Spring temperature drives phenotypic selection on plasticity of flowering time.

In seasonal environments, a high responsiveness of development to increasing temperatures in spring can infer benefits in terms of a longer growing season, but also costs in terms of an increased risk of facing unfavourable weather conditions. Still, we know little about how climatic conditions influence the optimal plastic response. Using 22 years of field observations for the perennial forest herb Lathyrus vernus, we assessed phenotypic selection on among-individual variation in reaction norms of flowering time to spring temperature, and examined if among-year variation in selection on plasticity was associated with spring temperature conditions. We found significant among-individual variation in mean flowering time and flowering time plasticity, and that plants that flowered earlier also had a more plastic flowering time. Selection favoured individuals with an earlier mean flowering time and a lower thermal plasticity of flowering time. Less plastic individuals were more strongly favoured in colder springs, indicating that spring temperature influenced optimal flowering time plasticity. Our results show how selection on plasticity can be linked to climatic conditions, and illustrate how we can understand and predict evolutionary responses of organisms to changing environmental conditions.

Impact of patch age and size on forest soil characteristics in European agricultural landscapes.

Many landscapes worldwide are characterized by the presence of a mosaic of forest patches with contrasting age and size embedded in a matrix of agricultural land. However, our understanding of the effects of these key forest patch features on the soil nutrient status (in terms of nitrogen, carbon, and phosphorus) and soil pH is still limited due to a lack of large-scale data. To address this research gap, we analyzed 830 soil samples from nearly 200 forest patches varying in age (recent versus ancient forests) and size (small versus larger patches) along a 2500-km latitudinal gradient across Europe. We also considered environmental covariates at multiple scales to increase the generality of our research, including variation in macroclimate, nitrogen deposition rates, forest cover in a buffer zone, basal area and soil type. Multiple linear mixed-effects models were performed to test the combined effects of patch features and environmental covariates on soil nutrients and pH. Recent patches had higher total soil phosphorus concentrations and stocks in the mineral soil layer, along with a lower nitrogen to phosphorus ratio within that layer. Small patches generally had a higher mineral soil pH. Mineral soil nitrogen stocks were lower in forest patches with older age and larger size, as a result of a significant interactive effect. Additionally, environmental covariates had significant effects on soil nutrients, including carbon, nitrogen, phosphorus, and their stoichiometry, depending on the specific covariates. In some cases, the effect of patch age on mineral soil phosphorus stocks was greater than that of environmental covariates. Our findings underpin the important roles of forest patch age and size for the forest soil nutrient status. Long-term studies assessing edge effects and soil development in post-agricultural forests are needed, especially in a context of changing land use and climate.

Maladaptive plastic responses of flowering time to geothermal heating.

Phenotypic plasticity might increase fitness if the conditions under which it evolved remain unaltered, but becomes maladaptive if the environment no longer provides reliable cues for subsequent conditions. In seasonal environments, timing of reproduction can respond plastically to spring temperature, maximizing the benefits of a long season while minimizing the exposure to unfavorable cold temperatures. However, if the relationship between early spring temperatures and later conditions changes, the optimal response might change. In geothermally heated ecosystems, the plastic response of flowering time to springtime soil temperature that has evolved in unheated areas is likely to be non-optimal, because soil temperatures are higher and decoupled from air temperatures in heated areas. We therefore expect natural selection to favor a lower plasticity and a delayed flowering in these areas. Using observational data along a natural geothermal warming gradient, we tested the hypothesis that selection on flowering time depends on soil temperature and favors later flowering on warmer soils in the perennial Cerastium fontanum. In both study years, plants growing in warmer soils began flowering earlier than plants growing in colder soils, suggesting that first flowering date (FFD) responds plastically to soil temperature. In one of the two study years, selection favored earlier flowering in colder soils but later flowering in warmer soils, suggesting that the current level of plastic advance of FFD on warmer soils may be maladaptive in some years. Our results illustrate the advantages of using natural experiments, such as geothermal ecosystems, to examine selection in environments that recently have undergone major changes. Such knowledge is essential to understand and predict both ecological and evolutionary responses to climate warming.

