Adsorption, uptake and toxicity of micro- and nanoplastics: Effects on terrestrial plants and aquatic macrophytes.

Plastic pollution is a new, pressing, environmental topic. Microplastics are considered contaminants of emerging concern and, consequently, microplastic research has grown exponentially in the last decade. Here, current knowledge regarding the impacts of micro- and nanoplastics on terrestrial plants and aquatic macrophytes is discussed, with a special focus on adsorption, uptake and toxicological effects. Our review reveals that a range of plants and macrophytes can adsorb or internalise plastic particles. Both processes depend on particle characteristics such as size and charge, as well as plant features including a sticky or hydrophobic surface layer. This finding is of concern given that plants and aquatic macrophytes are at the bottom of food webs and are a crucial component of the human diet. Therefore, there is a critical need for improved understanding of adsorption, uptake and impacts of micro- and nanoplastics, and the consequences thereof for trophic transfer, food safety and security. Also, a range of stress responses have been observed for many plant and macrophyte species after both short and long-term exposures to plastic particles. Given that some plastic particles can affect plant productivity, we surmise that plastic particles may potentially impact ecosystem productivity and function. Here we present a synthesis and a critical evaluation of the state of knowledge of micro- and nanoplastics and plants and macrophytes, identifying key questions for future research.

Beyond plastic microbeads - Short-term feeding of cellulose and polyester microfibers to the freshwater amphipod Gammarus duebeni.

Monitoring studies have revealed the presence of large numbers of natural as well as anthropogenic microfibers, plastic and non-plastic, in environmental samples. However, the interaction of organisms with microfibers is largely understudied. This is the first ecotoxicological study that compares short-term feeding of anthropogenic plastic and non-plastic microfibers on a consumer (leaf-shredding detritivores) species. The freshwater amphipod Gammarus duebeni was selected for this study as it is a model ecotoxicological species. After a 96-hour exposure, 58.3% and 41.7% of the amphipods contained cellulose or polyester fibers in their digestive tracts, respectively. Microfiber ingestion was analysed per polymers in presence or absence of food. The G. duebeni group exposed to 'polyester fibers in presence of food' accumulated highest numbers of microfibers in their digestive tracts (5.2 ±â€¯3.4 MFs/amphipod) followed by those exposed to 'cellulose in presence of food' (2.5 ±â€¯0.9 MFs/amphipod). A significantly (Three-way ANOVA, p-value <0.05) higher number of microfibers was found in the midgut-hindgut (posterior) sections, compared to the foregut (anterior) section. Microfiber uptake had no apparent short-term negative effect on amphipod survival at 96 h. Yet, as amphipods are both predators and prey, and therefore are key species in the aquatic food web, the rapid accumulation of anthropogenic microfibers in their digestive system has potentially further ecological implications. Future studies need to consider the possible transfer of ingested anthropogenic microfibers to higher trophic levels in freshwater communities.

Rapid fragmentation of microplastics by the freshwater amphipod Gammarus duebeni (Lillj.).

Microplastics have become ubiquitous in all environments. Yet, their environmental fate is still largely unknown. Plastic fragmentation is a key component of plastic degradation, which is mostly caused by abiotic processes over prolonged time scales. Here, it is shown that the freshwater amphipod Gammarus duebeni can rapidly fragment polyethylene microplastics, resulting in the formation of differently shaped and sized plastic fragments, including nanoplastics. Fragments comprised 65.7% of all observed microplastic particles accumulated in digestive tracts. Higher numbers of fragments were found in response to longer exposure times and/or higher microplastic concentrations. Furthermore, the proportion of smaller plastic fragments was highest when food was present during the depuration process. It is concluded that G. duebeni can rapidly fragment polyethylene microplastics and that this is closely associated with the feeding process. These results highlight the crucial role, currently understudied, that biota may play in determining the fate of microplastics in aquatic ecosystems.

Functional shifts in bird communities from semi-natural oak forests to conifer plantations are not consistent across Europe.

