Proteome of urticating setae of Ochrogaster lunifer, a processionary caterpillar of medical and veterinary importance, including primary structures of putative toxins.

Ochrogaster lunifer (Lepidoptera: Notodontidae) is an Australian processionary caterpillar with detachable urticating setae that have a defensive function. These true setae induce inflammation when they contact human skin, and equine foetal loss syndrome if they are accidentally ingested by gravid horses. We used transcriptomics and proteomics to identify proteins and peptides present in and on urticating setae, which may include toxins that contribute to inflammation and/or foetal loss syndromes. This process identified 37 putative toxins, including multiple homologues of the honeybee venom peptide secapin, and proteins with similarity to odorant binding proteins, arylphorins, and the insect immune modulator Diedel. This work identifies candidate molecules that may contribute to the adverse effects of processionary caterpillar setae on human and animal health.

Population genetics of a recent range expansion and subsequent loss of migration in monarch butterflies.

Range expansions-whether permanent or transient-strongly influence the distribution of genetic variation in space. Monarch butterflies are best known for long-distance seasonal migration within North America but are also established as nonmigratory populations around the world, including on Pacific Islands. Previous research has highlighted stepwise expansion across the Pacific, though questions remain about expansion timing and the population genetic consequences of migration loss. Here, we present reduced-representation sequencing data for 275 monarchs from North America (n = 85), 12 Pacific Islands (n = 136) and three locations in Australia (n = 54), with the goal of understanding (i) how the monarch's Pacific expansion has shaped patterns of population genetic variation and (ii) how loss of migration has influenced spatial patterns of differentiation. We find support for previously described stepwise dispersal across the Pacific and document an additional expansion from Hawaii into the Mariana Islands. Nonmigratory monarchs within the Mariana Islands show strong patterns of differentiation, despite their proximity; by contrast, migratory North American samples form a single genetically panmictic population across the continent. Estimates of Pacific establishment timing are highly uncertain (~100-1,000,000 years ago) but overlap with historical records that indicate a recent expansion. Our data support (i) a recent expansion across the Pacific whose timing overlaps with available historical records of establishment and (ii) a strong role for seasonal migration in determining patterns of spatial genetic variation. Our results are noteworthy because they demonstrate how the evolution of partial migration can drive population differentiation over contemporary timescales.

Current and future potential distributions of Helicoverpa punctigera (Lepidoptera: Noctuidae): is this the next FAW?

Helicoverpa punctigera (Wallengren), the native budworm, is an important highly polyphagous pest that has caused serious damage on a wide variety of crops in Australia. In Australia, its range overlaps that of its congener, Helicoverpa armigera (Hübner), a notorious invasive pest globally. We used CLIMEX, a bioclimatic niche modelling software package, to estimate the potential geographical distribution of H. punctigera under current and future climates (A1B scenario). Under both current and future climate conditions, the model indicates that H. punctigera could establish throughout the tropics and subtropics. Comparing the potential distributions under each climate scenario revealed that in the future its potential distribution is likely to shift poleward and into higher altitudes, into areas that are currently too cold as observed in the South of Brazil, Europe, North America, South East Asia, and South Pacific Islands including New Zealand. The projected potential distribution can inform pre- and post-border biosecurity strategies for the management of this pest in each country.

Malice at the Gates of Eden: current and future distribution of Agrilus mali threatening wild and domestic apples.

The apple buprestid, Agrilus mali Matsumura, that was widespread in north-eastern China, was accidently introduced to the wild apple forest ecosystem in mountainous areas of Xinjiang, China. This invasive beetle feeds on domesticated apples and many species of Malus and presents a serious threat to ancestral apple germplasm sources and apple production worldwide. Estimating the potential area at risk of colonization by A. mali is crucial for instigating appropriate preventative management strategies, especially under global warming. We developed a CLIMEX model of A. mali to project this pest's potential distribution under current and future climatic scenarios in 2100 using CSIRO-Mk 3.0 GCM running the SRES A1B emissions scenario. Under current climate, A. mali could potentially invade neighbouring central Asia and eventually the mid-latitude temperate zone, and some subtropical areas and Pampas Steppe in the Southern Hemisphere. This potential distribution encompasses wild apples species, the ancestral germplasm for domesticated apples. With global warming, the potential distribution shifts to higher latitudes, with the potential range expanding slightly, though the overall suitability could decline in both hemispheres. In 2100, the length of the growing season of this pest in the mid-latitude temperature zone could increase by 1-2 weeks, with higher growth rates in most sites compared with current climate in mid-latitudes, at least in China. Our work highlights the need for strategies to prevent the spread of this pest, managing the threats to wild apples in Tian Shan Mountain forests in Central Asia, and commercial apple production globally. We discuss practical management tactics to reduce the spread of this pest and mitigate its impacts.

