Dynamics of stored lipids in fall migratory monarch butterflies (Danaus plexippus): Nectaring in northern Mexico allows recovery from droughts at higher latitudes.

The eastern population of the North American monarch butterfly (Danaus plexippus) overwinters from November through March in the high-altitude (3000 m+) forests of central Mexico during which time they rely largely on stored lipids. These are acquired during larval development and the conversion of sugars from floral nectar by adults. We sampled fall migrant monarchs from southern Canada through the migratory route to two overwintering sites in 2019 (n = 10 locations), 2020 (n = 8 locations) and 2021 (n = 7 locations). Moderate to extreme droughts along the migratory route were expected to result in low lipid levels in overwintering monarchs but our analysis of lipid levels of monarchs collected at overwintering sites indicated that in all years most had high levels of lipids prior to winter. Clearly, a significant proportion of lipids were consistently acquired in Mexico during the last portion of the migration. Drought conditions in Oklahoma, Texas and northern Mexico in 2019 resulted in the lowest levels of lipid mass and wing loading observed in that year but with higher levels at locations southward in Mexico to the overwintering sites. Compared with 2019, lipid levels increased during the 2020 and 2021 fall migrations but were again higher during the Mexican portion of the migration than for Oklahoma and Texas samples, emphasizing a recovery of lipids as monarchs advanced toward the overwintering locations. In all 3 years, body water was highest during the Canada-USA phase of migration but then declined during the nectar foraging phase in Mexico before recovering again at the overwintering sites. The increase in mass and lipids from those in Texas to the overwintering sites in Mexico indicates that nectar availability in Mexico can compensate for poor conditions experienced further north. Our work emphasizes the need to maintain the floral and therefore nectar resources that fuel both the migration and storage of lipids throughout the entire migratory route.

Differences in δ13C and δ15N values for monarch butterflies raised on milkweeds from within and outside of crop fields: implications for inferences about natal origins.

Stable isotope (δ2H, δ13C) measurements of wing tissue have been used to determine the natal geographic origin of migrant monarch butterflies that overwinter in Mexico. This study examines the possibility of using δ13C and δ15N to identify the milkweed habitat used by monarchs in their natal region. Milkweeds were common in corn and soybean fields before herbicide use led to their extirpation around 2006, and the loss of those milkweeds has been proposed as a reason for the monarch population decline. If crop-field monarchs can be identified, then historical samples of monarchs could be examined to test that hypothesis. The δ15N and δ13C values of leaves from milkweeds growing in corn fields, soybean fields and non-agricultural habitats were examined as well as monarchs that were raised on those leaves. There were no δ15N values for leaves or monarchs that were distinctive for crop fields. Milkweeds in corn fields, and monarchs that were raised on those milkweeds, were found to have δ13C values distinctly lower than those of other habitats and unlike those of locations within the summer breeding range. Thus, it should be possible to identify monarchs that came from cornfields in samples of overwintering monarchs made before ca. 2006.

Monarch Butterfly Ecology, Behavior, and Vulnerabilities in North Central United States Agricultural Landscapes.

The North American monarch butterfly (Danaus plexippus) is a candidate species for listing under the Endangered Species Act. Multiple factors are associated with the decline in the eastern population, including the loss of breeding and foraging habitat and pesticide use. Establishing habitat in agricultural landscapes of the North Central region of the United States is critical to increasing reproduction during the summer. We integrated spatially explicit modeling with empirical movement ecology and pesticide toxicology studies to simulate population outcomes for different habitat establishment scenarios. Because of their mobility, we conclude that breeding monarchs in the North Central states should be resilient to pesticide use and habitat fragmentation. Consequently, we predict that adult monarch recruitment can be enhanced even if new habitat is established near pesticide-treated crop fields. Our research has improved the understanding of monarch population dynamics at the landscape scale by examining the interactions among monarch movement ecology, habitat fragmentation, and pesticide use.

Monarch butterfly population decline in North America: identifying the threatening processes.

The monarch butterfly (Danaus plexippus) population in North America has sharply declined over the last two decades. Despite rising concern over the monarch butterfly's status, no comprehensive study of the factors driving this decline has been conducted. Using partial least-squares regressions and time-series analysis, we investigated climatic and habitat-related factors influencing monarch population size from 1993 to 2014. Potential threats included climatic factors, habitat loss (milkweed and overwinter forest), disease and agricultural insecticide use (neonicotinoids). While climatic factors, principally breeding season temperature, were important determinants of annual variation in abundance, our results indicated strong negative relationships between population size and habitat loss variables, principally glyphosate use, but also weaker negative effects from the loss of overwinter forest and breeding season use of neonicotinoids. Further declines in population size because of glyphosate application are not expected. Thus, if remaining threats to habitat are mitigated we expect climate-induced stochastic variation of the eastern migratory population of monarch butterfly around a relatively stationary population size.

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.

Density estimates of monarch butterflies overwintering in central Mexico.

Given the rapid population decline and recent petition for listing of the monarch butterfly (Danaus plexippus L.) under the Endangered Species Act, an accurate estimate of the Eastern, migratory population size is needed. Because of difficulty in counting individual monarchs, the number of hectares occupied by monarchs in the overwintering area is commonly used as a proxy for population size, which is then multiplied by the density of individuals per hectare to estimate population size. There is, however, considerable variation in published estimates of overwintering density, ranging from 6.9-60.9 million ha-1. We develop a probability distribution for overwinter density of monarch butterflies from six published density estimates. The mean density among the mixture of the six published estimates was ∼27.9 million butterflies ha-1 (95% CI [2.4-80.7] million ha-1); the mixture distribution is approximately log-normal, and as such is better represented by the median (21.1 million butterflies ha-1). Based upon assumptions regarding the number of milkweed needed to support monarchs, the amount of milkweed (Asclepias spp.) lost (0.86 billion stems) in the northern US plus the amount of milkweed remaining (1.34 billion stems), we estimate >1.8 billion stems is needed to return monarchs to an average population size of 6 ha. Considerable uncertainty exists in this required amount of milkweed because of the considerable uncertainty occurring in overwinter density estimates. Nevertheless, the estimate is on the same order as other published estimates. The studies included in our synthesis differ substantially by year, location, method, and measures of precision. A better understanding of the factors influencing overwintering density across space and time would be valuable for increasing the precision of conservation recommendations.

