Two ways to be a myrmecophilous butterfly: natural history and comparative immature‐stage morphology of two species of Theope (Lepidoptera: Riodinidae)

Symbiotic interactions between butterfly larvae and ants, termed myrmecophily, require a range of behavioural and morphological adaptations (ant‐organs). Here, using light and scanning electron microscopy, we describe the complete life cycle of two species of Theope (Lepidoptera: Riodinidae) that have contrasting ways of life. Theope thestias larvae are facultatively tended by several ant species, whereas Theope pieridoides have obligate symbiotic interactions with Azteca ants that inhabit a myrmecophytic tree. Morphological differences associated with their different degrees of intimacy with tending ants are visible from hatching. In T. thestias, the untended first‐instar larva has elongated bifurcated setae and a few tiny perforated cupola organs (PCOs), whereas in T. pieridoides, the ant‐tended first instar has short dendritic setae, larger and more numerous PCOs, and functional tentacle nectary organs (TNOs). Throughout ontogeny, T. pieridoides always shows more conspicuous ant‐organs than T. thestias, with the exception of balloon setae, which are larger and more numerous in T. thestias. In addition, mature T. pieridoides larvae have an anterior set of ant‐organs, including a new type, here described and termed anterior glandular openings (AGOs). Based on the behavioural responses of ants in contact with these structures, a new interpretation for the mechanism whereby Theope larvae can manipulate the behaviour of their tending ants is proposed. Until now, three ecological syndromes can be defined among Theope species: (1) oligophagous larvae with facultative myrmecophily; (2) monophagous larvae with obligate myrmecophily; and (3) polyphagous larvae with obligate myrmecophily. These results suggest that differences in the degree of specificity in the ant–plant interactions may have an important role in the evolution of host‐plant use in Theope. © 2013 The Linnean Society of London     

Polyphagy and Obligate Myrmecophily in the Butterfly Hallonympha paucipuncta (Lepidoptera: Riodinidae) in the Neotropical Cerrado Savanna

This study analyzes host plant use and obligate myrmecophily in Hallonympha paucipuncta, an endemic butterfly of the Cerrado. Larvae of H. paucipuncta are polyphagous, using at least 19 species of plants in 10 families. All larvae found were being tended by Crematogaster ants. Spatial distribution of larvae and tending ants were strongly aggregated, suggesting an influence of ants on oviposition and/or larval survival. Implications of obligate myrmecophily in the evolution of polyphagy in Riodinidae are discussed.     

Behavioral strategy of a lycaenid (Lepidoptera) caterpillar against aggressive ants in a Brazilian savanna

Myrmecophily is widespread in lycaenid butterflies, in which ants receive food resources and, in turn, protect caterpillars against natural enemies. This interaction ranges from obligate myrmecophily, in which immatures are invariably associated with ants and are dependent on ants for survival, and facultative myrmecophily, in which larvae are not dependent on ants for survival, but the presence of the latter may increase larvae survival. Lycaenids also include non-myrmecophilous butterflies, which do not have positive associations with ants and have developed strategies to avoid being attacked or preyed upon by them. In this study, we examined the relationship between the lycaenid Michaelus ira and two ant species associated with Distictella elongata (Bignoniaceae). This plant has extrafloral nectaries and is patrolled by Camponotus crassus and Ectatomma tuberculatum. Morphological analyses revealed that M. ira larvae have ant organs, such as dorsal nectary organs and perforated cupolas, structures associated with myrmecophily. We performed larval exposure experiments in the field, predicting that, in the absence of myrmecophily, the butterfly larva would present strategies to avoid ant attack. Results showed that larvae were attacked by both ant species. To escape ant molestation, larvae lived and fed inside silk-sealed D. elongata flower buds. We concluded that the M. ira bud-sheltering behavior was a defensive strategy against these ant species, while the dorsal nectary organs were apparently nonfunctional. Nonetheless, myrmecophily, in general, cannot be excluded in M. ira since relationships with other ant species may exist.     

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.     

