Influence of Host Plants and Tending Ants on the Cuticular Hydrocarbon Profile of a Generalist Myrmecophilous Caterpillar.

In myrmecophilous organisms, which live in symbiosis with ants, cuticular hydrocarbons (CHCs) play a pivotal role in interspecific communication and defense against chemical-oriented predators. Although these interactions form complex information webs, little is known about the influence of biotic environmental factors on the CHC profiles of myrmecophiles. Here, we analyzed the effect of different host plants and tending ants on the larval CHC profile of Synargis calyce (Lepidoptera: Riodinidae), a polyphagous species with facultative myrmecophily. Groups of caterpillars were fed individually with three host plant species (without tending ants), and with two tending ant species. Through gas chromatography analysis, we compared the cuticular profiles of treatments and found a high similarity between plants and caterpillars (65-82%), but a low similarity between caterpillars and their tending ants (30 - 25%). Cluster analysis showed that caterpillars, ants, and plants form distinct groups, indicating that S. calyce caterpillars have their own chemical profile. These results are similar to those observed for Lycaenidae caterpillars indicating that there is functional convergence in the chemical strategies used by myrmecophilous caterpillar species with similar ecology. Also, the results suggest that the cuticular compounds of S. calyce are primarily influenced by their host plants rather than their tending ants. Thus, we propose that these caterpillars present a trade-off between camouflage and directly informing their presence to ants, maintaining their unique chemical profile, though slightly affected by biotic environmental factors.

Chemical Resemblance of Egg Surface Compounds and Dufour's Gland in Two Neotropical Polistinae Wasps Polistes versicolor (Olivier) and Mischocyttarus metathoracicus (de Saussure, 1854).

Chemical communication plays a major role in regulating social dynamics in social insect colonies. The most studied class of chemical compounds are the cuticular hydrocarbons (CHCs), compounds with high molecular weight that cover the insect body. CHCs are used in nestmate recognition and to signal reproductive status. Brood, in the form of larvae and eggs, is known to participate in chemical communication and social dynamics by performing hunger behaviour and inducing interaction with adults and conferring nest and maternity identity. CHCs of adults and egg surface compounds are similar in composition in social insect species. The main source of egg compounds is proposed to be Dufour's gland, an accessory reproductive gland found in several Hymenoptera females. There is still a lack of information about the level of similarity among CHCs, compounds of egg surface and Dufour's gland for several wasp species, which could provide correlational evidence about the origins of egg-marking compounds. Thus, we investigated whether egg surface compounds were more similar to CHCs or Dufour's gland secretions in two Neotropical primitively eusocial wasp species, Polistes versicolor (Olivier) and Mischocyttarus metathoracicus (de Saussure, 1854). As expected, there was a higher chemical similarity between eggs and Dufour's gland secretions in both studied species, supporting the hypothesis that this gland is the source of chemical compounds found over the eggs in these two primitively eusocial species.

A predatory social wasp does not avoid nestmates contaminated with a fungal biopesticide.

Fungus-based biopesticides have been used worldwide for crop pest control as a safer alternative to chemical pesticides such as neonicotinoids. Both agrochemicals can be lethal and may also trigger side effects on the behavioral traits of non-target social insects, which play a crucial role in providing essential biological pest control services in agroecosystems. Here, we evaluated whether a commercial formulation of the entomopathogenic fungus Beauveria bassiana or the neonicotinoid imidacloprid causes mortality in foragers of Mischocyttarus metathoracicus. These social wasps are natural enemies of caterpillars and other herbivorous insects and inhabit both urban and agricultural environments in Brazil. We also tested whether wasps discriminate between biopesticide-exposed and unexposed conspecifics. Through a combination of laboratory (survival assay) and field experiments (lure presentation), along with chemical analyses (cuticular hydrocarbon profiles), we showed that topic exposure to the label rate of each pesticide causes a lethal effect, with the biopesticide exhibiting a slower effect. Moreover, wasps do not discriminate biopesticide-exposed from unexposed conspecifics, likely because of the similarity of their cuticular chemical profiles 24 h after exposure. Overall, the delayed lethal time at the individual level, combined with the indistinctive chemical cues of exposure and the lack of discrimination by conspecifics suggests that the fungal biopesticide may ultimately pose a threat to the colony survival of this predatory wasp.

Sex recognition does not modulate aggression toward nest intruders in a paper wasp.

