Opposing global change drivers counterbalance trends in breeding North American monarch butterflies.

Many insects are in clear decline, with monarch butterflies (Danaus plexippus) drawing particular attention as a flagship species. It is well documented that, among migratory populations, numbers of overwintering monarchs have been falling across several decades, but trends among breeding monarchs are less clear. Here, we compile >135,000 monarch observations between 1993 and 2018 from the North American Butterfly Association's annual butterfly count to examine spatiotemporal patterns and potential drivers of adult monarch relative abundance trends across the entire breeding range in eastern and western North America. While the data revealed declines at some sites, particularly the US Northeast and parts of the Midwest, numbers in other areas, notably the US Southeast and Northwest, were unchanged or increasing, yielding a slightly positive overall trend across the species range. Negative impacts of agricultural glyphosate use appeared to be counterbalanced by positive effects of annual temperature, particularly in the US Midwest. Overall, our results suggest that population growth in summer is compensating for losses during the winter and that changing environmental variables have offsetting effects on mortality and/or reproduction. We suggest that density-dependent reproductive compensation when lower numbers arrive each spring is currently able to maintain relatively stable breeding monarch numbers. However, we caution against complacency since accelerating climate change may bring growing threats. In addition, increases of summer monarchs in some regions, especially in California and in the south, may reflect replacement of migratory with resident populations. Nonetheless, it is perhaps reassuring that ubiquitous downward trends in summer monarch abundance are not evident.

Parasite dynamics in North American monarchs predicted by host density and seasonal migratory culling.

Insect-pathogen dynamics can show seasonal and inter-annual variations that covary with fluctuations in insect abundance and climate. Long-term analyses are especially needed to track parasite dynamics in migratory insects, in part because their vast habitat ranges and high mobility might dampen local effects of density and climate on infection prevalence. Monarch butterflies Danaus plexippus are commonly infected with the protozoan Ophryocystis elektroscirrha (OE). Because this parasite lowers monarch survival and flight performance, and because migratory monarchs have experienced declines in recent decades, it is important to understand the patterns and drivers of infection. Here we compiled data on OE infection spanning 50 years, from wild monarchs sampled in the United States, Canada and Mexico during summer breeding, fall migrating and overwintering periods. We examined eastern versus western North American monarchs separately, to ask how abundance estimates, resource availability, climate and breeding season length impact infection trends. We further assessed the intensity of migratory culling, which occurs when infected individuals are removed from the population during migration. Average infection prevalence was four times higher in western compared to eastern subpopulations. In eastern North America, the proportion of infected monarchs increased threefold since the mid-2000s. In the western region, the proportion of infected monarchs declined sharply from 2000 to 2015, and increased thereafter. For both eastern and western subpopulations, years with greater summer adult abundance predicted greater infection prevalence, indicating that transmission increases with host breeding density. Environmental variables (temperature and NDVI) were not associated with changes in the proportion of infected adults. We found evidence for migratory culling of infected butterflies, based on declines in parasitism during fall migration. We estimated that tens of millions fewer monarchs reach overwintering sites in Mexico as a result of OE, highlighting the need to consider the parasite as a potential threat to the monarch population. Increases in infection among eastern North American monarchs post-2002 suggest that changes to the host's ecology or environment have intensified parasite transmission. Further work is needed to examine the degree to which human practices, such as mass caterpillar rearing and the widespread planting of exotic milkweed, have contributed to this trend.

Revisiting geographic variation in melanism of monarch butterfly larvae in North America using iNaturalist photos.

Monarch butterflies in North America have an exceptionally large breeding distribution, occupying regions west of the Rocky Mountains and throughout the eastern seaboard. An experimental study conducted 17 years ago and published in this journal appeared to show that western monarch larvae tend to have smaller black stripes than those from eastern parents, which at the time was thought to be an adaptation to higher solar exposure in California. Here, we revisit this question by measuring melanism of eastern and western larvae from online photographs submitted to iNaturalist by members of the public. We downloaded over 500 photos of larvae from 10 different states (4 in the west, 6 in the east) and used image analysis to quantify the size of each larva's black stripes (% of surface covered by black). We compared average melanism level between east and west, while also accounting for temperature (at the photo location), seasonal effects, and latitude. Results showed larvae tended to be darker with lower development temperatures, and later in the season, though there was no significant difference between eastern and western larvae in their degree of melanism. It is not yet clear why findings here were different from the prior experimental study. Also, the wild larvae appeared to be notably lighter in color than the captive-reared larvae of the original study, suggesting that the captive environment does not truly replicate the developmental experience of wild larvae, at least in terms of cuticular development. Thus, if there truly are innate tendencies for western larvae to differ (morphologically) from eastern, they are not observable in nature.