Myrmecophilous butterflies utilise ant–treehopper associations as visual cues for oviposition

1. Selection of a safe oviposition site is important for herbivorous insects whose immature stages have limited mobility. Female herbivores rely on environmental cues for this choice, and presence of natural enemies or mutualistic partners may be important in this process. 2. Some butterflies have mutualistic interactions with ants (myrmecophily), in which caterpillars offer a nutritional liquid and gain protection against natural enemies. Participants in butterfly–ant mutualisms may utilise signals to initiate interactions, but the use of visual cues by ovipositing myrmecophilous butterflies remains uncertain. 3. Larvae of facultatively myrmecophilous Parrhasius polibetes (Lycaenidae) feed on Schefflera vinosa, and females prefer to oviposit near aggregations of the ant‐tended treehopper Guayaquila xiphias, where caterpillars survive better due to increased ant attendance. Given the conspicuousness of ant–treehopper associations, it was investigated whether butterflies use them as visual cues for oviposition and, if so, which participants of the association are used as cues: ants, treehoppers, or both. 4. Experiments using dried insects on paired branches revealed that females visually recognise ants and ant–treehopper associations, using them for egg‐laying decisions. However, presence of a treehopper aggregation alone had no effect on oviposition choices. 5. This is a first insight into the importance of visual discrimination for ovipositing myrmecophilous butterflies. The results show that facultative mutualisms can be important enough to promote a behavioural adaptation (visual detection of ants) reinforcing the interaction. Our research highlights the importance of the behavioural interface within complex multispecies systems.     

A single lycaenid caterpillar gets an ant‐constructed shelter and uninterrupted ant attendance

Ant‐lycaenid associations range from mutualism to parasitism and the caterpillars of some species of lycaenids are reported to enter ant nests for shelter, diapause, or pupation. The present study aimed to examine the nature of the association between Euchrysops cnejus (Fabricius) (Lepidoptera: Lycaenidae) and Camponotus compressus (Fabricius) (Hymenoptera: Formicidae) worker ants on the extrafloral nectary‐bearing cowpea plant, Vigna unguiculata (L.) Walp. (Fabaceae). The abundance patterns of the ants and the lycaenid caterpillars together with the spatial patrolling patterns of the ants on the plants revealed that ant abundance increased with the occurrence of the lycaenid caterpillars and the ants preferred the lycaenids over the extrafloral nectar. Camponotus compressus worker ants constructed a shelter at the cowpea plant base after interaction with one or more lycaenid caterpillar(s) and tended the caterpillars and pupae till the emergence of the butterfly. The ant‐constructed shelters (ACSs) inhabited by the minor caste workers (13 ± 1.3 ants per ACS), were utilized by the caterpillars to undergo pupation. The ants confined their activities predominantly to tending the pod‐feeding caterpillars and the solitary pupa within each ACS. It appears that the behavior of the tending worker ants is modulated by the lycaenid vulnerable stages.     

Behavioural ecology of defence in a risky environment: caterpillars versus ants in a Neotropical savanna

1. Predatory ants may reduce infestation by herbivorous insects, and slow‐moving Lepidopteran larvae are often vulnerable on foliage. We investigate whether caterpillars with morphological or behavioural defences have decreased risk of falling prey to ants, and if defence traits mediate host plant use in ant‐rich cerrado savanna. 2. Caterpillars were surveyed in four cerrado localities in southeast Brazil (70–460 km apart). The efficacy of caterpillar defensive traits against predation by two common ant species (Camponotus crassus, C. renggeri) was assessed through experimental trials using caterpillars of different species and captive ant colonies. 3. Although ant presence can reduce caterpillar infestation, the ants' predatory effects depend on caterpillar defence traits. Shelter construction and morphological defences can prevent ant attacks (primary defence), but once exposed or discovered by ants, caterpillars rely on their size and/or behaviour to survive (secondary defence). 4. Defence efficiency depends on ant identity: C. renggeri was more aggressive and lethal to caterpillars than C. crassus. Caterpillars without morphological defences or inside open shelters were found on plants with decreased ant numbers. No unsheltered caterpillar was found on plants with extrafloral nectaries (EFNs). Caterpillars using EFN‐bearing plants lived in closed shelters or presented morphological defences (hairs, spines), and were less frequently attacked by ants during trials. 5. The efficiency of defences against ants is thus crucial for caterpillar survival and determines host plant use by lepidopterans in cerrado. Our study highlights the effect of EFN‐mediated ant‐plant interactions on host plant use by insect herbivores, emphasizing the importance of a tritrophic viewpoint in risky environments.     

