Black Garden Ant Lasius Niger Research Articles

No evidence that recruitment pheromone modulates olfactory, visual, or spatial learning in the ant Lasius niger

Pheromones are perhaps the most common form of intraspecific communication in the animal kingdom and used in various contexts. Their modulatory potential on cognitive processes has been demonstrated in both vertebrates and invertebrates. Particularly interesting in this regard are social insects, due to their extensive use of pheromones to organise collective behaviour. Recruitment pheromones might be expected to encourage learning, but could also hinder learning due to a blocking effect, whereby the pheromone already partially predicts the reward, hindering further cues being associated with the reward. Here, we use free-running learning assays using realistic pheromone strength to test for a modulation effect on learning in the black garden ant Lasius niger. We found no evidence that learning in three modalities (olfactory, visual, and spatial) is affected by the presence of a realistic pheromone trail. Interestingly, this is in contrast to findings in honeybees. The fact that associative learning does not seem to be influenced by recruitment pheromone in L. niger and reportedly the Argentine ant, while it is in honeybees, the possibly best-studied social insect species, is noteworthy. We speculate that a species-specific importance of social information use could drive modulatory effects of pheromones on a wide range of cognitive processes.Significance statementPheromones have been shown to modulate associative learning in a variety of animals. Among social insects, attractive pheromone has been found to enhance associative olfactory learning in honeybees but not in ants. In ants, recruitment pheromone predicts a food source; therefore, it might hinder learning of a new cue for a food reward. We use a free-running learning assay to test for an effect of trail pheromone on associative learning in three different modalities—olfactory, spatial, visual—in Lasius niger, but find no evidence of any effect. Our learning assay demonstrated fast olfactory learning, moderate spatial learning, and no visual learning after only one training visit. Based on our findings, and findings in two other ant species, we speculate that the ecological foraging conditions of mass-recruiting ants, i.e. following a trail, have not favoured a modulation potential of recruitment pheromone opposed to attractive pheromone in honeybees.

Urban abiotic stressors drive changes in the foraging activity and colony growth of the black garden ant Lasius niger

Changes in habitat characteristics are known to have profound effects on biotic communities and their functional traits. In the context of an urban-rural gradient, urbanisation drastically alters abiotic characteristics, e.g., by increasing environmental temperatures and through light pollution. These abiotic changes significantly impact the functional traits of organisms, particularly insects. Furthermore, changes in habitat characteristics also drive changes in the behavioural traits of animals, allowing them to adapt and thrive in new environments. In our study, we focused on the synanthropic ant species Lasius niger as a model organism. We conducted nocturnal field observations and complemented them with laboratory experiments to investigate the influence of night warming (NW) associated with Urban Heat Islands (UHI), light pollution (ALAN), and habitat type on ant foraging behaviour. In addition, we investigated the influence of elevated temperatures on brood development and worker mortality. Our findings revealed that urban populations of L. niger were generally more active during the night compared to their rural counterparts, although the magnitude of this difference varied with specific city characteristics. In laboratory settings, higher temperatures and continuous illumination were associated with increased activity level in ants, again differing between urban and rural populations. Rural ants exhibited more locomotion compared to their urban counterparts when maintained under identical conditions, which might enable them to forage more effectively in a potentially more challenging environment. High temperatures decreased the developmental time of brood from both habitat types and increased worker mortality, although rural colonies were more strongly affected. Overall, our study provides novel insights into the influence of urban environmental stressors on the foraging activity pattern and colony development of ants. Such stressors can be important for the establishment and spread of synanthropic ant species, including invasive ones, and the biotic homogenization of anthropogenic ecosystems.

Claustral colony founding is limited by body condition: experimental feeding increases brood size of Lasius niger queens (Hymenoptera: Formicidae)

Abstract Ants can found colonies in different ways. One is by claustral colony foundation, when fertilized queens typically have enough nutrients reserved in their bodies to raise the first ‘nanitic’ worker generation without any foraging activity. This colony-foundation strategy evolved in response to selective pressures (e.g. predators), but it limits the number of nanitic workers. We, therefore, assumed that fed queens might initially rear more nanitic workers, although feeding might also be associated with risks and stress. In this study, we analysed the effects of stress and different diets during the colony-foundation period on the success of colony founding by claustral black garden ant Lasius niger (Hymenoptera: Formicidae) queens. Our findings confirm that regularly fed claustral ant queens raise more pupae and workers, reflecting that limited brood size is a cost of safety behaviour for claustral ant queens. There is another notable aspect of our findings for laboratories: in order to maximize the size of the nanitic worker generation, they can feed claustral ant queens with crickets.

