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The Earth Hologenome Initiative (EHI) is a global collaboration to generate and analyse hologenomic data from wild animals and associated microorganisms using standardised methodologies underpinned by open and inclusive research principles. Initially focused on vertebrates, it aims to re-examine ecological and evolutionary questions by studying host–microbiota interactions from a systemic perspective.

Invasive alien species have widespread impacts on native biodiversity and ecosystem services. Since the number of introductions worldwide is continuously rising, it is essential to prevent the entry, establishment and spread of new alien species through a systematic examination of future potential threats. Applying a three-step horizon scanning consensus method, we evaluated non-established alien species that could potentially arrive, establish and cause major ecological impact in Spain within the next 10 years. Overall, we identified 47 species with a very high risk (e.g. Oreochromis niloticus, Popillia japonica, Hemidactylus frenatus, Crassula helmsii or Halophila stipulacea), 61 with high risk, 93 with moderate risk, and 732 species with low risk. Many of the species categorized as very high or high risk to Spanish biodiversity are either already present in Europe and neighbouring countries or have a long invasive history elsewhere. This study provides an updated list of potential invasive alien species useful for prioritizing efforts and resources against their introduction. Compared to previous horizon scanning exercises in Spain, the current study screens potential invaders from a wider range of terrestrial, freshwater, and marine organisms, and can serve as a basis for more comprehensive risk analyses to improve management and increase the efficiency of the early warning and rapid response framework for invasive alien species. We also stress the usefulness of measuring agreement and consistency as two different properties of the reliability of expert scores, in order to more easily elaborate consensus ranked lists of potential invasive alien species.

The genus Karyolysus was originally proposed to accommodate blood parasites of lacertid lizards in Western Europe. However, recent phylogenetic analyses suggested an inconclusive taxonomic position of these parasites of the order Adeleorina based on the available genetic information. Inconsistencies between molecular phylogeny, morphology, and/or life cycles can reflect lack of enough genetic informa- tion of the target group. We therefore surveyed 28 localities and collected blood samples from 828 lizards of 23 species including lacertids, skinks, and geckoes in the western Mediterranean, North Africa, and Macaronesia, where species of Karyolysus and other adeleorine parasites have been described. We com- bined molecular and microscopic methods to analyze the samples, including those from the host type species and the type locality of Karyolysus bicapsulatus. The phylogenetic relationship of these parasites was analyzed based on the 18S rRNA gene and the co-phylogenetic relationship with their vertebrate hosts was reconstructed. We molecularly detected adeleorine parasites in 37.9% of the blood samples and found 22 new parasite haplotypes. A phylogenetic reconstruction with 132 sequences indicated that 20 of the newly detected haplotypes clustered in a well-supported clade with another 18 sequences that included Karyolysus galloti and Karyolysus lacazei. Morphological evidence also supported that K. bicapsu- latus clustered in this monophyletic clade. These results supported the taxonomic validity of the genus. In addition, we found some parasite haplotypes that infected different lizard host genera with ancient diverging histories, which suggested that Karyolysus is less host-specific than other blood parasites of lizards in the region. A co-phylogenetic analysis supported this interpretation because no significant co-speciation signal was shown between Karyolysus and lizard hosts.

Rapid human-induced transformation of habitats is presenting organisms with challenging novel environmental conditions. Nowadays, extensive areas of the world are covered by exotic plantations, and among these, eucalypt plantations stand out for their worldwide occurrence. These novel environments probably threaten the persistence of native species, with several studies reporting biodiversity losses in these transformed areas. However, the effects of such habitat alteration on life-history traits that potentially constrain the ecology of individuals have been poorly explored. 2. In a mesocosm experiment, we exposed tadpoles of three anuran species (Rana temporaria, Alytes obstetricans, and Pelophylax perezi) to the leachates of natural oak trees and exotic eucalypt trees to explore how the transformation of natural forests into eucalypt plantation affects the expression of behavioural and morphological traits and immune response. 3. While anti-predatory responses appeared not to be impacted by leachate type, tadpoles exposed to eucalypt leachates differed in morphology and growth, and exhibited weaker immune responses and smaller size at metamorphosis than those exposed to oak leachates. Some of the responses were also species-specific, suggesting that some species are likely to be more sensitive than others to novel chemicals. 4. We conclude that the poor quality of eucalypt leaf litter (e.g., its low nutrient content), low amount of food (e.g., periphyton), and the presence of toxic detritus and leachates can have important stress-related consequences for the critical early stages of development in anurans

