Comparing the Fate and Transport of MS2 Bacteriophage and Sodium Fluorescein in a Karstic Chalk Aquifer.

Groundwater flow and contaminant migration tracing is a vital method of identifying and characterising pollutant source-pathway-receptor linkages in karst aquifers. Bacteriophages are an attractive alternative tracer to non-reactive fluorescent dye tracers, as high titres (>1012 pfu mL-1) can be safely released into the aquifer, offering improved tracer detectability. However, the interpretation of bacteriophage tracer breakthrough curves is complicated as their fate and transport are impacted by aquifer physicochemical conditions. A comparative tracer migration experiment was conducted in a peri-urban catchment in southeast England to characterise the behaviour of MS2 bacteriophage relative to sodium fluorescein dye in a karstic chalk aquifer. Tracers were released into a stream sink and detected at two abstraction boreholes located 3 km and 10 km away. At both sites, the loss of MS2 phage greatly exceeded that of the solute tracer. In contrast, the qualitative shape of the dye and phage breakthrough curves were visually very similar, suggesting that the bacteriophage arriving at each site was governed by comparable transport parameters to the non-reactive dye tracer. The colloid filtration theory was applied to explain the apparent contradiction of comparable tracer breakthrough patterns despite massive phage losses in the subsurface. One-dimensional transport models were also fitted to each breakthrough curve to facilitate a quantitative comparison of the transport parameter values. The model results suggest that the bacteriophage migrates through the conduit system slightly faster than the fluorescent dye, but that the former is significantly less dispersed. These results suggest that whilst the bacteriophage tracer cannot be used to predict receptor concentrations from transport via karstic flow paths, it can provide estimates for groundwater flow and solute contaminant transit times. This study also provides insight into the attenuation and transport of pathogenic viruses in karstic chalk aquifers.

Emerging organic contaminants in springs of the highly karstified Dinaric region.

Emerging organic contaminants (EOCs) have become of increasing interest due to concerns about their impact on humans and the wider environment. Karst aquifers are globally widespread, providing critical water supplies and sustaining rivers and ecosystems, and are particularly susceptible to pollution. However, EOC distributions in karst remain quite poorly understood. This study looks at the occurrence of EOCs in the Croatian karst, which is an example of the "classical" karst, a highly developed type of karst that occurs throughout the Dinaric region of Europe. Samples were collected from 17 karst springs and one karst lake used for water supply in Croatia during two sampling campaigns. From a screen of 740 compounds, a total of 65 compounds were detected. EOC compounds from the pharmaceutical (n = 26) and agrochemical groups (n = 26) were the most frequently detected, while industrials and artificial sweeteners had the highest concentrations (range 8-440 ng/L). The number of detected compounds and the frequency of detection demonstrate the vulnerability of karst to EOC pollution. Concentrations of 5 compounds (acesulfame, sucralose, perfluorobutane sulfonate, emamectin B1b, and triphenyl phosphate) exceeded EU standards and occurred at concentrations that are likely to be harmful to ecosystems. Overall, most detections were at low concentrations (50 % <1 ng/L). This may be due to high dilution within the exceptionally large springs of the Classical karst, or due to relatively few pollution sources within the catchments. Nevertheless, EOC fluxes are considerable (10 to 106 ng/s) due to the high discharge of the springs. Temporal differences were observed, but without a clear pattern, reflecting the highly variable nature of karst springs that occurs over both seasonal and short-term timescales. This research is one of a handful of regional EOC investigations in karst groundwater, and the first regional study in the Dinaric karst. It demonstrates the need for more frequent and extensive sampling of EOCs in karst to protect human health and the environment.

Unexpectedly deep diving in an albatross.

Albatrosses are the iconic aerial wanderers of the oceans, supremely adapted for long-distance dynamic soaring flight. Perhaps because of this they are considered poorly adapted for diving1, in contrast to many smaller shearwater and petrel relatives, despite having amphibious eyes2, and an a priori mass advantage for oxygen-storage tolerance3. Modern biologging studies have largely confirmed this view4,5, casting doubt on earlier observations using capillary tube maximum depth gauges1, which may exaggerate depths, and emphasising albatrosses' reliance on near-surface feeding. Nevertheless, uncertainty about albatross diving remains an important knowledge gap since bycatch in human fisheries (e.g. birds becoming hooked when diving for longline bait fish) is thought to be driving many population declines in this most threatened group of birds6. Here we show, using miniature electronic depth loggers (TDRs), that black-browed albatross, Thalassarche melanophris, can dive to much greater depths (19 m) and for much longer (52 s) than previously thought - three times the maxima previously recorded for this species (6 m and 15 s), and more than twice the maxima reliably recorded previously for any albatross (from 113.7 bird-days of tracking4,5,7). Further evidence that diving may be a significant behavioural adaptation in some albatrosses comes from co-deployed 3-axis accelerometers showing that these deeper dives, which occur in most individuals we tracked, involve active under-water propulsion without detectable initial assistance from momentum, sometimes with bottom phases typical of active prey pursuit. Furthermore, we find (from co-deployed GPS) that diving occurs primarily in the distal portions of long-distance foraging trips, with deeper dives occurring exclusively during daylight or civil twilight, confirming the importance of visual guidance.

Shearwaters know the direction and distance home but fail to encode intervening obstacles after free-ranging foraging trips.