While the area of plantation forest increased globally between 2010 and 2015, more than twice the area of natural forests was lost over the same period (6.5 million ha natural forest lost per year versus 3.2 million ha plantation gained per year). Consequently, there is an increasing need to understand how plantation land use affects biodiversity. The relative conservation value of plantation forests is context dependent, being influenced by previous land use, management regimes and landscape composition. What is less well understood, and of importance to conservation management, is the consistency of diversity patterns across regions, and the degree to which useful generalisations can be provided within and among bioregions. Here, we analyse forest birds in Ireland, France and Portugal, representing distinct regions across the Atlantic biogeographic area of Europe. We compared taxonomic, functional and phylogenetic diversity of bird communities among conifer plantations and semi-natural oak forests, and assessed correlations between species traits and forest type across these regions. Although bird composition (assessed with NMDS ordination) differed consistently between plantation and oak forests across all three regions, species richness and Shannon diversity did not show a consistent pattern. In Ireland and France, metrics of taxonomic diversity (richness and Shannon diversity), functional diversity, functional dispersion and phylogenetic diversity were greater in oak forests than plantations. However, in Portugal taxonomic and phylogenetic diversity did not differ significantly between forest types, while functional diversity and dispersion were statistically significantly greater in plantations. No single bird trait-forest type association correlated in a consistent direction across the three study regions. Trait associations for the French bird communities appeared intermediate between those in Ireland and Portugal, and when trait correlations were significant in both Ireland and Portugal, the direction of the correlation was always opposite. The variation in response of bird communities to conifer plantations indicates that care is needed when generalising patterns of community diversity and assembly mechanisms across regions.

Orthophosphate modulates the phytotoxicity of nano-ZnO to Lemna minor (L.).

Because of their applications in large numbers of products, Zinc Oxide nanoparticles (nano-ZnO) will inevitably enter into the environment. Nano-ZnO released into the environment will be present in a complex matrix which can cause various chemical and physical transformations and modulate the biological reactivity of these particles. Due to their rapid growth and small size, Lemna minor is recommended by OECD for toxicological testing. Here, we tested how nano-ZnO reactivity is modulated by the suite of macro- and micronutrients that are present in Lemna minor growth media. Specifically, we measured ex situ Reactive Oxygen Species (ROS) formation by nano-ZnO, and subsequent in planta toxicity. The data show how orthophosphate can modulate both ex situ ROS formation, and in planta toxicity. This has ramifications for phytotoxicity testing, which is commonly performed under controlled conditions and on media containing orthophosphate.

Effects of development of wind energy and associated changes in land use on bird densities in upland areas.

Wind energy development is the most recent of many pressures on upland bird communities and their habitats. Studies of birds in relation to wind energy development have focused on effects of direct mortality, but the importance of indirect effects (e.g., displacement, habitat loss) on avian community diversity and stability is increasingly being recognized. We used a control-impact study in combination with a gradient design to assess the effects of wind farms on upland bird densities and on bird species grouped by habitat association (forest and open-habitat species). We conducted 506 point count surveys at 12 wind-farm and 12 control sites in Ireland during 2 breeding seasons (2012 and 2013). Total bird densities were lower at wind farms than at control sites, and the greatest differences occurred close to turbines. Densities of forest species were significantly lower within 100 m of turbines than at greater distances, and this difference was mediated by habitat modifications associated with wind-farm development. In particular, reductions in forest cover adjacent to turbines was linked to the observed decrease in densities of forest species. Open-habitat species' densities were lower at wind farms but were not related to distance from turbines and were negatively related to size of the wind farm. This suggests that, for these species, wind-farm effects may occur at a landscape scale. Our findings indicate that the scale and intensity of the displacement effects of wind farms on upland birds depends on bird species' habitat associations and that the observed effects are mediated by changes in land use associated with wind-farm construction. This highlights the importance of construction effects and siting of turbines, tracks, and other infrastructure in understanding the impacts of wind farms on biodiversity.