Effects of Salinity on Hatchling Diamond-Backed Terrapin (Malaclemys terrapin) Growth, Behavior, and Stress Physiology.

Diamond-backed Terrapins inhabit coastal salt marshes along the eastern and Gulf coasts of North America. Terrapins are adapted to intermediate salinities yet frequently face saltwater-inundated marsh habitat exceeding 25 ppt (or grams/kilogram). We investigated the effect of salinity on the growth of hatchling terrapins and on their compensatory responses to salinity stress. We randomly assigned 30 terrapin hatchlings each to one of five salinity treatments (1, 5, 10, 20, or 35 ppt). Over 75 d, we regularly monitored behavior, appetite, and changes in growth; and calculated ratios of heterophils to lymphocytes (H:L ratio) to assess responses to prolonged salinity stress. Consistent with prior studies, chronic exposure to high salinity significantly reduced hatchling growth. Hatchlings in 20-ppt and 35-ppt salinities exhibited appetite suppression and saltwater avoidance and were more likely to show freshwater-seeking behaviors. H:L ratios were higher among hatchlings in 20-and 35-ppt salinities, consistent with a corticosterone-driven stress response to sustained high-salinity exposure, which may play a role in limiting growth. Our findings suggest hatchling growth and distribution among local habitats will vary spatially depending on habitat salinity and freshwater accessibility. The growth-limiting effects of chronically high salinity or limited access to freshwater could therefore increase hatchling mortality and be an important driver of spatial variation in terrapin demography and abundance. However, when freshwater sources are available, compensatory behaviors might reduce growth-limiting effects. Terrapin recruitment is likely to be impacted as rising sea levels, increased human water use, land development, and other anthropogenic changes alter freshwater inputs to coastal marshes.

A poor substitute for the real thing: captive-reared monarch butterflies are weaker, paler and have less elongated wings than wild migrants.

For many animals and insects that are experiencing dramatic population declines, the only recourse for conservationists is captive rearing. To ensure success, reared individuals should be biologically indistinct from those in the wild. We tested if this is true with monarch butterflies, Danaus plexippus, which are increasingly being reared for release by citizens and commercial breeders. Since late-summer monarchs should be as migration capable as possible for surviving the arduous long-distance migration, we evaluated four migration-relevant traits across two groups of captive-reared monarchs (n = 41 and 42) and one group of wild-caught migrants (n = 41). Monarchs (descendants of wild individuals) were reared from eggs to adulthood either in a warm indoor room next to a window, or in an incubator that mimicked late-summer conditions. Using an apparatus consisting of a perch mounted to an electronic force gauge, we assessed 'grip strength' of all groups, then used image analysis to measure forewing size, pigmentation and elongation. In three of the four traits, reared monarchs underperformed compared to wild ones, even those reared under conditions that should have produced migration-ready individuals. The average strength of reared monarchs combined was 56% less than the wild group, even when accounting for size. Their orange wing colour was paler (an indicator of poor condition and flight ability) and their forewings were less elongated (elongation is associated with migration propensity) than wild monarchs. The reason(s) behind these effects is unknown but could stem from the frequent disturbance and/or handling of reared monarchs, or the fact that rearing removes the element of natural selection from all stages. Regardless, these results explain prior tagging studies that showed reared monarchs have lower migratory success compared to wild.

The sicker the better: nematode-infected passalus beetles provide enhanced ecosystem services.

