Non-Lethal Detection of Ranavirus in Fish.

Emergent infectious diseases have an increasing impact on both farmed animals and wildlife. The ability to screen for pathogens is critical for understanding host-pathogen dynamics and informing better management. Ranavirus is a pathogen of concern, associated with disease outbreaks worldwide, affecting a broad range of fish, amphibian, and reptile hosts, but research has been limited. The traditional screening of internal tissues, such as the liver, has been regarded as the most effective for detecting and quantifying Ranavirus. However, such methodology imposes several limitations from ethical and conservation standpoints. Non-lethal sampling methods of viral detection were explored by comparing the efficacy of both buccal swabbing and fin clipping. The study was conducted on two Iberian, threatened freshwater fish (Iberochondrostoma lusitanicum and Cobitis paludica), and all samples were screened using qPCR. While for C. paludica both methods were reliable in detecting Ranavirus, on I. lusitanicum, there was a significantly higher detection rate in buccal swabs than in fin tissue. This study, therefore, reports that fin clipping may yield false Ranavirus negatives when in small-bodied freshwater fish. Overall, buccal swabbing is found to be good as an alternative to more invasive procedures, which is of extreme relevance, particularly when dealing with a threatened species.

Reproduction of the Blue Jack Mackerel, Trachurus picturatus, in Western Portugal: Microscopic Gonad Analysis Reveals Indeterminate Fecundity and Skipped Spawning Patterns.

Blue jack mackerel, Trachurus picturatus, is the fifth most landed fish species in mainland Portugal, but information on its reproductive biology is scarce. From September 2018 to August 2019, 626 specimens were collected from commercial vessels to clarify the reproductive strategy of the T. picturatus population off the west coast of Portugal. The proportion and length range of males and females were similar. Only three of the specimens collected were categorized as immature, indicating that the fish caught in the fishery are primarily mature. The spawning season lasted from late January until the end of March, with gonadosomatic indices being similar for males and females. Fecundity was indeterminate, and estimated batch fecundity ranged between 6,798 (at 25.4 cm TL) and 302,358 oocytes (at 33.8 cm TL). The low number of females showing direct evidence of imminent or recent spawning suggests a low number of spawning events. In addition, 12.7% of females were considered non-reproductive due to ovary abnormalities including parasitic infection by Kudoa species, atretic structures and skipped spawning events. This study highlights the importance of accounting for skipped spawning events and ovary abnormalities in the management of species fisheries.

New insights on patterns of genetic admixture and phylogeographic history in Iberian high mountain populations of midwife toads.

Multiple Quaternary glacial refugia in the Iberian Peninsula, commonly known as "refugia within refugia", allowed diverging populations to come into contact and admix, potentially boosting substantial mito-nuclear discordances. In this study, we employ a comprehensive set of mitochondrial and nuclear markers to shed light onto the drivers of geographical differentiation in Iberian high mountain populations of the midwife toads Alytes obstetricans and A. almogavarii from the Pyrenees, Picos de Europa and Guadarrama Mountains. In the three analysed mountain regions, we detected evidence of extensive mito-nuclear discordances and/or admixture between taxa. Clustering analyses identified three major divergent lineages in the Pyrenees (corresponding to the eastern, central and central-western Pyrenees), which possibly recurrently expanded and admixed during the succession of glacial-interglacial periods that characterised the Late Pleistocene, and that currently follow a ring-shaped diversification pattern. On the other hand, populations from the Picos de Europa mountains (NW Iberian Peninsula) showed a mitochondrial affinity to central-western Pyrenean populations and a nuclear affinity to populations from the central Iberian Peninsula, suggesting a likely admixed origin for Picos de Europa populations. Finally, populations from the Guadarrama Mountain Range (central Iberian Peninsula) were depleted of genetic diversity, possibly as a consequence of a recent epidemic of chytridiomycosis. This work highlights the complex evolutionary history that shaped the current genetic composition of high mountain populations, and underscores the importance of using a multilocus approach to better infer the dynamics of population divergence.

Seasonal study of the nutritional composition of unexploited and low commercial value fish species from the Portuguese coast.

