Gustatory synergism in ants mediates a species-specific symbiosis with lycaenid butterflies

Here we show that larvae of the lycaenid butterfly Niphanda fusca secrete droplets containing trehalose and glycine. These droplets attract the larva's host ants Camponotus japonicus, which collect and protect the larvae. We comparatively investigated gustatory preference for trehalose, glycine or a mixture of the two between host (C. japonicus) and non-host (Camponotus obscuripes) species of ants in behavioral and electrophysiological experiments. Glycine itself induced no taste sensation in either host or non-host ants. The mixture of trehalose plus glycine was chosen as much as pure trehalose by non-host ants. However, the host ants clearly preferred the mixture of trehalose plus glycine to trehalose alone. When we used sucrose instead of trehalose, the mixture of sucrose plus glycine was chosen as much as sucrose alone, in both species. These behavioral data are supported by the electrophysiological responsiveness to sugars and/or glycine in the sugar-taste receptor cells of the ants. Considering that lycaenid butterflies' secretions have species-specific compositions of sugar and amino acid; our results clearly showed that such species-specific compositions of larval secretions are precisely tuned to the feeding preferences of their host ant species, in which the feeding preferences are synergistically enhanced by amino acid.     

Chemical mimicry and host specificity in the butterfly Maculinea rebeli,a social parasite of Myrmica ant colonies

Although it has always been assumed that chemical mimicry and camouflage play a major role in the penetration of ant societies by social parasites, this paper provides the first direct evidence for such a mechanism between the larvae of the parasitic butterfly Maculinea rebeli and its ant host Myrmica schencki. In the wild, freshly moulted fourth-instar caterpillars, which have no previous contact with ants, appear to be recognized as ant larvae by foraging Myrmica workers, which return them to their nest brood chambers. Three hypotheses concerning the mechanism controlling this behaviour were tested: (i) the caterpillars produce surface chemicals that allow them to be treated as ant larvae; (ii) mimetic compounds would include hydrocarbons similar to those employed by Myrmica to recognize conspecifics and brood; and (iii) the caterpillars'' secretions would more closely mimic the profile of their main host in the wild, M. schencki, than that of other species of Myrmica. Results of behavioural bioassays and chemical analyses confirmed all three hypotheses, and explained the high degree of host specificity found in this type of highly specialized myrmecophile. Furthermore, although caterpillars biosynthesized many of the recognition pheromones of their host species (chemical mimicry), they later acquired additional hydrocarbons within the ant nest (chemical camouflage), making them near-perfect mimics of their individual host colony''s odour.     

Do Maculinea rebeli caterpillars provide vestigial mutualistic benefits to ants when living as social parasites inside Myrmica ant nests?

Caterpillars of the lycaenid butterfly Maculinea rebeli Hirschke (Lepidoptera: Lycaenidae) live for 11–23 months as social parasites in Myrmica (Hymenoptera: Formicidae) red ant nests, a trait that is believed to have evolved from mutualistic myrmecophilous ancestry. Although Maculinea rebeli caterpillars harm Myrmica larvae, they simultaneously produce copious secretions which the adult worker ants imbibe, perhaps representing a vestige of the ancestral mutualism. We report the results of laboratory experiments designed to test alternative hypotheses: (i) Maculinea rebeli caterpillars provide a beneficial source of sugar in return for being tended by Myrmicaworkers; (ii) Maculinea rebeli harms its host by stressing the workers by competing for available sugar. Comparisons were made of Myrmica worker fitness after 90–450 days under all possible combinations of three experimental treatments: ± M. rebeli caterpillars, ± sucrose and ± ant brood. Caterpillars always reduced the survival of both ant workers and their larvae, even when sugar was not provided, suggesting that M. rebeli is wholly parasitic on all stages in its host colony. The results also confirmed the importance of sucrose in the diet of Myrmica, and showed that M. rebeli caterpillars which eat ant brood to supplement their normal trophallactic feeding by workers develop more quickly - but have the same survival and pupal weights – as caterpillars that are fed solely by worker ants.     