Efectos del Desarrollo de la Energía Eólica y los Cambios Asociados al Uso de Suelo sobre las Densidades de Aves en Tierras Altas Resumen El desarrollo de la energía eólica es la más reciente de muchas presiones ejercidas sobre las comunidades de aves de tierras altas y sus hábitats. Los estudios sobre aves en relación con el desarrollo de la energía eólica se han enfocado en los efectos de la mortalidad directa, pero la importancia de los efectos indirectos (p. ej.: desplazamiento, pérdida de hábitat) sobre la diversidad y estabilidad de las comunidades aviares cada vez se reconoce más. Usamos un estudio de control-impacto combinado con un diseño de gradiente para evaluar los efectos de los campos eólicos sobre las densidades de aves de tierras altas y sobre las especies de aves agrupadas por asociación de hábitat (especies de bosque y de hábitat abierto). Realizamos 506 censos de conteo por puntos en 12 sitios de campos eólicos y 12 sitios control en Irlanda durante dos temporadas de reproducción (2012 y 2013). Las densidades de aves totales fueron más bajas en los campos eólicos que en los sitios control, con las diferencias más importantes ocurriendo cerca de las turbinas. Las densidades de las especies de bosque fueron significativamente más bajas a 100 m de las turbinas que a distancias mayores y esta diferencia estuvo mediada por modificaciones asociadas con el desarrollo de campos eólicos. De manera particular, las reducciones en la cobertura de bosque adyacente a las turbinas estuvieron vinculadas con la disminución observada en las densidades de las especies de bosque. Las densidades de las especies de hábitat abierto fueron más bajas en los campos eólicos pero no estuvieron relacionadas con la distancia a las turbinas y tuvieron una relación negativa con el tamaño del campo eólico. Lo anterior sugiere que, para estas especies, los efectos del campo eólico pueden ocurrir a la escala de paisaje. Nuestros hallazgos indican que la escala y la intensidad de los efectos de desplazamiento de los campos eólicos sobre las aves de tierras altas dependen de las asociaciones de hábitat de las especies de aves y que los efectos observados están mediados por cambios en el uso de suelo asociados con la construcción de campos eólicos. Esto remarca la importancia de los efectos de construcción y el sitiado de las turbinas, pistas y demás infraestructura en el entendimiento de los impactos que tienen los campos eólicos sobre la biodiversidad.

Natural succession and clearcutting as drivers of environmental heterogeneity and beta diversity in North American boreal forests.

Clear-cutting alters natural ecosystem processes by reducing landscape heterogeneity. It is the dominant harvesting technique across the boreal zone, yet understanding of how environmental heterogeneity and beta diversity are structured in forest ecosystems and post-clear cut is lacking. We use ground-dwelling arthropods as models to determine how natural succession (progression from deciduous to mixed to coniferous cover types) and clear-cutting change boreal forests, exploring the role of environmental heterogeneity in shaping beta diversity across multiple spatial scales (between-cover types and between-stands of the same cover type (1600 to 8500 m), between-plots (100 to 400 m) and within-plots (20 to 40 m)). We characterise environmental heterogeneity as variability in combined structural, vegetational and soil parameters, and beta diversity, as variability in assemblage composition. Clear-cutting homogenised forest environments across all spatial scales, reducing total environmental heterogeneity by 35%. Arthropod beta diversity reflected these changes at larger scales suggesting that environmental heterogeneity is useful in explaining beta diversity both between-cover types and between-stands of the same cover type. However, at smaller scales, within- and between-plots spider beta diversity reflected the lower environmental heterogeneity in regenerating stands, whereas staphylinid and carabids assemblages were not homogenised 12 years post-harvest. Differences in environmental heterogeneity and staphylinid beta diversity between cover types were also important at small scales. In regenerating stands, we detected a subtle yet notable effect of pre-felling cover type on environmental heterogeneity and arthropods, where pre-felling cover type accounted for a significant amount of variance in beta diversity, indicating that biological legacies (e.g. soil pH reflecting pre-harvest conditions) may have a role in driving beta diversity even 12 years post-harvest. This study highlights the importance of understanding site history when predicting impacts of change in forest ecosystems. Further, to understand drivers of beta diversity we must identify biological legacies shaping community structure.

Breeding phenology, provisioning behaviour, and unusual patterns of life history variation across an anthropogenic heterogeneous landscape.

Understanding how resource use and life history variation influence a population's response to modified, fragmented landscapes is a major challenge for ecologists. We investigated the phenology, life history decisions and provisioning behaviour of a generalist passerine-the great tit-across a heavily managed woodland landscape. Contrary to most previous studies on this species, reproductive investment and success were lower in deciduous than in coniferous woodland fragments. This could not be explained by differences in provisioning behaviour; instead population density was considerably higher in deciduous woodlands, suggesting birds did not follow an ideal free distribution. Clutch size declined with lay date amongst populations breeding in coniferous woodland fragments, but these populations also displayed pronounced seasonal declines in the proportion of fledglings produced per egg and fledgling mass. In contrast, and against patterns observed in other similar study systems, clutch size did not change with lay date in mixed-species deciduous woodland fragments. Furthermore, the proportion of young fledged and fledgling condition remained stable throughout the season, even though the quality of food provisioned to nestlings increased over the season. Local recruitment was negligible, suggesting that plasticity rather than natural selection played a key role in driving the patterns observed. The unusual patterns we report are likely explained by the fragmented nature of the landscape, and unreliable phenological cues in a mixed-species tree community coupled with low food availability. They contrast with those reported from most other populations situated in continuous woodland, demonstrating that caution is needed when generalising across different contexts and ecosystems.