Selection against early flowering in geothermally heated soils is associated with pollen but not prey availability in a carnivorous plant.

PREMISE: In high-latitude environments, plastic responses of phenology to increasing spring temperatures allow plants to extend growing seasons while avoiding late frosts. However, evolved plasticity might become maladaptive if climatic conditions change and spring temperatures no longer provide reliable cues for conditions important for fitness. Maladaptative phenological responses might be related to both abiotic factors and mismatches with interacting species. When mismatches arise, we expect selection to favor changes in phenology. METHODS: We combined observations along a soil temperature gradient in a geothermally heated area with pollen and prey supplementation experiments and examined how phenotypic selection on flowering time in the carnivorous plant Pinguicula vulgaris depends on soil temperature, and pollen and prey availability. RESULTS: Flowering advanced and fitness decreased with increasing soil temperature. However, in pollen-supplemented plants, fitness instead increased with soil temperature. In heated soils, there was selection favoring later flowering, while earlier flowering was favored in unheated soils. This pattern remained also after artificially increasing pollen and prey availability. CONCLUSIONS: Plant-pollinator mismatches can be an important reason why evolved plastic responses of flowering time to increasing spring temperatures become maladaptive under novel environmental conditions, and why there is selection to delay flowering. In our study, selection for later flowering remained after artificially increasing pollen availability, suggesting that abiotic factors also contribute to the observed selection. Identifying the factors that make evolved phenological responses maladaptive under novel conditions is fundamental for understanding and predicting evolutionary responses to climate warming.

Plant-animal interactions mediate climatic effects on selection on flowering time.

Selection on flowering time in plants is often mediated by multiple agents, including climatic conditions and the intensity of mutualistic and antagonistic interactions with animals. These selective agents can have both direct and indirect effects. For example, climate might not only influence phenotypic selection on flowering time directly by affecting plant physiology, but it can also alter selection indirectly by modifying the seasonal activity and relative timing of animals interacting with plants. We used 21 yr of data to identify the drivers of selection on flowering time in the perennial herb Lathyrus vernus, and to examine if antagonistic plant-animal interactions mediate effects of climate on selection. We examined the fitness consequences of vertebrate grazing and predispersal seed predation, and how these effects varied among years and among individuals within years. Although both antagonistic plant-animal interactions had important negative effects on plant fitness, only grazing intensity was consistently related to plant phenology, being higher in early-flowering individuals. Spring temperature influenced the intensity of both plant-animal interactions, as well as the covariance between seed predation and plant phenology. However, only differences in grazing intensity among years were associated with differences in selection on flowering time; the strength of selection for early flowering being stronger in years with lower mean intensity of grazing. Our results illustrate how climatic conditions can influence plant-animal interactions that are important selective agents for plant traits. A broader implication of our findings is that both ecological and evolutionary responses to climatic changes might be indirect, and largely mediated by species interactions.

Climate drives among-year variation in natural selection on flowering time.

To predict long-term responses to climate change, we need to understand how changes in temperature and precipitation elicit both immediate phenotypic responses and changes in natural selection. We used 22 years of data for the perennial herb Lathyrus vernus to examine how climate influences flowering phenology and phenotypic selection on phenology. Plants flowered earlier in springs with higher temperatures and higher precipitation. Early flowering was associated with a higher fitness in nearly all years, but selection for early flowering was significantly stronger in springs with higher temperatures and lower precipitation. Climate influenced selection through trait distributions, mean fitness and trait-fitness relationships, the latter accounting for most of the among-year variation in selection. Our results show that climate both induces phenotypic responses and alters natural selection, and that the change in the optimal phenotype might be either weaker, as for spring temperature, or stronger, as for precipitation, than the optimal response.

Multimodal analgesia for treatment of allodynia and hyperalgesia after major trauma in a cat.