Pollen Alone or a Mixture of Pollen Types? Assessing Their Suitability for Mass Rearing of Neoseiulus cucumeris (Acari: Phytoseiidae) Over 20 Generations.

The generalist predatory mite, Neoseiulus cucumeris (Oudemans) is known as one of the most effective natural enemies on many pests. This economically important biocontrol agent was reared for 20 generations on date palm and castor bean pollen as well as a mixture of pollen types, including date palm, castor bean, and almond. The performance of this predator was evaluated by comparing its life table parameters after different generations (G1-G20) fed on each diet in a laboratory at 25 ± 1°C, 60 ± 5% RH, and a photoperiod of 16:8 (L: D) h. The development time of the predator reared on all tested diets had no significant difference in G20. The intrinsic rate of increase (r) of N. cucumeris by feeding on the mixed pollen (0.197 day-1) was significantly higher than that on castor bean, or date palm pollen (0.146 or 0.152 day-1 in G1, respectively). Our results indicated that the predator's performance was not affected by long-term feeding on the pollen diets, as well as there was no considerable difference between pollen alone and pollen mixture diets. Furthermore, mites reared on pollen diets had higher quality than those reared on natural prey, Tetranychus urticae Koch.

Disentangling thermal effects using life cycle simulation modelling on the biology and demographic parameters of Dolichogenidea gelechiidivoris, a parasitoid of Tuta absoluta.

Dolichogenidea gelechiidivoris (Marsh) (Syn. Apanteles gelechiidivoris) (Hymenoptera: Braconidae), a konoibiont larval endoparasitoid of the South American tomato pinworm Tuta absoluta (Meyrick), was imported into Kenya in 2017 for the first classical biological control of T. absoluta in Africa. We determined the thermal thresholds of D. gelechiidivoris, with T. absoluta as host, using life cycle simulation modelling. Life-table data of D. gelechiidivoris were generated at six constant temperatures (10, 15, 20, 25, 30, and 35 °C, 65 ± 5% RH, and 12L: 12D photoperiod). Multiple non-linear functions were fitted to model some aspects of the biology of the parasitoid, including its longevity, mortality, reproduction, and senescence using Insect Life Cycle Modelling (ILCYM) software. The phenology models established were used to estimate life table parameters. Except at 35 °C, D. gelechiidivoris completed its development (i.e., egg-larval-pupa-adult) at all tested temperatures. The minimum temperature threshold from egg to cocoon was 7.0 °C and 5.9 °C from cocoon to adult wasp, while the maximum temperature thresholds were 34 °C and 36 °C, respectively. The optimal temperature for immature survival was 20-25 °C, while fecundity was optimum at 22.5 °C, with 69.5 eggs per female. Simulations of the population growth parameters indicated that the intrinsic rate of increase (rm) was maximum at 20 °C with 15 daughters per female per generation. Based on our findings the release of D. gelechiidivoris for inoculative augmentation in countries with temperatures ranging between 15 and 30 °C could be considered to control the pest.

Seasonal spatial dynamics of butterfly migration.

Understanding the seasonal movements of migratory species underpins ecological studies. Several hundred butterfly species show migratory behaviour, yet the spatial pattern of these migrations is poorly understood. We developed climatic niche models for 405 migratory butterfly species globally to estimate patterns of seasonal movement and the distribution of seasonal habitat suitability. We found strong seasonal variation in habitat suitability for most migratory butterflies with >75% of pixels within their distributions showing seasonal switching in predicted occupancy for 85% of species. The greatest rate of seasonal switching occurred in the tropics. Several species showed extreme range fluctuations between seasons, exceeding 10-fold for 53 species (13%) and more than 100-fold for nine species (2%), suggesting that such species may be at elevated extinction risk. Our results can be used to search for the ecological processes that underpin migration in insects, as well as to design conservation interventions for declining migratory insects.