Target species diversification is essential for fisheries sustainability and fish market revitalization. Fish discards are a widely recognized problem resulting from fisheries worldwide, and are of major concern for all sector players, from administrations, to fishermen, and scientists. However, non-target species are seldom studied, and information on nutritional profiles and seasonal changes in nutritional properties is generally lacking. This study assessed the seasonal nutritional composition of two unexploited (Serranus cabrilla, Capros aper) and three low commercial value fish species (Trachurus picturatus, Spondyliosoma cantharus, and Trigla lyra), captured on the Portuguese coast over 1 year. Significant seasonal variations were observed in the nutritional composition of all the species studied. Moisture and ash contents varied from 70% to 81% and from 5% to 13%, respectively. The maximum fat contents were 5% for C. aper and 4% for T. picturatus, allowing to classify all studied fishes as lean. The highest protein contents were recorded for C. aper (25%) and S. cantharus (20%). The unexploited and low commercial value fish species studied were shown to be good fat and protein sources, comparable to commonly consumed species, such as cod and salmon, having a great potential to become commonly consumed fish in Portugal.

Contrasting patterns from two invasion fronts suggest a niche shift of an invasive predator of native bees.

Background: The accuracy of predictions of invasive species ranges is dependent on niche similarity between invasive and native populations and on our ability to identify the niche characteristics. With this work we aimed to compare the niche dynamics of two genetically related invasive populations of Vespa velutina (an effective predator of honeybees and wild pollinators), in two distinct climatic regions, one in central Europe and another one in the north-western Iberian Peninsula, and hence to identify uninvaded regions susceptible to invasion. Methods: Niche dynamics and shifts of V. velutina were assessed by comparing the environmental niches of the native and of the two invasive populations, using climatic, topographic and land use variables. We also ran reciprocal distribution models using different algorithms and records from both native and invasive ranges to compare model predictions and estimate which regions are at a greater risk of being invaded. Results: An apparent niche shift was detected in the population of the NW of Iberian Peninsula, where the species is living under environmental conditions different from the native niche. In central Europe, large suitable areas remain unoccupied. The fact that both invasive populations are well established, despite occupying environmentally distinct regions indicates that V. velutina has a high ability to successfully invade different environmental envelopes from those existing in its native range. For example, in north-western Iberian Peninsula the species is now thriving out of its native niche limits. Moreover, the large extent of still unoccupied environmental space with similar conditions to those used by the species in its native range suggests that there is still a large area of central and eastern Europe that can be potentially invaded by the species.

Trophic niche changes associated with the eradication of invasive mammals in an insular lizard: an assessment using isotopes.

Invasive species are a major threat to island biodiversity, and their eradications have substantially contributed to the conservation of island endemics. However, the consequences of eradications on the trophic ecology of native taxa are largely unexplored. Here, we used the eradication of invasive black rats Rattus rattus and European rabbits Oryctolagus cuniculus from the Berlenga Island, in the western coast of Portugal, as a whole-ecosystem experiment to investigate the effects of the eradication of invasive mammals on the trophic niche and body dimensions of the island-restricted Berlenga wall lizard Podarcis carbonelli berlengensis over a 2-year period. Our results suggest an expansion of the isotopic niche and an intensification of the sexual dimorphism of the lizard following mammal eradication. Additionally, we found considerable variability in isotopic niche across the island and detected evidence of sex-specific and season-modulated nutritional requirements of this threatened reptile. Our findings support that the eradication of 2 of the planet's most problematic invasive vertebrates led to changes in the lizard trophic niche and sexual dimorphism in just 2 years. This suggests that the ecological pressures-for example, prey availability and habitat structure-to which lizards are exposed have substantially changed post-eradication. Our study emphasizes the scientific value of island eradications as experiments to address a wide range of ecological questions and adds to the increasing body of evidence supporting substantial conservation gains associated with these restoration interventions.

Enhancing knowledge on low-value fishing species: the distinct reproductive strategy of two gurnard species.