Variation in behavior of lycaenid larvae when attended by different ant species

Partner quality can be crucial for the outcome of a mutualistic interaction. In multi-species associations, the characteristics of potential partners can vary substantially and thus the associated benefits. As a consequence of such variation, one might expect strategies of adjusting investments to the characteristics of a partner. Lycaenid butterfly larvae often interact mutualistically with several ant species of different size and aggressiveness and thereby different ability to protect the larvae. Attending ants are rewarded with nutritious secretions. Both ant behavior and a larva's need for protection are known to influence larval investment in the relationship. This study investigates the effect of six ant species on larval behavior in the lycaenid butterfly, Glaucopsyche lygdamus. The overall level of secretion, as well as the response to varying number of attending ants, were found to be influenced by ant species. This revised version was published online in November 2006 with corrections to the Cover Date.     

Does ant predation favour leaf beetle specialization on toxic host plants?

Generalist predators are frequently seen as evolutionary forces that narrow the host range in herbivorous insects. Predators may favour specialization of herbivores on host plants containing toxic chemicals (which can be used by herbivores for their own defence) if host plant‐derived defences provide better protection from enemies than do autogenously produced defences. We compared the effectiveness of these two defensive strategies in the larvae of six species of leaf beetle (Chrysomelidae) against wood ants (Formica rufa group) in field experiments. Ants were more strongly repelled by larvae with host plant‐derived, salicylaldehyde‐containing secretions than by larvae with various autogenous secretions, but collectively foraging ants ultimately overcame any type of chemical defence by social interactions, chemical signalling, and olfactory learning. As a result, ants killed all larvae of Chrysomela lapponica defended by salicylaldehyde‐containing secretions within 2 days of their introduction to willows within 15 m of ant nests. We conclude that in the field neither type of chemical defence provides complete protection against wood ants in the vicinity of their nests, and that evolutionary shifts from autogenous production of secretion to sequestration of plant allelochemicals in leaf beetles may be favoured mostly at low ant densities on the periphery of ant foraging areas.     

Mutualism between Thisbe irenea butterflies and ants, and the role of ant ecology in the evolution of larval-ant associations

A facultative mutualism between the riodinid butterfly Thisbe irenea and the ponerine ant Ectatomma ruidum is described from Panama. Ants protect larvae against attacks of predatory wasps, but not against tachinid parasitoids. Several potential sources of ecological variation affecting the larval survival of Thisbe irenea are noted. A preliminary means of testing the ability of larvae to appease ants is described that may be applied to all butterfly-ant systems. Observations and literature records indicate that ant taxa which tend butterfly larvae are the same taxa that tend extrafloral nectaries and Homoptera. A general hypothesis for the evolution of myrmecophily among butterflies suggests that ant taxa which utilize secretions in their diet are major selective agents for the evolution of the larval ant-organs, and hence, ant-larval mutualisms. This idea is extended to suggest how appeasement of predaceous ant taxa through the use of larval ant-organs can influence an ant-larval relationship, eventually leading to mutualism.     

Myrmecophily in Hesperiidae. The case of Vettius tertianus in ant gardens

The larvae of the hesperiid butterfly Vettius tertianus develop by eating the leaves of Aechmea mertensii, a bromeliad epiphyte restricted to ant gardens. The relationships between ants and V. tertianus larvae highlight the preferential association of the caterpillars with Pachycondyla goeldii (Ponerinae), an ant-garden initiator. The oviposition strategy of V. tertianus may thus imply the identification of the inhabiting ant species and not only the identification of the host plant. The caterpillars neither provide secretions to the ants, nor possess defensive devices (i.e. hairs or appendices) against ants. Their activity rhythm does not isolate them from foraging workers of P. goeldii and their shelters are also attainable by the ants. Moreover, as the cuticular lipid profiles of V. tertianus larvae are clearly different from those of the ants and also from the leaf-surface of A. mertensii, acceptance is not due to mimicry between larvae and plants or ants. However, the caterpillars deposit, on the leaf they eat, silk containing a mixture of substances very similar to those found on their own cuticle. No interaction with ants was recorded during observations, even though the ant gardens were patrolled by numerous P. goeldii individuals during their activity period. But when confronted with the caterpillar, none of the tested ant species reacted aggressively. These results suggest the existence of compounds, other than cuticular lipids, responsible for the absence of aggressiveness in the ants. The case of V. tertianus is relatively new as myrmecophily within Hesperiidae has been described only once. Moreover, it preferentially involves a member of the Ponerinae, a subfamily in which interactions with other arthropods are exceptional.     

