Challenges and a call to action for protecting European red wood ants.

Red wood ants (RWAs) are a group of keystone species widespread in temperate and boreal forests of the Northern Hemisphere. Despite this, there is increasing evidence of local declines and extinctions. We reviewed the current protection status of RWAs throughout Europe and their International Union for the Conservation of Nature (IUCN) threat classification. Only some RWA species have been assessed at a global scale, and not all national red lists of the countries where RWAs are present include these species. Different assessment criteria, inventory approaches, and risk categories are used in different countries, and data deficiency is frequent. Legislative protection is even more complex, with some countries protecting RWAs implicitly together with the wildlife fauna and others explicitly protecting the whole group or particular species. This complexity often occurs within countries, for example, in Italy, where, outside of the Alps, only the introduced species are protected, whereas the native species, which are in decline, are not. Therefore, an international, coordinated framework is needed for the protection of RWAs. This first requires that the conservation target should be defined. Due to the similar morphology, complex taxonomy, and frequent hybridization, protecting the entire RWA group seems a more efficient strategy than protecting single species, although with a distinction between autochthonous and introduced species. Second, an update of the current distribution of RWA species is needed throughout Europe. Third, a protection law cannot be effective without the collaboration of forest managers, whose activity influences RWA habitat. Finally, RWA mounds offer a peculiar microhabitat, hosting a multitude of taxa, some of which are obligate myrmecophilous species on the IUCN Red List. Therefore, RWAs' role as umbrella species could facilitate their protection if they are considered not only as target species but also as providers of species-rich microhabitats.

Las hormigas rojas de la madera (HRM) conforman un grupo de especies clave con amplia distribución en los bosques templados y boreales del Hemisferio Norte. A pesar de lo anterior, cada vez hay más evidencia de su declinación y extinción local. Revisamos el estado actual de protección de las HRM en toda Europa y su clasificación en la Lista Roja de la Unión Internacional para la Conservación de la Naturaleza (UICN). Sólo se han evaluado algunas especies de HRM a escala mundial y no todas las listas rojas nacionales de los países con presencia de HRM incluyen a estas especies. Los diferentes países usan criterios de evaluación, estrategias de inventario y categorías de riesgo distintos, además de que la información deficiente es habitual. La protección legislativa es todavía más compleja pues algunos países protegen implícitamente a las HRM junto con la fauna silvestre y otros protegen explícitamente a todo el grupo o a una especie particular. Esta complejidad ocurre a menudo en los países (por ejemplo: Italia) en donde, fuera de los Alpes, sólo se protege a las especies introducidas, mientras a las especies nativas, que están declinando, no se les protege. Por lo tanto, se requiere un marco de trabajo internacional y coordinado para proteger a las HRM. Esto necesita primero que se defina el objetivo de conservación. Ya que las HRM tienen similitudes morfológicas, una taxonomía compleja e hibridación frecuente, la protección del grupo completo, con la distinción entre las especies autóctonas y las introducidas, parece ser una estrategia más eficiente que la protección de una sola especie. Segundo, se debe actualizar la distribución actual de las HRM en Europa. Tercero, una ley de protección no puede ser efectiva sin la colaboración de los gestores forestales, cuya actividad influye sobre el hábitat de las HRM Finalmente, los montículos de las HRM ofrecen un microhábitat peculiar pues hospedan a una multitud de taxones, algunos de los cuales son especies mirmecófilas obligadas presentes en la Lista Roja de la UICN. Así, el papel de las HRM como especie paraguas podría facilitar su protección si se les considera no sólo como especies diana sino también como proveedoras de microhábitats con riqueza de especies.

Combined effects of temperature and interspecific competition on the mortality of the invasive garden ant, Lasius neglectus: A laboratory study.

The invasive garden ant, Lasius neglectus, is a dominant species due to its capacity to form large supercolonies. This species was assumed to possess a wide thermal niche since it is able to adapt to cold climates, which is a factor that boosted its rapid expansion from south to many central-northern European Countries. However, the effect of variations in environmental temperatures on its competitive ability against other species has still not been investigated. In this paper, we analyzed the change in survival ability of Lasius neglectus during encounters with two Mediterranean dominant ants (Crematogaster scutellaris and Tapinoma nigerrimum) at four different temperatures (15, 20, 25 and 30°C). Firstly, control tests were performed to provide the baseline survival ability of the three species at different temperatures. Secondly, competition tests were carried out at the same temperatures. Lasius neglectus survival was negatively affected by high temperature (30°C) in control tests, and this impairment was much more pronounced in competition tests. On the contrary, the two opponent species were only marginally affected by temperatures in control tests. Crematogaster scutellaris was a better competitor than L. neglectus, particularly at high temperatures. Tapinoma nigerrimum was a weaker competitor and was always outcompeted by L. neglectus, particularly at low temperatures. This result could suggest that L. neglectus is at a disadvantage during interspecific encounters when temperatures are high and that the predicted future increase in environmental temperatures may potentially enhance this handicap.