The depletion and overexploitation of several fish stock demands for a valorisation of non-target and discarded species. Nonetheless, such species are often poorly studied, and information on their biological parameters must be gathered for effective population management. For 1 year, the reproductive strategy of the piper gurnard Trigla lyra and the red gurnard Chelidonichthys cuculus was studied by monthly samples obtained from commercial boats operating on western Portuguese coast. Both species showed a biased sex ratio towards females, especially for larger length classes. Length at first maturity could be estimated only for red gurnard (22.1 and 19.9 cm for females and males, respectively) because all piper gurnard individuals caught were mature. Piper gurnard showed determinate fecundity and a short spawning season, from November to February with a peak in January, whereas red gurnard showed indeterminate fecundity and a wide spawning season, from late December to May. The relative annual fecundity estimated for red gurnard (1893 ± 728 oocytes × g-1 eviscerated weight [EW]) was higher than the one estimated for piper gurnard (1018 ± 250 oocytes×g-1 EW). Although important information for understanding the species dynamics is presented in this study, additional information on other life-history parameters and of species landings is required.

Questioning the proverb 'more haste, less speed': classic versus metabarcoding approaches for the diet study of a remote island endemic gecko.

Dietary studies can reveal valuable information on how species exploit their habitats and are of particular importance for insular endemics conservation as these species present higher risk of extinction. Reptiles are often neglected in island systems, principally the ones inhabiting remote areas, therefore little is known on their ecological networks. The Selvagens gecko Tarentola (boettgeri) bischoffi, endemic to the remote and integral reserve of Selvagens Archipelago, is classified as Vulnerable by the Portuguese Red Data Book. Little is known about this gecko's ecology and dietary habits, but it is assumed to be exclusively insectivorous. The diet of the continental Tarentola species was already studied using classical methods. Only two studies have used next-generation sequencing (NGS) techniques for this genus thus far, and very few NGS studies have been employed for reptiles in general. Considering the lack of information on its diet and the conservation interest of the Selvagens gecko, we used morphological and DNA metabarcoding approaches to characterize its diet. The traditional method of morphological identification of prey remains in faecal pellets collected over a longer period was compared with metabarcoding of samples collected during rapid surveys. Molecular results revealed that this species is a generalist, feeding on invertebrate, plant and vertebrate items, whereas the morphological approaches were unable to detect the latter two. These results opened up new questions on the ecological role of the Selvagens gecko that deserves to be further explored, such as the possible predation on seabirds, plant services or trophic competition with the sympatric Madeira lizard Teira dugesii. Metabarcoding identified a greater diversity of dietary items at higher taxonomic resolution, but morphological identification enabled calculation of relative abundances and biomasses of ingested arthropods, and detected a dietary shift on invertebrate preys between seasons. Results of this study highlight the global applicability of rapid metabarcoding surveys for understudied taxa on remote islands that are difficult to access. We recommend using the metabarcoding approach, even if 'speedy' sampling only is possible, but we must highlight that disregarding long-term ecological data may lead to 'hasty' conclusion.

Adding Value to Bycatch Fish Species Captured in the Portuguese Coast-Development of New Food Products.

We live in a world of limited biological resources and ecosystems, which are essential to feed people. Consequently, diversifying target species and considering full exploitation are essential for fishery sustainability. The present study focuses on the valorization of three low commercial value fish species (blue jack mackerel, Trachurus picturatus; black seabream, Spondyliosoma cantharus; and piper gurnard, Trigla lyra) and of two unexploited species (comber, Serranus cabrilla and boarfish, Capros aper) through the development of marine-based food products with added value. A preliminary inquiry with 155 consumers from Região de Lisboa e Vale do Tejo (Center of Portugal) was conducted to assess fish consumption, the applicability of fish product innovation, and the importance of valorizing discarded fish. Five products (black seabream ceviche, smoked blue jack mackerel pâté, dehydrated piper gurnard, fried boarfish, and comber pastries) were developed and investigated for their sensory characteristics and consumer liking by hedonic tests to 90 consumers. The most important descriptors were identified for each product (texture, flavor, color, and appearance). Comber pastries had the highest purchase intention (88%), followed by black seabream ceviche (85%) and blue jack mackerel pâté (76%). Sensory evaluations showed a clear tendency of consumers to accept reformulated products, with the introduction of the low-value and unexploited species under study.