There is growing appreciation for the role that parasites have in ecosystems and food webs, though the possibility that they could improve an ecosystem service has never been considered. In forest ecosystems, fallen trees naturally decay over time and slowly return their nutrients to the soil. Beetles in the family Passalidae play a key role by excavating tunnels and consuming wood from these logs, thereby breaking down the wood into smaller debris. In the eastern United States, the horned passalus ( Odontotaenius disjunctus) is host to a naturally occurring nematode, Chondronema passali, which appears to cause little harm to the beetles. We suspected this was due to compensatory food consumption by parasitized individuals, which we tested here. We collected and housed 113 adult beetles in individual containers with wood for three months, then determined the amount of wood each beetle had processed into fine debris and frass. We then assessed beetles for C. passali and compared wood processing rates between parasitized and non-parasitized groups. Results showed the average daily processing rate of parasitized beetles ([Formula: see text] = 0.77 g d-1) was 15% greater than that of unparasitized ones ([Formula: see text] = 0.67 g d-1). Parasitized beetles were 6% larger, and this may explain some of this pattern, though the effect of parasitism was still significant in our analysis. By extrapolating the daily rates, we estimate that 10 adult beetles without nematodes would break down approximately 2.4 kg of wood in a single year, while a group of 10 parasitized beetles would break down 2.8 kg. While our data are consistent with the idea of compensatory feeding, because these results are based on natural infections, we cannot rule out the possibility that beetles with heightened wood consumption are simply more likely to acquire the parasite. At an ecosystem level, it may not matter which is the case; parasitized beetles provide a more effective ecosystem service.

Effects of simulated highway noise on heart rates of larval monarch butterflies, Danaus plexippus: implications for roadside habitat suitability.

Developed countries around the world are criss-crossed with vast networks of roadways. Conservationists have recently focused attention on roadsides as possible locations for establishing pollinator habitat, with the monarch butterfly (Danaus plexippus) featuring prominently in such discussions. However, roadsides are inherently loud, which could negatively affect developing larvae. We conducted a series of experiments testing if simulated highway noise stresses monarch larvae, which we gauged by non-destructive monitoring of heart rates. In two replicated experiments, larvae exposed for 2 h experienced a significant increase in heart rate (16 and 17% elevation), indicating they perceive traffic noise as a stressor. Meanwhile, experiments exposing larvae for either 7 or 12 days to continuous traffic noise both showed no heart rate elevation at the end of larval development, suggesting chronic noise exposure leads to habituation or desensitization. Habituation to stress as larvae may impair reactions to real-world stressors as adults, which could be problematic for a butterfly that undertakes an annual two-month migration that is fraught with dangers. More generally, these results could have far-reaching implications for the billions of insects worldwide that develop near roadways, and argue that further study is needed before promoting roadside habitat for butterfly conservation.

Cryptic infection of a broad taxonomic and geographic diversity of tadpoles by Perkinsea protists.

The decline of amphibian populations, particularly frogs, is often cited as an example in support of the claim that Earth is undergoing its sixth mass extinction event. Amphibians seem to be particularly sensitive to emerging diseases (e.g., fungal and viral pathogens), yet the diversity and geographic distribution of infectious agents are only starting to be investigated. Recent work has linked a previously undescribed protist with mass-mortality events in the United States, in which infected frog tadpoles have an abnormally enlarged yellowish liver filled with protist cells of a presumed parasite. Phylogenetic analyses revealed that this infectious agent was affiliated with the Perkinsea: a parasitic group within the alveolates exemplified by Perkinsus sp., a "marine" protist responsible for mass-mortality events in commercial shellfish populations. Using small subunit (SSU) ribosomal DNA (rDNA) sequencing, we developed a targeted PCR protocol for preferentially sampling a clade of the Perkinsea. We tested this protocol on freshwater environmental DNA, revealing a wide diversity of Perkinsea lineages in these environments. Then, we used the same protocol to test for Perkinsea-like lineages in livers of 182 tadpoles from multiple families of frogs. We identified a distinct Perkinsea clade, encompassing a low level of SSU rDNA variation different from the lineage previously associated with tadpole mass-mortality events. Members of this clade were present in 38 tadpoles sampled from 14 distinct genera/phylogroups, from five countries across three continents. These data provide, to our knowledge, the first evidence that Perkinsea-like protists infect tadpoles across a wide taxonomic range of frogs in tropical and temperate environments, including oceanic islands.