Environmental conditions experienced by a species during its evolutionary history may shape the signals it uses for communication. Consequently, rapid environmental changes may lead to less effective signals, which interfere with communication between individuals, altering life history traits such as predator detection and mate searching. Increased temperature can reduce the efficacy of scent marks released by male lizards, but the extent to which this negative effect is related to specific biological traits and evolutionary histories across species and populations have not been explored. We experimentally tested how increased temperature affects the efficacy of chemical signals of high‐ and low‐altitude populations of three lizard species that differ in their ecological requirements and altitudinal distributions. We tested the behavioral chemosensory responses of males from each species and population to male scent marks that had been incubated at one of two temperatures (cold 16°C or hot 20°C). In high‐altitude populations of a mountain species (Iberolacerta monticola), the efficacy of chemical signals (i.e., latency time and number of tongue flicks) was lower after scent marks had been exposed to a hot temperature. The temperature that scent marks were incubated at did not affect the efficacy of chemical signals in a ubiquitous species (Podarcis muralis) or another mountain species (I. bonalli). Our results suggest that specific ecological traits arising through local adaptation to restricted distributions may be important in determining species vulnerability to climatic change.

Previous studies suggest that immune-challenged individuals need to allocate resources to the immune system to combat infection, reducing escape ability and increasing the vulnerability of infected individuals to predators. Such behavioural responses might change in anthropogenic habitats where the balance between predation risk and countering infections can differ from that in natural habitats. We experimentally explored whether fighting an infection has an effect on hiding behaviour in a frog (Pelophylax perezi), and whether frogs from urban ponds respond differently from those from natural forested ponds. Independently of habitat type, we found no evidence for changes in hiding behaviour of male frogs before and after they were immune challenged (injected with lipopolysaccharide) or not (injected with phosphate-buffered saline). We also explored whether immune response (phytohaemagglutinin assay) or morphological traits (head width, body condition, body size and length) were related to variation in hiding time, but we did not find a relationship between the measured traits and behavioural response for males from either habitat. In conclusion, we show that frog anti-predator behaviour is not always affected by an immune challenge and the effect of urban habitats can be weaker than expected, at least in species that could be pre-adapted to the novel environmental conditions.

Chemical communication in aquatic species can affect many key life history traits, such as prey and predator detection and mate searching. However, changes in the environment can disrupt the effectiveness of signals and the ability of individuals to detect these signals. Many studies have examined the effect of secondary compounds from exotic plants on the ecology and physiology of a range of taxa, but whether the replacement of natural forests with exotic trees influences the behavioural responses of animals by disrupting chemical communication has rarely been investigated. We experimentally tested how eucalypt tree chemicals influenced three key aspects of chemical communication in adult male palmate newts. We tested for effects of both exposure to eucalypt water (i.e. extracts obtained by soaking leaves in mineral water) and the origin of newts (eucalypt plantations and natural oak forests). We examined whether exposure to eucalypt water altered the chemosensory ability of males to detect pools containing females or conspecific alarm cues and to find food. We found that eucalypt leachates had different effects on each behavioural trait. Fewer males detected female chemicals when exposed to the eucalypt than the oak water treatment, independent of the males' habitat of origin. Newts from oak forest were less able to detect conspecific alarm cues signalling predatory events when exposed to eucalypt water than when exposed to oak water, or than newts from eucalypt plantations for either water treatment. The ability of males to find food using chemical cues was similar in oak and eucalypt treatments. Our results suggest that chemical compounds not previously encountered during the evolutionary history of the species can influence the ability to respond to predators and locate mates. Future studies should explore the fitness costs associated with a reduced ability to respond to predators or detect mates.

