Metapopulations in temporary streams - the role of drought-flood cycles in promoting high genetic diversity in a critically endangered freshwater fish and its consequences for the future.

Genetic factors have direct and indirect impacts in the viability of endangered species. Assessing their genetic diversity levels and population structure is thus fundamental for conservation and management. In this paper we use mitochondrial and nuclear markers to address phylogeographic and demographic data on the critically endangered Anaecypris hispanica, using a broad sampling set which covered its known distribution area in the Iberian Peninsula. Our results showed that the populations of A. hispanica are strongly differentiated (high and significant ФST and FST values, corroborated by the results from AMOVA and SAMOVA) and genetically diversified. We suggest that the restricted gene flow between populations may have been potentiated by ecological, hydrological and anthropogenic causes. Bayesian skyline plots revealed a signal for expansion for all populations (tMRCA between 68kya and 1.33Mya) and a genetic diversity latitudinal gradient was detected between the populations from the Upper (more diversified) and the Lower (less diversified) Guadiana river basin. We postulate a Pleistocenic westwards colonization route for A. hispanica in the Guadiana river basin, which is in agreement with the tempo and mode of paleoevolution of this drainage. The colonization of River Guadalquivir around 60kya with migrants from the Upper Guadiana, most likely by stream capture, is also suggested. This study highlights the view that critically endangered species facing range retreats (about 47% of its known populations have disappeared in the last 15years) are not necessarily small and genetically depleted. However, the extinction risk is not negligible since A. hispanica faces the combined effect of several deterministic and stochastic negative factors and, moreover, recolonization events after localized extinctions are very unlikely to occur due to the strong isolation of populations and to the patchily ecologically-conditioned distribution of fish. The inferred species distribution models highlight the significant contribution of temperature seasonality and isothermality to A. hispanica occurrence in Guadiana environments and emphasize the importance of stable climatic conditions for the preservation of this species. Given the strong population structure, high percentage of private haplotypes and virtual absence of inter-basin gene flow we suggest that each A. hispanica population should be considered as an independent Operational Conservation Unit and that ex-situ and in-situ actions should be conducted in parallel to allow for the long-term survival of the species and the preservation of the genetic integrity of its populations.

Evaluation of online information sources on alien species in Europe: the need of harmonization and integration.

Europe is severely affected by alien invasions, which impact biodiversity, ecosystem services, economy, and human health. A large number of national, regional, and global online databases provide information on the distribution, pathways of introduction, and impacts of alien species. The sufficiency and efficiency of the current online information systems to assist the European policy on alien species was investigated by a comparative analysis of occurrence data across 43 online databases. Large differences among databases were found which are partially explained by variations in their taxonomical, environmental, and geographical scopes but also by the variable efforts for continuous updates and by inconsistencies on the definition of "alien" or "invasive" species. No single database covered all European environments, countries, and taxonomic groups. In many European countries national databases do not exist, which greatly affects the quality of reported information. To be operational and useful to scientists, managers, and policy makers, online information systems need to be regularly updated through continuous monitoring on a country or regional level. We propose the creation of a network of online interoperable web services through which information in distributed resources can be accessed, aggregated and then used for reporting and further analysis at different geographical and political scales, as an efficient approach to increase the accessibility of information. Harmonization, standardization, conformity on international standards for nomenclature, and agreement on common definitions of alien and invasive species are among the necessary prerequisites.

Invasive alien species of policy concerns show widespread patterns of invasion and potential pressure across European ecosystems.

Animals, plants, and other organisms unintentionally or deliberately brought into a natural environment where they are not normally found, and where they cause harmful effects on that environment, are known also as invasive alien species (IAS). They represent a major threat to native biodiversity and ecosystem functioning, and can affect negatively human health and the economy. We assessed the presence and potential pressure by IAS on terrestrial and freshwater ecosystems across 27 European countries, for 66 IAS of policy concern. We computed a spatial indicator that accounts for the number of IAS present in an area and the extent of the ecosystems affected; for each ecosystem, we also looked at the pattern of invasions in the different biogeographical regions. We found disproportionally greater invasion in the Atlantic region, followed by Continental and Mediterranean regions, possibly related to historical patterns of first introductions. Urban and freshwater ecosystems were the most invaded (nearly 68% and ca. 52% of their extent respectively), followed by forest and woodland (nearly 44%). The average potential pressure of IAS was greater across cropland and forests, where we also found the lowest coefficient of variation. This assessment can be repeated over time to derive trends and monitor progress towards environmental policy objectives.