Temperature variability and moisture synergistically interact to exacerbate an epizootic disease.

Climate change is altering global patterns of precipitation and temperature variability, with implications for parasitic diseases of humans and wildlife. A recent study confirmed predictions that increased temperature variability could exacerbate disease, because of lags in host acclimation following temperature shifts. However, the generality of these host acclimation effects and the potential for them to interact with other factors have yet to be tested. Here, we report similar effects of host thermal acclimation (constant versus shifted temperatures) on chytridiomycosis in red-spotted newts (Notophthalmus viridescens). Batrachochytrium dendrobatidis (Bd) growth on newts was greater following a shift to a new temperature, relative to newts already acclimated to this temperature (15°C versus 25°C). However, these acclimation effects depended on soil moisture (10, 16 and 21% water) and were only observed at the highest moisture level, which induced greatly increased Bd growth and infection-induced mortality. Acclimation effects were also greater following a decrease rather than an increase in temperature. The results are consistent with previous findings that chytridiomycosis is associated with precipitation, lower temperatures and increased temperature variability. This study highlights host acclimation as a potentially general mediator of climate-disease interactions, and the need to account for context-dependencies when testing for acclimation effects on disease.

Neurofunctional effects of quetiapine in patients with bipolar mania.

OBJECTIVES: Several lines of evidence suggest that abnormalities within portions of the extended limbic network involved in affective regulation and expression contribute to the neuropathophysiology of bipolar disorder. In particular, portions of the prefrontal cortex have been implicated in the appearance of manic symptomatology. The effect of atypical antipsychotics on activation of these regions, however, remains poorly understood. METHODS: Twenty-two patients diagnosed with bipolar mania and 26 healthy subjects participated in a baseline functional magnetic resonance imaging scan during which they performed a continuous performance task with neutral and emotional distractors. Nineteen patients with bipolar disorder were treated for eight weeks with quetiapine monotherapy and then rescanned. Regional activity in response to emotional stimuli was compared between healthy and manic subjects at baseline; and in the subjects with bipolar disorder between baseline and eight-week scans. RESULTS: At baseline, functional activity did not differ between subjects with bipolar disorder and healthy subjects in any region examined. After eight weeks of treatment, subjects with bipolar disorder showed a significant decrease in ratings on the Young Mania Rating Scale (YMRS) (p < 0.001), and increased activation in the right orbitofrontal cortex (OFC) (p = 0.002); there was a significant association between increased right OFC activity and YMRS improvement (p = 0.003). CONCLUSIONS: These findings are consistent with suggestions that mania involves a loss of emotional modulatory activity in the prefrontal cortex--restoration of the relatively greater elevation in prefrontal activity widely observed in euthymic patients is associated with clinical improvement. It is not clear, however, whether changes are related to quetiapine treatment or represent a non-specific marker of affective change.

Evidence of ectoparasite-induced endocrine disruption in an imperiled giant salamander, the eastern hellbender (Cryptobranchus alleganiensis).

Parasitic leeches and trypanosomes release chemical signals into their hosts to evade immuno-detection, but it is unknown whether these compounds manipulate host behavior or endocrine physiology. We determined whether parasitic infections with leeches and/or trypanosomes affected the immune and stress response of an imperiled giant species of amphibian, the eastern hellbender (Cryptobranchus alleganiensis Daudin). We monitored corticosterone and white blood cell counts in response to restraint and injection with adrenocorticotropin hormone (ACTH) or saline for up to 50 h. The presence of leeches dampened hellbender corticosterone responses to restraint and reduced diel patterns of plasma corticosterone. Injection with ACTH restored the normal inter-renal responses of hellbenders, suggesting that leeches, possibly through neurotransmitters in leech saliva, cause down-regulation of corticosterone release at the level of the pituitary or hypothalamus. Infection with leeches also increased the relative abundance of eosinophils, white blood cells often recruited into circulation in response to parasitic infection. Lastly, neutrophil to lymphocyte (N:L) ratios increased in all animals after 24 h of capture and remained elevated for up to 50 h, but these temporal dynamics did not differ with parasite infection. Trypanosome infection did not affect any aspect of hellbender physiology that we measured. Our findings reveal a previously undocumented host-parasite dynamic. While the functional significance to the parasite is unclear, the physiological and behavioral implications for the host are great, given the important role of glucocorticoids in regulating physiology and behavior.