Host-Specific Myrmecophily and Myrmecophagy in the Tropical Coccinellid Diomus thoracicus in French Guiana

A variety of arthropods, particularly insects, have developed myrmecophilous interactions with ants to gain access to resources and/or for protection. Among these myrmecophiles, only a few examples have been documented in the Coccinellidae, most of them involving species able to feed on ant-tended Hemiptera. We report here a new case of obligate myrmecophily in the coccinellid Diomus thoracicus. Larvae are invariably and exclusively found in the nests of the ant Wasmannia auropunctata and seem to rely on ant brood as their only food source. Not only do ant workers show no aggressiveness toward the D. thoracicus larvae in their behavioral interactions at the colonial level, but also at the species level; while coccinellid adults are always attacked. The integration of the larvae inside of the ant nests is based on their chemical mimicry of the host's cuticular cues. Therefore, given the presence of the D. thoracicus larvae inside of the ant's nest, their predation on Wasmannia brood and their chemical mimicry, this species can be considered a specific parasite of W. auropunctata. Overall, this new case of myrmecophily not only specifically involves a highly invasive ant species, but also provides insights into the evolution of myrmecophily and myrmecophagy in coccinellids.     

Ants andPolyommatus icarus immatures (Lycaenidae) —sex-related developmental benefits and costs of ant attendance

Summary Third and fourth instar larvae and pupae of the facultatively myrmecophilous Palaearctic blue butterflyPolyommatus icarus showed no alteration in developmental time when reared in the presence of two species ofLasius ants. Sex differences were observed in larval growth and pupal weight, with males growing larger and faster. Sex-related differences also occurred in the costs and benefits of ant-attendance. Male pupal masses tended to be larger in individuals associated with ants, and their pupal weight loss was not enhanced by ant attendance. This positive developmental effect of myrmecophily is tentatively attributed to a stimulating influence of ants on caterpillar feeding behavior. In contrast, females associated with ants tended to lose more weight during the pupal stage. Hence there is evidence for developmental benefits, rather than costs, of myrmecophily in maleP. icarus immatures, whereas ant attendance appears to be more costly for females during the pupal stage. These findings are discussed in relation to data on other myrmecophilous lycaenid species. It is suggested that maintaining low-level myrmecophily and its related organs is a comparatively inexpensive evolutionary stable strategy among this butterfly group.     

Ant-Homoptera Interactions in a Neotropical Savanna: The Honeydew-Producing Treehopper,Guayaquila xiphias (Membracidae), and its Associated Ant Fauna on Didymopanax vinosum (Araliaceae)1

We investigate the ant fauna associated with the Neotropical treehopper, Guayaquila xiphias, on shrubs of Didymopanax vinosum in the cerrado (savanna) of SE Brazil. Treehoppers infested plants at the border of the cerrado almost exclusively and preferably fed near the apical meristem. During the reproductive phase of the host plant, however, the vast majority of the treehoppers aggregated on the inflorescences. We found 21 ant species harvesting honeydew at G. xiphias aggregations, the most frequent being Camponotus rufipes, Ectatomma edentatum, C. crassus, and C. renggeri. Such a taxonomic diversity of ants tending G. xiphias aggregations in the cerrado is far greater than that reported for any other ant-homopteran system. Daily turnover of ant species at a given treehopper aggregation was observed on 29 percent (64 out of 222) of the G. xiphias aggregations recorded on D. vinosum shrubs. Species replacements probably reflect distinct humidity and temperature ranges tolerated by the species, and may ultimately reduce interspecific competition at homopteran aggregations. Since predation and parasitism on G. xiphias can be severe, and tending ants protect the homopterans against predators and parasitoids, the round-the-clock activity by ants at G. xiphias aggregations is regarded as crucial for the survival of these treehoppers in the cerrado.     

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.     

