Wolves were well-established members of the Pleistocene (Ice Age) carnivore community in Europe but today, many surviving populations of these charismatic animals are endangered because of human persecution and environmental change. As keystone predators, wolves play a vital role in maintaining biodiversity, particularly in keeping mammalian herbivore and medium-sized carnivore numbers in check, thereby limiting over-browsing on vegetation and over-predation on small vertebrates respectively. In this regard, they are the most influential large predator in the northern Palaearctic. The ripples from their activity can therefore be felt in diverse positive ways throughout the ecosystem but serious concerns exist as to the viability of European wolf populations under different scenarios of environmental and climate change. A key goal is therefore to understand how wolves have adapted to changing circumstances so that current and future conservation policy can be appropriately tailored. One of the best ways to approach this issue is through the study of diet, since this is closely linked to climate and environment (determining which prey species are available) and to competition for resources from other carnivores. Our previous research into the British fossil wolf record revealed marked changes in the size and shape of the jaws and teeth over the last half a million years, which together with evidence from tooth breakage and wear, indicate that wolves modified their diet (consuming more/less meat versus non-meat foods) in response to changing environmental parameters. Such morphological change cannot readily be measured in the short time scales (years to decades) of modern ecological studies but the rich Pleistocene fossil record offers a chronologically well-resolved series of wolf specimens spanning tens to hundreds of thousands of years, allowing patterns of change to be fully tested against diverse variables such as changing climates, environments, carnivore competition and prey availability. We successfully tested these palaeodietary assumptions in two NERC-funded studies on fossil wolf remains from three different climatic episodes (glacial and interglacial), using direct measurements of bone chemistry through carbon and nitrogen stable isotope analysis in order to verify changing prey choice through time. We now propose to expand this study in what will be the most comprehensive and state-of-the-art examination of diet in modern and recent fossil (<250,000 years) European wolves by using a series of independent proxies operating on different temporal scales. Working with conservation biologists, a key aim is the integration of morphological and dietary evidence from modern wolves from Sweden, Poland and Croatia, using a combination of GPS data on radio-collared wolves to identify kill sites, analysis of the contents of wolf scats, and stable isotope evidence from recently culled or dead specimens. As well as revealing seasonal and geographical variation in wolf diet, our research will allow for the first time: (1) direct comparison of modern, Holocene and Pleistocene wolf diet; (2) investigation of the degree to which direct (stable isotope, dental microwear) and indirect (morphometric) measurements of diet are in step with real-time dietary evidence from scat analyses and kill sites; (3) evaluation of the influence of diet on the morphology of modern wolves and (4) the opportunity to "ground truth" the evidence generated by current palaeodietary approaches, by assessing whether it replicates that obtained from analyses of modern wolf diet. By understanding the ecological trajectory of past and current wolf populations, we will generate a new, evidence-based view of the impacts on European large carnivores of climate, prey choice and environment. As well as academic beneficiaries, we will reach new audiences through public outreach at the Wildwood Trust and art commissions for gallery and online display.

Insects represent a vast but underexploited food resource. They have a global distribution and are amongst the most abundant animals in the world. Recent evaluations of their nutritional quality have also shown many of them to be comparable or superior as a source of nutrition to many of our current livestock animals. Although not widely used in European societies, insects are already traditionally consumed in two thirds of countries worldwide, with over 2000 species being eaten around the globe. As a widespread resource that can be collected without cultivation, the sustainable harvesting and preservation of insects offer solutions to food security problems, particularly in impoverished communities within developing world nations. Whilst this potential has been realised in some places (e.g. mopane worm in South Africa), insects have been underutilized in many areas where food security is poor. Our project will address food security issues in Sub-Saharan Africa. Many areas, such as the region of Benin where we work, experience seasonal food shortages. These arise as a consequence of extended dry seasons when crops will not grow. Limited development of food storage and preservation at the community level compounds this problem, as periods of plenty cannot be used to improve food security in famine times. This project aims to bridge this food security challenge through developing termites as a sustainable and locally available food source. Each year, a single termite mound will produce thousands of winged individuals which will disperse from the mound, and can be collected locally. These termites, which are already eaten traditionally in the region by some communities, are highly nutritious. They are naturally 'overproduced' - of the thousands who fly from a mound, 99% are eaten by birds or reptiles. As such, alate termites offer great prospects as a sustainable food product. We will assess the efficacy of different methods for the collection of termites, and combine this with an assessment of the total potential yield of termites regionally to determine the magnitude of natural capital represented in alate termites. We will then improve harvesting techniques, and develop preservation techniques that allow the product to be traded commercially/used locally over famine periods. Together, these data and techniques will allow us to determine the value-chain of a termite based food product. Within community use and local marketability will be examined as two means of maintaining nutrition through famine periods. Our project will work in collaboration with communities in Northern Benin, and supply them with the direct means to enhance food security using termites as a food product. In doing so, combining existing regional traditions of eating insects with modern advances in food preservation and production, food security can be targeted using an entirely local approach. The project aims further to be a proof of concept to establish more widespread use of termite as a food source across sub Saharan Africa.

