Occupational and environmental hazard assessments for the isolation, purification and toxicity testing of cyanobacterial toxins

Cyanobacteria can produce groups of structurally and functionally unrelated but highly potent toxins. Cyanotoxins are used in multiple research endeavours, either for direct investigation of their toxicologic properties, or as functional analogues for various biochemical and physiological processes. This paper presents occupational safety guidelines and recommendations for personnel working in field, laboratory or industrial settings to produce and use purified cyanotoxins and toxic cyanobacteria, from bulk harvesting of bloom material, mass culture of laboratory isolates, through routine extraction, isolation and purification. Oral, inhalational, dermal and parenteral routes are all potential occupational exposure pathways during the various stages of cyanotoxin production and application. Investigation of toxicologic or pharmacologic properties using in vivo models may present specific risks if radiolabelled cyanotoxins are employed, and the potential for occupational exposure via the dermal route is heightened with the use of organic solvents as vehicles. Inter- and intra-national transport of living cyanobacteria for research purposes risks establishing feral microalgal populations, so disinfection of culture equipment and destruction of cells by autoclaving, incineration and/or chlorination is recommended in order to prevent viable cyanobacteria from escaping research or production facilities

Carbon isotope discrimination in leaves of the common paperbark tree, Melaleuca quinquenervia, as a tool for quantifying past tropical and subtropical rainfall

Quantitative reconstructions of terrestrial climate are highly sought after but rare, particularly in Australia. Carbon isotope discrimination in plant leaves (Δleaf) is an established indicator of past hydroclimate because the fractionation of carbon isotopes during photosynthesis is strongly influenced by water stress. Leaves of the evergreen tree Melaleuca quinquenervia have been recovered from the sediments of some perched lakes on North Stradbroke and Fraser Islands, south-east Queensland, eastern Australia. Here, we examine the potential for using M. quinquenervia ∆leaf as a tracer of past rainfall by analysing carbon isotope ratios (δ13C) of modern leaves. We firstly assess Δleaf variation at the leaf and stand scale and find no systematic pattern within leaves or between leaves due to their position on the tree. We then examine the relationships between climate and Δleaf for an 11 year timeseries of leaves collected in a litter tray. M. quinquenervia retains its leaves for 1-4 years; thus cumulative average climate data are used. There is a significant relationship between annual mean ∆leaf and mean annual rainfall of the hydrological year for 1-4 years (i.e. 365-1460 days) prior to leaf fall (r2=0.64, p=0.003, n=11). This relationship is marginally improved by accounting for the effect of pCO2 on discrimination (r2=0.67, p=0.002, n=11). The correlation between rainfall and Δleaf, and the natural distribution of Melaleuca quinquenervia around wetlands of eastern Australia, Papua New Guinea and New Caledonia offers significant potential to infer past rainfall on a wide range of spatial and temporal scales

The James Webb Space Telescope Mission

Twenty-six years ago a small committee report, building on earlier studies, expounded a compelling and poetic vision for the future of astronomy, calling for an infrared-optimized space telescope with an aperture of at least $4m$. With the support of their governments in the US, Europe, and Canada, 20,000 people realized that vision as the $6.5m$ James Webb Space Telescope. A generation of astronomers will celebrate their accomplishments for the life of the mission, potentially as long as 20 years, and beyond. This report and the scientific discoveries that follow are extended thank-you notes to the 20,000 team members. The telescope is working perfectly, with much better image quality than expected. In this and accompanying papers, we give a brief history, describe the observatory, outline its objectives and current observing program, and discuss the inventions and people who made it possible. We cite detailed reports on the design and the measured performance on orbit.Comment: Accepted by PASP for the special issue on The James Webb Space Telescope Overview, 29 pages, 4 figure

Morphological changes during akinete germination in Cylindrospermopsis raciborskii (Nostocales, Cyanobacteria)

The occurrence of the cyanobacterium Cylindrospermopsis raciborskii (Woloszynska) Seenayya and Subba Raju is a global water quality issue. The misidentification of C. raciborskii in the past is a major concern for water quality users, considering the reported cases of human and livestock poisonings associated with the cyanobacterium. Many of the available taxonomic descriptions for this species provide little or no detail of the morphology of early developmental phases that may assist with identification. Therefore, typifying the morphological changes throughout the entire life cycle for such a species requires urgent attention. In this study, five distinct morphological phases identified using a new culturing technique are reported for the process of akinete germination in C. raciborskii. Before the terminal emergence of three to four cell germlings through a ruptured akinete envelope (phase 3), mature akinetes (phase 1) elongated and the endospore separated from the akinete envelope (phase 2). After the association with the envelope was lost, four-cell germlings (phase 4a) matured into young trichomes of more than four cells (phase 4b). Throughout the process of germination, internal granular structures decreased in size and were irregular in shape in germlings and young trichomes. The culturing technique, which used a Sedgwick-Rafter cell, was successful in its application but was limiting in that the development of young trichomes after phase 4b could not be monitored

Freshwater planktic chroococcales (Cyanoprokaryota) from north-eastern Australia : a morphological evaluation