CASE SUMMARY: A 2-year-old polytraumatized male cat was admitted to a teaching hospital for correction of a defective inguinal herniorrhaphy. Upon arrival, the cat showed signs of neuropathic pain, including allodynia and hyperalgesia. Analgesic therapy was initiated with methadone and metamizole; however, 24 h later, the signs of pain continued. Reparative surgery was performed, and a multimodal analgesic regimen was administered (methadone, ketamine, wound catheter and epidural anesthesia). Postoperatively, the cat showed signs of severe pain, assessed using the UNESP-Botucatu multidimensional composite pain scale. Rescue analgesia was initiated, which included methadone, bupivacaine (subcutaneous wound-diffusion catheter) and transversus abdominis plane block. Because the response was incomplete, co-adjuvant therapy (pregabalin and electroacupuncture) was then implemented. Fourteen days after admission, the patient was discharged with oral tramadol and pregabalin for at-home treatment. RELEVANCE AND NOVEL INFORMATION: Neuropathic pain is caused by a primary lesion or dysfunction in the nervous system and is a well-described complication following trauma, surgical procedures such as hernia repair, and inadequate analgesia. The aims of this report are to: (1) describe a presentation of neuropathic pain to highlight the recognition of clinical signs such as allodynia and hyperalgesia in cats; and (2) describe treatment of multi-origin, severe, long-standing, 'mixed' pain (acute inflammatory with a neuropathic component). The patient was managed using multiple analgesic strategies (multimodal analgesia), including opioids, non-steroidal anti-inflammatory drugs, locoregional anesthesia, co-adjuvant drugs and non-pharmacological therapy (electroacupuncture).

Resource overlap and dilution effects shape host plant use in a myrmecophilous butterfly.

The effects of consumers on fitness of resource organisms are a complex function of the spatio-temporal distribution of the resources, consumer functional responses and trait preferences, and availability of other resources. The ubiquitous variation in the intensity of species interactions has important consequences for the ecological and evolutionary dynamics of natural populations. Nevertheless, little is known about the processes causing this variation and their operational scales. Here, we examine how variation in the intensity of a consumer-resource interaction is related to resource timing, resource density and abundance of other resources. Using the butterfly consumer Phengaris alcon and its two sequential resources, the host plant Gentiana pneumonanthe and the host ants Myrmica spp., we investigated how butterfly egg-laying depended on focal host plant phenology, density and phenology of neighbouring host plants and host ant abundance. Butterflies preferred plants that simultaneously maximized the availability of both larval resources in time and space, that is, they chose early-flowering plants that were of higher nutritional quality for larvae where host ants were abundant. Both the probability of oviposition and the number of eggs were lower in plant individuals with a high neighbour density than in more isolated plants, and this dilution effect was stronger when neighbours flowered early. Our results show that plant-herbivore interactions simultaneously depend on the spatio-temporal distribution of a focal resource and on the small-scale spatial variation in the abundance of other herbivore resources. Given that consumers have negative effects on fitness and prefer certain timing of the resource organisms, this implies that processes acting at the levels of individuals, populations and communities simultaneously contribute to variation in consumer-mediated natural selection.

A natural heating experiment: Phenotypic and genotypic responses of plant phenology to geothermal soil warming.

Under global warming, the survival of many populations of sedentary organisms in seasonal environments will largely depend on their ability to cope with warming in situ by means of phenotypic plasticity or adaptive evolution. This is particularly true in high-latitude environments, where current growing seasons are short, and expected temperature increases large. In such short-growing season environments, the timing of growth and reproduction is critical to survival. Here, we use the unique setting provided by a natural geothermal soil warming gradient (Hengill geothermal area, Iceland) to study the response of Cerastium fontanum flowering phenology to temperature. We hypothesized that trait expression and phenotypic selection on flowering phenology are related to soil temperature, and tested the hypothesis that temperature-driven differences in selection on phenology have resulted in genetic differentiation using a common garden experiment. In the field, phenology was related to soil temperature, with plants in warmer microsites flowering earlier than plants at colder microsites. In the common garden, plants responded to spring warming in a counter-gradient fashion; plants originating from warmer microsites flowered relatively later than those originating from colder microsites. A likely explanation for this pattern is that plants from colder microsites have been selected to compensate for the shorter growing season by starting development at lower temperatures. However, in our study we did not find evidence of variation in phenotypic selection on phenology in relation to temperature, but selection consistently favoured early flowering. Our results show that soil temperature influences trait expression and suggest the existence of genetically based variation in flowering phenology leading to counter-gradient local adaptation along a gradient of soil temperatures. An important implication of our results is that observed phenotypic responses of phenology to global warming might often be a combination of short-term plastic responses and long-term evolutionary responses, acting in different directions.