Genetic modification of the ant Lasius niger using CRISPR-Cas9 technology.

CRISPR-Cas9 has become one of the most prominent gene editing tools available and it has been utilized in various organisms from bacteria to fungi, plants, and animals. In this study, we developed a CRISPR-Cas9 protocol for the black garden ant Lasius niger, a common and easily available study species for lab and field experiments. To create indel mutations using CRISPR-Cas9 in L. niger, we targeted three different locations in a well-studied eye pigmentation gene cinnabar, generating several mutations that disrupt the ommochrome biosynthesis pathway and result in the lack of the pigment and therefore, abnormal eye coloration in adult workers. We also developed a protocol to collect L. niger eggs, inject them with CRISPR-Cas9 construct, and rear the eggs into mature adult workers with the assistance of nursing workers. We demonstrated for the first time in L. niger that CRISPR-Cas9 is an excellent tool to create targeted mutations for this species. Our protocol can be referred to when developing similar studies for other species of ants and eusocial insects.

Local coexistence of native and invasive ant species is associated with micro-spatial shifts in foraging activity

Invasive species often displace native species by outcompeting them. Yet, some native species can persist even in heavily invaded areas. The mechanisms mediating this local coexistence are still unclear. Fine-scale microclimatic heterogeneity could promote the local coexistence of native and invasive animal competitors. We tested if native ant species could coexist with a recent ant invader, Tapinoma magnum, by shifting their foraging activity in time or space to different microclimatic conditions. We compared the foraging activity of native epigeic ant species among invaded and uninvaded sites. We collected ants at baits in green spaces on the north, east, south and west sides of buildings in the morning, at noon and in the afternoon to test if native species foraged under different microclimatic conditions in invaded sites. Invaded sites had lower ant species richness, diversity, and relative abundance. The native black garden ant Lasius niger – one of the most widespread Palearctic ant species – persisted at high densities in invaded areas but foraged less on the east side of buildings and more on the west side. Microclimatic heterogeneity might promote native and invasive species coexistence by allowing some native species to shift their foraging behaviour to locally avoid or outcompete invasive competitors. Better understanding how fine-scale micro-environmental heterogeneity affects native species’ persistence in invaded areas could help to predict and locally mitigate the negative impacts of biological invasions.

Individual vs. Combined Short-Term Effects of Soil Pollutants on Colony Founding in a Common Ant Species.

Insects are integral to terrestrial life and provide essential ecosystem functions such as pollination and nutrient cycling. Due to massive declines in insect biomass, abundance, or species richness in recent years, the focus has turned to find their causes. Anthropogenic pollution is among the main drivers of insect declines. Research addressing the effects of pollutants concentrates on aquatic insects and pollinators, despite the apparent risk of contaminated soils. Pollutants accumulating in the soil might pose a significant threat because concentrations tend to be high and different pollutants are present simultaneously. Here, we exposed queens of the black garden ant Lasius niger at the colony founding stage to different concentrations and combinations of pollutants (brake dust, soot, microplastic particles and fibers, manure) to determine dose-dependent effects and interactions between stressors. As proxies for colony founding success, we measured queen survival, the development time of the different life stages, the brood weight, and the number of offspring. Over the course of the experiment queen mortality was very low and similar across treatments. Only high manure concentrations affected the colony founding success. Eggs from queens exposed to high manure concentrations took longer to hatch, which resulted in a delayed emergence of workers. Also, fewer pupae and workers were raised by those queens. Brake dust, soot and plastic particles did not visibly affect colony founding success, neither as single nor as multiple stressors. The application of manure, however, affected colony founding in L. niger negatively underlining the issue of excessive manure application to our environment. Even though anthropogenic soil pollutants seem to have little short-term effects on ant colony founding, studies will have to elucidate potential long-term effects as a colony grows.