Trade-offs underlying polyphagy in a facultative ant-tended florivorous butterfly: the role of host plant quality and enemy-free space

The underlying mechanisms mediating the use of multiple host plants were investigated in Parrhasius polibetes (Lycaenidae), a florivorous and facultative myrmecophilous butterfly. Plant traits such as presence of ant–treehopper associations as a source of enemy-free space, flower bud dimensions, toughness, thickness, trichomes, and the corresponding performance and wear of P. polibetes mandibles were examined for three natural hosts: Schefflera vinosa (Araliaceae), Pyrostegia venusta (Bignoniaceae) and Luehea grandiflora (Malvaceae). Parasitism levels of larvae found on the three hosts were also determined. Almost all Luehea had ant–treehopper associations, and all larvae found on this host were non-parasitized. Parasitism was low in larvae found on Schefflera, half of which hosted ant–treehopper associations. No ant–treehopper association was found on Pyrostegia, where parasitism was significantly higher compared to other hosts. In the laboratory, P. polibetes performed well on Schefflera, followed by Pyrostegia. No larvae survived when fed with Luehea. Flower buds of Luehea were thicker and tougher than those of Schefflera and Pyrostegia. Indeed, mandibles of larvae reared on Luehea showed substantial wear, whereas those reared either on Schefflera or Pyrostegia presented no significant damage. Additionally, we suggest that co-occurrence with ant–treehopper associations on a plant provides parasitoid-free space for P. polibetes larvae. Our results support the hypothesis that ecological trade-offs among host plants (i.e., food quality and enemy-free space) promote polyphagy in natural populations of P. polibetes. Host morphological traits seem to play a relevant role in P. polibetes performance. To our knowledge, this is the first report showing the costs of polyphagy in a myrmecophilous butterfly.     

Development of parasitic <Emphasis Type="Italic">Maculinea teleius</Emphasis> (Lepidoptera,Lycaenidae) larvae in laboratory nests of four <Emphasis Type="Italic">Myrmica</Emphasis> ant host species

Maculinea butterflies are social parasites of Myrmica ants. Methods to study the strength of host ant specificity in the Maculinea–Myrmica association include research on chemical and acoustic mimicry as well as experiments on ant adoption and rearing behaviour of Maculinea larvae. Here we present results of laboratory experiments on adoption, survival, development and integration of M. teleius larvae within the nests of different Myrmica host species, with the objective of quantifying the degree of specialization of this Maculinea species. In the laboratory, a total of 94 nests of four Myrmica species: M. scabrinodis, M. rubra, M. ruginodis and M. rugulosa were used. Nests of M. rubra and M. rugulosa adopted M. teleius larvae more readily and quickly than M. ruginodis colonies. No significant differences were found in the survival rates of M. teleius larvae reared by different ant species. Early larval growth of M. teleius larvae differed slightly among nests of four Myrmica host species. Larvae reared by colonies of M. rugulosa which were the heaviest at the beginning of larval development had the lowest mean larval body mass after 18 weeks compared to those reared by other Myrmica species. None of the M. teleius larvae was carried by M. scabrinodis or M. rubra workers after ant nests were destroyed, which suggests a lack of integration with host colonies. Results indicate that Myrmica species coming from the same site differ in their ability to adopt and rear M. teleius larvae but there was no obvious adaptation of this butterfly species to one of the host ant species. This may explain why, under natural conditions, all four ants can be used as hosts of this butterfly species. Slight advantages of particular Myrmica species as hosts at certain points in butterfly larval development can be explained by the ant species biology and colony structure rather than by specialization of M. teleius.     

Association between the African lycaenid,Anthene usamba,and an obligate acacia ant,Crematogaster mimosae