Wood ash residue causes a mixture of growth promotion and toxicity in Lemna minor.

The use of wood as a sustainable biofuel results in the generation of residual wood ash. The ash contains high amounts of plant macronutrients such as phosphorus, potassium, calcium as well as several micronutrients. To explore the potential use of wood ash as a fertiliser, the growth enhancing properties of Sitka spruce (Picea sitchensis Bong.) wood ash were contrasted with the potential toxic action, using common duckweed (Lemna minor L.) as a model test species. The growth of L. minor exposed to wood bottom and fly ash solids and corresponding leachates was assessed in ultra-oligotrophic and eutrophic media. Ash solids and leachates were also tested as neutralized preparations. Suspended ash solids promoted L. minor growth up to concentrations of 2.5-5g/L. Leachates promoted growth up to 10g ash equivalents per litre, but for bottom ash only. Beneficial effects of wood ash were most pronounced on ultra-oligotrophic medium. However, on such nutrient-deficient medium severe inhibition of L. minor biomass and frond growth was observed at relatively low concentrations of fly ash (EC50=14g/L). On standard, eutrophic medium, higher concentrations of fly ash (EC50=21g/L), or neutralized fly ash (EC50=37g/L) were required to impede growth. Bottom ash, or neutralized bottom ash retarded growth at concentrations of 51g/L and 74g/L (EC50), respectively, in eutrophic medium. It appears that phytotoxicity is due to the elemental composition of the ash, its alkaline character, and possible interactions between these two properties. Growth promotion was due to the substantial content of plant nutrients. This study underlines the importance of the receiving environment (nutrient status and pH) in determining the balance between toxicity and growth promotion, and shows that the margin between growth promoting and toxicity inducing concentrations can be enlarged through ash neutralization.

Analysis and comparative genomics of R997, the first SXT/R391 integrative and conjugative element (ICE) of the Indian Sub-Continent.

The aim of this study was to analyse R997, the first integrative and conjugative element (ICE) isolated from the Indian Sub-Continent, and to determine its relationship to the SXT/R391 family of ICEs. WGS of Escherichia coli isolate AB1157 (which contains R997) was performed using Illumina sequencing technology. R997 context was assessed by de novo assembly, gene prediction and annotation tools, and compared to other SXT/R391 ICEs. R997 has a size of 85 Kb and harbours 85 ORFs. Within one of the variable regions a HMS-1 ß-lactamase resistance gene is located. The Hotspot regions of the element contains restriction digestion systems and insertion sequences. R997 is very closely related to the SXT-like elements from widely dispersed geographic areas. The sequencing of R997 increases the knowledge of the earliest isolated SXT/R391 elements and may provide insight on the emergence of these elements on the Indian sub-continent.

Investigation of the genotoxic potential of the marine biotoxins azaspiracid 1-3.

Azaspiracids (AZAs) are the most recently discovered group of biotoxins and are the cause of azaspiracid shellfish poisoning (AZP) in humans. To date over thirty analogues have been identified. However, toxicological studies of AZAs are limited due to the lack of availability of toxins and toxin standards. Most data available are on acute toxicity and there are no data available on genotoxicity of AZAs. This study presents an integrated approach investigating the genotoxic potential of AZA1-3 in cell culture systems using the Comet assay combined with assays to provide information on possible apoptotic processes, cytotoxicity and changes in cell number. Results demonstrate a time and dose dependent increase in DNA fragmentation in most cell lines, indicating a genotoxic effect of AZA1-3. However, a significant reduction in cell number and a clear shift from early to late apoptosis was observed for all analogues in Jurkat T cells and HepG-2 cells; CaCo-2 cells did not show a clear apoptotic profile. Late apoptotic/necrotic cells correlate well with the percentage of tail DNA for all analogues in all three cell lines. All data taken together indicate that AZA1-3 is not genotoxic per se and demonstrate apoptotic/necrotic processes to be involved to some extent in AZAs toxicity. The sensitivities of cell lines and the different potencies of AZA1-3 are in agreement with the literature available. The order of sensitivity for all three AZAs tested in the present study is, in increasing order, CaCo-2 cells < HepG-2 cells < Jurkat T cells. The order of potency of AZA1-3 varies among the cell lines.