Non-native freshwater fauna in Portugal: A review.

We present the most updated list of non-native freshwater fauna established in Portugal, including the Azores and Madeira archipelagos. This list includes 67 species at national level but corresponds to 84 species records, of which 53 are in the mainland, 23 in the Azores and 8 in Madeira archipelagos. We also discuss the progression of the cumulative number of introductions since 1800 and identify the most probable vectors of introduction, main taxonomic groups and their regions of origin. Furthermore, we review the existing knowledge about ecological and economic impacts, invasion risk and potential distribution of invaders, under present and future climatic conditions, and the applied management actions, including the production of legislation. Along the 20th century the number of successful introductions increased at an approximate rate of two new species per decade until the beginning of 1970s. Since then, this rate increased to about 14 new species per decade. These introductions were mainly a result of fisheries, as contaminants or for ornamental purposes. Fish and mollusks are the taxonomic groups with more established species, representing more than half of the total. Most species (>70%) are native from other regions of Europe and North America. Studies about ecological or socioeconomic impacts are more common for fish, crustaceans and mollusks. Impacts for most amphibians, reptiles and mammals are not thoroughly studied. A few studies on the impacts and management actions of health-threatening mosquitoes are also available. The potential distribution in the Portuguese territory was modelled for 26 species. Only a minority of these models provides projections of distributions under scenarios of future climate change. A comparison of the Portuguese and EU legislation shows large discrepancies in the invasive species lists. Using the EU list and a ranking procedure for the national context, we identify freshwater species of high national concern for which actions are urgently needed.

Can heat waves change the trophic role of the world's most invasive crayfish? Diet shifts in Procambarus clarkii.

In the Mediterranean basin, the globally increasing temperatures are expected to be accompanied by longer heat waves. Commonly assumed to benefit cold-limited invasive alien species, these climatic changes may also change their feeding preferences, especially in the case of omnivorous ectotherms. We investigated heat wave effects on diet choice, growth and energy reserves in the invasive red swamp crayfish, Procambarus clarkii. In laboratory experiments, we fed juvenile and adult crayfish on animal, plant or mixed diets and exposed them to a short or a long heat wave. We then measured crayfish survival, growth, body reserves and Fulton's condition index. Diet choices of the crayfish maintained on the mixed diet were estimated using stable isotopes (13C and 15N). The results suggest a decreased efficiency of carnivorous diets at higher temperatures, as juveniles fed on the animal diet were unable to maintain high growth rates in the long heat wave; and a decreased efficiency of herbivorous diets at lower temperatures, as juveniles in the cold accumulated less body reserves when fed on the plant diet. Heat wave treatments increased the assimilation of plant material, especially in juveniles, allowing them to sustain high growth rates in the long heat wave. Contrary to our expectations, crayfish performance decreased in the long heat wave, suggesting that Mediterranean summer heat waves may have negative effects on P. clarkii and that they are unlikely to boost its populations in this region. Although uncertain, it is possible that the greater assimilation of the plant diet resulted from changes in crayfish feeding preferences, raising the hypotheses that i) heat waves may change the predominant impacts of this keystone species and ii) that by altering species' trophic niches, climate change may alter the main impacts of invasive alien species.

Effects of 6-months' Exercise on Cardiac Function, Structure and Metabolism in Female Hypertensive Rats-The Decisive Role of Lysyl Oxidase and Collagen III.