Effect of grazing intensity on the interaction between Shijimiaeoides divinus asonis (Matsumura, 1929) and its attendant ants

This study was carried out in the natural herbaceous grassland of Mt. Aso, which had been almost completely grazed, and which is subjected to routine grassland burning every spring (February or March) to conserve Shijimiaeoides divinus asonis (Matsumura, 1929). We clarify that ants protect the larvae of this butterfly and evaluate the effects of grazing intensity on the attendant ant population. The results obtained are summarized as follows: (i) Five species of attendant ant were identified, with the dominant species being Formica japonica (Motschulsky, 1866) and Camponotus japonicus (Mayr, 1866). (ii) The number of attendant ants decreased at night time; however, only one or two ants attended the larva until the following morning. (iii) Ten species of insect excluding ants and three species of spiders that approached the larva were recorded on the larval host‐plant. Formica japonica and C. japonicus fought off most newcomers of other insects including the natural enemies of these butterfly larvae. (iv) The number of ants in this butterfly habitat under regular grazing intensity was significantly higher than during low grazing intensity and non‐grazing periods. (v) A positive correlation was found between the number of attendant ants and the number of butterfly larvae on the host‐plant. We concluded that the interaction between this butterfly and attendant ants is one of facultative mutualism because the attendant ants protect the butterfly larva. Therefore, the numbers of this butterfly species may decrease if the number of attendant ants decreases due to the cessation of pasturage.     

Comparison of acoustical signals in Maculinea butterfly caterpillars and their obligate host Myrmica ants

An acoustical comparison between calls of parasitic butterfly caterpillars and their host ants is presented for the first time. Overall, caterpillar calls were found to be similar to ant calls, even though these organisms produce them by different means. However, a comparison of Maculinea caterpillars with those of Myrmica ants produced no evidence suggesting fine level convergence of caterpillar calls upon those of their species specific host ants. Factors mediating the species specific nature of the Maculinea-Myrmica system are discussed, and it is suggested that phylogenetic analysis is needed for future work.     

Adoption of lycaenid Niphanda fusca (Lepidoptera: Lycaenidae) caterpillars by the host ant Camponotus japonicus (Hymenoptera: Formicidae)

Caterpillars of the parasitic lycaenid butterfly are often adopted by host ants. It has been proposed that this adoption occurs because the caterpillars mimic the cuticular hydrocarbons of the host ant. This study aimed to examine whether caterpillars of the Japanese lycaenid butterfly Niphanda fusca induce adoption by mimicking their host ant Camponotus japonicus. Behavioral observations conducted in the laboratory showed that most second‐instar caterpillars were not adopted, whereas most third‐instar caterpillars were successfully adopted by host workers. A chemical comparison detected no characteristic differences in the cuticular hydrocarbon profiles between second‐ and third‐instar caterpillars. However, morphological features of the caterpillars differed between the second and third instars; third‐instar caterpillars developed exocrine glands (ant organs) such as tentacle organs and a dorsal nectary organ. These results suggest that multiple chemical signatures, not only cuticular hydrocarbons, may be important for invasion of the host ant nest.     