Spatial orientation of social caterpillars is influenced by polarized light.

Processionary caterpillars of Thaumetopoea pityocampa (in Europe) and Ochrogaster lunifer (in Australia) (Lepidoptera: Notodontidae) form single files of larvae crawling head-to-tail when moving to feeding and pupation sites. We investigated if the processions are guided by polarization vision. The heading orientation of processions could be manipulated with linear polarizing filters held above the leading caterpillar. Exposure to changes in the angle of polarization around the caterpillars resulted in corresponding changes in heading angles. Anatomical analysis indicated specializations for polarization vision of stemma I in both species. Stemma I has a rhabdom with orthogonal and aligned microvilli, and an opaque and rugged surface, which are optimizations for skylight polarization vision, similar to the dorsal rim of adult insects. Stemmata II-VI have a smooth and shiny surface and lobed rhabdoms with non-orthogonal and non-aligned microvilli; they are thus optimized for general vision with minimal polarization sensitivity. Behavioural and anatomical evidence reveal that polarized light cues are important for larval orientation and can be robustly detected with a simple visual system.

Behavioral and Physiological Plasticity Provides Insights into Molecular Based Adaptation Mechanism to Strain Shift in Spodoptera frugiperda.

How herbivorous insects adapt to host plants is a key question in ecological and evolutionary biology. The fall armyworm, (FAW) Spodoptera frugiperda (J.E. Smith), although polyphagous and a major pest on various crops, has been reported to have a rice and corn (maize) feeding strain in its native range in the Americas. The species is highly invasive and has recently established in China. We compared behavioral changes in larvae and adults of a corn population (Corn) when selected on rice (Rice) and the molecular basis of these adaptational changes in midgut and antennae based on a comparative transcriptome analysis. Larvae of S. frugiperda reared on rice plants continuously for 20 generations exhibited strong feeding preference for with higher larval performance and pupal weight on rice than on maize plants. Similarly, females from the rice selected population laid significantly more eggs on rice as compared to females from maize population. The most highly expressed DEGs were shown in the midgut of Rice vs. Corn. A total of 6430 DEGs were identified between the populations mostly in genes related to digestion and detoxification. These results suggest that potential adaptations for feeding on rice crops, may contribute to the current rapid spread of fall armyworm on rice crops in China and potentially elsewhere. Consistently, highly expressed DEGs were also shown in antennae; a total of 5125 differentially expressed genes (DEGs) s were identified related to the expansions of major chemosensory genes family in Rice compared to the Corn feeding population. These results not only provide valuable insight into the molecular mechanisms in host plants adaptation of S. frugiperda but may provide new gene targets for the management of this pest.

The genetics of monarch butterfly migration and warning colouration.

The monarch butterfly, Danaus plexippus, is famous for its spectacular annual migration across North America, recent worldwide dispersal, and orange warning colouration. Despite decades of study and broad public interest, we know little about the genetic basis of these hallmark traits. Here we uncover the history of the monarch's evolutionary origin and global dispersal, characterize the genes and pathways associated with migratory behaviour, and identify the discrete genetic basis of warning colouration by sequencing 101 Danaus genomes from around the globe. The results rewrite our understanding of this classic system, showing that D. plexippus was ancestrally migratory and dispersed out of North America to occupy its broad distribution. We find the strongest signatures of selection associated with migration centre on flight muscle function, resulting in greater flight efficiency among migratory monarchs, and that variation in monarch warning colouration is controlled by a single myosin gene not previously implicated in insect pigmentation.

Parasite-host specificity: A cross-infection study of the parasite Ophryocystis elektroscirrha.