The African lycaenid butterfly, Anthene usamba, is an obligate myrmecophile of the acacia ant, Crematogaster mimosae. Female butterflies use the presence of C. mimosae as an oviposition cue. The eggs are laid on the foliage and young branches of the host plant, Acacia drepanolobium. Larvae shelter in the swollen thorns (domatia) of the host tree, where they live in close association with the acacia ants, and each larva occupies a domatium singly. Anthene usamba are tended by ants that feed from the dorsal nectary organ at regular intervals. Larvae also possess tentacle organs flanking the dorsal nectary organ and appear to signal to ants by everting these structures. Larvae were observed to spend most of their time within the domatia. Stable isotope analysis of matched host plant–ant–butterfly samples revealed that Anthene usamba are δ¹⁵N enriched relative to the ants with which they associate. These data, based on the increase in δ¹⁵N through trophic levels, indicate that the caterpillars of these butterflies are aphytophagous and either exploit the ant brood of C. mimosae within the domatia, or are fed mouth to mouth by adult workers via trophallaxis. This is the first documented case of aphytophagy in African Anthene. Pupation occurs inside the domatium and the imago emerges and departs via the hole chewed by the larva. The adult females remain closely associated with their natal patch of trees, whereas males disperse more widely across the acacia savannah. Females prefer to oviposit on trees with the specific host ant, C. mimosae, an aggressive obligate mutualist, and avoid neighbouring trees with other ant species. Adult butterflies are active during most months of the year, and there are at least two to three generations each year. Observations made over a 5‐year period indicate that a number of different lycaenid species utilize ant‐acacias in East Africa, and these observations are summarized, together with comparisons from the literature. © 2013 The Linnean Society of London, Biological Journal of the Linnean Society, 2013 , 109 , 302–312.     

Not all ants are equal: obligate acacia ants provide different levels of protection against mega‐herbivores

In obligate ant–plant mutualisms, the asymmetric engagement of a single plant species with multiple ant species provides the opportunity for partners to vary in their behaviour. Variation in behaviour has implications for the interactions with third‐party species such as herbivores. This study assessed the effect of obligate ant‐mutualists (Crematogaster mimosae, Crematogaster nigriceps and Tetraponera penzigi) inhabiting the African ant‐acacia (Acacia drepanolobium) on three mega‐herbivore browsers: the Maasai giraffe (Giraffa camelopardalis tippelskirchi), the reticulated giraffe (Giraffa c. reticulata) and the black rhino (Diceros bicornis). Giraffes are abundant and wide‐ranging herbivores of the acacias, whereas black rhinos are localized and perennial herbivores of the acacias. Multiyear field studies comparing the ants’ aggressive behaviour and browsing by mega‐herbivores suggested differences between the tending abilities of the primary ant species inhabiting A. drepanolobium. Trees occupied by the aggressive ant species C. mimosae had significantly less browsing by giraffes and black rhino than trees occupied by other ant species. The results of this study provide evidence that ant‐mutualists on African acacias can serve as deterrents to mega‐herbivores and that different ant species vary in their tending abilities.     

Experimental tests of the mechanism for ant-enhanced growth in an ant-tended lycaenid butterfly

In a previous laboratory study, larvae of the ant-tended lycaenid butterfly Hemiargus isola developed into larger adults when reared with the ant Formica perpilosa than when reared without ants. Ants neither fed butterfly larvae nor significantly delayed developmental duration. We investigated two non-exclusive hypotheses for the mechanism of this effect: larvae tended by F. perpilosa (1) consume more food, and (2) digest the food they consume more efficiently, than those reared without ants. Larvae reared in the laboratory with F. perpilosa ants became significantly heavier adults but produced a significantly lighter fecal mass than their untended counterparts, suggesting that greater food consumption was not the primary mechanism for the higher growth rates of ant-tended larvae. Tended and untended larvae were equally proficient at digesting the contents of pollen (a major natural food source) throughout the tended portion of the life cycle. Taken together, the results suggest that neither greater consumption nor higher assimilation accounts for the larger size of F. perpilosa-tended larvae. We propose that tended larvae may expend less energy than their untended counterparts. Received: 3 January 1997 / Accepted: 18 June 1997     

Ant-partners play a minor role on occurrence of the myrmecophilous butterfly <Emphasis Type="Italic">Leptotes cassius</Emphasis> in its host plant