Purpose: According to the current therapeutic guidelines of the WHO physical activity and exercise are recommended as first-line therapy of arterial hypertension. Previous results lead to the conclusion, however, that hearts of spontaneously hypertensive rats (SHR) with established hypertension cannot compensate for the haemodynamic stresses caused by long-term exercise. The current study was initiated to investigate the effects of aerobic exercise on the cardiac remodeling as the sole therapeutic measure before and during hypertension became established. Methods: Beginning at their 6th week of life, six SHR were provided with a running wheel over a period of 6 months. Normotensive Wistar rats served as non-hypertensive controls. Results: In Wistar rats and SHR, voluntary exercise led to cardioprotective adaptation reactions that were reflected in increased mitochondrial respiration, reduced heart rate and improved systolic function. Exercise also had antioxidant effects and reduced the expression of maladaptive genes (TGF-ß1, CTGF, and FGF2). However, at the end of the 6-months' training, the echocardiograms revealed that SHR runners developed a restrictive cardiomyopathy. The induction of lysyl oxidase (LOX), which led to an increased network of matrix proteins and a massive elevation in collagen III expression, was identified as the underlying cause. Conclusions: Running-induced adaptive mechanisms effectively counteract the classic remodeling of hearts subject to chronic pressure loads. However, with sustained running stress, signaling pathways are activated that have a negative effect on left ventricular relaxation. Our data suggest that the induction of LOX may play a causative role in the diagnosed filling disorder in trained SHR.

Global realized niche divergence in the African clawed frog Xenopus laevis.

Although of crucial importance for invasion biology and impact assessments of climate change, it remains widely unknown how species cope with and adapt to environmental conditions beyond their currently realized climatic niches (i.e., those climatic conditions existing populations are exposed to). The African clawed frog Xenopus laevis, native to southern Africa, has established numerous invasive populations on multiple continents making it a pertinent model organism to study environmental niche dynamics. In this study, we assess whether the realized niches of the invasive populations in Europe, South, and North America represent subsets of the species' realized niche in its native distributional range or if niche shifts are traceable. If shifts are traceable, we ask whether the realized niches of invasive populations still contain signatures of the niche of source populations what could indicate local adaptations. Univariate comparisons among bioclimatic conditions at native and invaded ranges revealed the invasive populations to be nested within the variable range of the native population. However, at the same time, invasive populations are well differentiated in multidimensional niche space as quantified via n-dimensional hypervolumes. The most deviant invasive population are those from Europe. Our results suggest varying degrees of realized niche shifts, which are mainly driven by temperature related variables. The crosswise projection of the hypervolumes that were trained in invaded ranges revealed the south-western Cape region as likely area of origin for all invasive populations, which is largely congruent with DNA sequence data and suggests a gradual exploration of novel climate space in invasive populations.

Are invasive populations characterized by a broader diet than native populations?

BACKGROUND: Invasive species are among the most significant threats to biodiversity. The diet of invasive animal populations is a crucial factor that must be considered in the context of biological invasions. A broad dietary spectrum is a frequently cited characteristic of invasive species, allowing them to thrive in a wide range of environments. Therefore, empirical studies comparing diet in invasive and native populations are necessary to understand dietary requirements, dietary flexibility, and the associated impacts of invasive species. METHODS: In this study, we compared the diet of populations of the African clawed frog, Xenopus laevis in its native range, with several areas where it has become invasive. Each prey category detected in stomach contents was assigned to an ecological category, allowing a comparison of the diversity of ecological traits among the prey items in the diet of native and introduced populations. The comparison of diets was also performed using evenness as a niche breadth index on all sampled populations, and electivity as a prey selection index for three out of the six sampled populations. RESULTS: Our results showed that diet breadth could be either narrow or broad in invasive populations. According to diet and prey availability, zooplankton was strongly preferred in most cases. In lotic environments, zooplankton was replaced by benthic preys, such as ephemeropteran larvae. DISCUSSION: The relative proportions of prey with different ecological traits, and dietary variability within and between areas of occurrence, suggest that X. laevis is a generalist predator in both native and invasive populations. Shifts in the realized trophic niche are observed, and appear related to resource availability. Xenopus laevis may strongly impact aquatic ecosystems because of its near complete aquatic lifestyle and its significant consumption of key taxa for the trophic relationships in ponds.

Impact of asynchronous emergence of two lethal pathogens on amphibian assemblages.