Many parasites are constrained to only one or a few hosts, showing host specificity. It remains unclear why some parasites are specialists and other parasites are generalists. The parasite Ophryocystis elektroscirrha (OE) is a neogregarine protozoan thought to be restricted to monarch butterflies, Danaus plexippus (Nymphaliae) and D. gilippus. Recently, we found OE-like spores in other Lepidoptera, specifically in three noctuid moths: Helicoverpa armigera, H. assulta and H. punctigera, as well as another nymphalid, Parthenos sylvia. To our knowledge, this is the first report of OE-like parasite infections in species other than the genus Danaus. In sequencing 558 bp of 18S rRNA, we found the genetic similarity between OE from D. plexippus and OE-like parasite from the moths H. armigera and H. punctigera to be 95.2%. When we conducted cross-species infection experiments, we could not infect the moths with OE from D. plexippus, but OE-like parasite from H. armigera did infect D. plexippus and a closely related moth species Heliothis virescens. Interestingly, we did not find the OE-like parasite in the H. armigera population from Spain. Inter-population infection experiments with H. armigera demonstrated a higher sensitivity to OE-like infection in the population from Spain compared to the populations from Australia and China. These results suggest geographic variation in OE-like susceptibility and coevolution between parasite and host. Our findings give important new insights into the prevalence and host specificity of OE and OE-like parasites, and provide opportunities to study parasite transmission over spatial and temporal scales.

Reducing the fitness of an invasive weed, Parthenium hysterophorus: Complementing biological control with plant competition.

We studied the effects of a biological control agent, Epiblema strenuana Walker (Lepidoptera: Tortricidae) alone and together with a sown native grass, Astrebla squarrosa C.E. Hubb. and an introduced pasture plant, Clitoria ternatea L. on growth and seed production of Parthenium hysterophorus L. Astrebla squarrosa and C. ternatea individually reduced shoot dry biomass of P. hysterophorus by 30 and 42%, respectively; and by 48 and 70%, respectively in the presence of biological control agent, E. strenuana. Similarly, A. squarrosa and C. ternatea individually reduced weed seed production up to 48 and 64%, respectively; and by 73 and 81%, respectively in the presence of E. strenuana. In the presence of E. strenuana, the biomass of A. squarrosa and C. ternatea was increased by 13 and 10%, respectively. The biological control agent induced more galls per P. hysterophorus plant when either of the competing plants were present than when grown alone. The abundance of galls increased with pasture competition, but only for C. ternatea, and not for A. squarrosa. The biological control agent worked synergistically with the two competitive plants to reduce the growth and production of viable seed, which should lead to a decrease in the P. hysterophorus soil seed banks in the field, and eventually seedling recruitment in future generations of P. hysterophorus.

Moving to Keep Fit: Feeding Behavior and Movement of Helicoverpa armigera (Lepidoptera: Noctuidae) on Artificial Diet With Different Protein: Carbohydrate Ratios.

Insect herbivores can modify their foraging behavior to obtain a balanced food intake, and they tend to move between food sources with different nutrient values. We investigated this movement in early instar larvae of Helicoverpa armigera (Hübner) (Lepidoptera: Noctuidae) using a putative optimal artificial diet (OP) and high protein (HP) and high carbohydrate (HC) artificial diets based on protein (p) and carbohydrate (c) ratios. Larvae were allowed to choose between the same kind of diet cubes (effectively no-choice), or diet cubes with different p: c ratios. In no-choice tests, we found that first instar larvae remained longest on OP diet and spent the least time on HC diet, while third instar larvae remained longest on HC diet and spent least time on OP diet. First instar larvae moved the most when provided with HC diet, while third instar larvae moved most when provided with OP diet. However, both stages moved the least when allowed to choose between diet cubes with different p: c ratios. The relative growth rate decreased when larvae increased their movement, but this influence was not evident when larvae fed on HC diet. Larvae that fed only on HC diet had the highest relative growth rate, followed by larvae with access to all diets simultaneously, indicating a behavior to mix nutrient intake. We relate these findings to behavior of this major pest species under field conditions.

Variation in Morphology and Airborne Dispersal of the Urticating Apparatus of Ochrogaster lunifer (Lepidoptera: Notodontidae), an Australian Processionary Caterpillar, and Implications for Livestock and Humans.