Promoting biodiversity values of small forest patches in agricultural landscapes: Ecological drivers and social demand.

Small forest patches embedded in agricultural (and peri-urban) landscapes in Western Europe play a key role for biodiversity conservation with a recognized capacity of delivering a wide suite of ecosystem services. Measures aimed to preserve these patches should be both socially desirable and ecologically effective. This study presents a joint ecologic and economic assessment conducted on small forest patches in Flanders (Belgium) and Picardie (N France). In each study region, two contrasted types of agricultural landscapes were selected. Open field (OF) and Bocage (B) landscapes are distinguished by the intensity of their usage and higher connectivity in the B landscapes. The social demand for enhancing biodiversity and forest structure diversity as well as for increasing the forest area at the expenses of agricultural land is estimated through an economic valuation survey. These results are compared with the outcomes of an ecological survey where the influence of structural features of the forest patches on the associated herbaceous diversity is assessed. The ecological and economic surveys show contrasting results; increasing tree species richness is ecologically more important for herbaceous diversity in the patch, but both tree species richness and herbaceous diversity obtain insignificant willingness to pay estimates. Furthermore, although respondents prefer the proposed changes to take place in the region where they live, we find out that social preferences and ecological effectiveness do differ between landscapes that represent different intensities of land use. Dwellers where the landscape is perceived as more "degraded" attach more value to diversity enhancement, suggesting a prioritization of initiatives in these area. In contrast, the ecological analyses show that prioritizing the protection and enhancement of the relatively better-off areas is more ecologically effective. Our study calls for a balance between ecological effectiveness and welfare benefits, suggesting that cost effectiveness studies should consider these approaches jointly.

Habitat properties are key drivers of Borrelia burgdorferi (s.l.) prevalence in Ixodes ricinus populations of deciduous forest fragments.

BACKGROUND: The tick Ixodes ricinus has considerable impact on the health of humans and other terrestrial animals because it transmits several tick-borne pathogens (TBPs) such as B. burgdorferi (sensu lato), which causes Lyme borreliosis (LB). Small forest patches of agricultural landscapes provide many ecosystem services and also the disservice of LB risk. Biotic interactions and environmental filtering shape tick host communities distinctively between specific regions of Europe, which makes evaluating the dilution effect hypothesis and its influence across various scales challenging. Latitude, macroclimate, landscape and habitat properties drive both hosts and ticks and are comparable metrics across Europe. Therefore, we instead assess these environmental drivers as indicators and determine their respective roles for the prevalence of B. burgdorferi in I. ricinus. METHODS: We sampled I. ricinus and measured environmental properties of macroclimate, landscape and habitat quality of forest patches in agricultural landscapes along a European macroclimatic gradient. We used linear mixed models to determine significant drivers and their relative importance for nymphal and adult B. burgdorferi prevalence. We suggest a new prevalence index, which is pool-size independent. RESULTS: During summer months, our prevalence index varied between 0 and 0.4 per forest patch, indicating a low to moderate disservice. Habitat properties exerted a fourfold larger influence on B. burgdorferi prevalence than macroclimate and landscape properties combined. Increasingly available ecotone habitat of focal forest patches diluted and edge density at landscape scale amplified B. burgdorferi prevalence. Indicators of habitat attractiveness for tick hosts (food resources and shelter) were the most important predictors within habitat patches. More diverse and abundant macro- and microhabitat had a diluting effect, as it presumably diversifies the niches for tick-hosts and decreases the probability of contact between ticks and their hosts and hence the transmission likelihood. CONCLUSIONS: Diluting effects of more diverse habitat patches would pose another reason to maintain or restore high biodiversity in forest patches of rural landscapes. We suggest classifying habitat patches by their regulating services as dilution and amplification habitat, which predominantly either decrease or increase B. burgdorferi prevalence at local and landscape scale and hence LB risk. Particular emphasis on promoting LB-diluting properties should be put on the management of those habitats that are frequently used by humans. In the light of these findings, climate change may be of little concern for LB risk at local scales, but this should be evaluated further.