Species-specific effects of tending ants on the development of lycaenid butterfly larvae

The transfer of nutrients between organisms is a common feature of mutualism. The production of these food rewards is often assumed to be costly. Estimation of the costs of producing food rewards is important for understanding the overall effects of the interaction on fitness. When food rewards are harvested by several species differing in foraging behavior, costs to the producer may differ. The larvae of many species in the butterfly family Lycaenidae produce secretions consumed by tending ants. Here I report that three North American ant species, Formica perpilosa, Dorymyrmex sp. (smithi complex), and Forelius foetida, had no negative effect on the duration of development and adult size of the lycaenid Hemiargus isola. Moreover, tending by the ant Formica perpilosa significantly enhanced larval growth, resulting in butterflies that were 20% heavier than their untended counterparts. Tending by the ants Dorymyrmex sp. (smithi complex) and Forelius foetida had no effect on butterfly weight. Tended, nonfeeding larvae lost 69% more weight than untended, nonfeeding larvae. Taken together, the results suggest that, although ant tending imposes a physiological cost, H. isola larvae use behavioral or physiological mechanisms to compensate or overcompensate for nutrients lost to ants.     

Myrmica ants host highly diverse parasitic communities: from social parasites to microbes

Myrmica ants have been model species for studies in a variety of disciplines, including insect physiology, chemical communication, ant social dynamics, ant population, community ecology, and ant interactions with other organisms. Species belonging to the genus Myrmica can be found in virtually every habitat within the temperate regions of the northern hemisphere and their biology and systematics have been thoroughly studied. These ants serve as hosts to highly diverse parasitic organisms from socially parasitic butterfly caterpillars to microbes, and many Myrmica species even evolved into parasitizing species of their own genus. These parasites have various impacts both on the individuals and on the social structure of their hosts, ranging from morphological malformations to reduction in colony fitness. A comprehensive review of the parasitic organisms supported by Myrmica and the effects of these organisms on individuals and on whole ant colonies has not yet been compiled. Here, we provide a review of the interactions of these organisms with Myrmica ants by discussing host and parasite functional, behavioral or physiological adaptations. In addition, for all “symbiont groups” of Myrmica ants described in this paper, we examine the present limitations of the knowledge at present of their impact on individuals and host colony fitness. In conclusion, we argue that Myrmica ants serve as remarkable resource for the evolution of a wide variety of associated organisms.     

Mimetic host shifts in an endangered social parasite of ants

An emerging problem in conservation is whether listed morpho-species with broad distributions, yet specialized lifestyles, consist of more than one cryptic species or functionally distinct forms that have different ecological requirements. We describe extreme regional divergence within an iconic endangered butterfly, whose socially parasitic young stages use non-visual, non-tactile cues to infiltrate and supplant the brood in ant societies. Although indistinguishable morphologically or when using current mitochondrial and nuclear sequence-, or microsatellite data, Maculinea rebeli from Spain and southeast Poland exploit different Myrmica ant species and experience 100 per cent mortality with each other''s hosts. This reflects major differences in the hydrocarbons synthesized from each region by the larvae, which so closely mimic the recognition profiles of their respective hosts that nurse ants afford each parasite a social status above that of their own kin larvae. The two host ants occupy separate niches within grassland; thus, conservation management must differ in each region. Similar cryptic differentiation may be common, yet equally hard to detect, among the approximately 10 000 unstudied morpho-species of social parasite that are estimated to exist, many of which are Red Data Book listed.     

The influence of ants and water availability on oviposition behaviour and survivorship of a facultatively ant-tended herbivore

1. The lycaenid butterfly Hemiargus isola associates facultatively with the ant species Formica perpilosa in arid areas of south-western North America. Ants solicit liquid food rewards from butterfly larvae as larvae feed on the host plant, Acacia constricta . Previous studies have shown that tending by F. perpilosa enhances larval growth and pupal survivorship.
2. The effects of ants and plant water content on oviposition behaviour and survivorship to the last larval instar were tested by excluding ants and supplementing water to host plants in a two-way factorial experiment.
3. Butterflies, which lay eggs singly on host plant inflorescences, laid significantly higher egg numbers and densities (eggs/inflorescence) on plants with ants than on plants without ants. This is the first report of a facultative, generalized ant-associate using ants as oviposition cues. Water supplements increased the number, but not the density, of eggs laid on plants. Therefore, it appears that egg-laying butterflies responded to number of inflorescences, rather than plant tissue water per se .
4. Plants with ants had significantly greater numbers of inflorescences during the experiment than plants without ants. Water supplements increased number of inflorescences slightly, but not significantly.
5. Ants increased larval survivorship. Twice as many fourth-instar larvae survived per egg laid on plants with ants than on plants without ants. Ants did not reduce the number of predators present on acacias, but may have reduced predator effectiveness. Ants also did not reduce the numbers of potential H. isola competitors present.
6. Water supplementation affected neither the survivorship of H. isola larvae, nor the intensity of ant tending. Water supplementation did not affect the abundance of predators on plants, but did increase the abundance of several herbivorous insect taxa.     

