Differing Dietary Nutrients and Diet-Associated Bacteria Has Limited Impact on Spider Gut Microbiota Composition

Spiders are a key predator of insects across ecosystems and possess great potential as pest control agents. Unfortunately, it is difficult to artificially cultivate multiple generations of most spider species. Since gut bacterial flora has been shown to significantly alter nutrient availability, it is plausible that the spiders’ microbial community plays a key role in their unsuccessful breeding. However, both the gut microbial composition and its influencing factors in many spiders remain a mystery. In this study, the gut microbiota of Campanicola campanulata, specialists who prey on ants and are widely distributed across China, was characterized. After, the impact of diet and diet-associated bacteria on gut bacterial composition was evaluated. First, two species of prey ants (Lasius niger and Tetramorium caespitum) were collected from different locations and fed to C. campanulata. For each diet, we then profiled the nutritional content of the ants, as well as the bacterial communities of both the ants and spiders. Results showed that the protein and carbohydrate content varied between the two prey ant species. We isolated 682 genera from 356 families in the ants (dominant genera including Pseudomonas, Acinetobacter, Paraburkholderia, Staphylococcus, and Novosphingobium), and 456 genera from 258 families in the spiders (dominated by Pseudomonas). However, no significant differences were found in the gut microbiota of spiders that were fed the differing ants. Together, these results indicate that nutritional variation and diet-associated bacterial differences have a limited impact on the microbial composition of spider guts, highlighting that spiders may have a potentially stable internal environment and lay the foundation for future investigations into gut microbiota.

Combined Effect of Different Flower Stem Features on the Visiting Frequency of the Generalist Ant Lasius niger: An Experimental Study

In order to understand the effects of the morphology and surface texture of flower stems in Smyrnium rotundifolium on the visiting frequency of generalist ants, we conducted experiments with Lasius niger ants running on dry wooden sticks mimicking different types of stems: (1) intact (grooved) sticks; (2) sticks painted with slaked (hydrated) lime (calcium carbonate coverage) imitating plant epicuticular wax coverage; (3) intact sticks with smooth polyester plate-shaped cuffs imitating upper leaves; and (4) intact sticks bearing cuffs painted with slaked lime. Ants were attracted by the sweet sugar syrup droplets placed on a stick tip, and the number of ants visiting the drops was counted. Our data showed significant differences in the visiting frequencies between the different types of stem-mimicking samples. The number of recorded ants progressively decreased in the following order of samples: intact sticks—painted sticks—sticks with intact cuffs—sticks with painted cuffs. These results clearly demonstrated that micro/nanoscopic surface coverages and macroscopic physical barriers, especially if combined, have a negative impact on the attractiveness of stems to ants. This study provides further evidence for the hypothesis that having a diversity of plant stems in the field, generalist ants prefer substrates where their locomotion is less hindered by obstacles and/or surface slipperiness.

Potent Ant Deterrents Emitted from Nematode-Infected Insect Cadavers

Most known species of entomopathogenic nematodes (EPNs) are generalist obligate parasites of insects. They kill their hosts within days after infection and mortality is mainly caused by toxins produced by bacteria that co-infect the hosts and serve as food for the nematodes. EPNs can infect a very broad spectrum of insects and these insects can therefore be expected to have evolved strategies to avoid infection. Indeed, ants are known to avoid feeding on EPN-infected insect cadavers, most likely because they are repelled by semiochemicals that emanate from the cadavers. The source and nature of these repellents are so far unknown. In a series of behavioral and chemical analytical experiments we identified hexadecanal and 2-heptadecanone as two compounds that are emitted by insect larva that are infected by the EPN Steinernema feltiae, but not by uninfected larvae. When spiking honey water with the two semiochemicals, they were confirmed to be highly deterrent to the ant Lasius niger. The environmentally benign hexadecanal and 2-heptadecanone could be employed to ward off ants and possibly other pests. Additional experiments are needed to fully determine their application potential.

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.

Aphids harbouring different endosymbionts exhibit differences in cuticular hydrocarbon profiles that can be recognized by ant mutualists

Cuticular hydrocarbons (CHCs) have important communicative functions for ants, which use CHC profiles to recognize mutualistic aphid partners. Aphid endosymbionts can influence the quality of their hosts as ant mutualists, via effects on honeydew composition, and might also affect CHC profiles, suggesting that ants could potentially use CHC cues to discriminate among aphid lines harbouring different endosymbionts. We explored how several strains of Hamiltonella defensa and Regiella insecticola influence the CHC profiles of host aphids (Aphis fabae) and the ability of aphid-tending ants (Lasius niger) to distinguish the profiles of aphids hosting different endosymbionts. We found significant compositional differences between the CHCs of aphids with different infections. Some endosymbionts changed the proportions of odd-chain linear alkanes, while others changed primarily methyl-branched compounds, which may be particularly important for communication. Behavioural assays, in which we trained ants to associate CHC profiles of endosymbiont infected or uninfected aphids with food rewards, revealed that ants readily learned to distinguish differences in aphid CHC profiles associated with variation in endosymbiont strains. While previous work has documented endosymbiont effects on aphid interactions with antagonists, the current findings support the hypothesis that endosymbionts also alter traits that influence communicative interactions with ant mutualists.