During social interactions, the behavior of an individual often depends on the sex of its social partner. Many animal societies have males and females that play very different behavioral roles, although they coexist and interact non-sexually. At specific phases of the colony cycle, social wasp females and males are contemporaries within a nest, they often interact, although mating occurs mostly off the nest, therefore providing an opportunity to test sex discrimination in contexts other than classical sexual ones. We performed a lure presentation experiment to test if Mischocyttarus metathoracicus discriminate between conspecifics of the 2 sexes during on-nest social interactions. Female wasps discriminated conspecific sex during experimentally simulated nest intrusions. Visual and chemical cues may account for this sex discrimination. Despite sex discrimination (evidenced by differential inspective behavior from the nest females toward the female and the male lures), female wasps were as aggressive toward lures of both sexes. In the female-dominated hymenopteran societies, males are often subordinate and not aggressive on nest, resulting in females directing less aggression to them compared to other females. Instead, M. metathoracicus males and females are both aggressive toward nestmates, so they might be perceived as similar threat during on-nest social interactions.

Chemical signatures of egg maternity and Dufour's gland in Vespine wasps.

Cuticular hydrocarbons (CHCs) are often used in the chemical communication among social insects. CHCs can be used in nestmate recognition and as queen pheromones, the latter allows the regulation of the reproductive division of labor. In the common wasp Vespula vulgaris, CHCs and egg-marking hydrocarbons are caste-specific, being hydrocarbon queen pheromones and egg maternity signals. Whether these compounds are conserved among other Vespinae wasps remains unknown. Queens, virgin queens, reproductive workers, and workers belonging to four different wasp species, Dolichovespula media, Dolichovespula saxonica, Vespa crabro, and Vespula germanica, were collected and studied. The cuticular hydrocarbons, egg surface, and Dufour's gland composition were characterized and it was found that chemical compounds are caste-specific in the four species. Quantitative and qualitative differences were detected in the cuticle, eggs, and Dufour's gland. Some specific hydrocarbons that were shown to be overproduced in the cuticle of queens were also present in higher quantities in queen-laid eggs and in their Dufour's gland. These hydrocarbons can be indicated as putative fertility signals that regulate the division of reproductive labor in these Vespine societies. Our results are in line with the literature for V. vulgaris and D. saxonica, in which hydrocarbons were shown to be conserved queen signals. This work presents correlative evidence that queen chemical compounds are found not only over the body surface of females but also in other sources, such as the Dufour's gland and eggs.

Sex and lifestyle dictate learning performance in a neotropical wasp.

In contrast to extensive investigations on bee cognition, the cognitive capacities of wasps remain largely unexplored despite their key role as pollinators and predators of insect pests. Here we studied learning and memory in the neotropical wasp Mischocyttarus cerberus using a Pavlovian conditioning in which harnessed wasps respond with conditioned movements of their mouthparts to a learned odorant. We focused on the different castes, sexes, and ages coexisting within a nest and found that adults of M. cerberus learned and memorized efficiently the odor-sugar associations. In contrast, newly emerged females, but not males, were unable to learn odorants. This difference concurs with their different lifestyle as young males perform regular excursions outside the nest while young females remain in it until older age. Our results thus highlight the importance of socio-ecological constraints on wasp cognition and set the basis for mechanistic studies on learning differences across ages and castes.

Ingestion of polystyrene microparticles impairs survival and defecation in larvae of Polistes satan (Hymenoptera: Vespidae).

Microplastics (MPs) are widespread pollutants of emerging concern, and the risks associated with their ingestion have been reported in many organisms. Terrestrial environments can be contaminated with MPs, and terrestrial organisms, including arthropods, are predisposed to the risk of ingesting MPs. In the current study, the larvae of the paper wasp Polistes satan were fed two different doses (6 mg or 16 mg at once) of polystyrene MPs (1.43 mm maximum length), and the effects of these treatments on immature development and survival till adult emergence were studied. Ingestion of the two doses resulted in mortality due to impaired defecation prior to pupation. The survival of larvae that ingested 16 mg of MPs was significantly lower than that of the control. The ingestion of 16 mg of MPs also reduced the adult emergence (11.4%) in comparison to the control (44.4%). MPs were not transferred from the larvae to the adults that survived. These findings demonstrate that MP ingestion can be detrimental to P. satan, e.g. larval mortality can decrease colony productivity and thus the worker force, and that MPs can potentially affect natural enemies that occur in crops, such as predatory social wasps.