Rivers emit ~2-3 Pg of carbon as the greenhouse gas carbon dioxide (CO2) to the atmosphere, each year. This is equivalent to 20% of annual anthropogenic CO2 emissions and an important component of the global carbon cycle. Methane (CH4) emissions from river networks are very poorly understood. CH4 is a potent greenhouse gas, 34 times stronger than CO2 over a 100-year timeframe. Rivers are estimated to emit ~27 Tg of CH4 each year, equivalent to 8% of anthropogenic CH4 emissions. However, these CH4 emissions vary greatly both spatially and over time. Rivers, acting as conduits for terrestrial greenhouse gases, can thus influence ongoing climate change. Landscape disturbance, either through human activity or climate change, can enhance river carbon emissions adding substantially to an already overloaded atmospheric carbon pool. This may represent a feedback to the global climate system as river carbon emissions can be enhanced by the impact of climate change on the terrestrial carbon cycle. Characterising the magnitude and source of river carbon emissions across globally representative ecosystems is therefore urgently needed for us to understand and predict current and future climate change. Carbon emissions from rivers are primarily derived from the landscapes they drain. But sources within these landscapes can vary depending on the ecosystem. Carbon sources can include recent atmospheric CO2 fixed into biomass via photosynthesis, carbon that has accumulated in organic soils over millennia such as in Arctic, temperate and tropical peatlands, and even ancient geological carbon derived from erosion and weathering. With such a diverse range of potential carbon sources across ecosystems, it is vital to establish a framework from which to determine whether the source of carbon observed in river networks matches what would be expected from normal landscape function, or if it represents signals of a disturbed carbon cycle. I.e. are older and slower carbon cycles becoming shorter and faster? Isotopes, especially radiocarbon (14C), are a powerful tool for identifying disturbed carbon cycles. Through a network of leading researchers, this project will bring together novel techniques and study sites to serve as a foundation for in-depth investigations into river carbon emissions around the globe. The project will utilise low-cost sensors for measuring the magnitude of river carbon emissions developed by the international Project Partners. These will be combined with in-depth isotopic investigations using novel techniques developed by the UK investigators. A network of existing study site and measurement infrastructure will be established covering a diverse range of ecosystems. The project will therefore provide a springboard from which to constrain the magnitude and source of river carbon emissions through direct observations at globally representative scales. Rivers can drain large landscape areas and as such their water chemistry represents an integrated signal of landscape carbon loss. This project will provide the techniques to tease apart these signals and determine if they represent natural or disturbed carbon cycling. The project will build a database of existing observations of these signals. In addition, we will use the interacting, complimentary techniques brought together in this project to carry out a scoping project to provide preliminary observations of the magnitude and source of carbon emissions from a subset of disturbed landscapes. CONFLUENCE will also include planning for an international meeting of researchers in relevant fields to grow the network of people, techniques and sites beyond the lifetime of this project. CONFLUENCE will be used as a launchpad for consortium funding to use this unprecedented infrastructure to make a step-change in observational capability of freshwater carbon emissions at spatial and temporal scales that individual research groups alone cannot achieve.

