Crypsis versus intimidation—anti-predation defence in three closely related butterflies

Butterflies that hibernate exhibit particularly efficient defence against predation. A first line of defence is crypsis, and most hibernating butterflies are leaf mimics. When discovered, some species have a second line of defence; the peacock, I. io, when attacked by a predator flicks its wings open exposing large eyespots and performs an intimidating threat display. Here we test the hypothesis that butterflies relying solely on leaf mimicking and butterflies with an intimidating wing pattern, when attacked, exhibit different behavioural suites—because leaf mimicking is best implemented by immobility, whereas intimidating coloration is best implemented by intimidating behaviour. In laboratory experiments blue tits, Parus caeruleus, were allowed 40 min to attack single individuals of three species of butterfly: one relying solely on crypsis, the comma, Polygonia c-album; one relying on intimidating wing pattern in addition to crypsis, the peacock; and one intermediate species, the small tortoiseshell Aglais urticae. The results are in accordance with expectations and demonstrate that: (1) birds take longer to discover the leaf mimicking species, the comma, than the tortoiseshell and the peacock; (2) the comma remained motionless throughout experimental trials but small tortoiseshells and peacocks flicked their wings when attacked; (3) the most intimidating butterfly, the peacock, started flicking its wings at a greater distance from the attacking bird than the small tortoiseshell; and (4) the intimidating pattern and behaviour of peacocks was effective—when discovered, all peacocks survived interactions with blue tits, whereas only 22% of commas and 8% of small tortoiseshells survived.     

Auditory defence in the peacock butterfly (<Emphasis Type="Italic">Inachis io</Emphasis>) against mice (<Emphasis Type="Italic">Apodemus flavicollis</Emphasis> and <Emphasis Type="Italic">A. sylvaticus</Emphasis>)

Morphological and behavioural traits can serve as anti-predator defence either by reducing detection or recognition risks, or by thwarting initiated attacks. The latter defence is secondary and often involves a ‘startle display’ comprising a sudden release of signals targeting more than one sensory modality. A suggested candidate for employing a multimodal defence is the peacock butterfly, Inachis io, which, by wing-flicking suddenly, produces sonic and ultrasonic sounds and displays four large eyespots when attacked. The eyespots make small birds retreat, but whether the sounds produced thwart predator attacks is largely unknown. Peacocks hibernate as adults in dark wintering sites and employ their secondary defence upon encounter with small rodent predators during this period. In this study, we staged predator–prey encounters in complete darkness in the laboratory between wild mice, Apodemus flavicollis and Apodemus sylvaticus, and peacocks which had their sound production intact or disabled. Results show that mice were more likely to flee from sound-producing butterflies than from butterflies which had their sound production disabled. Our study presents experimental evidence that the peacock butterfly truly employs a multimodal defence with different traits targeting different predator groups; the eyespots target birds and the sound production targets small rodent predators.     

“An eye for an eye?”—on the generality of the intimidating quality of eyespots in a butterfly and a hawkmoth

Large eyespots on the wings of butterflies and moths have been ascribed generally intimidating qualities by creating a frightening image of a bird or mammal much larger than the insect bearing the eyespots. However, evidence for this anti-predator adaptation has been largely anecdotal and only recently were peacock butterflies, Inachis io, shown to effectively thwart attacks from blue tits, Parus caeruleus. Here, we test whether large eyespots on lepidopterans are generally effective in preventing attacks from small passerines and whether the size of insect or bird can influence the outcome of interactions. We staged experiments between the larger eyed hawkmoths, Smerinthus ocellatus, and the smaller peacock butterflies, I. io, and the larger great tits, Parus major, and the smaller blue tits, P. caeruleus. Survival differed substantially between the insect species with 21 of 24 peacock butterflies, but only 6 of 27 eyed hawkmoths, surviving attacks from the birds. Thus, surprisingly, the smaller prey survived to a higher extent, suggesting that factors other than insect size may be important. However, great tits were less easily intimidated by the insects’ eyespots and deimatic behaviour and consumed 16 of 26, but the blue tits only 8 of 25, of the butterflies and hawkmoths. Our results demonstrate that eyespots per se do not guarantee survival and that these two insects bearing equally large eyespots are not equally well protected against predation.     