European primary datasets of alien bacteria and viruses.

Bacteria and viruses are a natural component of Earth biodiversity and play an essential role in biochemical and geological cycles. They may also pose problems outside their native range, where they can negatively impact on natural resources, wildlife, and human health. To address these challenges and develop sustainable conservation strategies, a thorough understanding of their invasion related- factors is needed: origin, country and year of introduction, and pathways dynamics. Yet, alien bacteria and viruses are underrepresented in invasion ecology studies, which limits our ability to quantify their impacts and address future introductions. This study provides primary datasets of alien bacteria and viruses of plants and animals present in the European environment. The datasets contain expert-revised data on 446 taxa and their invasion related- factors across terrestrial and aquatic environments. Taxa information are complemented with spatial occurrences. The datasets provide a basis for collaborative initiatives to improve the collection of alien bacteria and viruses' data, and a starting point for data-driven conservation practices.

Spatial priorities for freshwater biodiversity conservation in light of catchment protection and connectivity in Europe.

Freshwater ecosystems host disproportionately high numbers of species relative to their surface area yet are poorly protected globally. We used data on the distribution of 1631 species of aquatic plant, mollusc, odonate and fish in 18,816 river and lake catchments in Europe to establish spatial conservation priorities based on the occurrence of threatened, range-restricted and endemic species using the Marxan systematic conservation planning tool. We found that priorities were highest for rivers and ancient lakes in S Europe, large rivers and lakes in E and N Europe, smaller lakes in NW Europe and karst/limestone areas in the Balkans, S France and central Europe. The a priori inclusion of well-protected catchments resulted in geographically more balanced priorities and better coverage of threatened (critically endangered, endangered and vulnerable) species. The a priori exclusion of well-protected catchments showed that priority areas that need further conservation interventions are in S and E Europe. We developed three ways to evaluate the correspondence between conservation priority and current protection by assessing whether a cathment has more (or less) priority given its protection level relative to all other catchments. Each method found that priority relative to protection was high in S and E Europe and generally low in NW Europe. The inclusion of hydrological connectivity had little influence on these patterns but decreased the coverage of threatened species, indicating a trade-off between connectivity and conservation of threatened species. Our results suggest that catchments in S and E Europe need urgent conservation attention (protected areas, restoration, management, species protection) in the face of imminent threats such as river regulation, dam construction, hydropower development and climate change. Our study presents continental-scale conservation priorities for freshwater ecosystems in ecologically meaningful planning units and will thus be important in freshwater biodiversity conservation policy and practice, and water management in Europe.

Alien plants of Europe: introduction pathways, gateways and time trends.

The Convention on Biological Diversity (CBD) pathways classification framework used in the implementation of the European Union's (EU) Regulation 1143/2014 on invasive alien species (IAS Regulation) has recently been adopted by the European Alien Species Information Network (EASIN), the official information system supporting the implementation of the IAS Regulation. In the current paper, the result of an alignment of the primary introduction pathways of all alien plants in Europe included in the EASIN catalogue is presented, based on the CBD framework. In total, 6,250 alien plant taxa (marine plants excluded), both alien to Europe (native range outside Europe) and alien in Europe (native range partially in Europe) are reported. Altogether 5,175 plant taxa had their primary introduction pathway aligned based on the CBD framework, while for the rest the pathway remains unknown. In addition, the taxonomy, year and country of its first record in the wild are provided for each taxon. Our analyses reveal that the main primary introduction pathways of alien plants into Europe are linked to accidental escapes from ornamental and horticultural activities. Northwestern European countries seem to act as the main gateway areas of alien plants into Europe. Recent first observations of new alien taxa growing spontaneously exhibit a contemporary accelerating trend for plants alien to Europe, particularly linked to ornamental and horticultural activities. On the other hand, the number of new plants alien in Europe seems to have stabilized over the last few decades. The present work can assist in the prioritization of introduction pathways control, with the target of slowing down the rate of alien plants introductions into Europe, following also the requirements of the IAS Regulation.