Planktic cyanoprokaryotes ofthe order Chroococcales are described from 192 sites in north-eastern Australia. Despite the diversity of flrehwater biotypes, and increasing evidence of seasonal cynaoprokeryote dominance in planktic habitats, particularly man-made reservoirs, this is the first detailed study documenting the cyanoprokaryote microflora from this region. Over five thousand samples were observed from which 54 Chroococcalean species were recorded representing four families. Sampling locations included natural lakes, man made reservoirs and weir pools, rivers and streams. The majority of the taxa could be regarded as cosmopolitan, followed by tropical/subtropical and temperate taxa. The most common genera recorded included Aphanocapsa, Cyanodictyon, Myxobaktron, Merismopedia and Microcystis.The highest taxon richness occurred in man-made lakes and reservoirs, followed by natural lakes, and streams and rivers. Fifty three taxa including three newly described species (Aphanocapsa annulata sp. nova, Gloeothece lacustrus sp. nova, Gloeocapsa planktonica sp. nova) are presented including information on their morphological variability, ecology and distribution

REV1 accumulates in DNA damage-induced nuclear foci in human cells and is implicated in mutagenesis by benzo[a]pyrenediolepoxide

The REV1 gene encodes a Y-family DNA polymerase that has been postulated to have both catalytic and structural functions in translesion replication past UV photoproducts in mammalian cells. To examine if REV1 is implicated in DNA damage tolerance mechanisms after exposure of human cells to a chemical carcinogen, we generated a plasmid expressing REV1 protein fused at its C-terminus with green fluorescent protein (GFP). In transient transfection experiments, virtually all of the transfected cells had a diffuse nuclear pattern in the absence of carcinogen exposure. In contrast, in cells exposed to benzo[a]pyrenediolepoxide, the fusion protein accumulated in a focal pattern in the nucleus in 25% of the cells, and co-localized with PCNA. These data support the idea that REV1 is present at stalled replication forks. We also examined the mutagenic response at the HPRT locus of human cells that had greatly reduced levels of REV1 mRNA due to the stable expression of gene-specific ribozymes, and compared them to wild-type cells. The mutant frequency was greatly reduced in the ribozyme-expressing cells. These data indicate that REV1 is implicated in the mutagenic DNA damage tolerance response to BPDE and support the development of strategies to target this protein to prevent such mutations

Subtle variability in water quality structures tropical diatom assemblages in streams of Cape York Peninsula, Australia

Contemporary benthic diatom assemblages were examined from 52 riverine and palustrine wetlands on Cape York Peninsula, Australia, to determine their environmental sensitivities and develop inference models. Multivariate analyses identified strong relationships between nine environmental variables and the diatom assemblage composition, with the aim to select variables for developing models. Total alkalinity, bicarbonate concentration, pH, electrical conductivity (EC) and latitude were most consistently and strongly correlated with diatom composition. The river basins sampled generally have an east-west orientation, so latitude potentially represents biogeographic differences between basins. Comparison of diatom assemblages between river basins showed significant differences, but substantial overlap in species. Diatom-based transfer functions were developed for each environmental variable and tested using the relationships between measured values and values predicted by the transfer functions. These were significant, and had low root mean square errors. An independent validation dataset for EC was analysed and applied to the EC transfer function. Results showed good predictions, giving confidence in its relevance beyond the training dataset. These understandings and models of the environmental effects on diatom assemblages allow for their application to future monitoring programs and reconstruction of past water quality conditions using fossilised diatoms in layered aquatic sediments of Cape York

Aquatic biota in hot water: thermal gradients in rheocrene hot spring discharges as analogues for the effects of climate warming

Hot springs are characterised by water temperatures above 36.7 °C. Temperature decreases with distance in flow away from spring vents; this natural gradient provides a unique opportunity to investigate the influence of water temperature on aquatic biota. This study investigated the relationship between water temperature and the aquatic invertebrates and benthic diatoms in outflows from a hot spring complex in tropical north Queensland, Australia. Water temperature ranged from 62.7 °C at the vents to 26.0 °C at the location furthest downstream. Richness of benthic diatoms and aquatic invertebrates increased linearly in response to decreasing temperature, with no species present in the hot vents. Multivariate analysis showed that both community assemblages had a response to the temperature gradient. A drop in aquatic invertebrate richness and a change in assemblage composition occurred between 40 °C and 42 °C, indicating a threshold at this temperature. The nearby Einasleigh River has experienced several contemporary peaks in water temperature over 40 °C, which corresponds to this threshold level. The relationships indicate that consistent increases in water temperature expected under climate change could decrease biological richness and precipitate changes in the aquatic invertebrate and benthic diatom taxa of tropical aquatic ecosystems

RAD18 and associated proteins are immobilized in nuclear foci in human cells entering S-phase with ultraviolet light-induced damage.

Proteins required for translesion DNA synthesis localize in nuclear foci of cells with replication-blocking lesions. The dynamics of this process were examined in human cells with fluorescence-based biophysical techniques. Photobleaching recovery and raster image correlation spectroscopy experiments indicated that involvement in the nuclear foci reduced the movement of RAD18 from diffusion-controlled to virtual immobility. Examination of the mobility of REV1 indicated that it is similarly immobilized when it is observed in nuclear foci. Reducing the level of RAD18 greatly reduced the focal accumulation of REV1 and reduced UV mutagenesis to background frequencies. Fluorescence lifetime measurements indicated that RAD18 and RAD6A or poleta only transferred resonance energy when these proteins colocalized in damage-induced nuclear foci, indicating a close physical association only within such foci. Our data support a model in which RAD18 within damage-induced nuclear foci is immobilized and is required for recruitment of Y-family DNA polymerases and subsequent mutagenesis. In the absence of damage these proteins are not physically associated within the nucleoplasm