Ciliated Epibionts Modify the Cardiac Stress Reaction to Perceived Predation in Daphnia.

When animals perceive an acute stressor like a predator, they typically undergo a suite of physiological changes that function to improve survival during the encounter, such as elevation in cardiac output, to supply more energy to muscles. If bodily energy is limited, such as by parasites or infections, these functions could become less efficient and lessen host survival. In the aquatic world of microorganisms, individuals can become colonized by other organisms on their surface (epibionts), which could sap energy from their host from their weight, or even compete with the host for food. Here, we tested if one epibiont (a ciliated protozoan, Vorticella spp.) affects its hosts' ability to mount a physiological stress reaction. We collected wild daphnia (Daphnia ambigua) that had varying burdens of these on their bodies and exposed them to a simulated stressor (crushed daphnia, to simulate nearby predation) under a microscope while monitoring for changes in their heart rates in real time. Out of 121 daphnia, those with no Vorticella epibionts showed no meaningful changes in their heart rate after exposure, but those with light or heavy burdens showed immediate elevations (within 5 min). Moreover, the heart rates of heavily burdened daphnia continued to rise for 1.5 h thereafter, to as much as 17% higher than at baseline. These patterns were unexpected, as they suggest that the ciliated epibionts act to elevate their hosts' physiological reaction, rather than dampen it, perhaps by churning the water column around the host, thereby enhancing the chemical alarm cue. The procedures used in this study may be useful for future investigations into the acute stress reactions of daphnia or other microorganisms.

Do Invasive Joro Spiders (Trichonephila clavata) from Asia Avoid Eating Unpalatable Monarch Butterflies (Danaus plexippus) in North America?

An invasive spider from East Asia has established in the U.S. southeast (the "joro spider," Trichonephila clavata) and is rapidly expanding its range. Studies assessing the impact of this species are needed, including how expansive its diet is. An open question is whether monarch butterflies, Danaus plexippus, are a potential prey item for this spider, given that joro spiders do not coexist with monarchs in their native range. Since monarch larvae feed on milkweed, they sequester cardiac glycosides into their adult tissues, rendering them unpalatable to many predators. At sites within northeast Georgia, we staged a series of trials (n = 61) where we tossed monarchs into joro spider webs and, for comparison, performed similar trials with another aposematic species, gulf fritillary (Agraulis vanilla), and a palatable species, tiger swallowtail (Papilio glaucus). We recorded the outcome of the trials, which included whether the spider attacked or did not attack the prey. We also conducted a visual survey during the same fall season to look for evidence of joro spiders consuming monarchs naturally. Our findings revealed that joro spiders avoided eating monarchs; spiders only attacked monarchs 20% of the time, which was significantly less than the attack rates of similarly sized or larger butterflies: 86% for gulf fritillaries and 58% for tiger swallowtails. Some joro spiders even removed monarchs from their webs. From our visual surveys of the surrounding area, we found no evidence of natural monarch consumption and, in general, butterflies made up only a fraction of the joro spider diet. We conclude that joro spiders appear to recognize monarch butterflies as being unpalatable, even without having a prior history with the species. This invokes questions about how these spiders can immediately recognize their unpalatability without touching the butterflies.

The habitat quality paradox: loss of riparian forest cover leads to decreased risk of parasitism and improved body condition in an imperiled amphibian.