Antennal responses to floral scents in the butterflies <Emphasis Type="Italic">Inachis io</Emphasis>, <Emphasis Type="Italic">Aglais urticae</Emphasis> (Nymphalidae), and <Emphasis Type="Italic">Gonepteryx rhamni</Emphasis> (Pieridae)

Summary. To better understand the biological role of floral scents for butterflies, electrophysiological responses to floral scents were investigated using combined gas chromatography and electroantennographic detection (GC-EAD). The antennal responses of three butterfly species, Aglais urticae L. (Nymphalidae), Inachis io L. (Nymphalidae), and Gonepteryx rhamni L. (Pieridae) to floral scent compounds from both natural and synthetic mixtures were examined. Floral scents were collected from the butterfly nectar plants Cirsium arvense (L.) (Asteraceae), and Buddleja davidii Franchet cv. (Loganicaeae) with dynamic head-space methods on Tenax-GR and eluted with pentane. These eluates, composed of natural floral scent blends, represent an array of compounds in their natural state. In the GC-EAD analyses eleven compounds were identified from C. arvense with the benzenoid compound phenylacetaldehyde in highest abundance. Seventeen compounds were identified from B. davidii with the irregular terpene oxoisophorone in highest abundance. Thirty-nine synthetic floral scent compounds were mixed in pentane, in equal amounts; about 35 ng were allowed to reach the antennae. The butterflies showed antennal responses to most of the floral scent compounds from both natural and synthetic blends except to the highly volatile monoterpene alkenes. Certain benzenoid compounds such as phenylacetaldehyde, monoterpenes such as linalool, and irregular terpenes such as oxoisophorone, were emitted in relatively large amounts from C. arvense and B. davidii, and elicited the strongest antennal responses. These compounds also elicited strong antennal responses when present in the synthetic scent blends. Thus, the butterflies seem to have many and /or sensitive antennal receptors for these compounds, which points to their biological importance. Moreover, these compounds are exclusively of floral scent origin. For B. davidii, which depends highly on butterflies for pollination, the exclusive floral scent compounds emitted in high abundance could be the result of an adaptive pressure to attract butterflies. Received 2 Septemter 2001; accepted 9 September 2002.     

Smooth-billed ani (Crotophaga ani) predation on butterflies in Mato Grosso, Brazil: risk decreases with increased group size

Although most insects are vulnerable to predation by a variety of predators, including birds, there are few direct observational studies in the wild of avian predation on adult butterflies. We examined the predatory behavior of smooth-billed anis (Crotophaga ani) on butterflies, and the spacing behavior of the butterflies which were concentrated on a mineral-rich beach on the Cristalino River, in Mato Grosso, Brazil. We studied eight of the most regularly occurring butterfly species which came each morning to engage in "puddling." Most species of butterfly were closely associated with conspecifics, although nearest-neighbor distance varied among species. The pierids - "yellows" (Aphrissa statira, Phoebis trite), "oranges" (Phoebis argante), and sometimes "whites" (Daptoneura leucadia) - formed very dense groups (or clusters) of up to 1,000 individuals occasionally joined by a few kite swallowtails (Eurytides spp.). Most other butterfly species formed small groups (e.g., daggerwings, Marpesia spp.) or were dispersed individually and non-clumped over the beach (e.g., dingy purplewing, Eunica monima). Anis foraged using two strategies: rapid frontal attack on dense groups of butterflies (yellows, oranges, whites), and a stealthy approach to solitary butterflies (mainly purplewings) or those in small groups. For yellows, the most common butterfly caught by anis, the capture rate reached over 6 per 15 min per ani, and about 8% of those captured managed to escape. Capture rates were much lower for the other species. Time of day, age of the ani (adult or young-of-the-year), and total number of each butterfly species present accounted for variation in the number of each species captured by anis. The number of butterflies captured per 15 min increased as the number of butterflies present increased, but reached a threshold beyond which the capture rate did not increase. The capture rate per individual butterfly (individual risk) decreased with group size up to a group of 40 individuals and remained low with further increases. Thus a butterfly in a group of 100 was no less likely to be eaten than one in a group of 40. For individual ani forays into dense groups of pierids, an individual ani was unable to catch more than 16, regardless of group size. These data confirm the dilution effect of group size for butterflies; each individual yellow or orange was less at risk from ani predation when in a group.     

