting in 1981. The global long-term temperature trend below 2000 m, representing the time period 1991–2010, is equivalent to a mean heat flux of 0.065 ± 0.040 W m?2 applied over the Earth's surface area. The strongest warming rates are found in the abyssal layer (4000–6000 m), which contributes to one third of the total heat uptake with the largest contribution from the Southern and Pacific Oceans. A similar regional pattern is found in the deep layer (2000–4000 m), which explains the remaining two thirds of the total heat uptake yet with larger uncertainties. The global average warming rate did not change within uncertainties pre-2000 versus post-2000, whereas ocean average warming rates decreased in the Pacific and Indian Oceans and increased in the Atlantic and Southern Oceans