Ant nests as a microbial hot spots in a long-term heavy metal-contaminated soils

Interactions between soil fauna and soil microorganisms are not fully recognized, especially in extreme environments, such as long-term metal-polluted soils. The purpose of the study was to assess how the presence of Lasius niger ants affected soil microbial characteristics in a long-term metal-polluted area (Upper Silesia in Poland). Paired soil samples were taken from bulk soil and from ant nests and analysed for a range of soil physicochemical properties, including metal content (zinc, cadmium, and lead). Microbial analysis included soil microbial activity (soil respiration rate), microbial biomass (substrate-induced respiration rate), and bacteria catabolic properties (Biolog® ECO plates). Soil collected from ant nests was drier and was characterized by a lower content of organic matter, carbon and nitrogen contents, and also lower metal content than bulk soil. Soil microbial respiration rate was positively related to soil pH (p = 0.01) and negatively to water-soluble metal content, integrated into TIws index (p = 0.01). Soil microbial biomass was negatively related to TIws index (p = 0.04). Neither soil microbial activity and biomass nor bacteria catabolic activity and diversity indices differed between bulk soil and ant nests. Taken together, ant activity reduced soil contamination by metals in a microscale which support microbial community activity and biomass but did not affect Biolog® culturable bacteria.

Experimental increase of worker diversity benefits brood production in ants

Background The reproductive division of labor of eusocial insects, whereby one or several queens monopolize reproduction, evolved in a context of high genetic relatedness. However, many extant eusocial species have developed strategies that decrease genetic relatedness in their colonies, suggesting some benefits of the increased diversity. Multiple studies support this hypothesis by showing positive correlations between genetic diversity and colony fitness, as well as finding effects of experimental manipulations of diversity on colony performance. However, alternative explanations could account for most of these reports, and the benefits of diversity on performance in eusocial insects still await validation. In this study, we experimentally increased worker diversity in small colonies of the ant Lasius niger while controlling for typical confounding factors. Results We found that experimental colonies composed of workers coming from three different source colonies produced more larvae and showed more variation in size compared to groups of workers coming from a single colony. Conclusions We propose that the benefits of increased diversity stemmed from an improved division of labor. Our study confirms that worker diversity enhances colony performance, thus providing a possible explanation for the evolution of multiply mated queens and multiple-queen colonies in many species of eusocial insects.

Early social context does not influence behavioral variation at adulthood in ants

Early experience can prepare offspring to adapt their behaviors to the environment they are likely to encounter later in life. In several species of ants, colonies show ontogenic changes in the brood-to-worker ratio that are known to have an impact on worker morphology. However, little information is available on the influence of fluctuations in the early social context on the expression of behavior in adulthood. Using the ant Lasius niger, we tested whether the brood-to-worker ratio during larval stages influenced the level of behavioral variability at adult stages. We raised batches of 20 or 180 larvae in the presence of 60 workers until adulthood. We then quantified the activity level and wall-following tendency of callow workers on ten successive trials to test the prediction that larvae reared under a high brood-to-worker ratio should show greater behavioral variations. We found that manipulation of the brood-to-worker ratio influenced the duration of development and the size of individuals at emergence. We detected no influence of early social context on the level of between- or within individual variation measured for individual activity level or on wall-following behavior. Our study suggests that behavioral traits may be more canalized than morphological traits.

Differing Dietary Nutrients and Diet-associated Bacteria has Limited Impact on Spider Gut Microbiota Composition

Spiders are a key predator of insects across ecosystems and possess great potential as pest control agents. Unfortunately, it is difficult to artificially cultivate multiple generations of most spider species. Since gut bacterial flora has been shown to significantly alter nutrient availability, it is plausible that the spiders’ microbial community play a key role in their unsuccessful breeding. However, both the gut microbial composition and its influencing factors in many spiders remain a mystery. In this study, the gut microbiota of Campanicola campanulata, specialist prey on ants and are widely distributed across China, was characterized. After, the impact of diet and diet-associated bacteria on gut bacterial composition was evaluated. First, two species of prey ants (Lasius niger and Tetramorium caespitum) were collected from different locations and fed to C. campanulata. For each diet, we then profiled the nutritional content of the ants, as well as the bacterial communities of both the ants and spiders. Results showed that the protein and carbohydrate content varied between the two prey ant species, and that the bacterial communities of the ants were clearly delineated by collection site. However, no significant differences were found in the gut microbiota of spiders that were fed the differing ants. Together, these results indicate that nutritional variation and diet-associated bacterial differences have a limited impact on the microbial composition of spider guts, suggesting that spiders have a mechanism keeping their gut bacterial community stable to ensure normal physiological function and development.

Appetitive olfactory learning suffers in ants when octopamine or dopamine receptors are blocked

Associative learning relies on the detection of coincidence between a stimulus and a reward or punishment. In the insect brain, this process is carried out in the mushroom bodies under control of octopaminergic and dopaminergic neurons. It was assumed that appetitive learning is governed by octopaminergic neurons, while dopamine is required for aversive learning. This view has been recently challenged: Both neurotransmitters are involved in both types of learning in bees and flies. Here, we test which neurotransmitters are required for appetitive learning in ants. We trained Lasius niger workers to discriminate two mixtures of linear hydrocarbons and to associate one of them with a sucrose reward. We analysed the walking paths of the ants using machine learning and found that the ants spent more time near the rewarded odour than the other, a preference that was stable for at least 24 hours. We then treated the ants before learning with either epinastine, an octopamine receptor blocker, or with flupentixol, a dopamine receptor blocker. Ants with blocked octopamine receptors did not prefer the rewarded odour. Octopamine signalling is thus necessary for appetitive learning of olfactory cues, likely because it signals information about odours or reward to the mushroom body. In contrast, ants with blocked dopamine receptors initially learned the rewarded odour but failed to retrieve this memory 24 hours later. Dopamine is thus likely required for long-term memory consolidation, independent of short-term memory formation. Our results show that appetitive olfactory learning depends on both octopamine and dopamine signalling in ants.