Foraging responses in the butterflies <Emphasis Type="Italic">Inachis io</Emphasis>, <Emphasis Type="Italic">Aglais urticae</Emphasis> (Nymphalidae), and <Emphasis Type="Italic">Gonepteryx rhamni</Emphasis> (Pieridae) to floral scents

Summary. For butterflies to be efficient foragers, they need to be able to recognize rewarding flowers. Flower signals such as colours and scents assist this recognition process. For plant species to attract and keep butterflies as pollinators, species-specific floral signals are crucial. The aim of this study is to investigate foraging responses to floral scents in three temperate butterfly species, Inachis io L. (Nymphalidae), Aglais urticae L. (Nymphalidae), and Gonepteryx rhamni L. (Pieridae), in behavioural choice bioassays. The butterflies were allowed to choose bet-ween flower models varying in scent and colour (mauve or green). Flowers or vegetative parts from the plants Centaurea scabiosa L. (Asteraceae), Cirsium arvense (L.) (Asteraceae), Knautia arvensis (L.) (Dipsacaceae), Buddleja davidii Franchet (Loganicaeae), Origanum vulgareL. (Lamiaceae), Achillea millefolium L. (Asteraceae), and Philadelphus coronarius L. (Hydrangiaceae) were used as scent sources. All visits to the models — those that included probing and those that did not — were counted, as was the duration of these behaviours. Both flower-naive and flower-experienced (conditioned to sugar-water rewards, the colour mauve, and specific floral scents) butterflies were tested for their preference for floral versus vegetative scents, and to floral scent versus colour. The butterflies were also tested for their ability to switch floral scent preferences in response to rewards. Flower-naive butterflies demonstrated a preference for the floral scent of the butterfly-favourable plants C. arvense and K. arvensis over the floral scent of the non-favourable plants Achillea millefolium (Asteraceae), and Philadelphus coronarius cv. (Hydrangiaceae). Most of the butterflies that were conditioned to floral scents of either C. arvense, K. arvensis, or B. davidii readily switched theirfloral scent preferences to the one most recently associated with reward, thus demonstrating that floral scent constancy is a result from learning. These findings suggest that these butterflies use floral scent as an important cue signal to initially identify and subsequently recognize and distinguish among rewarding plants. Received 2 September 2001; accepted 9 September 2002.     

Body morphologies shape territorial dominance in the satyrine butterfly <Emphasis Type="Italic">Lethe diana</Emphasis>

Male butterflies compete over mating territories via aerial interactions. It has often been suggested that flight performance, and thus flight-related traits, would play an important role in butterflies’ contest resolution; however, most studies have not demonstrated a clear relationship between these traits and territory ownership in butterflies. Males of the satyrine butterfly Lethe diana compete over territories via linear chases, a behavior that is uncommon among butterflies: rather, contests of most butterfly species consist of circling flights. In the present study, some morphological and physiological traits of L. diana were compared between territory owners and intruders to search for traits correlated with resource holding power (RHP). Territory owners had higher flight-muscle ratio (FMR) and were heavier than intruders, indicating that flight performance plays a key role in their contest resolution. Contest duration was not affected by the owners’ FMR or body mass, and thus I obtained no evidence supporting the hypothesis that contestants assess their relative RHP before making a decision to retreat. I speculate that the fact that these traits had an effect on territorial status in L. diana was a result of the characteristics of the contest behavior of this butterfly. During their contests, males L. diana fly faster in their linear flights than males of most other butterflies, which engage in slower circling flights. Therefore, males of L. diana probably need higher flight performance capabilities in order to win their contests.     

Recognising one’s enemies: a functional approach to risk assessment by prey

Little has been done to compare the relative importance of various mechanisms through which prey assess the potential risk from natural enemies. We used predator-naive spider mites (Tetranychus urticae, Tetranychidae) to (1) compare the responses of prey to chemical cues from enemy and non-enemy species and (2) investigate the source of these cues. In the laboratory, we observed the distribution of T. urticae in response to cues from nine mite species, including (1) predators of spider mites, (2) predators/parasites of other animals, and (3) fungivores/pollen-feeders. When given a choice over 24 h, spider mites foraged and oviposited in fewer numbers on leaf discs that were previously exposed to predatory or parasitic mites (including species incapable of attacking spider mites) than on clean leaf discs (unexposed to mites). Interestingly, previous exposure of leaf arenas to fungivores and pollen-feeders had no significant effect on spider mite distribution. We then observed the response of T. urticae to cues from two species of predator that had been reared on a diet of either spider mites or pollen. T. urticae showed stronger avoidance of leaf discs that were previously exposed to spider-mite-fed predators than of discs exposed to pollen-fed predators. Nevertheless, for one predator species (Amblyseius andersoni), T. urticae still preferred to forage and oviposit on clean (unexposed) discs than on discs exposed to pollen-fed predators. Protein-derived metabolic wastes of predatory or parasitic mites may provide a general cue about potential predation risk for T. urticae. However, T. urticae also avoided areas exposed to pollen-fed predators, suggesting there may be other sources of enemy recognition by the spider mites. We discuss the ecological and evolutionary mechanisms that may influence the scope of information through which animals assess predation risk. Received: 11 January 1999 / Received in revised form: 25 October 1999 / Accepted: 20 November 1999     