The life cycle of Amblyomma auricularium (Acari: Ixodidae) using rabbits (Oryctolagus cuniculus) as experimental host.

The life cycle of Amblyomma auricularium (Conil) is reported for the first time, using rabbits as experimental host. Developmental periods of free-living stages were observed in an incubator at 27 +/- 1 degrees C, 80 +/- 10% RH and 24 h darkness. The complete life cycle, including pre-feeding periods for each parasitic stage, ranged from 97 to 162 days. The overall sex ratio was 1.16:1 (M:F). Feeding and premolt periods, molting success, and engorgement weight of nymphs were statistically different between males and females (P < 0.01), but because their ranges overlapped, they cannot be used to predict the sex with accuracy. The potential role of rabbits as experimental hosts for rearing A. auricularium in the laboratory is discussed.

Assessing invasive alien species in European catchments: Distribution and impacts.

Invasive alien species (IAS) induce changes to community structure and functions which lead to a decline of endemic species and major irreversible changes to the local physical habitat. The distribution and the impacts of multiple freshwater IAS are not well known, and they have not been investigated simultaneously at catchment and at European scales. This study provides an assessment of the distribution and cumulative impact of freshwater IAS over European catchments. IAS occurrences were retrieved from the European Alien Species Information Network geospatial dataset and updated with the most recent records from the literature. The Cumulative Impact Index of Invasive Alien Species (CIMPAL) was derived by aggregating the impacts of species and their occupied area at catchment level by following three steps: i) IAS were scored by both the magnitude of impacts on freshwater ecosystems and the strength of evidence in the literature, ii) scores were mapped over the catchment area, and iii) scores were summed across IAS over the catchment. The distribution of CIMPAL in the river ecological classes of the Water Framework Directive was examined and increasing/decreasing patterns identified across ecological statuses. Results showed strong spatial variation in the documented distribution and impacts of IAS in Europe. Catchments with CIMPAL scores >40 (range 0-55) clustered in Western European countries (e.g. Belgium and France) were characterised by plant, invertebrate and vertebrate IAS that had both a large impact in magnitude and colonisation at local (catchment level) and large scale (across catchments). CIMPAL showed statistically significant and increasing values from high to bad ecological classes in eight countries only (Belgium, the Czech Republic, Germany, France, Hungary, Italy, Luxemburg, Poland). This study provides comprehensive evidence of the distribution and impact of IAS within freshwater environments that could be used to improve understanding of the ecological pressures at catchment scale.

Impacts of multiple stressors on freshwater biota across spatial scales and ecosystems.

Climate and land-use change drive a suite of stressors that shape ecosystems and interact to yield complex ecological responses (that is, additive, antagonistic and synergistic effects). We know little about the spatial scales relevant for the outcomes of such interactions and little about effect sizes. These knowledge gaps need to be filled to underpin future land management decisions or climate mitigation interventions for protecting and restoring freshwater ecosystems. This study combines data across scales from 33 mesocosm experiments with those from 14 river basins and 22 cross-basin studies in Europe, producing 174 combinations of paired-stressor effects on a biological response variable. Generalized linear models showed that only one of the two stressors had a significant effect in 39% of the analysed cases, 28% of the paired-stressor combinations resulted in additive effects and 33% resulted in interactive (antagonistic, synergistic, opposing or reversal) effects. For lakes, the frequencies of additive and interactive effects were similar for all spatial scales addressed, while for rivers these frequencies increased with scale. Nutrient enrichment was the overriding stressor for lakes, with effects generally exceeding those of secondary stressors. For rivers, the effects of nutrient enrichment were dependent on the specific stressor combination and biological response variable. These results vindicate the traditional focus of lake restoration and management on nutrient stress, while highlighting that river management requires more bespoke management solutions.

Management priorities for marine invasive species.