The toxicity of zinc oxide nanoparticles to Lemna minor (L.) is predominantly caused by dissolved Zn.

Nano-ZnO particles have been reported to be toxic to many aquatic organisms, although it is debated whether this is caused by nanoparticles per sé, or rather dissolved Zn. This study investigated the role of dissolved Zn in nano-ZnO toxicity to Lemna minor. The technical approach was based on modulating nano-ZnO dissolution by either modifying the pH of the growth medium and/or surface coating of nano-ZnO, and measuring resulting impacts on L. minor growth and physiology. Results show rapid and total dissolution of nano-ZnO in the medium (pH 4.5). Quantitatively similar toxic effects were found when L. minor was exposed to nano-ZnO or the "dissolved Zn equivalent of dissolved nano-ZnO". The conclusion that nano-ZnO toxicity is primarily caused by dissolved Zn was further supported by the observation that phytotoxicity was absent on medium with higher pH-values (>7), where dissolution of nano-ZnO almost ceased. Similarly, the reduced toxicity of coated nano-ZnO, which displays a slower Zn dissolution, is also consistent with a major role for dissolved Zn in nano-ZnO toxicity.

Assessment of emerging biotoxins (pinnatoxin G and spirolides) at Europe's first marine reserve: Lough Hyne.

Active and passive sampling methods were employed over a four-month period, at a site off the South-West coast of Ireland, to characterise the occurrence of cyclic imines in the water column. The marine toxins 13-desmethyl-SPXC, 20-methyl SPXG toxins and pinnatoxin G were detected using active sampling from Diaion HP-20 resin. Seven water depths were sampled to determine stratification of the toxins in the water column using Solid Phase Adsorption and Toxin Tracking (SPATT). Both 13-desmethyl-SPXC and pinnatoxin G were detected using two different resin types; Diaion HP-20 and Amberlite XAD761. HP-20 proved more effective at accumulating the toxins, with a higher percentage of positive samples and a higher ratio of toxin adsorbed relative to XAD761. No temporal variation in toxin-quantities was detected, indicating that there was no change in density of causative algal species in the water column. Pinnatoxin G was detected more frequently from surface to 30 m depth, with a similar pattern observed for 13-desmethyl-SPXC occurrence using XAD761. No difference in the occurrence of 13-desmethyl-SPXC was observed between depths using HP-20 resin. This is the first reported incidence of pinnatoxin G in Irish waters and highlights cyclic imines as emerging toxins in European waters.

Forest Cover Estimation in Ireland Using Radar Remote Sensing: A Comparative Analysis of Forest Cover Assessment Methodologies.

Quantification of spatial and temporal changes in forest cover is an essential component of forest monitoring programs. Due to its cloud free capability, Synthetic Aperture Radar (SAR) is an ideal source of information on forest dynamics in countries with near-constant cloud-cover. However, few studies have investigated the use of SAR for forest cover estimation in landscapes with highly sparse and fragmented forest cover. In this study, the potential use of L-band SAR for forest cover estimation in two regions (Longford and Sligo) in Ireland is investigated and compared to forest cover estimates derived from three national (Forestry2010, Prime2, National Forest Inventory), one pan-European (Forest Map 2006) and one global forest cover (Global Forest Change) product. Two machine-learning approaches (Random Forests and Extremely Randomised Trees) are evaluated. Both Random Forests and Extremely Randomised Trees classification accuracies were high (98.1-98.5%), with differences between the two classifiers being minimal (<0.5%). Increasing levels of post classification filtering led to a decrease in estimated forest area and an increase in overall accuracy of SAR-derived forest cover maps. All forest cover products were evaluated using an independent validation dataset. For the Longford region, the highest overall accuracy was recorded with the Forestry2010 dataset (97.42%) whereas in Sligo, highest overall accuracy was obtained for the Prime2 dataset (97.43%), although accuracies of SAR-derived forest maps were comparable. Our findings indicate that spaceborne radar could aid inventories in regions with low levels of forest cover in fragmented landscapes. The reduced accuracies observed for the global and pan-continental forest cover maps in comparison to national and SAR-derived forest maps indicate that caution should be exercised when applying these datasets for national reporting.

