The price of defence: toxins, visual signals and oxidative state in an aposematic butterfly.

In a variety of aposematic species, the conspicuousness of an individual's warning signal and the quantity of its chemical defence are positively correlated. This apparent honest signalling is predicted by resource competition models which assume that the production and maintenance of aposematic defences compete for access to antioxidant molecules that have dual functions as pigments and in protecting against oxidative damage. To test for such trade-offs, we raised monarch butterflies (Danaus plexippus) on different species of their milkweed host plants (Apocynaceae) that vary in quantities of cardenolides to test whether (i) the sequestration of cardenolides as a secondary defence is associated with costs in the form of oxidative lipid damage and reduced antioxidant defences; and (ii) lower oxidative state is associated with a reduced capacity to produce aposematic displays. In male monarchs conspicuousness was explained by an interaction between oxidative damage and sequestration: males with high levels of oxidative damage became less conspicuous with increased sequestration of cardenolides, whereas those with low oxidative damage became more conspicuous with increased levels of cardenolides. There was no significant effect of oxidative damage or concentration of sequestered cardenolides on female conspicuousness. Our results demonstrate a physiological linkage between the production of coloration and oxidative state, and differential costs of sequestration and signalling in monarch butterflies.

Interpreting surveys to estimate the size of the monarch butterfly population: Pitfalls and prospects.

To assess the change in the size of the eastern North American monarch butterfly summer population, studies have used long-term data sets of counts of adult butterflies or eggs per milkweed stem. Despite the observed decline in the monarch population as measured at overwintering sites in Mexico, these studies found no decline in summer counts in the Midwest, the core of the summer breeding range, leading to a suggestion that the cause of the monarch population decline is not the loss of Midwest agricultural milkweeds but increased mortality during the fall migration. Using these counts to estimate population size, however, does not account for the shift of monarch activity from agricultural fields to non-agricultural sites over the past 20 years, as a result of the loss of agricultural milkweeds due to the near-ubiquitous use of glyphosate herbicides. We present the counter-hypotheses that the proportion of the monarch population present in non-agricultural habitats, where counts are made, has increased and that counts reflect both population size and the proportion of the population observed. We use data on the historical change in the proportion of milkweeds, and thus monarch activity, in agricultural fields and non-agricultural habitats to show why using counts can produce misleading conclusions about population size. We then separate out the shifting proportion effect from the counts to estimate the population size and show that these corrected summer monarch counts show a decline over time and are correlated with the size of the overwintering population. In addition, we present evidence against the hypothesis of increased mortality during migration. The milkweed limitation hypothesis for monarch decline remains supported and conservation efforts focusing on adding milkweeds to the landscape in the summer breeding region have a sound scientific basis.

Migration distance as a selective episode for wing morphology in a migratory insect.

BACKGROUND: Selective pressures that occur during long-distance migration can influence morphological traits across a range of taxa. In flying insects, selection should favour individuals that have wing morphologies that increase energy efficiency and survival. In monarch butterflies, differences in wing morphology between migratory and resident populations suggest that migratory populations have undergone selection for larger (as measured by length and area) and more elongated (as measured by roundness and aspect ratio) forewings. However, selection on wing morphology may also occur within migratory populations, particularly if individuals or populations consistently migrate different distances. RESULTS: Using 613 monarch butterflies that were collected on the Mexican wintering grounds between 1976 - 2014, we tested whether monarch wing traits were associated with migratory distance from their natal areas in eastern North America (migration range: 774-4430 km), as inferred by stable-hydrogen (δ2H) and -carbon (δ13C) isotopic measurements. Monarchs that migrated farther distances to reach their overwintering sites tended to have longer and larger wings, suggesting positive selective pressure during migration on wing length and area. There was no relationship between migration distances and either roundness or aspect ratio. CONCLUSIONS: Our results provide correlative evidence that the migratory period may act as a selective episode on monarch butterfly wing morphology, although selection during other portions of the annual cycle, as well as extensive mixing of individuals from various natal locations on the breeding grounds, likely counteracts directional selection of migration on morphology.