Polymorphism of the beta-casein gene and adaptability of Sindhi cows to semiarid conditions.

This study aimed to investigate the effect of beta-casein genotypes (A1A2 and A2A2) in three different thermal comfort conditions on the adaptability of Sindhi cows and as a tool for selecting thermotolerant cattle. Twelve Sindhi cows were used in a completely randomized design in a 2 × 3 factorial arrangement, with six replicates, with two genotypes, and three thermal comfort conditions. The climatic variables were recorded, while black globe temperature, humidity index, and radiant heat load were calculated. We measured respiratory rate, surface temperature, and rectal temperature while the heat tolerance coefficient was calculated. Genotype had no significant effect (p > 0.05) on any of the parameters measured. However, there was a thermal comfort condition effect (p < 0.05) on rectal temperature, surface temperature, and thermal gradients. The respiratory rate and heat tolerance coefficient were not significantly affected (p > 0.05). Therefore, although the results indicate substantial adaptability of Sindhi cows under any thermal conditions, the tested genotypes should not be used as a tool for selecting thermotolerant Sindhi cows.

Close-range cues used by males of Polistes dominula in sex discrimination.

Sexual pheromones are chemical molecules responsible for mediating sex recognition and mating events. Long- and close-range sexual pheromones act differently. The first type is released to attract potential partners, whereas the second coordinates the interactions after potential mating partners encounter each other. Cuticular hydrocarbons (CHCs) have been suggested to be important cues in the mating systems of several Hymenoptera species, although empirical data are still lacking for many species. Here, we evaluated whether males of the model species Polistes dominula can differentiate the sex of individuals based on their CHC composition. In August 2019, several post-worker emergent nests (n = 19) were collected in the vicinity of Leuven (Belgium) and taken to the lab (KU Leuven), where newly emerged females and males were sampled, marked individually, and kept in plastic boxes for at least a week before being used in the mating trials. Focal males were paired with females and males from different nests and subjected to five different conditions: (I) alive, (II) dead, (III) CHCs washed, (IV) CHCs partially returned, and (V) CHCs from the opposite sex. We videotaped the interactions for 10 min and analysed the duration and different behavioural interactions of the focal male. Our results indicate that CHCs may be used by males as cues to recognise a potential mating partner in P. dominula, since the focal males displayed specific courtship behaviours exclusively toward females. Although we cannot exclude that visual cues could also be used in combination with the chemical ones, we empirically demonstrate that CHCs may be important to convey sexual information at close range in mating systems, allowing fast decisions toward potential sexual partners or rivals.

Meliponamycins: Antimicrobials from Stingless Bee-Associated Streptomyces sp.

Social insects establish complex interactions with microorganisms, some of which play defensive roles in colony protection. The important role of pollinators such as the stingless bee Melipona scutellaris in nature encouraged us to pursue efforts to study its associated microbiota. Here we describe the discovery of two novel cyclic hexadepsipeptides, meliponamycin A (1) and meliponamycin B (2), from Streptomyces sp. ICBG1318 isolated from M. scutellaris nurse bees. Their structures were established by interpretation of NMR and MS data, and the absolute configuration of the constituent amino acids was determined by the advanced Marfey's method. Compounds 1 and 2 showed strong activity against the entomopathogen Paenibacillus larvae and human pathogens Staphylococcus aureus and Leishmania infantum.

Paenibacillus polymyxa Associated with the Stingless Bee Melipona scutellaris Produces Antimicrobial Compounds against Entomopathogens.

Social insects are frequently observed in symbiotic association with bacteria that produce antimicrobial natural products as a defense mechanism. There is a lack of studies on the microbiota associated with stingless bees and their antimicrobial compounds. To the best of our knowledge, this study is the first to report the isolation of Paenibacillus polymyxa ALLI-03-01 from the larval food of the stingless bee Melipona scutellaris. The bacterial strain was cultured under different conditions and produced (L)-(-)-3-phenyllactic acid and fusaricidins, which were active against entomopathogenic fungi and Paenibacillus larvae. Our results indicate that such natural products could be related to colony protection, suggesting a defense symbiosis between P. polymyxa ALLI-03-01 and Melipona scutellaris.