Sub-lethal doses of imidacloprid alter food selection in the invasive garden ant Lasius neglectus.

Despite several restrictions to their use, neonicotinoid insecticides are still widely employed worldwide. Residual sub-lethal amounts of these chemicals can have detrimental effects on the behavior of non-target insects. Toxic effects on economically important species such as bees have been widely documented, but less is known about their toxic action on other social insects, such as ants. In this study, we assessed the effect of different sub-lethal doses of the neonicotinoid imidacloprid on the ability of colonies of the invasive ant Lasius neglectus to select the most profitable resource. We used Y-shaped mazes having an imidacloprid-polluted or an unpolluted sucrose solution on the two branches. Two sucrose (0.1 M, 0.5 M) and two imidacloprid (1 µg/ml, 10 µg/ml) concentrations were used. In parallel, we evaluated the marking activity of foragers who fed on the same solutions. We found that the 0.1 M sugar solution polluted with 1 µg/ml imidacloprid was significantly more frequently selected in binary choices experiments than the unpolluted resource. Moreover, the ingestion of the same combination of sugar and imidacloprid significantly increased the marking rate of foragers. The higher concentration of the pollutant had lower effects, probably because of the hormesis phenomenon. Results suggest that the lower sub-lethal dose of imidacloprid can lead ants to select again the polluted resource. This "active" selection of the pollutant may magnify the negative effects on the colonies. Due to their ecological role, any impairment of ant survival or behavior may have detrimental cascade effects on the whole ecosystem.

Keep Your Eggs Away: Ant Presence Reduces Ceratitis capitata Oviposition Behaviour through Trait-Mediated Indirect Interactions.

Alternative methods to achieve sustainable agricultural production while reducing the use of chemical pesticides, such as biological control, are increasingly needed. The exploitation of trait-mediated indirect interactions (TMIIs), in which pests modify their behavior in response to some cues (e.g., pheromones and other semiochemicals) to avoid predation risk, may be a possible strategy. In this study, we tested the effect of TMIIs of two Mediterranean ant species, Crematogaster scutellaris and Tapinoma nigerrimum, on the oviposition behaviour of Ceratitis capitata (Diptera: Tephritidae), one of the world's most economically damaging pests, which attacks fruits. For each ant species, we performed choice experiments using ant-scented and control plums, counting the time spent by medflies on fruits and the number of pupae emerging from them. Results of both ant species tests showed a significantly shorter time spent by ovipositing medflies on ant-exposed plums and a lower number of pupae, when compared to the control group. Our findings highlighted that the semiochemicals released by ants on plums triggered an avoidance behaviour by medfly females, leading to lower oviposition rates. This study contributes to the understanding of indirect ant-pest interactions in Mediterranean agricultural settings and points out the potential of utilising ant-borne semiochemicals in sustainable IPM strategies.

Comparative Analysis of Facial Coloration between Introduced and Source Populations of the Red Wood Ant Formica paralugubris.

The variation in the typical black-reddish color of red wood ants (Formica rufa group) has been recently suggested as a good indicator of habitat quality, being dependent on environmental conditions. However, the relative contribution of external factors and heritability in shaping this trait is poorly investigated. In this study, we compared the facial coloration of workers from four introduced populations of Formica paralugubris with those of the two Alpine populations from which they had been taken. We used a Relative Warp Analysis to describe the variations in the shape of this trait. We expected each introduced population to be more similar to its population of origin if the color pattern was predominantly genetically determined. On the contrary, due to the considerable differences in habitat type and climate between the Alps and the Apennines, we expected to observe differences between the introduced population and their origin population if the coloration was mostly environmentally determined. With one exception that we discuss, the results showed that ants from the two source populations had different phenotypes, and that the introduced populations had a shape similar to the population of origin, suggesting a stable genetic background. Surprisingly, the habitat type seems to have a less clear effect, even if within-population differences suggest the influence of very localized environmental factors. Finally, we found that the facial coloration shape is affected by the ant's size, a result in line with previous studies.

Intraspecific Relationships and Nest Mound Shape Are Affected by Habitat Features in Introduced Populations of the Red Wood Ant Formica paralugubris.