Environmental drivers of Ixodes ricinus abundance in forest fragments of rural European landscapes.

BACKGROUND: The castor bean tick (Ixodes ricinus) transmits infectious diseases such as Lyme borreliosis, which constitutes an important ecosystem disservice. Despite many local studies, a comprehensive understanding of the key drivers of tick abundance at the continental scale is still lacking. We analyze a large set of environmental factors as potential drivers of I. ricinus abundance. Our multi-scale study was carried out in deciduous forest fragments dispersed within two contrasting rural landscapes of eight regions, along a macroclimatic gradient stretching from southern France to central Sweden and Estonia. We surveyed the abundance of I. ricinus, plant community composition, forest structure and soil properties and compiled data on landscape structure, macroclimate and habitat properties. We used linear mixed models to analyze patterns and derived the relative importance of the significant drivers. RESULTS: Many drivers had, on their own, either a moderate or small explanatory value for the abundance of I. ricinus, but combined they explained a substantial part of variation. This emphasizes the complex ecology of I. ricinus and the relevance of environmental factors for tick abundance. Macroclimate only explained a small fraction of variation, while properties of macro- and microhabitat, which buffer macroclimate, had a considerable impact on tick abundance. The amount of forest and the composition of the surrounding rural landscape were additionally important drivers of tick abundance. Functional (dispersules) and structural (density of tree and shrub layers) properties of the habitat patch played an important role. Various diversity metrics had only a small relative importance. Ontogenetic tick stages showed pronounced differences in their response. The abundance of nymphs and adults is explained by the preceding stage with a positive relationship, indicating a cumulative effect of drivers. CONCLUSIONS: Our findings suggest that the ecosystem disservices of tick-borne diseases, via the abundance of ticks, strongly depends on habitat properties and thus on how humans manage ecosystems from the scale of the microhabitat to the landscape. This study stresses the need to further evaluate the interaction between climate change and ecosystem management on I. ricinus abundance.

Caterpillar seed predators mediate shifts in selection on flowering phenology in their host plant.

Variation in selection among populations and years has important implications for evolutionary trajectories of populations. Yet, the agents of selection causing this variation have rarely been identified. Selection on the time of reproduction within a season in plants might differ both among populations and among years, and selection can be mediated by both mutualists and antagonists. We investigated if differences in the direction of phenotypic selection on flowering phenology among 20 populations of Gentiana pneumonanthe during 2 yr were related to the presence of the butterfly seed predator Phengaris alcon, and if butterfly incidence was associated with the abundance of the butterfly's second host, Myrmica ants. In plant populations without the butterfly, phenotypic selection favored earlier flowering. In populations where the butterfly was present, caterpillars preferentially attacked early-flowering individuals, shifting the direction of selection to favoring later flowering. Butterfly incidence in plant populations increased with ant abundance. Our results demonstrate that antagonistic interactions can shift the direction of selection on flowering phenology, and suggest that such shifts might be associated with differences in the community context.

Yield and Textural Characteristics of Panela Cheeses Produced with Dairy-Vegetable Protein (Soybean or Peanut) Blends Supplemented with Transglutaminase.