Emerging diseases have been increasingly associated with population declines, with co-infections exhibiting many types of interactions. The chytrid fungus (Batrachochytrium dendrobatidis) and ranaviruses have extraordinarily broad host ranges, however co-infection dynamics have been largely overlooked. We investigated the pattern of co-occurrence of these two pathogens in an amphibian assemblage in Serra da Estrela (Portugal). The detection of chytridiomycosis in Portugal was linked to population declines of midwife-toads (Alytes obstetricans). The asynchronous and subsequent emergence of a second pathogen - ranavirus - caused episodes of lethal ranavirosis. Chytrid effects were limited to high altitudes and a single host, while ranavirus was highly pathogenic across multiple hosts, life-stages and altitudinal range. This new strain (Portuguese newt and toad ranavirus - member of the CMTV clade) caused annual mass die-offs, similar in host range and rapidity of declines to other locations in Iberia affected by CMTV-like ranaviruses. However, ranavirus was not always associated with disease, mortality and declines, contrasting with previous reports on Iberian CMTV-like ranavirosis. We found little evidence that pre-existing chytrid emergence was associated with ranavirus and the emergence of ranavirosis. Despite the lack of cumulative or amplified effects, ranavirus drove declines of host assemblages and changed host community composition and structure, posing a grave threat to all amphibian populations.

Impacts of Climate Change on the Global Invasion Potential of the African Clawed Frog Xenopus laevis.

By altering or eliminating delicate ecological relationships, non-indigenous species are considered a major threat to biodiversity, as well as a driver of environmental change. Global climate change affects ecosystems and ecological communities, leading to changes in the phenology, geographic ranges, or population abundance of several species. Thus, predicting the impacts of global climate change on the current and future distribution of invasive species is an important subject in macroecological studies. The African clawed frog (Xenopus laevis), native to South Africa, possesses a strong invasion potential and populations have become established in numerous countries across four continents. The global invasion potential of X. laevis was assessed using correlative species distribution models (SDMs). SDMs were computed based on a comprehensive set of occurrence records covering South Africa, North America, South America and Europe and a set of nine environmental predictors. Models were built using both a maximum entropy model and an ensemble approach integrating eight algorithms. The future occurrence probabilities for X. laevis were subsequently computed using bioclimatic variables for 2070 following four different IPCC scenarios. Despite minor differences between the statistical approaches, both SDMs predict the future potential distribution of X. laevis, on a global scale, to decrease across all climate change scenarios. On a continental scale, both SDMs predict decreasing potential distributions in the species' native range in South Africa, as well as in the invaded areas in North and South America, and in Australia where the species has not been introduced. In contrast, both SDMs predict the potential range size to expand in Europe. Our results suggest that all probability classes will be equally affected by climate change. New regional conditions may promote new invasions or the spread of established invasive populations, especially in France and Great Britain.

Unequal contribution of native South African phylogeographic lineages to the invasion of the African clawed frog, Xenopus laevis, in Europe.

Due to both deliberate and accidental introductions, invasive African Clawed Frog (Xenopus laevis) populations have become established worldwide. In this study, we investigate the geographic origins of invasive X. laevis populations in France and Portugal using the phylogeographic structure of X. laevis in its native South African range. In total, 80 individuals from the whole area known to be invaded in France and Portugal were analysed for two mitochondrial and three nuclear genes, allowing a comparison with 185 specimens from the native range. Our results show that native phylogeographic lineages have contributed differently to invasive European X. laevis populations. In Portugal, genetic and historical data suggest a single colonization event involving a small number of individuals from the south-western Cape region in South Africa. In contrast, French invasive X. laevis encompass two distinct native phylogeographic lineages, i.e., one from the south-western Cape region and one from the northern regions of South Africa. The French X. laevis population is the first example of a X. laevis invasion involving multiple lineages. Moreover, the lack of population structure based on nuclear DNA suggests a potential role for admixture within the invasive French population.

Evaluating green infrastructure in urban environments using a multi-taxa and functional diversity approach.