Sexual ornaments reveal the strength of melanization immune response and longevity of male paper wasps.

It has been recently suggested that female mate choice, based on sexually selected ornaments, is an important component of social wasps' reproductive biology. The correlates of male ornaments that could be of a female's interest, however, remain to be investigated. Males of the Neotropical paper wasp Polistes simillimus have sexually dimorphic melanin-based black spots on their faces. In this species, male spots work like sexual ornaments, as it has been experimentally demonstrated that females prefer sexual partners with a higher proportion of black pigment on their faces. We have shown that, under laboratory conditions, male sexual ornamentation positively predicts the strength of the melanization immune response and longevity. Therefore, in P. simillimus, melanin-based facial patterns (ornaments) seem to be honest indicators of male quality.

Cuticular Hydrocarbons of Orchid Bees Males: Interspecific and Chemotaxonomy Variation.

Recent studies have investigated the composition of compounds that cover the cuticle in social insects, but few studies have focused on solitary bees. Cuticular hydrocarbons may provide a tool for chemotaxonomy, and perhaps they can be used as a complement to morphology and genetic characters in phylogenetic studies. Orchid bees (Tribe Euglossini) are a highly diverse group of Neotropical bees with more than 200 species. Here, the cuticular hydrocarbons of 17 species were identified and statistical analysis revealed 108 compounds, which allowed for the taxonomic classification according to the genera. The most significant compounds discriminating the four genera were (Z)-9-pentacosene, (Z,Z)-pentatriacontene-3, (Z)-9-tricosene, and (Z)-9-heptacosene. The analyses demonstrated the potential use of CHCs to identify different species.

Exoskeleton formation in Apis mellifera: cuticular hydrocarbons profiles and expression of desaturase and elongase genes during pupal and adult development.

Cuticular hydrocarbons (CHCs) are abundant in the superficial cuticular layer (envelope) of insects where they play roles as structural, anti-desiccation and semiochemical compounds. Many studies have investigated the CHC composition in the adult insects. However, studies on the profiles of these compounds during cuticle formation and differentiation are scarce and restrict to specific stages of a few insect species. We characterized the CHCs developmental profiles in the honeybee workers during an entire molting cycle (from pupal-to-adult ecdyses) and in mature adults (forager bees). Gas chromatography/mass spectrometry (GC/MS) analysis revealed remarkable differences in the relative quantities of CHCs, thus discriminating pupae, developing and newly-ecdysed adults, and foragers from each other. In parallel, the honeybee genome database was searched for predicted gene models using known amino acid sequences of insect enzymes catalyzing lipid desaturation (desaturases) or elongation (elongases) as queries in BLASTP analysis. The expression levels of six desaturase genes and ten elongase genes potentially involved in CHC biosynthesis were determined by reverse transcription and real time polymerase chain reaction (RT-qPCR) in the developing integument (cuticle and subjacent epidermis). Aiming to predict roles for these genes in CHC biosynthesis, the developmental profiles of CHCs and desaturase/elongase transcript levels were evaluated using Spearman correlation coefficient. This analysis pointed to differential roles for these gene products in the biosynthesis of certain CHC classes. Based on the assumption that homologous proteins may share a similar function, phylogenetic trees were reconstructed as an additional strategy to predict functions and evolutionary relationships of the honeybee desaturases and elongases. Together, these approaches highlighted the molecular complexity underlying the formation of the lesser known layer of the cuticular exoskeleton, the envelope.

Sexy faces in a male paper wasp.

Sexually selected signals are common in many animals, though little reported in social insects. We investigated the occurrence of male visual signals mediating the dominance relationships among males and female choice of sexual partner in the paper wasp Polistes simillimus. Males have three conspicuous, variable and sexually dimorphic traits: black pigmentation on the head, a pair of yellow abdominal spots and body size differences. By conducting behavioral assays, we found that none of the three visual traits are associated with male-male dominance relationship. However, males with higher proportion of black facial pigmentation and bigger yellow abdominal spots are more likely chosen as sexual partners. Also, after experimentally manipulating the proportion of black pigment on males' face, we found that females may evaluate male facial coloration during the choice of a sexual partner. Thus, the black pigmentation on P. simillimus male's head appears to play a role as a sexually selected visual signal. We suggest that sexual selection is a common force in Polistes and we highlight the importance of this group as a model for the study of visual communication in insects.