Cardenolide content,emetic potency,and thin-layer chromatography profiles of monarch butterflies,Danaus plexippus,and their larval host-plant milkweed,Asclepias humistrata,in Florida

Summary This paper is the fourth in a series on cardenolide fingerprints of monarch butterflies (Danaus plexippus, Danainae) and their host-plant milkweeds (Asclepiadaceae) in the eastern United States. Cardenolide concentrations ofAsclepias humistrata plants from north central Florida ranged from 71 to 710 µg/0.1 g dry weight, with a mean of 417 µg/0.1 g. Monarchs reared individually on these plants contained cardenolide concentrations ranging from 243 to 575 µg/0.1 g dry weight, with a mean of 385 µg/0.1 g. Cardenolide uptake by butterflies was independent of plant concentration, suggesting that sequestration saturation occurs in monarchs fed cardenolide-rich host plants. Thinlayer chromatography resolved 19 cardenolides in the plants and 15 in the butterflies. In addition to humistratin,A. humistrata plants contained several relatively non-polar cardenolides of the calotropagenin series which are metabolized to more polar derivatives in the butterflies. These produced a butterfly cardenolide fingerprint clearly distinct from those previously established for monarchs reared on otherAsclepias species. In emetic assays with the blue jay,Cyanocitta cristata, the 50% emetic dose (ED₅₀) per jay was 57.1 µg, and the average number of ED₅₀ units per butterfly was 13.8, establishing that this important south eastern milkweed produces highly emetic, chemically defended monarchs. Our data provide further support for the use of cardenolide fingerprints of wild-caught monarchs to make ecological predictions concerning defence against natural enemies, seasonal movement and larval host-plant utilization by monarch butterflies during their annual cycle of migration, breeding and overwintering.     

The downfall of mating: the effect of mate-carrying and flight muscle ratio on the escape ability of a pierid butterfly

In many species, the ability to evade predators is known to be periodically impaired by increased weight loads due to feeding and reproduction. Not only may extra weight reduce escape speed, but feeding and mating can also make the prey more noticeable to visually hunting predators. A number of butterfly species mate for hours, and if a mating couple is disturbed, one of the butterflies is responsible for flying, whereas its partner remains still. This study investigated the ability of male Pieris napi butterflies to fly while mating, with the prediction that mate carrying impairs flight ability compared to single flying males and that males with relatively high flight muscle ratios (FMR; male thorax mass/male + female body mass) will have better flight performance in copula. Our results clearly show that whereas single males always take off at steep angles and fly upwards, couples invariably have a negative take-off angle and rarely gain height. Moreover, landing height of the couples is positively associated with higher FMR. Hence, male flight ability when in copula is positively associated with a high relative thorax mass. Butterfly pairs may thus be at greater risk of predation as a consequence of their impaired flight ability, especially couples with critically low FMRs (<16%).     

Energy conservation in hibernating endotherms: Why “suboptimal” temperatures are optimal

Many endotherms use facultative heterothermic responses of torpor or hibernation to conserve energy during periods of low energy availability. A common assumption when estimating winter energy budgets is that endotherms should hibernate at the ambient temperature (T_a) that minimizes torpid metabolic rate (TMR) and maximizes the duration of torpor bouts. However, previous studies of the energetic benefits of hibernation have assumed constant T_a within hibernacula. Here we use an individual-based energetic model to estimate overwinter energy expenditure of mammals hibernating at T_as that vary temporally. We show that, in accordance with the principles of Jenson's inequality, hibernators can conserve energy by selecting microclimates warmer than the single T_a value that minimizes TMR (T_min). As temporal variation in T_a increases, endotherms should choose microclimates with mean T_as progressively warmer than T_min. Further, as thermal conductance decreases, as it does with increasing body mass and use of social thermoregulation, the mean T_a that minimizes overwinter energy expenditure approaches, but never equals, T_min. We suggest that the commonly held assumption of stable microclimates in hibernacula has skewed the interpretation of the optimal expression of hibernation for energy conservation. Our results contradict much of the accepted understanding of hibernation energetics and add to a growing body of literature proposing that hibernating at a T_a warmer than T_min is optimal.     