Ants belonging to the Formica rufa group build large nest mounds, which aid their survival during severe winters. We investigated whether different environmental features of the habitats affected the nest mound shape and the population structure. We assessed the shape of all the nest mounds and mapped inter-nest trails connecting mounds for three imported populations of Formica paralugubris in three forest habitats: fir-dominated, beech-dominated, and a mixture of fir and beech. Single-nest mounds were averagely smaller and flatter in the beech-dominated forest, probably because of lighter building materials. Nonetheless, by summing the volumes of all interconnected nests, the size was similar among all three sites. In fir- and beech-dominated forests, large nests were also central in the networks, suggesting a central place foraging model with these nests as reference. We finally performed aggression tests, and found that aggressiveness was significantly higher among nests belonging to the same population than between populations. The results highlight the plasticity of the species to adapt nest and colony structure to different environments. Additionally, it appears that none of these populations is unicolonial, as observed in various alpine sites, there and the observed patterns of aggression are coherent with the 'nasty neighbor' effect.

Assessing Molecular Diversity in Native and Introduced Populations of Red Wood Ant Formica paralugubris.

The Formica rufa group comprises several ant species which are collectively referred to as "red wood ants" and play key roles in boreal forest ecosystems, where they are ecologically dominant and greatly influence habitat dynamics. Owing to their intense predatory activity, some of these species are used as biocontrol agents against several forest insect pests and for this aim in Italy, nearly 6000 ant nests were introduced from their native areas in the Alps to several Appeninic sites during the last century. In this work, we assessed and compared the genetic variability and structure of native and introduced populations of F. paralugubris, thus evaluating the extent of genetic drift that may have occurred since the time of introduction, using amplified fragment length polymorphism (AFLP) markers. PCR amplification with a fam_EcoRI-TAC/MseI-ATG primers combination produced a total of 147 scorable bands, with 17 identified as outlier loci. The genetic variation was higher in the introduced population compared to the native ones that, on the other hand, showed a higher diversity between nests. AMOVA results clearly pointed out that the overall genetic structure was dominated by among-worker variation, considering all populations, the Alpine vs. Apennine groups and the comparison among native and related introduced populations (all ranging between 77.84% and 79.84%). Genetic analyses unveiled the existence of six main different groups that do not entirely mirror their geographic subdivision, pointing towards a wide admixture between populations, but, at the same time, rapid diversification of some Apennine populations. Future studies based on high-throughput genomic methods are needed to obtain a thorough understanding of the effects of environmental pressure on the genetic structure and mating system of these populations.

Small-scale drivers on plant and ant diversity in a grassland habitat through a multifaceted approach.

Semi-natural grasslands are characterized by high biodiversity and require multifaceted approaches to monitor their biodiversity. Moreover, grasslands comprise a multitude of microhabitats, making the scale of investigation of fundamental importance. Despite their wide distribution, grasslands are highly threatened and are considered of high conservation priority by Directive no. 92/43/EEC. Here, we investigate the effects of small-scale ecological differences between two ecosites present within the EU habitat of Community Interest of semi-natural dry grasslands on calcareous substrates (6210 according to Dir. 92/43/EEC) occurring on a Mediterranean mountain. We measured taxonomic and functional diversity of plant and ant communities, evaluating the differences among the two ecosites, how these differences are influenced by the environment and whether vegetation affects composition of the ant community. Our results show that taxonomic and functional diversity of plant and ant communities are influenced by the environment. While vegetation has no effect on ant communities, we found plant and ant community composition differed across the two ecosites, filtering ant and plant species according to their functional traits, even at a small spatial scale. Our findings imply that small-scale monitoring is needed to effectively conserve priority habitats, especially for those that comprise multiple microhabitats.

Palatability of glyphosate in ants: a field experiment reveals broad acceptance of highly polluted solutions in a Mediterranean ant.

Glyphosate is a systemic herbicide still used in many countries, though there are several known detrimental effects on animals. Previous studies concerning its effects on social insects are available, but they are primarily focused on honeybees; little is known about the interactions of this compound with ants. Here, we assessed whether different concentrations of glyphosate can be perceived by ant workers and to what extent. As a model species, we used the Mediterranean ant Crematogaster scutellaris, commonly found in agroecosystems. We performed 3000 individual tests of acceptance using ten different solutions of various concentrations of the herbicide. Half of the solutions contained added sucrose in order to test the possible masking effect of the sugar taste on glyphosate. We used comparable glyphosate concentrations to those previously used in other studies on social insects or suggested by the producer. We found that the acceptance of the solutions decreased as the concentration of the herbicide increased. However, a significant percentage of ants drank the solutions with concentrations up to dozens of times higher than those inducing toxic effects in bees. In light of these results, we urge further assessment of the effects of glyphosate on ants, particularly because the food ingested by workers is transferred to the brood and queens, posing a potential threat to the health of the entire colony. Surprisingly, we did not record any difference in acceptance between solutions with and without sugar; this point is discussed regarding drought stress.