Non-consumptive effects between predators depend on the foraging mode of intraguild prey.

Predator non-consumptive effects (NCE) are a suite of phenotypic responses of prey to avoid predation that cascade down food webs and can have a stronger impact than predation itself. However, the role of NCE in intraguild interactions remains understudied. Thus, multi-species experiments based on species traits are needed to improve our understanding of the impact of NCE on community structure. We investigated NCE of ants on 17 spider species as intraguild prey (8 cursorial and 9 sedentary) to test the influence of spider hunting mode on the strength of NCE. Sedentary spiders select a habitat patch and wait for prey, whereas cursorial spiders roam around searching for prey. Because actively foraging organisms have higher encounter rates with predators, cursorial spiders should show stronger reactions to ant presence. We collected cues (footprint hydrocarbons) of the black garden ant Lasius niger on filter papers. Then, we placed spiders individually on the papers and filmed their behaviour once on a paper with cues and once on a cue-free one. We used the increase in walking activity in the presence of cues as an indicator of avoidance behaviour towards ants. Cursorial spiders spent a longer time moving and being mobile, and had a higher overall mean mobility when exposed to ant cues compared with blank filter papers, whereas sedentary spider species did not react to ant cues at all. The avoidance of ant cues by cursorial spiders corresponds to their higher risk of encountering ants compared with sedentary species, in accordance with the threat-sensitivity hypothesis. Our multi-species study shows the relevance of NCE for intraguild interactions and highlights the importance of experimental trait-based approaches to improve predictability of species interactions and the role of NCE in ecological communities.

Different preferences for new versus old nestboxes by birds, rodents and social insects

The use of artificial nestboxes has improved the understanding of the ecology and behaviour of secondary cavity-nesting animals. Although earlier investigations have shown that nestbox characteristics can influence where animals roost or breed, the impact of ageing of nestboxes on occupation rates has rarely been investigated with experiments based on choice. In this study, the occupation rate of new and old wooden nestboxes set up side by side on the same tree (n = 140 pairs) was investigated in mixed coniferous-deciduous forest in Lithuania. Great tits Parus major and pied flycatchers Ficedula hypoleuca evidently preferred to breed in new nestboxes, while hazel dormice Muscardinus avellanarius and yellow-necked mice Apodemus flavicollis did not show any preference to a certain nestbox type and equally occupied old and new nestboxes. Saxon wasps Dolichovespula saxonica also occupied both nestbox types equally and a similar tendency was observed in European hornets Vespa crabro. By contrast, black garden ants Lasius niger showed a tendency to occupy old nestboxes. The results of the present study are important when considering the use of nestboxes in monitoring small hole-nesting birds and hazel dormice. The preference of birds for new nestboxes may influence long-term monitoring results when nestboxes become older and when old nestboxes are replaced with new ones. This important limitation has not been emphasised in review papers on the inadequacies of nestboxes in studies of hole-nesting birds. However, as ageing of nestboxes does not influence the occupation rates by hazel dormice, the use of nestboxes does not bias the results of long-term monitoring of this species.

Differential responses to chemical cues correlate with task performance in ant foragers