Regional climate on the breeding grounds predicts variation in the natal origin of monarch butterflies overwintering in Mexico over 38 years.

Addressing population declines of migratory insects requires linking populations across different portions of the annual cycle and understanding the effects of variation in weather and climate on productivity, recruitment, and patterns of long-distance movement. We used stable H and C isotopes and geospatial modeling to estimate the natal origin of monarch butterflies (Danaus plexippus) in eastern North America using over 1000 monarchs collected over almost four decades at Mexican overwintering colonies. Multinomial regression was used to ascertain which climate-related factors best-predicted temporal variation in natal origin across six breeding regions. The region producing the largest proportion of overwintering monarchs was the US Midwest (mean annual proportion = 0.38; 95% CI: 0.36-0.41) followed by the north-central (0.17; 0.14-0.18), northeast (0.15; 0.11-0.16), northwest (0.12; 0.12-0.16), southwest (0.11; 0.08-0.12), and southeast (0.08; 0.07-0.11) regions. There was no evidence of directional shifts in the relative contributions of different natal regions over time, which suggests these regions are comprising the same relative proportion of the overwintering population in recent years as in the mid-1970s. Instead, interannual variation in the proportion of monarchs from each region covaried with climate, as measured by the Southern Oscillation Index and regional-specific daily maximum temperature and precipitation, which together likely dictate larval development rates and food plant condition. Our results provide the first robust long-term analysis of predictors of the natal origins of monarchs overwintering in Mexico. Conservation efforts on the breeding grounds focused on the Midwest region will likely have the greatest benefit to eastern North American migratory monarchs, but the population will likely remain sensitive to regional and stochastic weather patterns.

The influence of eastern North American autumnal migrant monarch butterflies (Danaus plexippus L.) on continuously breeding resident monarch populations in southern Florida.

In Florida, the eastern North American population of the monarch butterfly exhibits geographic variability in population structure and dynamics. This includes the occurrence of migrants throughout the peninsula during the autumnal migration, occasional overwintering clusters that form along the Gulf Coast, remigrants from Mexico that breed in north-central Florida during the spring, and what have been assumed to be year-round, resident breeding populations in southern Florida. The work reported here focused on two monarch populations west of Miami and addressed four questions: Are there permanent resident populations of monarchs in southern Florida? Do these breed continuously throughout the year? Do they receive northern monarchs moving south during the autumn migration? Do they receive overwintered monarchs returning via Cuba or the Yucatan during the spring remigration from the Mexican overwintering area? Monthly collections and counts of spermatophores in the bursa copulatrices of females established that a resident population of continuously breeding monarchs exists year-round in southern Florida. It was determined through cardenolide fingerprinting that most of the butterflies had bred on the local southern Florida milkweed species, Asclepias curassavica. During the autumn migration period, however, some monarchs had fed on the northern milkweed, Asclepias syriaca. It appears that instead of migrating to Mexico, these individuals travel south through peninsular Florida, break diapause, mate with and become incorporated into the resident breeding populations. None of the monarchs captured in spring had the A. syriaca cardenolide fingerprint, which is evidence against the southern Florida populations receiving overwintered remigrants from Cuba, Central America or Mexico.

Fueling the fall migration of the monarch butterfly.