Does the marine biotoxin okadaic acid cause DNA fragmentation in the blue mussel and the pacific oyster?

Two bivalve species of global economic importance: the blue mussel, Mytilus edulis and the pacific oyster, Crassostrea gigas were exposed in vivo, to the diarrhoetic shellfish toxin okadaic acid (OA), and impacts on DNA fragmentation were measured. Shellfish were exposed using two different regimes, the first was a single (24 h) exposure of 2.5 nM OA (∼0.1 µg/shellfish) and algal feed at the beginning of the trial (T0), after which shellfish were only fed algae. The second was daily exposure of shellfish to two different concentrations of OA mixed with the algal feed over 7 days; 1.2 nM OA (∼0.05 µg OA/shellfish/day) and 50 nM OA (∼2 µg OA/shellfish/day). Haemolymph and hepatopancreas cells were extracted following 1, 3 and 7 days exposure. Cell viability was measured using the trypan blue exclusion assay and remained above 85% for both cell types. DNA fragmentation was examined using the single-cell gel electrophoresis (comet) assay. A significant increase in DNA fragmentation was observed in the two cell types from both species relative to the controls. This increase was greater in the pacific oyster at the higher toxin concentration. However, there was no difference in the proportion of damage measured between the two cell types, and a classic dose response was not observed, increasing toxin concentration did not correspond to increased DNA fragmentation.

Trophic ecology of the Grey Wagtail Motacilla cinerea before and during the breeding season in the region of Bejaia (Algeria).

The diet composition of the Grey Wagtail Motacilla cinerea of the Babor Range is followed by analysis of faecal samples (90 faeces) before and during the breeding season 2010. The Grey Wagtail's diet varies depending on the stage of the breeding cycle at the southern edge of their breeding area in North Africa (Bejaia). The diet consists predominantly of aquatic preys (51.79%), with Coleoptera being the most frequent constituent (n=331, 45.5%). During the pre-laying period (February-March), the diet was variable (91 prey-taxa and H'=3,36 bits) and preys of medium size (5 to 8mm) were most common. During the incubation period (April-May), preys were mainly aquatic (60%) and larger (20 to 32mm). At the end of the breeding season (June-July), there was a greater occurrence of terrestrial preys (31 aquatic versus 30 terrestrial taxa).

Application of passive (SPATT) and active sampling methods in the profiling and monitoring of marine biotoxins.

Solid phase adsorbent and toxin tracking (SPATT) enables temporally and spatially integrated monitoring of biotoxins in aquatic environments. Monitoring using two adsorbent resins was performed over a four-month period at Lough Hyne Marine Reserve, Ireland. A range of Diarhettic Shellfish Poisoning (DSP) toxins were detected from SPATT extracts throughout the study period. The majority of biotoxins were detected in the top 20-30 m of the water column and a spike in toxin accumulation was measured during August 2010. Phytoplankton analysis confirmed the presence of toxin-producing species Dinophysis acuta and Dinophysis acuminata during the bloom. SPATT has the potential to provide useful information on phycotoxin distribution in the water column; enabling evidence-based decisions regarding appropriate depths for obtaining phytoplankton and shellfish samples in marine biotoxin monitoring programmes. Active sampling was performed continuously over 7-days and high quantities of toxins were successfully accumulated in the HP-20 resin, okadaic acid (∼13 mg), dinophysis toxin-2 (∼29 mg), pectenotoxin-2 (∼20 mg) and pectenotoxin-2-seco acid (∼6 mg) proving this an effective method for accumulating DSP toxins from the marine environment. The method has potential application as a tool for assessing toxin profiles at proposed shellfish harvesting sites.

Invading and expanding: range dynamics and ecological consequences of the greater white-toothed shrew (Crocidura russula) invasion in Ireland.