Serpentine soils affect heavy metal tolerance but not genetic diversity in a common Mediterranean ant.

Natural habitats with serpentine soils are rich in heavy metal ions, which may significantly affect ecological communities. Exposure to metal pollutants results, for instance, in a reduction of population genetic diversity and a diffused higher tolerance towards heavy metals. In this study, we investigated whether chronic exposure to metals in serpentine soils affect accumulation patterns, tolerance towards metal pollutants, and genetic diversity in ants. In particular, we studied colonies of the common Mediterranean ant, Crematogaster scutellaris, along a contamination gradient consisting of two differently contaminated forests and a reference soil with no geogenic contamination. We first evaluated the metal content in both soil and ants' body. Then, we tested for tolerance towards metal pollutants by evaluating the mortality of ants fed with nickel (Ni) solutions of increasing concentrations. Finally, differences in genetic diversity among ants from different areas were assessed using eight microsatellite loci. Interestingly, a higher tolerance to nickel solutions was found in ants sampled in sites with intermediate levels of heavy metals. This may occur, because ants inhabiting strongly contaminated areas tend to accumulate higher amounts of contaminants. Additional ingestion of toxicants beyond the saturation threshold would lead to death. There was no difference in the genetic diversity among ant colonies sampled in different sites. This was probably the result of queen mediated gene flow during nuptial flights across uncontaminated and contaminated areas of limited geographical extent.

The Rules of Aggression: How Genetic, Chemical and Spatial Factors Affect Intercolony Fights in a Dominant Species, the Mediterranean Acrobat Ant Crematogaster scutellaris.

Nest-mate recognition plays a key role in the biology of ants. Although individuals coming from a foreign nest are, in most cases, promptly rejected, the degree of aggressiveness towards non nest-mates may be highly variable among species and relies on genetic, chemical and environmental factors. We analyzed intraspecific relationships among neighboring colonies of the dominant Mediterranean acrobat ant Crematogaster scutellaris integrating genetic, chemical and behavioral analyses. Colony structure, parental relationships between nests, cuticular hydrocarbons profiles (CHCs) and aggressive behavior against non nest-mates were studied in 34 nests located in olive tree trunks. Bayesian clustering analysis of allelic variation at nine species-specific microsatellite DNA markers pooled nests into 14 distinct clusters, each representing a single colony, confirming a polydomous arrangement of nests in this species. A marked genetic separation among colonies was also detected, probably due to long distance dispersion of queens and males during nuptial flights. CHCs profiles varied significantly among colonies and between nests of the same colony. No relationship between CHCs profiles and genetic distances was detected. The level of aggressiveness between colonies was inversely related to chemical and spatial distance, suggesting a 'nasty neighbor' effect. Our findings also suggest that CHCs profiles in C. scutellaris may be linked to external environmental factors rather than genetic relationships.

Modeling warfare in social animals: a "chemical" approach.

We present here a general method for modelling the dynamics of battles among social animals. The proposed method exploits the procedures widely used to model chemical reactions, but still uncommon in behavioural studies. We applied this methodology to the interpretation of experimental observations of battles between two species of ants (Lasius neglectus and Lasius paralienus), but this scheme may have a wider applicability and can be extended to other species as well. We performed two types of experiment labelled as interaction and mortality. The interaction experiments are designed to obtain information on the combat dynamics and lasted one hour. The mortality ones provide information on the casualty rates of the two species and lasted five hours. We modelled the interactions among ants using a chemical model which considers the single ant individuals and fighting groups analogously to atoms and molecules. The mean-field behaviour of the model is described by a set of non-linear differential equations. We also performed stochastic simulations of the corresponding agent-based model by means of the Gillespie event-driven integration scheme. By fitting the stochastic trajectories with the deterministic model, we obtained the probability distribution of the reaction parameters. The main result that we obtained is a dominance phase diagram, that gives the average trajectory of a generic battle, for an arbitrary number of opponents. This phase diagram was validated with some extra experiments. With respect to other war models (e.g., Lanchester's ones), our chemical model considers all phases of the battle and not only casualties. This allows a more detailed description of the battle (with a larger number of parameters), allowing the development of more sophisticated models (e.g., spatial ones), with the goal of distinguishing collective effects from the strategic ones.