Monarch butterflies in eastern North America accumulate lipids during their fall migration to central Mexico, and use them as their energy source during a 5 month overwintering period. When and where along their migratory journey the butterflies accumulate these lipids has implications for the importance of fall nectar sources in North America. We analyzed the lipid content of 765 summer breeding and fall migrant monarch butterflies collected at 1 nectaring site in central Virginia over 4 years (1998-2001), and compared them with 16 additional published and unpublished datasets from other sites, dating back to 1941. Virginia migrants store significantly more lipid than summer butterflies, and show significant intraseason and between-year variation. None of the Virginia samples, and none of the historical samples, with one exception, had lipid levels comparable with those found in migrants that had reached Texas and northern Mexico. This evidence suggests that upon reaching Texas, the butterflies undergo a behavioral shift and spend more time nectaring. The one exceptional sample led us to the discovery that monarchs that form roosts along their migratory routes have higher lipid contents than monarchs collected while nectaring at flowers. We propose that for much of their journey monarchs are opportunistic migrants, and the variation within and between samples reflects butterflies' individual experiences. The stored lipids appear to be of less importance as fuel for the butterflies' migration than for their survival during their overwintering period, in part because soaring on favorable winds reduces the energetic cost of flying. The conservation of nectar plants in Texas and northern Mexico is crucial to sustaining the monarch's migratory spectacle, and nectar abundance throughout eastern North America is also important. As generalists in their selection of nectar sources and nectaring habitats, monarchs are unlikely to be affected by small changes in plant communities. Agricultural transformations of natural communities in the eastern United States and Great Plains, however, and especially the extensive planting of genetically modified herbicide-resistant soybeans and corn, may be changing the availability of nectar for monarchs and other pollinators. This new technology is eliminating virtually all forbs in and surrounding agricultural fields, including the monarch's larval hostplants (milkweeds) and native and nonnative nectar sources. To evaluate whether changes in nectar availability are altering the butterflies' ability to accumulate energy, we recommend that monarchs' lipid contents be assayed annually at sites throughout eastern North America.

Behavioral and ecological interactions of foraging mice (Peromyscus melanotis) with overwintering monarch butterflies (Danaus plexippus) in México.

Mice (Peromyscus melanotis) immigrate extensively to overwintering colonies of monarch butterflies (Danaus plexippus) in México. There they feed on both live and dead butterflies that accumulate on the ground and in low vegetation. Through a series of feeding experiments, we examined the potential impact of mouse predation on these colonies, as well as how this predation was influenced by the accessibility and the degree of desiccation of the monarchs. Mice attacked on average 39.9 wet (freshlykilled) butterflies per night. We estimated that a population of mice (75-105 individuals) could kill approximately 0.40-0.57 million butterflies in a 1 ha colony (4-5.7% of the colony) over the 135-day overwintering season. In feeding experiments, mice fed disproportionately on: 1) wet (hydrated) monarchs close to the ground versus those perched higher; 2) wet monarchs, when both wet and dry (desiccated) monarchs were on the ground; and 3) wet monarchs on stakes versus dry monarchs on the ground. Mice commonly ate the entire abdomen of dry monarchs, whereas they fed selectively on the abdomen of wet monarchs by discarding the bitter, cardenolide-laden cuticle and eating the internal tissues. These results suggest that the monarchs' state of desiccation is more important than their accessibility in determining the feeding preferences of these mice. However, the monarchs' strong tendency to crawl up vegetation does appear to reduce their risk to mouse predation.

FORAGING DYNAMICS OF BIRD PREDATORS ON OVERWINTERING MONARCH BUTTERFLIES IN MEXICO.

By suspending nets within and adjacent to a 2.25 hectare overwintering colony of monarch butterflies in Mexico, we estimated that black-backed orioles and black-headed grosbeaks killed 4,550 to 34,300 and an average of 15,067 butterflies per day. A conservative calculation of mortality through the 135 day overwintering season was 2.034 million butterflies, or about 9% of the colony. The birds preyed selectively upon male butterflies, possibly because of a difference in fat content, or possibly because females contain higher concentrations and larger amounts of cardenolide or other defensive chemicals. The risk to individual monarchs of being killed was much greater on the colony periphery and in thinned areas of the forest. Bird predation thus is sufficient to have played a major role in shaping the evolution of the monarch's overwintering and aggregation behavior. Substantial daily variation in predation intensity occurred, 26% of which was attributable to the birds eating more butterflies on colder days, and 30% of which was attributable to a 7.85 day predation cycle. The hypothesis is put forward that the birds feed cyclically because they build up toxic levels of cardenolides or other defensive chemicals contained in the butterflies. The cyclic predation may reduce total predation on the colony by as much as 50%. Such chemical-based group protection is interpreted as a fortuitous by-product of the evolution of unpalatability through selective processes acting on other phases of the monarch's life history.