Managing invasive alien species is particularly challenging in the ocean mainly because marine ecosystems are highly connected across broad spatial scales. Eradication of marine invasive species has only been achieved when species were detected early, and management responded rapidly. Generalized approaches, transferable across marine regions, for prioritizing actions to control invasive populations are currently lacking. Here, expert knowledge was elicited to prioritize 11 management actions for controlling 12 model species, distinguished by differences in dispersion capacity, distribution in the area to be managed, and taxonomic identity. Each action was assessed using five criteria (effectiveness, feasibility, acceptability, impacts on native communities, and cost), which were combined in an 'applicability' metric. Raising public awareness and encouraging the commercial use of invasive species were highly prioritized, whereas biological control actions were considered the least applicable. Our findings can guide rapid decision-making on prioritizing management options for the control of invasive species especially at early stages of invasion, when reducing managers' response time is critical.

Protecting and restoring Europe's waters: An analysis of the future development needs of the Water Framework Directive.

The Water Framework Directive (WFD) is a pioneering piece of legislation that aims to protect and enhance aquatic ecosystems and promote sustainable water use across Europe. There is growing concern that the objective of good status, or higher, in all EU waters by 2027 is a long way from being achieved in many countries. Through questionnaire analysis of almost 100 experts, we provide recommendations to enhance WFD monitoring and assessment systems, improve programmes of measures and further integrate with other sectoral policies. Our analysis highlights that there is great potential to enhance assessment schemes through strategic design of monitoring networks and innovation, such as earth observation. New diagnostic tools that use existing WFD monitoring data, but incorporate novel statistical and trait-based approaches could be used more widely to diagnose the cause of deterioration under conditions of multiple pressures and deliver a hierarchy of solutions for more evidence-driven decisions in river basin management. There is also a growing recognition that measures undertaken in river basin management should deliver multiple benefits across sectors, such as reduced flood risk, and there needs to be robust demonstration studies that evaluate these. Continued efforts in 'mainstreaming' water policy into other policy sectors is clearly needed to deliver wider success with WFD goals, particularly with agricultural policy. Other key policy areas where a need for stronger integration with water policy was recognised included urban planning (waste water treatment), flooding, climate and energy (hydropower). Having a deadline for attaining the policy objective of good status is important, but even more essential is to have a permanent framework for river basin management that addresses the delays in implementation of measures. This requires a long-term perspective, far beyond the current deadline of 2027.

Managing aquatic ecosystems and water resources under multiple stress--an introduction to the MARS project.

Water resources globally are affected by a complex mixture of stressors resulting from a range of drivers, including urban and agricultural land use, hydropower generation and climate change. Understanding how stressors interfere and impact upon ecological status and ecosystem services is essential for developing effective River Basin Management Plans and shaping future environmental policy. This paper details the nature of these problems for Europe's water resources and the need to find solutions at a range of spatial scales. In terms of the latter, we describe the aims and approaches of the EU-funded project MARS (Managing Aquatic ecosystems and water Resources under multiple Stress) and the conceptual and analytical framework that it is adopting to provide this knowledge, understanding and tools needed to address multiple stressors. MARS is operating at three scales: At the water body scale, the mechanistic understanding of stressor interactions and their impact upon water resources, ecological status and ecosystem services will be examined through multi-factorial experiments and the analysis of long time-series. At the river basin scale, modelling and empirical approaches will be adopted to characterise relationships between multiple stressors and ecological responses, functions, services and water resources. The effects of future land use and mitigation scenarios in 16 European river basins will be assessed. At the European scale, large-scale spatial analysis will be carried out to identify the relationships amongst stress intensity, ecological status and service provision, with a special focus on large transboundary rivers, lakes and fish. The project will support managers and policy makers in the practical implementation of the Water Framework Directive (WFD), of related legislation and of the Blueprint to Safeguard Europe's Water Resources by advising the 3rd River Basin Management Planning cycle, the revision of the WFD and by developing new tools for diagnosing and predicting multiple stressors.

Assessing the ecological status in the context of the European Water Framework Directive: where do we go now?