Amphibian declines are a global phenomenon but responses of populations to specific threats are often context dependent and mediated by individual physiological condition. Habitat degradation due to reduced riparian forest cover and parasitism are two threats facing the hellbender salamander (Cryptobranchus alleganiensis), but their potential to interact in nature remains largely unexplored. We investigated associations between forest cover, parasitic infection and physiology of hellbenders to test the hypotheses that physiological condition responds to infection and/or habitat degradation. We sampled 17 stream reaches in southwest Virginia, USA, on a year-round basis from 2013 to 2016 and recorded 841 captures of 405 unique hellbenders. At each capture we documented prevalence of two blood-associated parasites (a leech and trypanosome) and quantified up to three physiological condition indices (body condition, hematocrit, white blood cell [WBC] differentials). We used generalized linear mixed models to describe spatiotemporal variation in parasitic infection and each condition index. In general, living in the most heavily forested stream reaches, where hellbender density was highest, was associated with the greatest risk of parasitism, elevated neutrophil-to-lymphocyte (N:L) ratios and eosinophils, slightly lower hematocrit and lower mean body condition in hellbenders. All condition indices fluctuated temporally in a manner consistent with seasonal variation in hellbender metabolic demands and breeding phenology and were associated with land use during at least part of the year. Paradoxically, relatively low levels of forest cover appeared to confer a potential advantage to individuals in the form of release from parasites and improved body condition. Despite improved body condition, individuals from less forested areas failed to exhibit fluctuating body condition in response to spawning, which was typical in hellbenders from more forested habitats. We postulate this lack of fluctuation could be due to reduced conspecific competition or reproductive investment and/or high rates of filial cannibalism in response to declining forest cover.

Widespread trypanosome infections in a population of eastern hellbenders (Cryptobranchus alleganiensis alleganiensis) in Virginia, USA.

Eastern hellbender salamanders (Cryptobranchus alleganiensis alleganiensis) are declining in North America and because of this the health status of individuals in several populations is closely monitored by researchers. During a health survey of hellbenders from a stream in Smyth County, VA, USA, we examined Giemsa-stained blood smears of 71 animals captured during 2011 for the presence of blood parasites. We discovered an unknown species of trypanosome that was apparently widespread within this population; 40 of the 71 individuals (56.3 %) were infected. There are seven known trypanosome species of caudate amphibians; based on microscopic examination, the parasite we observed appeared most similar to Trypanosoma cryptobranchi, which was reported in this species only once before, 76 years ago, from a single animal apparently captured in Iowa. Given that some trypanosomes can adversely affect the health of their hosts, we recommend further monitoring be done in this and other hellbender populations to determine the geographic extent of the parasite and its effects on its increasingly rare host.

Measuring the strength of the horned passalus beetle, Odontotaenius disjunctus: revisiting an old topic with modern technology.

Over a century ago, a pioneering researcher cleverly devised a means to measure how much weight the horned passalus beetle, Odontotaenius disjunctus (Illiger) (Coleoptera: Passalidae), could pull using a series of springs, pulleys, and careful observation. The technology available in modern times now allows for more rigorous data collection on this topic, which could have a number of uses in scientific investigations. In this study, an apparatus was constructed using a dynamometer and a data logger in an effort to ascertain the pulling strength of this species. By allowing beetles to pull for 10 min, each beetle's mean and maximum pulling force (in Newtons) were obtained for analyses, and whether these measures are related was determined. Then, whether factors such as body length, thorax size, horn size, or gender affect either measure of strength was investigated. Basic body measurements, including horn size, of males versus females were compared. The measurements of 38 beetles (20 females, 18 males) showed there was no difference in overall body length between sexes, but females had greater girth (thorax width) than males, which could translate into larger muscle mass. A total of 21 beetles (10 females, 11 males) were tested for pulling strength. The grand mean pulling force was 0.14 N, and the grand mean maximum was 0.78 N. Despite the fact that beetles tended to pull at 20% of their maximum capacity most of the time, and that maximum force was over 5 times larger than the mean force, the 2 measures were highly correlated, suggesting they may be interchangeable for research purposes. Females had twice the pulling strength (both maximum and mean force) as males in this species overall, but when the larger thorax size of females was considered, the effect of gender was not significant. Beetle length was not a significant predictor of pulling force, but horn size was associated with maximum force. The best predictor of both measures of strength appeared to be thorax size. There are a multitude of interesting scientific questions that could be addressed using data on beetle pulling strength, and this project serves as a starting point for such work.

Assessing Leukocyte Profiles of Salamanders and Other Amphibians: A Herpetologists' Guide.