Ants that associate with Lycaeninae butterfly larvae: diversity, ecology and biogeography

Abstract. Field data on associations of butterfly larvae with ants were collated for 435 species of the subfamily Lycaeninae to analyse patterns in the identity, diversity and ecology of attendant ants. Worldwide, members of at least fifty-three ant genera from six subfamilies associate with Lycaeninae larvae. All these ant genera also forage for other liquid carbohydrate food sources. Species of Crematogaster and Camponotus constitute by far the most important ant associates of Lycaeninae larvae. Diversity of butterfly–ant associations, as measured by Hurlbert rarefaction, is highest in the Oriental, Australian and Nearctic faunal regions, and lower in Africa and the Palaearctic. The Neotropical region could not be assessed separately due to paucity of data. Multiple regression analyses revealed that the number of Lycaeninae species associated with a particular ant genus in a facultatively mutualistic manner is largely determined by the ecological prevalence and geographical distribution, but does not correlate with species richness, of an ant genus. This opportunism is suggested to be the key variable supporting the geographically and taxonomically widespread occurrence of ant–caterpillar mutualism in the Lycaeninae. Obligate interactions are much more specific and almost exclusively involve ecologically dominant ants with long-lived colonies. Neither species richness nor geographical distribution of an ant genus are significant predictors. In such associations, the role of ants shifts from visitors to hosts, and patterns of host use and specificity resemble those known from inquiline ant guests.     

Use of Dufour's gland secretion in nest defence and brood nutrition by hover wasps (Hymenoptera, Stenogastrinae)

Social wasps of the subfamily Stenogastrinae produce an abdominal secretion that is used in two distinct biological contexts. First, the secretion plays an important role in larval nutrition where it serves as a substrate in which food is placed by the adults for eventual consumption by the larvae. Second, in several species, females apply the same secretion to the substrate on which their nests are constructed, where it constitutes a sticky barrier that defends the immature brood from predation by ants. This paper describes for the first time ant guard construction behaviour of three species of stenogastrine wasps belonging to the genera Eustenogaster and Liostenogaster. The identification of compounds making up these secretions was also performed by gas chromatography-mass spectrometry. Ant guards and brood secretions were similar, with saturated and unsaturated long chain hydrocarbons and alcohols as major components. We further confirm that the glandular source of abdominal secretion is the Dufour's gland. This gland contains the same hydrocarbons, and in the same proportions as ant guards and brood secretion. We discuss the fundamental importance of Dufour's gland secretion in the social life of these wasps by comparing species with and without ant guards within the subfamily.     

Chemical disguise as particular caste of host ants in the ant inquiline parasite Niphanda fusca (Lepidoptera: Lycaenidae)

The exploitation of parental care is common in avian and insect 'cuckoos' and these species engage in a coevolutionary arms race. Caterpillars of the lycaenid butterfly Niphanda fusca develop as parasites inside the nests of host ants (Camponotus japonicus) where they grow by feeding on the worker trophallaxis. We hypothesized that N. fusca caterpillars chemically mimic host larvae, or some particular castes of the host ant, so that the caterpillars are accepted and cared for by the host workers. Behaviourally, it was observed that the host workers enthusiastically tended glass dummies coated with the cuticular chemicals of larvae or males and those of N. fusca caterpillars living together. Cuticular chemical analyses revealed that N. fusca caterpillars grown in a host ant nest acquired a colony-specific blend of cuticular hydrocarbons (CHCs). Furthermore, the CHC profiles of the N. fusca caterpillars were particularly close to those of the males rather than those of the host larvae and the others. We suggest that N. fusca caterpillars exploit worker care by matching their cuticular profile to that of the host males, since the males are fed by trophallaxis with workers in their natal nests for approximately ten months.     