Selection can favour phenotypic plasticity in mate choice in response to environmental factors that alter the costs and benefits of being choosy, or of choosing specific mates. Human-induced environmental change could alter sexual selection by affecting the costs of mate choice, or by impairing the ability of individuals to identify preferred mates. For example, variation in mate choice could be driven by environmentally induced differences in body condition (e.g. health) that change the cost of choosiness, or by environmental effects on the ability to detect or discriminate sexual signals. We teased apart these possibilities experimentally, by comparing female mate choice in the palmate newt Lissotriton helveticus between environments that mimic water from either native oak forests or exotic eucalypt plantations. In laboratory two-choice mate trials in clean water, females with prolonged exposure (21 days) to waterborne chemicals leached from eucalypt leaves did not preferentially associate with the male with a stronger immune response, but females exposed to water with chemicals from oak leaves did. In contrast, female choice was unaffected by the immediate presence or absence of eucalypt leachates during mate choice (using only females previously held in oak-treated water). The habitat-related change in female choice we observed is likely to be driven by effects of eucalypt leachates on female physiology, rather than immediate inhibition of pheromone transmission or blocking of pheromone reception.

Chemical communication in aquatic species can affect many key life history traits, such as prey and predator detection and mate searching. However, changes in the environment can disrupt the effectiveness of signals and the ability of individuals to detect these signals. Many studies have examined the effect of secondary compounds from exotic plants on the ecology and physiology of a range of taxa, but whether the replacement of natural forests with exotic trees influences the behavioural responses of animals by disrupting chemical communication has rarely been investigated. We experimentally tested how eucalypt tree chemicals influenced three key aspects of chemical communication in adult male palmate newts. We tested for effects of both exposure to eucalypt water (i.e. extracts obtained by soaking leaves in mineral water) and the origin of newts (eucalypt plantations and natural oak forests). We examined whether exposure to eucalypt water altered the chemosensory ability of males to detect pools containing females or conspecific alarm cues and to find food. We found that eucalypt leachates had different effects on each behavioural trait. Fewer males detected female chemicals when exposed to the eucalypt than the oak water treatment, independent of the males’ habitat of origin. Newts from oak forest were less able to detect conspecific alarm cues signalling predatory events when exposed to eucalypt water than when exposed to oak water, or than newts from eucalypt plantations for either water treatment. The ability of males to find food using chemical cues was similar in oak and eucalypt treatments. Our results suggest that chemical compounds not previously encountered during the evolutionary history of the species can influence the ability to respond to predators and locate mates. Future studies should explore the fitness costs associated with a reduced ability to respond to predators or detect mates.

Background The optimal allocation of resources to sexual signals and other life history traits is usually dependent on an individual’s condition, while variation in the expression of sexual traits across environments depends on the combined effects of local adaptation, mean condition, and phenotypic responses to environment-specific cues that affect resource allocation. A clear contrast can often be drawn between natural habitats and novel habitats, such as forest plantations and urban areas. In some species, males seem to change their sexual signals in these novel environments, but why this occurs and how it affects signal reliability is still poorly understood. Results The relative size of sexual traits and level of immune responses were significantly lower for male palmate newts Lissotriton helveticus caught in pine and eucalyptus plantations compared to those caught in native forests, but there was no habitat-dependent difference in body condition (n = 18 sites, 382 males). The reliability with which sexual traits signalled body condition and immune responses was the same in all three habitats. Finally, we conducted a mesocosm experiment in which males were maintained in pine, eucalypt or oak infused water for 21 days. Males in plantation-like water (pine or eucalypt) showed significantly lower immune responses but no change in body condition. This matches the pattern seen for field-caught males. Unlike field-caught males, however, there was no relationship between water type and relative sexual trait size. Conclusions Pine and eucalyptus plantations are likely to be detrimental to male palmate newt because they are associated with reduced immune function and smaller sexual traits. This could be because ecological aspects of these novel habitats, such as high water turbidity or changes in male-male competition, drive selection for reduced investment into sexual traits. However, it is more probable that there are differences in the ease of acquisition, hence opti