Hibernation clustering and its consequences for associative nest foundation in<Emphasis Type="Italic"> Polistes dominulus</Emphasis> (Hymenoptera Vespidae)

Various studies indicate that cuticular hydrocarbons (CHCs) act as cues for nestmate recognition in paper wasps and other social insects. A recent study showed that associative nest foundation in Polistes dominulus is mainly performed by foundresses coming from the same locality. In the present study, we induced future foundresses of P. dominulus collected in two different localities to hibernate in the laboratory in aggregates of individuals from the same or different localities. After 2 months of hibernation, foundresses from different localities but from the same experimental cluster did not show any preference to associate, at the time of nest foundation, with individuals coming from the same original locality. The cuticular chemical profiles of individuals from the mixed hibernation clusters were quite similar and significantly different from those of individuals which hibernated with other foundresses from the same locality. These findings suggest that, in this species, mechanisms other than nestmate chemical recognition play a major role in the spring association of gynes during the foundation of a new nest.Communicated by R.F.A. Moritz     

Sequestration of pyrrolizidine alkaloids by larvae ofTellervo zoilus (Lepidoptera: Ithomiinae) and their role in the chemical protection of adults against the spiderNephila maculata (Araneidae)

Summary Life stages of the primitive Australian ithomiine butterflyTellervo zoilus and its larval hostplant, the apocynaceous vineParsonsia straminea, were quantitatively assayed for pyrrolizidine alkaloids (PAs). PAs were found in all stages, mainly as N-oxides, being most concentrated in larvae and freshly-emerged adults. Although adults feed at various confirmed PA sources this probably does not compensate for losses, as wild-caught adults had considerably lower concentrations of PAs. The main alkaloid present in both freshly-emerged adults and in leaves of the host-plant was lycopsamine (1b), stored by butterflies in the N-oxide form. Its presence in higher proportion, in relation to intermedine (1a), in larvae, pupae and adults ofTellervo in relation to the host-plants suggests the inversion of intermedine to lycopsamine by the insects. No 14-member ring macrocyclic PAs were detected in either food-plant or butterflies. Several other PAs were found in wild-caught adults reflecting visits to other PA sources. PAs were also found in high concentrations in freshly-emerged individuals of the danaineEuploea core bred onParsonsia straminea. Wild-caughtDanaus affinis had high PA levels acquired from adult feeding. Freshly emergedEuploea raised onIschnocarpus frutescens andDanaus raised onIschnostemma carnosum (both PA-free) were preyed on by the orb weaving spiderNephila maculata, and showed no PAs. In all cases where PAs were present, most butterflies were liberated, usually cut out of the web unharmed, byNephila. The spider's response was not closely linked to PA concentration, however, and may also depend on hunger levels and previous experience with PA-containing butterflies. All control and other non-PA containing butterflies were consumed although rejection of some body parts of freshly-emergedDanaus affinis suggests that compounds other than PAs may be involved.     

Seed choice by rodents: learning or inheritance?

Learning plays a central role in animal life, and it has received special attention in the context of foraging. In this study, we have tested whether learning operates in seed choices by rodents using the Algerian mouse (Mus spretus) and Holm oak (Quercus ilex) acorns as a model. At the laboratory, those rodents captured in the field during the acorn fall period (experienced individuals) rejected weevil-infested acorns, presumably because of their lower energetic value. By contrast, rodents born in captivity and reared without any contact with acorns (naïves) predated weevil-infested acorns at similar rates than sound ones. After exposing naïves to infested and sound acorns during 15 days, they rejected infested acorns as the experienced individuals. In the field, predation on weevil-infested acorns was lower than on sound ones. Predation rates on infested acorns were lowest at the end of the acorn fall season, whereas predation rates of sound acorns increased along the same period. This might be explained by the improved ability to reject infested acorns because of the accumulated experience acquired by the initially naïve rodents along the acorn fall season. We show that learning shapes strongly seed choices by rodents, and it may be advantageous over inherited behaviors in variable unpredictable situations, such as acorn infestation rates that vary strongly between years and trees. We consider that the role of learning has to be taken into account in future studies on seed predation by rodents.     