The Water Framework Directive (WFD) is now well established as the key management imperative in river basins across Europe. However, there remain significant concerns with the way WFD is implemented and there is now a need for water managers and scientists to communicate better in order to find solutions to these concerns. To address this, a Science-Policy Interface (SPI) activity was launched in 2010 led by Directorate-General for Research and Innovation and Onema (the French national agency for water and aquatic ecosystems), which provided an interactive forum to connect scientists and WFD end-users. One major aim of the SPI activity was to establish a list of the most crucial research and development needs for enhancing WFD implementation. This paper synthesises the recommendations from this event highlighting 10 priority issues relating to ecological status. For lakes, temporary streams and transitional and coastal waters, WFD implementation still suffers from a lack of WFD-compliant bioassessment methods. For rivers, special attention is required to assess the ecological impacts of hydromorphological alterations on biological communities, notably those affecting river continuity and riparian covering. Spatial extrapolation tools are needed in order to evaluate ecological status for water bodies for which no data are available. The need for more functional bioassessment tools as complements to usual WFD-compliant tools, and to connect clearly good ecological state, biodiversity and ecosystem services when implementing WFD were also identified as crucial issues.

Toward a holistic and risk-based management of European river basins.

The European Union Water Framework Directive (WFD) requires a good chemical and ecological status of European surface waters by 2015. Integrated, risk-based management of river basins is presumed to be an appropriate approach to achieve that goal. The approach of focusing on distinct hazardous substances in surface waters together with investment in best available technology for treatment of industrial and domestic effluents was successful in significantly reducing excessive contamination of several European river basins. The use of the concept of chemical status in the WFD is based on this experience and focuses on chemicals for which there is a general agreement that they should be phased out. However, the chemical status, based primarily on a list of 33 priority substances and 8 priority hazardous substances, considers only a small portion of possible toxicants and does not address all causes of ecotoxicological stress in general. Recommendations for further development of this concept are 1) to focus on river basin-specific toxicants, 2) to regularly update priority lists with a focus on emerging toxicants, 3) to consider state-of-the-art mixture toxicity concepts and bioavailability to link chemical and ecological status, and 4) to add a short list of priority effects and to develop environmental quality standards for these effects. The ecological status reflected by ecological quality ratios is a leading principle of the WFD. While on the European scale the improvement of hydromorphological conditions and control of eutrophication are crucial to achieve a good ecological status, on a local and regional scale managers have to deal with multiple pressures. On this scale, toxic pollution may play an important role. Strategic research is necessary 1) to identify dominant pressures, 2) to predict multistressor effects, 3) to develop stressor- and type-specific metrics of pressures, and 4) to better understand the ecology of recovery. The concept of reference conditions to define the ecological status is hard to apply and tends to ignore the fact that ecosystems can be highly dynamic. A better understanding of ecosystem responses to changes as well as early warning systems and concepts sensitive to various stressors to discriminate disturbances from natural variation are required. Because ecosystems are closely interconnected, an integrated monitoring, diagnosis, and stressors-based management of the whole water, sediment, groundwater, soil, and air system is required considering land use and the interaction with a changing climate. Extending this holistic approach beyond a consideration of existing pressures by anticipating on future ones to use and protect the aquatic environment in a sustainable way is one of the big challenges.

The effect of humidity on the oviposition and larval mortality of Haemaphysalis leporispalustris (Packard, 1869) (Acari: Ixodidae) under laboratory conditions.

Oviposition, reproductive and nutritional efficiency indices (REI and NEI), eclosion and larvae mortality of Haemaphysalis leporispalustris were studied under controlled conditions, inside desiccators, at constant temperatures of 25+/-1 degrees C and relative humidity (RH) levels adjusted at 30, 50, 70 and 95%. Fourth generation tick specimens, originated from a colony fed on rabbits (Oryctolagus cuniculus), were used. The experiment was conducted between Jul/1999 and Jan/2001. The pre-oviposition and oviposition periods decreased and increased, respectively, according to the increase in RH. The egg mass weight, REI and NEI, increased according to the increase in RH. The females' residual weight, under high humidity levels, was probably small due to a better energetic use for egg production (51.77 +/- 11.06 mg at 95% RH). The incubation period was inversely proportional to RH increase (45.10+/- 2.47 days; 40.33+/- 1.45 days and 36.85+/- 0.37 days at 50% 70% and 95% RH, respectively). There was no eclosion at 30% RH. Period and percentage of eclosion were directly proportional to the increase of the other RH levels. A diminished formation of embryonic points as well as the desiccation of the already formed eggs were observed. Larval longevity was negatively influenced by 50% RH, with death occurring concomitant to eclosion.