Unpredictability in the facultative association between larvae of Euphilotes enoptes (Lepidoptera: Lycaenidae) and ants

Larvae of the lycaenid butterfly, Euphilotes enoptes , are facultatively associated with ants. To determine whether variation in this association is predictable, I documented the proportion of larvae tended by ants and the identity of those ants over a three-year period in two study areas in central Washington state. Each study area contained numerous patches of Eriogonum compositum , the sole host plant of E. enoptes in this region. Using complete censuses and random subsampling, I estimated the dispersion and densities of E. enoptes larvae occupying each patch. The results of these observations indicated significant spatial and temporal variation in the identities of ants tending larvae and the frequency of ant attendance. In addition, early instar larvae were tended less frequently and by smaller ant species, than fourth instar larvae. Although larvae were aggregated at two spatial scales: individual plants within a patch and individual inflorescences on a plant, attendance rates did not vary consistently with aggregation size at either scale. Similarly, attendance rates were independent of the patch-wide density of larvae. I discuss the potential sources of such unpredictable variation and its implications for the evolution of further specialization in this facultative association.     

Effects of group size and pine defence chemicals on Diprionid sawfly survival against ant predation

The defence chemicals and behavioural adaptations (gregariousness and active defensive behaviour) of pine sawfly larvae may be effective against ant predation. However, previous studies have tested their defences against very few species of ants, and few experiments have explored ant predation in nature. We studied how larval group size (groups of 5 and 20 in Neodiprion sertifer and 10, 20 and 40 in Diprion pini) and variation in levels of defence chemicals in the host tree (Scots pine, Pinus sylvestris) affect the survival of sawfly larvae. Food preference experiments showed that ants do eat sawfly larvae, although they are not their most preferred food item. According to our results, ant predation significantly increases the mortality rate of sawfly larvae. Larval mortality was minor on pine tree branches where ant traffic was excluded. We also found that a high resin acid concentration in the host tree significantly decreased the mortality of D. pini larvae when ants were present. However, there was no such relationship between the chemical concentrations of the host tree and larval mortality for N. sertifer. Surprisingly, grouping did not help sawfly larvae against ant predation. Mortality risk was the same for all group sizes. The results of the study seemingly contradict previous understanding of the effectiveness of defence mechanisms of pine sawfly against ant predation, and suggest that ants (Formica exsecta in particular) are effective predators of sawfly larvae.An erratum to this article can be found at     

Tuned protection of aphids by ants against a predatory hoverfly

1. Aphid‐tending ants that feed on honeydew have evolved strategies against aphidophagous insects and tune their aggressive behaviour according to the level of danger for their trophobionts. Here we investigate how Lasius niger Linnaeus (Hymenoptera: Formicidae) ants react to different instars of Episyrphus balteatus De Geer (Diptera: Syrphidae) hoverflies which vary in their voracity and defensive abilities. 2. During pairwise encounters, early syrphid instars (eggs, L1, and L2 larvae) elicited lower aggression scores compared to third larval instars (L3), which was intensively bitten by ants. L3 tried to escape from ants by releasing a sticky and toxic secretion over biting ants that died or underwent severe morbidity. 3. In a standardised system including the host plant, aphid, tending ant, and hoverfly, the ability of ants to protect an Aphis fabae Scopoli (Hemiptera: Aphididae) colony was evaluated. Early E. balteatus instars placed onto the plant elicited no mobilisation of ants, which often removed the hoverfly successfully. Eggs and early instars appeared as the weak links for integrated pest management by hoverfly auxiliaries. 4. In contrast, L3 induced the number of ant patrollers to increase at a local scale without any further recruitment from inside the ant nest. L3 syrphids were quite efficient at gluing ants with defensive secretions and at resisting to removal attempts by ants. 5. While supporting the assumption that ants tune their defensive response to the aphidophagous predator, the present results also showed a lack of efficient protection of their trophobionts from the most voracious late syrphid instar.