Tropane and pyrrolizidine alkaloids in the ithomiinesPlacidula euryanassa andMiraleria cymothoe (Lepidoptera: Nymphalidae)

Summary Larvae of the ithomiine butterflyPlacidula euryanassa sequester tropane alkaloids (TAs) from the host plantBrugmansia suaveolens and pass them through the pupae to freshly emerged adults. Wild caught adults also show in their tissues, variable amounts of pyrrolidizine alkaloids (PAs), probably sequestered from variable plant sources and subject to dynamics of incorporation, accumulation and utilization of PAs by ithomiine butterflies. The ratio TAs/PAs is also variable between different populations.Miraleria cymothoe, another ithomiine that feeds onB. suaveolens as larvae, does not sequester TAs from the host-plant, but sequesters PAs from plant sources visited by the adult butterflies. The main alkaloid found in both butterflies is lycopsamine, which also is the principal PA found in all genera of Ithomiinae.     

Prevalence, patterns, and effects of shell damage on Geukensia demissa in South Carolina estuarine habitats

Shelled molluscs frequently exhibit a record of damage on exterior surfaces that can evidence past predation attempts and may affect survival and growth. In South Carolina populations of the ribbed marsh mussel, Geukensia demissa, >90% of the individuals and up to 60% of the total shell area are damaged. A trend toward greater amounts of damage occurred on mid-marsh compared to oyster reef mussels from the barrier beach side of inlets. Shell damage effects on survivorship and shell and tissue growth were assessed seasonally during multi- and single-season field experiments. Mussels from a common mid-marsh site were divided into size classes (~50 or 70 mm), treated to create two damage levels (undamaged and damaged), and replaced within mid-marsh exclusion cages to minimize additional shell damage. In both multi- and single-season experiments increased shell damage resulted in significantly greater mortality. Linear shell growth was unaffected by increased damage, but 50 mm mussels grew twice as fast. Shell mass increased 16–50% in the multi-season and single-season winter period, but decreased 7–12% during the single-season summer period. Tissue mass significantly decreased 31–43% in 50 mm damaged mussels, but increased by 33% for 70 mm mussels in both multi-season and the single-season winter period experiments. Shell damage did reduce tissue mass 43% in 70 mm single-season summer mussels. Experimental results indicate shell damage from a simulated increase in predation can affect negatively both survival and growth of marsh mussels. Seasonal timing of shell damage and initial mussel size also influenced the effects of sublethal predation on shell and tissue growth. The previously unrecognized importance of sublethal predation and the resultant significant negative effects of shell damage on survival and growth will affect the distribution and population dynamics of G. demissa in coastal marshes and will influence the overall contribution of ribbed mussels to estuarine ecosystems.     

To flee or not to flee: predator avoidance by cheetahs at kills

Mammalian carnivores are unusual because their primary competitors for food are often their primary predators. This relationship is most evident at persistent kills where dominant competitors are attracted to both the carcass (as a free meal) and to the killers (as potential prey). Cheetahs (Acinonyx jubatus) are frequent victims of kleptoparasitism, and cubs, and sometimes adults, are killed by lions (Panthera leo) or spotted hyenas (Crocuta crocuta). Between 1980 and 2002, we observed 639 kills made by cheetahs in Serengeti National Park, Tanzania. These kills were often visited by scavengers, including relatively innocuous species such as vultures and jackals and potentially dangerous species, like spotted hyenas and lions. We used cheetah behavior at kills to test a number of predictions about how cheetahs should minimize risk at kill sites given they face an increased risk of predation of themselves or their cubs. In particular, we examined the propensity of cheetahs of different age/sex classes to hide carcasses after making a kill, vigilance at kills, and the delay in leaving after finishing feeding with respect to ecological factors and scavenger presence. The behavior of single females at kills did not suggest that they were trying to avoid being killed, but the behavior of males, often found in groups, was in line with this hypothesis. In contrast, the behavior of mother cheetahs at kills appeared to be influenced greatly by the risk of cubs being killed. Our results suggest that cheetahs use several behavioral counterstrategies to avoid interspecific predation of self or cubs.     