True setae borne on the abdominal tergites of Ochrogaster lunifer Herrich-Schӓffer caterpillars are the agents of an irritating contact dermatitis, osteomyelitis, ophthalmia, and severe allergic reactions in humans, and are the cause of Equine Amnionitis and Fetal Loss in Australia. The setae are detached and readily dislodge from the integument whereby they disperse throughout the environment. To better understand the true setae of O. lunifer as agents of medical and veterinary concern, we studied their characteristics and distance dispersed. Whereas members of the European Thaumetopoeinae have been widely studied, their southern-hemisphere counterparts such as O. lunifer are not well known despite their harmfulness and known medical and veterinary importance. The caterpillar's investment in true setae increased with age and size, and two distinct size classes co-occurred in setae fields. A previously undescribed morphological type of true seta was found on the first abdominal segment. All true setae were calculated to travel long distances in the air even under light breeze conditions. Our results show there is a high risk of exposure to airborne urticating setae within 100 m of elevated caterpillar activity, and a likely risk of exposure for some kilometers in the direction of the prevailing breeze. This information should be used to inform management strategies in areas where urticating processionary caterpillars are active, and especially during periods of an outbreak.

Temporal Resource Partitioning and Interspecific Correlations in a Warm, Temperate Climate Assemblage of Dung Beetles (Coleoptera: Scarabaeidae).

Guilds of dung dwelling and tunneling dung beetles coexist in local assemblages in warm temperate regions, despite the tendency of dwellers to be inferior competitors. A field experiment on the Black Sea coast of Turkey examined the role of temporal resource partitioning in their coexistence. Standardized dung pads deposited at 4 h intervals through a 24 h period in summer were collected 12, 24, or 48 h later. Adults from 10 tunneling and seven dung dwelling species were collected. The tunnelers contributed a high proportion of both total abundance and biomass. There was a significant effect of dung deposition time and exposure period on mean tunneler abundance. Mean tunneler abundance was nearly seven times higher in dung deposited at 06:00 than at 18:00. The dwellers reduced the potential for competitive interactions with tunnelers by relatively uniform dispersal across the six dung deposition times. The distinctly different dung use patterns by dwellers and tunnelers demonstrated temporal resource partitioning. Interspecific correlation coefficients were also determined because interspecific relationships are at the core of resource partitioning. Total tunneler and dweller abundances were not correlated. Overall, there were strong positive correlations between tunneling species and low correlations between tunneling and dwelling species, and between dwelling species. The five most abundant tunnelers, from two tribes and three genera, were strongly positively correlated. There were substantial size differences among the four most abundant tunnelers that probably facilitate their coexistence.

Serial founder effects and genetic differentiation during worldwide range expansion of monarch butterflies.

Range expansions can result in founder effects, increasing genetic differentiation between expanding populations and reducing genetic diversity along the expansion front. However, few studies have addressed these effects in long-distance migratory species, for which high dispersal ability might counter the effects of genetic drift. Monarchs (Danaus plexippus) are best known for undertaking a long-distance annual migration in North America, but have also dispersed around the world to form populations that do not migrate or travel only short distances. Here, we used microsatellite markers to assess genetic differentiation among 18 monarch populations and to determine worldwide colonization routes. Our results indicate that North American monarch populations connected by land show limited differentiation, probably because of the monarch's ability to migrate long distances. Conversely, we found high genetic differentiation between populations separated by large bodies of water. Moreover, we show evidence for serial founder effects across the Pacific, suggesting stepwise dispersal from a North American origin. These findings demonstrate that genetic drift played a major role in shaping allele frequencies and created genetic differentiation among newly formed populations. Thus, range expansion can give rise to genetic differentiation and declines in genetic diversity, even in highly mobile species.

Long-distance aerial dispersal modelling of Culicoides biting midges: case studies of incursions into Australia.

BACKGROUND: Previous studies investigating long-distance, wind-borne dispersal of Culicoides have utilised outbreaks of clinical disease (passive surveillance) to assess the relationship between incursion and dispersal event. In this study, species of exotic Culicoides and isolates of novel bluetongue viruses, collected as part of an active arbovirus surveillance program, were used for the first time to assess dispersal into an endemic region. RESULTS: A plausible dispersal event was determined for five of the six cases examined. These include exotic Culicoides specimens for which a possible dispersal event was identified within the range of two days--three weeks prior to their collection and novel bluetongue viruses for which a dispersal event was identified between one week and two months prior to their detection in cattle. The source location varied, but ranged from Lombok, in eastern Indonesia, to Timor-Leste and southern Papua New Guinea. CONCLUSIONS: Where bluetongue virus is endemic, the concurrent use of an atmospheric dispersal model alongside existing arbovirus and Culicoides surveillance may help guide the strategic use of limited surveillance resources as well as contribute to continued model validation and refinement. Further, the value of active surveillance systems in evaluating models for long-distance dispersal is highlighted, particularly in endemic regions where knowledge of background virus and vector status is beneficial.