The study evaluated panela cheeses made from dairy-plant protein blends, using soybean or peanut protein isolates, supplemented with transglutaminase (TG). Plant proteins were isolated using an alkaline extraction method followed by acid precipitation, and added to the dairy system in order to increase 50% or 100% the protein concentration. The total protein extraction for peanut and soybean isolates was 30.3% and 54.6%, respectively (based on initial protein content of sources), and no impairment of their essential amino acid profile was detected. Cheeses supplemented with TG and soybean showed the highest moisture and crude yield (>67.8% and 20.7%, respectively), whereas protein content was higher in the peanut isolate--added samples without TG (>67.4%). Cheese solids yield (ratio between final and initial solids) was higher for treatments with TG and 100% of plant protein addition (>50.7%). Regarding texture, 4 parameters were measured: hardness, cohesiveness, chewiness, and springiness. All cheeses containing soybean isolates and TG presented the highest chewiness and cohesiveness values, similar to those of the control treatment. Springiness was similar for all treatments, but hardness was higher in cheeses prepared with the peanut protein isolate added with TG. From these results it can be concluded that panela cheeses can be elaborated following a traditional procedure, but with the addition of soybean or peanut protein to the dairy ingredients. Cheeses containing these protein isolates showed higher protein than the milk control cheese and similar textural characteristics.

[Presence of Bartonella henselae in cats: natural reservoir quantification and human exposition risk of this zoonoses in Chile]. / Presencia de Bartonella henselae en gatos: cuantificación del reservorio natural y riesgo de exposición humana de esta zoonosis en Chile.

BACKGROUND: The availability of a serologic test for cat scratch disease in humans has allowed the diagnosis of an increasing number of cases of this disease in Chile. AIM: To perform a serological survey for Bartonella henselae among cats in Chile. MATERIAL AND METHODS: Blood samples from 187 cats living in three Chilean cities were obtained. IgG antibodies against Bartonella henselae were measured using indirect immunofluorescence. Blood cultures were done in 60 samples. The presence of Bartonella henselae in positive cultures was confirmed by restriction fragment length polymorphism polymerase chain reaction (RFLP-PCR). RESULTS: The general prevalence of IgG antibodies against Bartonella henselae was 85.6%. No differences in this prevalence were found among cats younger or older than 1 year, or those infested or not infested with fleas. However domestic cats had a lower prevalence when compared with stray cats (73 and 90% respectively, p <0.01). Bartonella henselae was isolated in 41% of blood cultures. All the isolated were confirmed as Bartonella henselae by RFLP-PCR. CONCLUSIONS: This study found an important reservoir of Bartonella henselae in Chilean cats and therefore a high risk of exposure in humans who have contact with them.

Detección de antígenos de interés diagnóstico en infecciones animales por fasciola hepática / Antigen detection with diagnostic potential in fasciola hepatica animal infections

Se realizó una caracterización e identificación de los antígenos de relevancia diagnóstica de fasciola hepatica empleando electroforesis en geles de poliacrilamida (SDS-PAGE), e inmunoelectrotransferencia. Se estudiaron tres grupos de antígenos: somáticos de fasciolas adultas (FhA), de ejemplares juveniles (FhJ) y productos de excreción-secreción (Fh E-S) del parásito. ellos fueron enfrentados a sueros de conejos hiperinmunes policlonales así como a sueros de 10 ovinos antes y después de ser expuestos a la infección natural, durante el período prepatente y patente de la misma. Se incluyeron como controles muestras de ovinos infectados con quistes hidatídicos y con Thysanosoma actinoides procedentes de áreas libres de fasciolosis. Empleando antígenos A y J se detectó una importante banda de 64 kDa así como bandas de 20-21, 26, 30-31 y 38 kDa con antígenos E-S al ser enfrentados a los sueron hiperinmunes de conejos. En el caso de los ovinos infectados se detectaron por su frecuencia y especificidad las fracciones de 16, 26-28, 35-36 y 56-58 kDa. Los polipéptidos de 26-28 y 35-38 kDa fueron reconocidos por nueve de los diez animales durante la etapa prepatente, siendo todos ellos detectados en la etapa patente de la fasciolosis. Estas fracciones se costituirían en las de mayor potencial diagnóstico de la fasciolosis ovina, siendo por lo tanto recomendable su posterior semipurificación