Forested areas within cities host a large number of species, responsible for many ecosystem services in urban areas. The biodiversity in these areas is influenced by human disturbances such as atmospheric pollution and urban heat island effect. To ameliorate the effects of these factors, an increase in urban green areas is often considered sufficient. However, this approach assumes that all types of green cover have the same importance for species. Our aim was to show that not all forested green areas are equal in importance for species, but that based on a multi-taxa and functional diversity approach it is possible to value green infrastructure in urban environments. After evaluating the diversity of lichens, butterflies and other-arthropods, birds and mammals in 31 Mediterranean urban forests in south-west Europe (Almada, Portugal), bird and lichen functional groups responsive to urbanization were found. A community shift (tolerant species replacing sensitive ones) along the urbanization gradient was found, and this must be considered when using these groups as indicators of the effect of urbanization. Bird and lichen functional groups were then analyzed together with the characteristics of the forests and their surroundings. Our results showed that, contrary to previous assumptions, vegetation density and more importantly the amount of urban areas around the forest (matrix), are more important for biodiversity than forest quantity alone. This indicated that not all types of forested green areas have the same importance for biodiversity. An index of forest functional diversity was then calculated for all sampled forests of the area. This could help decision-makers to improve the management of urban green infrastructures with the goal of increasing functionality and ultimately ecosystem services in urban areas.

Rapid evolution of constitutive and inducible defenses against an invasive predator.

Invasive alien predators can impose strong selection on native prey populations and induce rapid evolutionary change in the invaded communities. However, studies on evolutionary responses to invasive predators are often complicated by the lack of replicate populations differing in coexistence time with the predator, which would allow the determination of how prey traits change during the invasion. The red swamp crayfish Procambarus clarkii has invaded many freshwater areas worldwide, with negative impacts for native fauna. Here, we examined how coexistence time shapes antipredator responses of the Iberian waterfrog (Pelophylax perezi) to the invasive crayfish by raising tadpoles from five populations differing in historical exposure to P. clarkii (30 years, 20 years, or no coexistence). Tadpoles from non-invaded populations responded to the presence of P. clarkii with behavioral plasticity (reduced activity), whereas long-term invaded populations showed canalized antipredator behavior (constant low activity level). Tadpoles from one of the long-term invaded populations responded to the crayfish with inducible morphological defenses (deeper tails), reflecting the use of both constitutive and inducible antipredator defenses against the exotic predator by this population. Our results suggest that, while naive P. perezi populations responded behaviorally to P. clarkii, the strong predation pressure imposed by the crayfish has induced the evolution of qualitatively different antipredator defenses in populations with longer coexistence time. These responses suggest that strong selection by invasive predators may drive rapid evolutionary change in invaded communities. Examining responses of prey species to biological invasions using multiple populations will help us better forecast the impact of invasive predators in natural communities.

Morphological and life-history responses of anurans to predation by an invasive crayfish: an integrative approach.

Predator-induced phenotypic plasticity has been widely documented in response to native predators, but studies examining the extent to which prey can respond to exotic invasive predators are scarce. As native prey often do not share a long evolutionary history with invasive predators, they may lack defenses against them. This can lead to population declines and even extinctions, making exotic predators a serious threat to biodiversity. Here, in a community-wide study, we examined the morphological and life-history responses of anuran larvae reared with the invasive red swamp crayfish, Procambarus clarkii, feeding on conspecific tadpoles. We reared tadpoles of nine species until metamorphosis and examined responses in terms of larval morphology, growth, and development, as well as their degree of phenotypic integration. These responses were compared with the ones developed in the presence of a native predator, the larval dragonfly Aeshna sp., also feeding on tadpoles. Eight of the nine species altered their morphology or life history when reared with the fed dragonfly, but only four when reared with the fed crayfish, suggesting among-species variation in the ability to respond to a novel predator. While morphological defenses were generally similar across species (deeper tails) and almost exclusively elicited in the presence of the fed dragonfly, life-history responses were very variable and commonly elicited in the presence of the invasive crayfish. Phenotypes induced in the presence of dragonfly were more integrated than in crayfish presence. The lack of response to the presence of the fed crayfish in five of the study species suggests higher risk of local extinction and ultimately reduced diversity of the invaded amphibian communities. Understanding how native prey species vary in their responses to invasive predators is important in predicting the impacts caused by newly established predator-prey interactions following biological invasions.