Afloxina M1 em queijo prato e parmesäo determinada por coluna de imunoafinidade e cromatografia líquida / Aflatoxin M1 in soft and parmesan chesse by immunoaffinity column and liquid chromatography

Produtos derivados de leite, como o queijo, podem estar com aflatoxina M1 (AFM1) quando o gado leiteiro consome raçäo contendo aflatoxina B1 (AFB1). Amostras de queijo tipo prato e tipo parmesäo ralado foram coletadas na cidade de Belo Horizonte pela Vigilância Sanitária de Minas Gerais - Brasil. Foi obtido um extrato purificado através de extraçäo com diclorometano, seguido de lavagem com n-hexano e purificaçäo em coluna de imunoafinidade. A quantificaçäo da AFM1 foi feita por cromatografia líquida de alta eficiência (CLAE) usando detetor de fluorescência. AFM1 foi detectada em todas as marcas de queijo tipo prato analisadas, em uma faixa de concentraçäo de 0,02 a 0,54 ng/g e média de 0,15 ng/g. Em queijo ralado, tipo parmesäo, AFM1 foi detectada em 13 das 14 marcas de amostras analisadas (93 por cento), em faixa de concentraçäo de 0,04 a 0,30 ng/g e média de 0,14 ng/g. (AU)

Aflatoxin M1 was determined in Soft and Parmesan cheese by immunoaffinity columnand liquid chromatography. Milk products such as cheese may be contaminated by aflatoxin M1 (AFM1)when dairy cattle have consumed feeds contaminated with aflatoxin B1 (AFB1). Samples of Soft andParmesan cheese were collected in Belo Horizonte city, by the Inspection Service of Minas Gerais ­Brazil. A purified extract was obtained by extraction with dichloromethane followed by a washing withn-hexane and immunoaffinity column clean-up. The quantification of AFM1 was done by highperformance liquid chromatography (HPLC) using a fluorescence detector. AFM1 was detected in allsoft cheese brands analysed, in concentrations ranging between 0.02 and 0.54 ng/g and mean level of0.15 ng/g. In grated cheese, Parmesan variety, AFM1 was detected in 13 of 14 brands analysed (93%),in the range 0.04-0.30 ng/g with mean level of 0.14 ng/g. (AU)

Biologia da fase parasitária e de vida livre de adultos de Amblyomma cajennense (Fabricius, 1787) (Acari: Ixodidae) em cäes (Canis familiaris L) sob condiçöes experimentais / Biology of the parasitic and free living phases of Amblyomma cajennense (Fabricius, 1787) (Acari: Ixodidae) adults on dogs (Canis familiaris L) under experimentaly conditions

Com o objetivo de ampliar as informaçöes sobre a relaçäo parasito-hospedeiro e avaliar o desenvolvimento das fases parasitária e näo-parasitária de adultos de Amblyomma cajennense alimentados em cäes, foram realizadas infestaçöes, em cinco cäes mestiços, utilizando-se dez casais por pavilhäo auricular. A fase näo-parasitária foi estudada sob condiçöes controladas de laboratório com temperatura de 27 ñ 1§C, umidade relativa do ar superior a 70 por cento e escotofase. O período parasitário médio foi de 14,93 ñ 5,36 dias. Do total de 100 casais de Amblyomma cajennense utilizados, 46 fêmeas foram recuperadas. O peso médio das fêmeas ingurgitadas foi de 324,98 ñ 207,47mg e o período médio de pré-postura e postura foram, respectivamente, de 8,71 ñ 2,40 e 16,79 ñ 6,84 dias. O peso médio da massa de ovos foi de 110,29 ñ 100,79mg e os índices de eficiência reprodutiva e nutricional foram de 33,67 por cento e 52,22 por cento, respectivamente. O período de incubaçäo médio foi de 36,79 ñ 6,13 dias. O percentual de eclosäo oscilou em torno de 40 por cento e o período médio de eclosäo foi de 14,36 ñ 9,24 dias.