Quasi-extinction risk and population targets for the Eastern, migratory population of monarch butterflies (Danaus plexippus).

The Eastern, migratory population of monarch butterflies (Danaus plexippus), an iconic North American insect, has declined by ~80% over the last decade. The monarch's multi-generational migration between overwintering grounds in central Mexico and the summer breeding grounds in the northern U.S. and southern Canada is celebrated in all three countries and creates shared management responsibilities across North America. Here we present a novel Bayesian multivariate auto-regressive state-space model to assess quasi-extinction risk and aid in the establishment of a target population size for monarch conservation planning. We find that, given a range of plausible quasi-extinction thresholds, the population has a substantial probability of quasi-extinction, from 11-57% over 20 years, although uncertainty in these estimates is large. Exceptionally high population stochasticity, declining numbers, and a small current population size act in concert to drive this risk. An approximately 5-fold increase of the monarch population size (relative to the winter of 2014-15) is necessary to halve the current risk of quasi-extinction across all thresholds considered. Conserving the monarch migration thus requires active management to reverse population declines, and the establishment of an ambitious target population size goal to buffer against future environmentally driven variability.

Ecophysiological consequences of non-random leaf orientation in the prairie compass plant, Silphium laciniatum.

The prairie compass plant (Silphium laciniatum L.) has vertical leaves that are characteristically oriented in a north-south plane (i.e., the flat surfaces of the lamina face east and west). We explored the consequences of this orientation by determining basic photosynthetic and water use characteristics in response to environmental factors and by determining total daily photosynthesis and water use of leaves held in different orientations. Average maximum CO2 exchange rate (CER) of leaves near Ames, IA was constant at 22 micromol m-2 s-1 from May through August and then declined. CER did not exhibit a distinct lightsaturation point. CER at photon flux densities near full sunlight was constant from 22 to 35°C leaf temperature but declined at higher temperatures. However, leaf temperatures rarely exceed 35°C during the growing season. There was no change in the pattern of response of CER to temperature over the growing season. We constrained leaves to face east-west (EW,=natural), to face north-south (NS), or to be horizontal (HOR) on eight days in 1986-1988. EW leaves had the highest light interception, leaf temperatures, CER, and transpiration early and late in the day, whereas HOR leaves had the highest values in the middle of the day. Integrations of CER and transpiration over the eight daytime periods showed EW and HOR leaves to have equivalent carbon gain, higher than that of NS leaves. HOR leaves had the highest daily transpiration. Daily water use efficiency (WUE, carbon gained/water lost) was always highest in EW leaves, with the HOR leaves having 16% lower WUE and NS leaves having 33% lower WUE. The natural orientation of compass plant leaves results in equivalent or higher carbon gain and in increased WUE when compared to leaves with other possible orientations; this is likely to have a selective advantage in a prairie environment.

The distribution of standing crop of nectar: what does it really tell us?

Brink (1982) characterizes the distribution of standing crop of nectar for Delphinium nelsonii as bonanzablank, based on comparison with a Poisson. He then discusses possible effects of standing crop variability on pollinator foraging behavior. We disagree with the use of the Poisson and the resulting conclusions. The expected distribution should not be based on doling out random amounts of nectar to flowers, but based on random return times to flowers by pollinators (elapsed time=nectar accumulated). When this model is used, standing crop variance does not differ markedly from expectation. What differences do exist can be accounted for by variability in nectar production rates of individual plants. We also take issue with the use of the bonanza-blank terminology. As originally formulated this refers to nectar production differences within a plant rather than standing crop differences among plants.

Nectar production rates of Asclepias quadrifolia: causes and consequences of individual variation.

The causes and reproductive consequences of individual variation in nectar production rates within a population of Asclepias quadrifolia were investigated. Two parameters were correlated with nectar production rate per flower: the root weight of the plant and the number of flowers in the inflorescence (umbel). Nectar production increased with increasing root weight but levelled off after a root size of about 3 g was reached. Nectar production decreased with increasing umbel size, but only for umbels that were greater than average size. A total of 57% of the variance in individual nectar production could be explained by these two variables with root weight accounting for 67% of the explained variance. Root weight is a good indicator of a plant's energetic status, indicating the importance of available energy in determining quantity of nectar produced. About 30% of the energy devoted to flowering is utilized in nectar production. Nectar production was significantly correlated with the male component of reproductive fitness, pollinaria removal, but not with the female component, pollinia insertion. Since pod production is limited by resource availability rather than the number of pollinia insertions, nectar production in A. quadrifolia is most closely associated with the maximization of the male function.

Patchiness in the dispersion of nectar resources: Evidence for hot and cold spots.

The dispersion pattern of resources can have a significant effect on foraging behavior. We examined the dispersion pattern of standing crop of nectar in a population of Delphinium nelsonii. The nectar content of flowers was measured for subject inflorescences and their two nearest neighbors. Inflorescences were divided into those which had some nectar (hot plants) and those which had none (cold plants). Tests for independence showed that subject plants and their neighbors were likely to have the same "temperature", indicating that resources were patchy. The implications of this pattern for the foraging movements of bumblebees are discussed.