While displacement experiments have been powerful for determining the sensory basis of homing navigation in birds, they have left unresolved important cognitive aspects of navigation such as what birds know about their location relative to home and the anticipated route. Here, we analyze the free-ranging Global Positioning System (GPS) tracks of a large sample (n = 707) of Manx shearwater, Puffinus puffinus, foraging trips to investigate, from a cognitive perspective, what a wild, pelagic seabird knows as it begins to home naturally. By exploiting a kind of natural experimental contrast (journeys with or without intervening obstacles) we first show that, at the start of homing, sometimes hundreds of kilometers from the colony, shearwaters are well oriented in the homeward direction, but often fail to encode intervening barriers over which they will not fly (islands or peninsulas), constrained to flying farther as a result. Second, shearwaters time their homing journeys, leaving earlier in the day when they have farther to go, and this ability to judge distance home also apparently ignores intervening obstacles. Thus, at the start of homing, shearwaters appear to be making navigational decisions using both geographic direction and distance to the goal. Since we find no decrease in orientation accuracy with trip length, duration, or tortuosity, path integration mechanisms cannot account for these findings. Instead, our results imply that a navigational mechanism used to direct natural large-scale movements in wild pelagic seabirds has map-like properties and is probably based on large-scale gradients.

Patterns of at-sea behaviour at a hybrid zone between two threatened seabirds.

Patterns of behavioural variation and migratory connectivity are important characteristics of populations, particularly at the edges of species distributions, where processes involved in influencing evolutionary trajectories, such as divergence, mutual persistence, and natural hybridization, can occur. Here, we focused on two closely related seabird species that breed in the Mediterranean: Balearic shearwaters (Puffinus mauretanicus) and Yelkouan shearwaters (Puffinus yelkouan). Genetic and phenotypic evidence of hybridization between the two species on Menorca (the eastern and westernmost island in the breeding ranges of the two shearwaters, respectively) has provided important insights into relationships between these recently diverged species. Nevertheless, levels of behavioural and ecological differentiation amongst these populations remain largely unknown. Using geolocation and stable isotopes, we compared the at-sea movement behaviour of birds from the Menorcan 'hybrid' population with the nearest neighbouring populations of Balearic and Yelkouan shearwaters. The Menorcan population displayed a suite of behavioural features intermediate to those seen in the two species (including migration strategies, breeding season movements and limited data on phenology). Our findings provide new evidence to support suggestions that the Menorcan population is admixed, and indicate a role of non-breeding behaviours in the evolutionary trajectories of Puffinus shearwaters in the Mediterranean.

The ancient Britons: groundwater fauna survived extreme climate change over tens of millions of years across NW Europe.

Global climate changes during the Cenozoic (65.5-0 Ma) caused major biological range shifts and extinctions. In northern Europe, for example, a pattern of few endemics and the dominance of wide-ranging species is thought to have been determined by the Pleistocene (2.59-0.01 Ma) glaciations. This study, in contrast, reveals an ancient subsurface fauna endemic to Britain and Ireland. Using a Bayesian phylogenetic approach, we found that two species of stygobitic invertebrates (genus Niphargus) have not only survived the entire Pleistocene in refugia but have persisted for at least 19.5 million years. Other Niphargus species form distinct cryptic taxa that diverged from their nearest continental relative between 5.6 and 1.0 Ma. The study also reveals an unusual biogeographical pattern in the Niphargus genus. It originated in north-west Europe approximately 87 Ma and underwent a gradual range expansion. Phylogenetic diversity and species age are highest in north-west Europe, suggesting resilience to extreme climate change and strongly contrasting the patterns seen in surface fauna. However, species diversity is highest in south-east Europe, indicating that once the genus spread to these areas (approximately 25 Ma), geomorphological and climatic conditions enabled much higher diversification. Our study highlights that groundwater ecosystems provide an important contribution to biodiversity and offers insight into the interactions between biological and climatic processes.

Using boreholes as windows into groundwater ecosystems.

Groundwater ecosystems remain poorly understood yet may provide ecosystem services, make a unique contribution to biodiversity and contain useful bio-indicators of water quality. Little is known about ecosystem variability, the distribution of invertebrates within aquifers, or how representative boreholes are of aquifers. We addressed these issues using borehole imaging and single borehole dilution tests to identify three potential aquifer habitats (fractures, fissures or conduits) intercepted by two Chalk boreholes at different depths beneath the surface (34 to 98 m). These habitats were characterised by sampling the invertebrates, microbiology and hydrochemistry using a packer system to isolate them. Samples were taken with progressively increasing pumped volume to assess differences between borehole and aquifer communities. The study provides a new conceptual framework to infer the origin of water, invertebrates and microbes sampled from boreholes. It demonstrates that pumping 5 m(3) at 0.4-1.8 l/sec was sufficient to entrain invertebrates from five to tens of metres into the aquifer during these packer tests. Invertebrates and bacteria were more abundant in the boreholes than in the aquifer, with associated water chemistry variations indicating that boreholes act as sites of enhanced biogeochemical cycling. There was some variability in invertebrate abundance and bacterial community structure between habitats, indicating ecological heterogeneity within the aquifer. However, invertebrates were captured in all aquifer samples, and bacterial abundance, major ion chemistry and dissolved oxygen remained similar. Therefore the study demonstrates that in the Chalk, ecosystems comprising bacteria and invertebrates extend from around the water table to 70 m below it. Hydrogeological techniques provide excellent scope for tackling outstanding questions in groundwater ecology, provided an appropriate conceptual hydrogeological understanding is applied.