Establishing how invasive species impact upon pre-existing species is a fundamental question in ecology and conservation biology. The greater white-toothed shrew (Crocidura russula) is an invasive species in Ireland that was first recorded in 2007 and which, according to initial data, may be limiting the abundance/distribution of the pygmy shrew (Sorex minutus), previously Ireland's only shrew species. Because of these concerns, we undertook an intensive live-trapping survey (and used other data from live-trapping, sightings and bird of prey pellets/nest inspections collected between 2006 and 2013) to model the distribution and expansion of C. russula in Ireland and its impacts on Ireland's small mammal community. The main distribution range of C. russula was found to be approximately 7,600 km2 in 2013, with established outlier populations suggesting that the species is dispersing with human assistance within the island. The species is expanding rapidly for a small mammal, with a radial expansion rate of 5.5 km/yr overall (2008-2013), and independent estimates from live-trapping in 2012-2013 showing rates of 2.4-14.1 km/yr, 0.5-7.1 km/yr and 0-5.6 km/yr depending on the landscape features present. S. minutus is negatively associated with C. russula. S. minutus is completely absent at sites where C. russula is established and is only present at sites at the edge of and beyond the invasion range of C. russula. The speed of this invasion and the homogenous nature of the Irish landscape may mean that S. minutus has not had sufficient time to adapt to the sudden appearance of C. russula. This may mean the continued decline/disappearance of S. minutus as C. russula spreads throughout the island.

Pseudovibrio axinellae sp. nov., isolated from an Irish marine sponge.

A Gram-negative, motile, rod-shaped bacterial strain, designated Ad2(T), was isolated from a marine sponge, Axinella dissimilis, which was collected from a semi-enclosed marine lake in Ireland. Strain Ad2(T) grew optimally at 24 °C, at pH 7.0 and in the presence of 3 % (w/v) NaCl. Phylogenetic analysis based on 16S rRNA gene sequences showed that strain Ad2(T) clustered with members of the genus Pseudovibrio, and showed 97.3-98.2 % sequence similarity to the type strains of recognized Pseudovibrio species. DNA-DNA relatedness values between strain Ad2(T) and the type strains of other Pseudovibrio species were <27 %. The DNA G+C content of strain Ad2(T) was 50.5 mol%. The major fatty acid was 18 : 1ω7c. Differences in phenotypic properties, together with phylogenetic and DNA-DNA hybridization analyses, indicated that strain Ad2(T) represented a novel species of the genus Pseudovibrio. The name Pseudovibrio axinellae sp. nov. is proposed, with Ad2(T) (= DSM 24994(T) = NCIMB 14761(T)) as the type strain.

Elucidation of the mass fragmentation pathways of the polyether marine toxins, dinophysistoxins, and identification of isomer discrimination processes.

RATIONALE: Most of the liquid chromatography/mass spectrometry (LC/MS) methods that have been developed for the analysis of Diarrhetic Shellfish Poisoning (DSP) toxins in shellfish and algae samples have been unable to differentiate the isomers okadaic acid (OA) and dinophysistoxin-2 (DTX2), unless separated by chromatography. Since there are many bioconversion products of these compounds it is imperative to determine characteristic product ions, which can provide unequivocal identification of OA and DTX2 and their analogs. METHODS: Using electrospray ionization, the fragmentation processes for two types of precursor ions, [M+Na](+) and [M-H](-), of the polyether marine toxins, dinophysistoxins (DTXs), were studied using a hybrid linear ion trap Orbitrap mass spectrometer which provided high mass accuracy data in combination with multiple tandem mass (MS(n)) spectra. Three structurally related toxins were compared; okadaic acid (OA), dinophysistoxin-2 (DTX2) and dinophysistoxin-1 (DTX1). A quick multiple reaction monitoring (MRM) LC/MS/MS method was developed utilizing the characteristic precursor/product ion mass transitions. RESULTS: Comparison of the high-resolution product ion, [M-H](-), spectra of these toxins featured dominant signals that resulted from two six-centered rearrangements and previously proposed fragmentation pathways for the ion of m/z 321 and 293 have been corrected and identified. By contrast, the [M+Na](+) product ion spectra only revealed distinctive ions for the isomers, OA (m/z 595, 443 and 151) and DTX2 (m/z 581, 429 and 165). To illustrate the benefits of this study, a mass selective LC/MS/MS method was developed in which the isomers OA and DTX2 co-eluted but were distinguished using the mass transitions, m/z 827/595, 827/443 (OA) and m/z 827/581, 827/429 (DTX2). CONCLUSIONS: Comparison of OA, DTX2 and DTX1 led to the correction of proposed negative ion mode fragmentation pathways. Through extensive study and comparison of the [M+Na](+) product ion spectra, distinctive product ions were identified which allowed for these compounds to be identified and distinguished without separation for the first time.