Assessing numbers of leukocytes in salamanders and other amphibians can be useful metrics for understanding health or stress levels of individuals in a population. In this chapter we describe the procedures for obtaining blood samples from amphibians, preparing blood films for microscopy, counting, and identifying cells. We also provide reference values for amphibian leukocytes for use in interpreting leukocyte data. From our assessment of the published and unpublished literature, "non-stressed" salamanders would have a leukocyte profile where 60-70% of cells are lymphocytes, 17-30% are neutrophils, 1-4% are eosinophils, 4-12% are basophils, and 2-6% are monocytes. In Ambystoma spp., the eosinophil abundance can be notably higher (30% of all white blood cells), for reasons unknown. Finally, the neutrophil-lymphocyte ratio of most non-stressed salamanders tends to be between 0.3 and 0.4 (sometimes less), while the ratios of stressed salamanders tend to be over 1.0.

How the monarch got its spots: Long-distance migration selects for larger white spots on monarch butterfly wings.

Elucidating the adaptations that promote flight in animals can aid the understanding of evolution and species divergence, and/or provide inspiration for aerospace engineering and the design of better aerial vehicles. The famed long-distance migration of monarch butterflies in North America still holds many questions and opportunities for inspiration. For example, there is little research on whether the monarch's primary wing colors themselves (black, orange, or white) have any aerodynamic or migration function. Dark colors on wings of other animals have recently been shown to aid flight by enhancing solar absorption, which reduces drag forces. However, too much black surface could be problematic for monarchs, which are exposed to increasing amounts of solar energy along their flightpath. This paper describes the results of two related investigations that attempt to elucidate the importance of wing color to the monarch migration. By measuring the color proportions of nearly 400 monarch wings collected at different stages of their journey, we found, surprisingly, that successful migrants tended to have less black on their wings (about 3% less), but also more white pigment (about 3% more); monarchs have a band of light-colored marginal wing spots. Second, image analysis of museum specimens revealed migratory monarchs had significantly larger white spots, proportional to the wing area, than most non-migratory, New World Danaid butterflies, which argues spot size has evolved along with migratory behavior. Combined, these findings strongly suggest that the long-distance migration itself selects for larger white spots every fall, so that only those individuals with large spots will survive to pass on their genes. Further experimental work is needed to elucidate how the spots aid the migration, but it is possible that they enhance aerodynamic efficiency; other work by the authors demonstrates how alternating white and black pigment on wings can reduce drag. These results will serve as a useful starting point for such endeavors, which should improve understanding of one of the world's most fascinating animal migrations, and also provide practical knowledge for the field of aerospace engineering.

Sex-specific effects of a parasite on stress-induced freezing behavior in a natural beetle-nematode system.

Some animals react to predation threats or other stressors by adopting a freezing posture in an attempt to avoid detection, and the duration of this behavior usually corresponds with individual personality, such that timid individuals freeze longer. Despite decades of research on this or related behaviors (thanatosis), never has the impact of parasitism been considered. Parasites could prolong the duration, if hosts are less motivated to move (i.e. lethargic), or they could reduce it, if hosts are motivated to forage more to compensate for energy drain. We examined this behavior within a natural beetle-nematode system, where hosts (horned passalus beetles, Odontotaenius disjunctus) are parasitized by a nematode, Chondronema passali. We exposed beetles (n = 238) to four stressors in our lab, including noise, vibration, light and inversion, and recorded how long they adopt a frozen stance. Afterward, we determined nematode burdens, which can range from dozens to hundreds of worms. Beetles tended to freeze for 20 seconds on average, with some variation between stressors. We detected no effect of beetle mass on the duration of freezing, and this behavior did not differ in beetles collected during the breeding or non-breeding season. There was a surprising sex-based difference in the impact of nematodes; unparasitized females remained frozen twice as long as unparasitized males, but for beetles with heavy nematode burdens, the opposite was true. From this we infer that heavily parasitized females are more bold, while males with heavy burdens would be more timid. The explanation for this finding remains elusive, though we can rule out many possibilities based on prior work on this host-parasite system.

Monarchs Reared in Winter in California Are Not Large Enough to Be Migrants. Comment on James et al. First Population Study on Winter Breeding Monarch Butterflies, Danaus plexippus (Lepidoptera: Nymphalidae) in the Urban South Bay of San Francisco, California. Insects 2021, 12, 946.

A recent study in this journal aimed to understand certain changes in the wintering behavior of monarch butterflies, specifically in the western subpopulation of North America [...].