Division of labor in social insects has been explained by response threshold models which are based on differential responses to task-specific stimuli. In the present study, we argue that other types of stimuli, such as location-related cues, which are correlated with but not directly linked to task performance, may be significant. Using the black garden ant Lasius niger as a model, we focused on three groups of workers that perform extranidal tasks: (1) scouts, which explore new and unmarked areas remote from the nest; (2) patrollers, which are not recruited by other ants, but nonetheless walk outside in the nest vicinity; and (3) recruits, which are temporary foragers whose exit is triggered by recruitment. We used standardized tests to investigate, in a context-independent way, whether differences in task performance by these three groups could be correlated to intrinsic differences in their responsiveness to trail pheromone or to nest-related stimuli such as the presence of nestmates or colony odor. Overall, we found that the task profile of workers was correlated neither with their tendency to explore unmarked areas nor with their social attraction to nestmates. Scouts showed a lower attraction to colony odor and lower scores of trail following than recruits. Conversely, recruits were more attracted to colony chemical cues and showed lower response threshold to trail pheromone. Patrollers displayed behaviors between those of recruits and scouts. Our study thus shows that differences in ant responsiveness to location-related cues and recruitment trails contribute to the regulation of extranidal tasks in ants. Work organization in insect societies has been explained by models of differential response thresholds to stimuli that are directly linked to the performance of specific tasks. However, the task performance of individuals could also depend on their responsiveness to social and location-related cues that are correlated but not directly linked to these tasks. Using standardized context-independent tests, we found that scout ants, which explore unknown areas, display a lower responsiveness to colony area marking and trail pheromone. Conversely, recruits, which forage outside the nest only during food exploitation, are the most attracted to nestmates, colony area marking and trail pheromone. Patrollers display intermediate levels of attraction. Location-related cues could therefore play a role in determining the level of task performance by ants and should be taken into consideration in threshold models of task allocation.

Phenotypic correlation between queen and worker brood care supports the role of maternal care in the evolution of eusociality.

Cooperative brood care by siblings, a defining feature of eusociality, is hypothesized to be evolutionarily derived from maternal care via shifts in the timing of the expression of genes underlying maternal care. If sibling and maternal care share a genetic basis, the two behaviors are expected to be genetically and phenotypically correlated. We tested this prediction in the black garden ant Lasius niger by quantifying the brood retrieval rate of queens and their first and later generation worker offspring. Brood retrieval rate of queens was positively phenotypically correlated with the brood retrieval rate of first generation but not with later generation workers. The difference between first and later generation workers could be due to the stronger similarity in care behavior provided by queens and first generation workers compared to later generations. Furthermore, we found that queen retrieval rate was positively correlated with colony productivity, suggesting that natural selection is acting on maternal care. Overall, our results support the idea of a shared genetic basis between maternal and sibling care as well as queen and worker traits more generally, which has implications for the role of intercaste correlations in the evolution of queen and worker traits and eusociality.

Prediction of the Ligands of the CYP9e Subfamily of Ant Cytochrome P450 with the ChEBI Ontologies of Chemical and Biological Characteristics

Cytochromes P450 are involved in the metabolism of various endogenous and exogenous compounds, and their role in the detoxification of xenobiotics has been extensively studied. CYP9e, one of the subfamilies of cytochromes P450, whose functions have been poorly studied, is amplified in the black garden ant Lasius niger. We have performed molecular modeling of 23 proteins of this family belonging to L. niger and other ant species, as well as molecular docking and virtual screening of suspected ligands. The substances used as ligands have been annotated with ChEBI ontologies to predict the chemical and biological properties of molecules forming complexes with CYP9e of ants. It has been shown that, among the ligands forming energetically favorable complexes, ChEBI ontologies of mycotoxins, phytotoxins, steroids, glycosides and terpenoids are overrepresented. Nevertheless, it has been demonstrated that in carrying out a large number of inaccurate simulations, the results of function predictions can be correlated with molecular docking and the evolutionary history of a protein family.

Genomic Signatures of Selection between Urban and Rural Populations of Black Garden Ant Lasius niger

Black garden ant L. niger is known as a dominant species of urban ant fauna in Europe. Successful propagation of L. niger in transformed ecosystems was associated with behavioral traits in many surveys; however, molecular and genetic basis of such adaptation was never studied. In the present study, genomes of populations from the city of Moscow and natural habitats of Moscow oblast are compared. Pooled samples from rural and urban habitats are collected and sequenced with Illumina HiSeq. SNP frequency, Tajima’s D, and fixation index are estimated with PoPoolation and Popoolation2 software. SNP frequencies are significantly different in 64 genes according to Fisher’s exact test. Some of these genes are detected as affected by recent selection. Out of 64 genes, 26 encode retrotransposon proteins. The genes of immune response to viral and fungal infections, fatty acid synthases, and elements of the Hippo/Fat pathway have different SNP frequencies between populations. Certain retrotransposon genes also can be under selection. It is important to note that repressors and mediators of RNA polymerase II are significantly different between urban and rural populations of L. niger. These genes can influence retrotransposon mobility.