Background: The optimal allocation of resources to sexual signals and other life history traits is usually dependent on an individual's condition, while variation in the expression of sexual traits across environments depends on the combined effects of local adaptation, mean condition, and phenotypic responses to environment-specific cues that affect resource allocation. A clear contrast can often be drawn between natural habitats and novel habitats, such as forest plantations and urban areas. In some species, males seem to change their sexual signals in these novel environments, but why this occurs and how it affects signal reliability is still poorly understood. Results: The relative size of sexual traits and level of immune responses were significantly lower for male palmate newts Lissotriton helveticus caught in pine and eucalyptus plantations compared to those caught in native forests, but there was no habitat-dependent difference in body condition (n= 18 sites, 382 males). The reliability with which sexual traits signalled body condition and immune responses was the same in all three habitats. Finally, we conducted a mesocosm experiment in which males were maintained in pine, eucalypt or oak infused water for 21 days. Males in plantation-like water (pine or eucalypt) showed significantly lower immune responses but no change in body condition. This matches the pattern seen for field-caught males. Unlike field-caught males, however, there was no relationship between water type and relative sexual trait size. Conclusions: Pine and eucalyptus plantations are likely to be detrimental to male palmate newt because they are associated with reduced immune function and smaller sexual traits. This could be because ecological aspects of these novel habitats, such as high water turbidity or changes in male-male competition, drive selection for reduced investment into sexual traits. However, it is more probable that there are differences in the ease of acquisition, hence op

Chemical signals play an important role in intraspecific communication and social organization of many animals, but they also may be useful in interspecific recognition. In lizards, chemical signals are often contained in femoral gland secretions, of which composition may vary between species and populations. This may be especially important in recognition and reproductive isolation between closely related species. We analyzed by gas chromatography-mass spectrometry (GC-MS) the lipophilic fraction of femoral gland secretions of two closely related wall lizard species, Podarcis bocagei and Podarcis carbonelli to test for possible interspecific differences in chemical composition. We found 56 lipophilic compounds in femoral gland secretions of male P. bocagei and 60 in P. carbonelli. The main compounds were steroids and waxy esters, but we also found carboxylic acids and their esters, alcohols, amydes, aldehydes, squalene, ketones and furanones. There were significant differences between species with respect to the number and relative proportions of compounds. Differences in chemical composition might be a consequence of phylogenetic differences per se, but they could also be explained by ecological adaptation to different microclimatic conditions. These differences in chemical profiles may explain the known chemosensory interspecific recognition between these two lizards, contributing to their reproductive isolation.

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Background The optimal allocation of resources to sexual signals and other life history traits is usually dependent on an individual’s condition, while variation in the expression of sexual traits across environments depends on the combined effects of local adaptation, mean condition, and phenotypic responses to environment-specific cues that affect resource allocation. A clear contrast can often be drawn between natural habitats and novel habitats, such as forest plantations and urban areas. In some species, males seem to change their sexual signals in these novel environments, but why this occurs and how it affects signal reliability is still poorly understood., Results The relative size of sexual traits and level of immune responses were significantly lower for male palmate newts Lissotriton helveticus caught in pine and eucalyptus plantations compared to those caught in native forests, but there was no habitat-dependent difference in body condition (n = 18 sites, 382 males). The reliability with which sexual traits signalled body condition and immune responses was the same in all three habitats. Finally, we conducted a mesocosm experiment in which males were maintained in pine, eucalypt or oak infused water for 21 days. Males in plantation-like water (pine or eucalypt) showed significantly lower immune responses but no change in body condition. This matches the pattern seen for field-caught males. Unlike field-caught males, however, there was no relationship between water type and relative sexual trait size., Conclusions Pine and eucalyptus plantations are likely to be detrimental to male palmate newt because they are associated with reduced immune function and smaller sexual traits. This could be because ecological aspects of these novel habitats, such as high water turbidity or changes in male-male competition, drive selection for reduced investment into sexual traits. However, it is more probable that there are differences in the ease of acquisition, hence optimal allocation, of resources among habitats. Our mesocosm experiment also provides some evidence that water toxicity is a causal factor. Our findings offer insights into how plantations affect amphibian life histories, and how novel habitats might generate long-term selection for new resource allocation strategies in native species.