Catastrophic disease events can be devastating for the survival of threatened species, and can reverse years of conservation effort. When populations are already small and vulnerable, due to poaching or habitat loss, disease can be the final straw. Examples of disease as a conservation issue include the Ethiopian wolf, which is susceptible to distemper and rabies from domestic dogs, and rinderpest, which decimated the wild and domestic ungulate populations of Africa in the 19th century. Recently it has been recognised that disease is best understood and tackled in a wider context than just the individual species of host and pathogen; resilient ecosystems are better able to accommodate disease outbreaks, and human-caused environmental change can make species more vulnerable. The saiga antelope is a migratory ungulate which gathers to give birth in large aggregations. It is critically endangered due to a >95% decline in population size over <10 years due to poaching for its horns and meat. However one population (Betpak-dala) has recovered well, to about 250,000 individuals in April 2015. Mass die-offs from disease occur regularly in this species, but the causes and contributing factors have never been properly investigated. In May 2015, 120,000 saigas died within a few days in the Betpak-dala population, about half this population, and >1/3 of the global population. For the first time, a rapid response team was able to attend and collect samples from the affected saigas and their environment. Initial observations suggest that the deaths were a result of a complex interaction between particular environmental conditions (wet weather, lush grass) and the weakness of females which had just given birth, causing pathogens which were present but latent within the saigas to take hold. This is not the whole story, however, which may also include toxins in plants or water, an insect-carried disease, or a directly-transmitted virus. In this project, we will analyse the already-collected samples to diagnose the causes of and contributing factors to this mass die-off. We will run an urgent mission to the field, to collect supplementary information which will help us to home in on the triggers for this disease. We will visit both affected and unaffected areas, to understand what the differences are. We will talk to local herders, and get weather records for the days leading up to the deaths. Next, we will compile everything we know about this outbreak, and about previous outbreaks (recent and historical, in saigas and similar species), to get an overall picture of the pattern of events and environmental conditions which leads to mass saiga deaths. Combining this understanding with projections of future environmental conditions (e.g. climate change) and emerging infectious diseases (e.g. peste des petits ruminants, which is entering Central Asia from Africa), we will explore scenarios of risk from a range of diseases to both saigas and livestock, and how risks could be mitigated (e.g. through vaccination or changes in land use practices). Having assembled this evidence, we will help the Government of Kazakhstan to prepare for future disease outbreaks; we will design surveillance protocols so they can have early warning of potential triggers for mortality, and help them to examine whether, and which, interventions might reduce the risk of outbreaks, or mitigate them. We will run a technical workshop at the upcoming meeting of the UN Convention on Migratory Species' Memorandum of Understanding on saiga conservation, and support signatories (governments and NGOs) to develop and ratify an action plan. This project is a unique chance to investigate a dramatic and complex disease event of huge conservation importance, which will also shed light on the relationship between environmental change and disease. This makes it of wide general interest for ecologists, and an opportunity which it is vital to take while there is still time to act.

Biodiversity is declining at an alarming rate. Multiple stressors are driving many of these declines with freshwater (FW) ecosystems particularly impacted. Ephemeral FWs (e.g. marshes, ponds) are exceptionally biodiverse and highly exposed to varied environmental stressors but are generally overlooked within academia and regulation. Amphibians have been a major faunal component of these habitats for at least 350 million years, being highly evolved to these ecosystems. Amphibians and wetlands are some of the most highly threatened Phyla/ecosystems globally, with wetland health key to the climate crisis, due to the high methane levels emitted from human impacted systems. Using both field and laboratory approaches, here we will investigate the environmental stressor combinations driving negative impacts in amphibians (common frog, Rana temporaria) and seek to develop a biomonitoring approach to assess the health of these vital ecosystems. As amphibians are the most highly threatened vertebrate Phyla, this project is highly relevant to conservation priorities. General health, disease status, stress markers and global gene expression in wild and caged tadpoles will be measured. The use of toxicogenomics and alterations to physiology to assess impacts on tadpoles allows both the anchoring of molecular initiating events to downstream physiological endpoints and resulting adversity, as well as mapping these responses to stressor combinations. This mapping presents a highly novel approach, allowing the identification of specific stressors and their combinations that are driving negative impacts, and is widely applicable across biota. Catchment-scale eco-epidemiological studies between wild taxa and the presence/severity of stressors often rank pollution as amongst the most important variables driving negative effects in FWs. However, studies on effects of pollution at environmentally relevant levels and mixture combinations are scarce, particularly in the context of multiple stressors. Here pollutant mixture formulations will be based directly on measured levels in ephemeral FWs and combined with other ubiquitous stressors (salinity, heat wave and/or invasive crayfish - Pacifasticus leniusculus cue), all at environmentally relevant levels and combinations. These laboratory exposures will be highly novel and of vital importance to understand the true impacts of multiple stressors on iconic amphibian biota that inhabit vital ephemeral FWs. It will be tested how best to utilise data from single stressor exposures, to predict effects using theoretical models. For this, we will apply novel theoretical paradigms to the data - dominance (few stressors contribute disproportionately to observed effects) and burden (total stressor load determines effects) - which have huge potential for wide applicability for multi-stressor science. In contrast to the single-endpoint approach, here we propose to use ecological modelling to investigate effects on whole organisms and their populations in order to drastically improve the utility of these data for conservation. Finally, by transplanting spawn and sampling both caged and native tadpoles, the utility of naïve/locally adapted tadpoles as a biomonitoring tool to assess the health of FW wetlands will be assessed. This work will address an important gap in the literature between field-based catchment-level evidence demonstrating the importance of multiple stressors and the current limited laboratory-based evidence/understanding; as well as developing a new testing paradigm with practical application for conservation. The research team combines excellence in FW ecotoxicology, multiple stressors/mixture effect biology, FW ecology, ecological modelling, bioinformatics and chemistry needed for this project. In addition, the project partners and supporting organisations comprise a range of stakeholders that are focused on the health of FW ecosystems and reducing the impacts of pollution.