Understanding heliothine (Lepidoptera: Heliothinae) pests: what is a host plant?

Heliothine moths (Lepidoptera: Heliothinae) include some of the world's most devastating pest species. Whereas the majority of nonpest heliothinae specialize on a single plant family, genus, or species, pest species are highly polyphagous, with populations often escalating in size as they move from one crop species to another. Here, we examine the current literature on heliothine host-selection behavior with the aim of providing a knowledge base for research scientists and pest managers. We review the host relations of pest heliothines, with a particular focus on Helicoverpa armigera (Hübner), the most economically damaging of all heliothine species. We then consider the important question of what constitutes a host plant in these moths, and some of the problems that arise when trying to determine host plant status from empirical studies on host use. The top six host plant families in the two main Australian pest species (H. armigera and Helicoverpa punctigera Wallengren) are the same and the top three (Asteraceae, Fabaceae, and Malvaceae) are ranked the same (in terms of the number of host species on which eggs or larvae have been identified), suggesting that these species may use similar cues to identify their hosts. In contrast, for the two key pest heliothines in the Americas, the Fabaceae contains approximately 1/3 of hosts for both. For Helicoverpa zea (Boddie), the remaining hosts are more evenly distributed, with Solanaceae next, followed by Poaceae, Asteraceae, Malvaceae, and Rosaceae. For Heliothis virescens (F.), the next highest five families are Malvaceae, Asteraceae, Solanaceae, Convolvulaceae, and Scrophulariaceae. Again there is considerable overlap in host use at generic and even species level. H. armigera is the most widely distributed and recorded from 68 plant families worldwide, but only 14 families are recorded as a containing a host in all geographic areas. A few crop hosts are used throughout the range as expected, but in some cases there are anomalies, perhaps because host plant relation studies are not comparable. Studies on the attraction of heliothines to plant odors are examined in the context of our current understanding of insect olfaction, with the aim of better understanding the connection between odor perception and host choice. Finally, we discuss research into sustainable management of pest heliothines using knowledge of heliothine behavior and ecology. A coordinated international research effort is needed to advance our knowledge on host relations in widely distributed polyphagous species instead of the localized, piecemeal approaches to understanding these insects that has been the norm to date.

Ontogenetic Changes in the Feeding Behaviour of Helicoverpa armigera Larvae on Pigeonpea (Cajanus cajan) Flowers and Pods.

Despite substantial research examining caterpillar-plant interactions, changes in the feeding behaviour of lepidopteran larvae as they develop are poorly understood. In this study, we investigated ontogenetic changes in the behaviour of Helicoverpa armigera larvae feeding on reproductive structures of pigeonpea (Cajanus cajan). Specifically, we examined the preference for and avoidance of pigeonpea flowers and pods of first, second, third, and fourth instar H. armigera larvae. We also conducted a no-choice assay to compare the ability of third and fourth instar larvae to penetrate pigeonpea pod walls, which act as a physical defence against herbivory. When presented with a choice between pigeonpea pods and flowers, different instars behaved differently. First and second instar larvae largely avoided pigeonpea pods, instead feeding on flowers; third instar larvae initially avoided pods, but by 24 h, did not strongly discriminate between the structures; and fourth instars demonstrated a preference for pods. When initially placed on pods, first instars were slower than other instars to leave these structures, despite pods being suboptimal feeding sites for small caterpillars. We identified a clear instar-specific ability to penetrate through the pod wall to reach the seeds. Most third instar larvae were unable to penetrate the pod wall, whereas most fourth instars succeeded. Third instars suffered a physiological cost (measured by relative growth rate) when boring through the pod wall, which was not observed in fourth instars. Our study further illuminates the insect-plant interactions of the H. armigera-pigeonpea system and provides evidence for the significant changes in feeding behaviour that may occur during lepidopteran larval development.