Molecular studies on the ouabain binding site of the Na+, K+-ATPase in milkweed butterflies

Summary. The Na⁺, K⁺-ATPase of the Monarch butterfly (Danaus plexippus) is insensitive to the inhibition by cardiac glycosides due to an amino acid replacement: histidine instead of asparagine at position 122 of the α-subunit representing the ouabain binding site. By PCR amplification of the DNA sequence of this site, a PCR product of 270 bp was obtained from DNA extracted from Danainae species (Danaus plexippus, D. chrysippus, D. gillipus, D. philene, D. genutia, Tirumala hamata, Euploea spp., Parantica weiskei, P. melusine), Sphingidae (Daphnis nerii) and mimics of milkweed butterflies (Hypolimnas missipus, Limenitis archippus and L. arthemis, Nymphalidae). Analysis of the nucleotide sequences revealed that the single point mutation in the ouabain binding domain (AAC-Asn for CAC-His) was present only in Danaus plexippus, but not in the other species investigated. Since these milkweed butterflies also store cardenolides, other structural modifications of the Na⁺, K⁺-ATPase may have occurred or other strategies of cardenolide tolerance have been developed. Received 15 May 2000; accepted 29 June 2000     

Effects of fertilization and elevated CO<Subscript>2</Subscript> on larval food and butterfly nectar amino acid preference in<Emphasis Type="Italic"> Coenonympha pamphilus</Emphasis> L.

The effects of larval diet on the nutritional preferences of butterflies has rarely been examined. This study investigates whether alterations in the larval diet result in changes in adult preferences for nectar amino acids. Larvae of Coenonympha pamphilus were raised on fertilized or unfertilized Festuca rubra, grown under ambient (350 ppm) or elevated (750 ppm) atmospheric CO ₂environments. Fertilization led to marked increases in leaf nitrogen concentration. In plants grown under elevated CO ₂conditions, leaf water and nitrogen concentrations were significantly lower, and the C/N-ratio increased significantly. Fertilization of the host plant shortened the development time of C. pamphilus larvae, and pupal weight increased. In contrast, larvae of C. pamphilus developed significantly slower on F. rubra grown under elevated CO ₂, but adult emergence weight was not affected by CO ₂treatment of the plant. C. pamphilus females showed a clear preference for nectar mimics containing amino acids, whereas males, regardless of treatment, either preferred the nectar mimic void of amino acids or showed no preference for the different solutions. Female butterflies raised on fertilized plants showed a significant decline in their preference for nectar mimics containing amino acids. A slight, but not significant, trend towards increased nectar amino acid preference was found in females raised on plants grown under elevated CO ₂. We clearly demonstrate that alterations in larval host quality led to changes in butterfly nectar preferences. The ability of the butterfly to either rely less on nectar uptake or compensate for poor larval conditions represents a trade-off between larval and adult butterfly feeding.     

Oviposition stimulant for the zebra swallowtail butterfly, Eurytides marcellus, from the foliage of pawpaw, Asimina triloba

Summary. We have isolated a caffeoylcyclohexane-1-carboxylic acid derivative, 3-caffeoyl-muco-quinic acid (3-CmQA), as a contact oviposition stimulant for the zebra swallowtail butterfly, Eruytides marcellus (Papilionidae), from the foliage of its primary host plant, Asimina triloba (Annonaceae). This compound alone was as active in stimulating oviposition by females as were the parent ethanolic plant extract and the host plant itself. Other tested isomers of 3-CmQA, including 5-caffeoylquinic acid (5-CQA or trans-chlorogenic acid), were inactive. We found, however, that experienced female butterflies responded strongly to host volatiles, which enhanced landing rates and hence oviposition.? This is the first report of an oviposition stimulant for a swallowtail butterfly of the tribe Graphiini. We found 3-CmQA to be the major caffeoylcyclohexane-1-carboxylic acid isomer in plants of the genus Asimina. These plants lack appreciable amounts of 5-CQA, which has been shown previously to be one of the oviposition stimulants for certain Rutaceae- or Apiaceae-feeding swallowtails of the related tribe Papilionini.? Our findings, along with earlier results from the tribes Troidini and Papilionini, suggest that responses by swallowtails to hydroxycinnamic acid derivatives as oviposition cues date back at least to the ancestor of the subfamily Papilioninae. Received 24 March 1998; accepted 27 May 1998.