Individual ant workers show self-control.

Often, the first option is not the best. Self-control can allow humans and animals to improve resource intake under such conditions. Self-control in animals is often investigated using intertemporal choice tasks-choosing a smaller reward immediately or a larger reward after a delay. However, intertemporal choice tasks may underestimate self-control, as test subjects may not fully understand the task. Vertebrates show much greater apparent self-control in more natural foraging contexts and spatial discounting tasks than in intertemporal choice tasks. However, little is still known about self-control in invertebrates. Here, we investigate self-control in the black garden ant Lasius niger We confront individual workers with a spatial discounting task, offering a high-quality reward far from the nest and a poor-quality reward closer to the nest. Most ants (69%) successfully ignored the closer, poorer reward in favour of the further, better one. However, when both the far and the close rewards were of the same quality, most ants (83%) chose the closer feeder, indicating that the ants were indeed exercising self-control, as opposed to a fixation on an already known food source.

Genomic exaptation enables Lasius niger adaptation to urban environments

BackgroundThe world is rapidly urbanizing, and only a subset of species are able to succeed in stressful city environments. Efficient genome-enabled stress response appears to be a likely prerequisite for urban adaptation. Despite the important role ants play in the ecosytem, only the genomes of ~13 have been sequenced so far. Here, we present the draft genome assembly of the black garden ant Lasius niger – the most successful urban inhabitant of all ants – and we compare it with the genomes of other ant species, including the closely related Camponotus floridanus.ResultsSequences from 272 M Illumina reads were assembled into 41,406 contigs with total length of 245 MB, and N50 of 16,382 bp, similar to other ant genome assemblies enabling comparative genomic analysis. Remarkably, the predicted proteome of L. niger is significantly enriched relative to other ant genomes in terms of abundance of domains involved in nucleic acid binding, DNA repair, and nucleotidyl transferase activity, reflecting transposable element proliferation and a likely genomic response. With respect to environmental stress, we note a proliferation of various detoxification genes, including glutatione-S-transferases and those in the cytochrome P450 families. Notably, the CYP9 family is highly expanded with 19 complete and 21 nearly complete members - over twice as many compared to other ants. This family exhibits the signatures of strong directional selection, with eleven positively selected positions in ligand-binding pockets of enzymes. Gene family contraction was detected for several components of the olfactory system, accompanied by instances of both directional selection and relaxation.ConclusionsOur results suggest that the success of L. niger in urbanized areas may be the result of fortuitous coincidence of several factors, including the expansion of the CYP9 cytochrome family due to coevolution with parasitic fungi, the diversification of DNA repair systems as an answer to proliferation of retroelements, and the reduction of olfactory system and behavioral preadaptations from non-territorial subdominant life strategies found in natural environments. Diversification of cytochromes and DNA repair systems along with reduced odorant communication are the basis of L. niger pollutant resistance and polyphagy, while non-territorial and mobilization strategies allows more efficient exploitation of large but patchy food sources.

The effects of juvenile hormone on Lasius niger reproduction

Reproduction has been shown to be costly for survival in a wide diversity of taxa. The resulting trade-off, termed the reproduction-survival trade-off, is thought to be one of the most fundamental forces of life-history evolution. In insects the pleiotropic effect of juvenile hormone (JH), antagonistically regulating reproduction and pathogen resistance, is suggested to underlie this phenomenon. In contrast to the majority of insects, reproductive individuals in many eusocial insects defy this trade-off and live both long and prosper. By remodelling the gonadotropic effects of JH in reproductive regulation, the queens of the long-lived black garden ant Lasius niger (living up to 27 years), have circumvented the reproduction-survival trade off enabling them to maximize both reproduction and pathogen resistance simultaneously. In this study we measure fertility, vitellogenin gene expression and protein levels after experimental manipulation of hormone levels. We use these measurements to investigate the mechanistic basis of endocrinological role remodelling in reproduction and determine how JH suppresses reproduction in this species, rather then stimulating it, like in the majority of insects. We find that JH likely inhibits three key aspects of reproduction both during vitellogenesis and oogenesis, including two previously unknown mechanisms. In addition, we document that juvenile hormone, as in the majority of insects, has retained some stimulatory function in regulating vitellogenin expression. We discuss the evolutionary consequences of this complex regulatory architecture of reproduction in L. niger, which might enable the evolution of similar reproductive phenotypes by alternate regulatory pathways, and the surprising flexibility regulatory role of juvenile hormone in this process.