Ant-related oviposition in facultatively myrmecophilous lycaenid butterflies is common, but not universal. In fact, our knowledge of ant-related oviposition in lycaenids is based on some common species (e.g., Rekoa marius, Allosmaitia strophius, Parrhasius polibetes), which limits generalizations about these systems. In this study, we experimentally investigated whether the oviposition pattern of the florivorous lycaenid Leptotes cassius was influenced by the presence of Camponotus ants and whether larvae were attended, rather than attacked, by ants. This might be evidence of myrmecophily. Both L. cassius and Camponotus ants occur on Bionia coriacea, an extrafloral nectaried legume shrub that grows in the Brazilian cerrado. Plants were randomly assigned to ant-present and ant-excluded treatments and were observed twice throughout the short reproductive season. Larvae of L. cassius were tended by ants, whose attendance was characterized by active antennation on the last body segments of the caterpillars. Therefore, Camponotus can be considered a partner of L. cassius. Lycaenid abundance was on average 1.9- and 1.21-fold higher in plants with ants in each sampling period, respectively, indicating a tendency of L. cassius to occur in plants with ants. Nonetheless, results were not statistically significant, suggesting that in this case ants are not a major cue for lycaenid oviposition. In many ant–lycaenid mutualisms, butterfly immatures benefit from reduced parasitism rates. However, no L. cassius immature, regardless of ant presence or absence, was parasitized. Furthermore, larvae may occur inside flower buds that may provide protection from natural enemies; thus, ants may not be required for immature protection.     

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.     

Introduced ants reduce interaction diversity in a multi‐species,ant–aphid mutualism

Mutualisms contribute in fundamental ways to the origin, maintenance and organization of biological diversity. Introduced species commonly participate in mutualisms, but how this phenomenon affects patterns of interactions among native mutualists remains incompletely understood. Here we examine how networks of interactions among aphid‐tending ants, ant‐tended aphids, and aphid‐attacking parasitoid wasps differ between 12 spatially paired riparian study sites with and without the introduced Argentine ant Linepithema humile in southern California. To resolve challenges in species identification, we used DNA barcoding to identify aphids and screen for parasitoid wasps (developing inside their aphid hosts) from 170 aphid aggregations sampled on arroyo willow Salix lasiolepis. Compared to uninvaded sites, invaded sites supported significantly fewer species of aphid‐tending ants and ant‐tended aphids. At invaded sites, for example, we found only two species of ant‐tended aphids, which were exclusively tended by L. humile, whereas at uninvaded sites we found 20 unique ant–aphid interactions involving eight species of ant‐tended aphids and nine species of aphid‐tending ants. Ant–aphid linkage density was thus significantly lower at invaded sites compared to uninvaded sites. We detected aphid parasitoids in 14% (28/198) of all aphid aggregations. Although the level of parasitism did not differ between invaded and uninvaded sites, more species of wasps were detected within uninvaded sites compared to invaded sites. These results provide a striking example of how the assimilation of introduced species into multi‐species mutualisms can reduce interaction diversity with potential consequences for species persistence.     

Mutualistic ants and parasitoid communities associated with a facultative myrmecophilous lycaenid,Arhopala japonica,and the effects of ant attendance on the avoidance of parasitism

Herbivorous insects have evolved various defensive strategies to avoid their primary enemies, parasitoids. Many species of Lycaenidae (Lepidoptera) have food‐for‐protection mutualism with ants in their larval stages, where larvae produce nectar for ants and in return ants exclude parasitoids as well as predators. Myrmecophilous relationships are divided into two categories, obligate and facultative, by degrees of myrmecophily. Although parasitoids attacking obligate lycaenids always encounter lycaenid‐specific ant species, parasitoids that use facultative lycaenids are likely to encounter diverse ant species showing various defense systems. However, we know little about the parasitoid community of facultative lycaenid larvae. In this study, we investigated the mutualistic ant and parasitoid communities of a facultative myrmecophilous species, Arhopala japonica, in seven localities in Japan. The present field observation newly recorded four ant species attending A. japonica larvae, and combined with the previous data, the number of attending ant species reached 16, which is nearly the maximum number of reported attending ant species among myrmecophilous lycaenids. However, the present study revealed that almost all parasitized A. japonica larvae were attacked by a single braconid species, Cotesia sp. near inducta. We also assessed the efficiency of facultative ant defense against the parasitoid in the laboratory and revealed that oviposition by Cotesia sp. near inducta females was almost completely hindered when A. japonica larvae were attended by ants. This suggests that the dominant parasitoid does not have effective traits to overcome defensive behavior of ants and that the female wasps oviposit mainly in A. japonica larvae without intensive attendance.