No effect of Zn-pollution on the energy content in the black garden ant.

Social insects may display a response to environmental pollution at the colony level. The key trait of an ant colony is to share energy between castes in order to maintain the existing adult population and to feed the brood. In the present study we calorimetrically measured the energy content per body mass (J/mg) of adults and pupae of workers, males and females of the black garden ant Lasius niger. The ants were sampled from 37 wild colonies originating from 19 sites located along the metal pollution gradient established in a post-mining area in Poland. The cost of metal detoxification seen as a possible reduction in energy content with increasing pollution was found neither for pupae nor adults. However, a considerable part of variance in energy content is explained by belonging to the same colony. These findings stress the importance of colony-specific factors and/or the interaction of these factors with specific site in shaping the response of ants to metal-pollution stress. Colony-related factors may constrain possible selfish decisions of workers over energy allocation in workers and sexual castes.

Pre-adaptive cadmium tolerance in the black garden ant

The black garden ant Lasius niger is a common component of habitats subjected to anthropological stress. The species can develop very abundant populations in metal-polluted areas. In this study, we raised the question of its tolerance to Cd pollution. Workers of L. niger were collected from 54 colonies, originating from 19 sites located along an increasing gradient of Cd pollution in Poland. Ants were exposed to a range of dietary Cd concentrations in a controlled 14-day laboratory experiment in order to test Cd-sensitivity in the investigated ants. The level of ant mortality was recorded as the endpoint of the experiment. We used much higher concentrations of dietary Cd than those the ants are most likely exposed to in field conditions. The investigated ants were highly Cd-tolerant; even a very high dietary Cd concentration of approx. 1300 mg/kg did not affect mortality of workers when compared to the control. Mortality was unrelated to Cd-pollution along the pollution gradient, meaning that high Cd-tolerance can be found even in ants from unpolluted areas. The results stress the importance of pre-adaptive mechanisms in the development of metal tolerance in ants.

Lasius niger(L.) Ants Invade the Web of an Agelenid Spider

ABSTRACT Black garden ants Lasius niger (L.) were observed to invade a web of an agelenid funnel spider (Agelena labyrinthica Clerck or Allagelena gracilens C. L. Koch) and to take the entangled prey away, probably after driving the resident spider out of the web. The observation adds to a few examples of ants invading spider webs and suggests complex interactions between ants and funnel spiders which are known to feed on rather than being ousted by ants.

Amplification of CYP9 genes as a preadaptation of the black garden ant Lasius niger to urban conditions

Amplification of CYP9 genes as a preadaptation of the black garden ant Lasius niger to urban conditions

The amplification of CYP9 genes as a preadaptation of the black garden ant Lasius niger to urban conditions

Ants are one of the most ancient and successful groups of eusocial animals and they are spread all over the world. The nucleotide sequences of the genomes of eight ant species were determined by the year 2014. In these species, the mechanisms of ecological success, cast differentiation, and social communication were studied at genomic level. In ants, the genes of the cytochromes P450 involved in metabolism of xenobiotics and various endogenic substances are amplified. Although the substrates for several cytochrome P450 families have been identified, the functions of the ninth family, which is one of the most amplified, remain unknown. The black garden ant Lasius niger is one of the spices that have successfully adapted to urban conditions. To study the mechanisms of adaptation, we have read and annotated the nucleotide sequence of the L. niger genome; we have predicted the functions of the CYP9 genes using virtual screening. The obtained data allow us to suggest that cytochromes P450 are involved in the metabolism of various xenobiotics such as phytotoxins, mycotoxins, and insecticides. We assume that the functional divergence of the new CYP9 duplications was initially aimed at developing resistance to various mycotoxins, in particular to those produced by Fusarium fungi and, subsequently, to other xenobiotics.