Flower colour and size-signals vary with altitude and resulting climate on the tropical-subtropical islands of Taiwan.

The diversity of flower colours in nature provides quantifiable evidence for how visitations by colour sensing insect pollinators can drive the evolution of angiosperm visual signalling. Recent research shows that both biotic and abiotic factors may influence flower signalling, and that harsher climate conditions may also promote salient signalling to entice scarcer pollinators to visit. In parallel, a more sophisticated appreciation of the visual task foragers face reveals that bees have a complex visual system that uses achromatic vision when moving fast, whilst colour vision requires slower, more careful inspection of targets. Spectra of 714 native flowering species across Taiwan from sea level to mountainous regions 3,300 m above sea level (a.s.l.) were measured. We modelled how the visual system of key bee pollinators process signals, including flower size. By using phylogenetically informed analyses, we observed that at lower altitudes including foothills and submontane landscapes, there is a significant relationship between colour contrast and achromatic signals. Overall, the frequency of flowers with high colour contrast increases with altitude, whilst flower size decreases. The evidence that flower colour signaling becomes increasingly salient in higher altitude conditions supports that abiotic factors influence pollinator foraging in a way that directly influences how flowering plants need to advertise.

Complementary use of visual and olfactory cues to assess capture of Bactrocera dorsalis (Hendel): Implementation and field verification via an IoT-based automatic monitoring system.

This study examined the effect of combining visual and olfactory cues to attract oriental fruit flies (OFFs). Six different colored light-emitting diodes (LEDs) served as a visual attractant and methyl eugenol served as olfactory bait to lure male flies. An internet of things (IoT)-based pest monitoring system, consisting of sensor nodes, a gateway, and automatic counting traps, was deployed in the field to automatically collect environmental data and pest counts. The results of the calibrated experiments indicated that green, yellow, or red LEDs exhibited better performance in attracting flies than white, purple, or blue LEDs or no LEDs. With an accurate combination of visual and olfactory cues, the proposed IoT-based pest monitoring system may be an effective tool in agricultural pest management, given its advantages for efficiently capturing OFFs in a labor and time saving manner, providing accurate information regarding increases in pest populations, and enabling long-term, real-time data collection.

Neuroplasticity in honey bee brains: An enhanced micro-computed tomography protocol for precise mushroom body volume measurement.

BACKGROUND: In insect brains, mushroom bodies are associated with memory and learning behavior. It has been demonstrated that the volume of the mushroom bodies in the brain of a worker honey bee changes during the adult stage. Changes in mushroom body volume imply high neuroplasticity in the brains and may be related to the age polyethism of honey bees. A suitable volume measurement method is needed to understand the correlation between behavioral changes and mushroom body volume changes in honey bees. NEW METHOD: We developed a new protocol for insect micro-computed tomography by modifying a previously reported method. Permount™ mounting medium was used as the embedding medium for micro-computed tomography scanning. RESULTS: This protocol can generate images with high contrast inside the brain and reduce the marked shape changes during specimen processing. From the resulting high-contrast images, we used freeware to generate a three-dimensional model and calculate the volumes of the mushroom bodies in honey bees. The measured volumes of the mushroom bodies were larger than the values reported in most previous studies. There was no significant difference between the left and right mushroom body volumes, but the volumes of honey bee mushroom bodies significantly increased with age. COMPARISON WITH EXISTING METHODS: Previous protocols for micro-computed tomography using dried samples would cause brain shrinkage; protocols using ethanol-preserved or resin-embedded samples generated images with lower contrast. CONCLUSIONS: The embedding protocol for micro-computed tomography is suitable for calculating volume of the mushroom bodies in honey bee brains.

Honey bee foraging ability suppressed by imidacloprid can be ameliorated by adding adenosine.

Honey bees are important pollinators in most ecosystem, but they are currently facing many threats, which have led to a reduction in their population. Previous studies have indicated that neonicotinoid pesticide can impair the memory and learning ability of honey bees, which can eventually lead to a decline in their foraging and homing abilities. In this study, we investigated the homing ability barrier from the perspective of energy supply. We believe that when worker bees experience stress, their energy supply may shift from pro-movement to pro-resistance; this will lead to inadequate energy provision to the flight muscles, causing a reduction in wingbeat frequency and impairing the flight ability of the worker bees. To test this, the worker bees were treated with imidacloprid, and wing beats between the treatment groups were compared. Their glucose, glycogen, trehalose, and ATP contents were also measured, and their genes for energy metabolism and resistance were analyzed. The addition of adenosine improved the ATP content and helped recover the wingbeat frequency of the worker bees. The preliminary results obtained showed that wingbeat frequency and glucose content in the worker bees treated with imidacloprid were significantly lower than those in the control group. This result is consistent with our hypothesis and demonstrates that energy supply imbalances can prevent worker bees from returning to their hives.

The impact of thigmotaxis deprivation on the development of the German cockroach (Blattella germanica).

Thigmotaxis is required in small animals. In this study, we examined how the shelter angle affects the development of German cockroaches, Blattella germanica. Groups and individual cockroaches showed a strong preference for shelters with an angle of ≤40° after 15 min or 24 h in shelter-selection trials. For cockroaches that developed in 90/180-degree shelters, survival and fecundity were low, and the nymphal stage lasted longer. Post-molting transcriptomes of second- and sixth-instar nymphs were analyzed at 12 h and 2 days post-molting. Upregulation was observed in genes related to ATP metabolism and cellular amide metabolism. Chitin-based cuticle development and postembryonic development-related genes were downregulated. The stress responses of cockroaches that developed in shelters with angles of 90° were similar to those of gregarious cockroaches experiencing social isolation. For German cockroaches, environmental tactile stimuli are crucial to development and homeostasis.

Efficiency of a Novel Light-Emitting Diode (LED) Trap for Trapping Rhyzopertha dominica (Coleoptera: Bostrichidae) in Paddy Rice Storehouses.

The lesser grain borer Rhyzopertha dominica is the major pest of stored paddy rice globally, including in Taiwan. It has strong phototaxis and is good at flying, suitable for developing a light-trapping method to monitor and control it. In the present study, a wavelength of light-emitting diodes (LEDs), i.e., 373 nm, was determined to be the most efficient to trap R. dominica using a dodecagon maze. Accordingly, an LED trap, named the Taiwan Agricultural Research Institute-LED (TARI-LED) trap, was invented, which comprised LEDs of two distinct wavelengths (373 and 408 nm), a wavelength switch, a suction fan, and an insect collector. The trapping efficiency was assessed in a 4-m3 laboratory arena and two paddy rice storehouses. An initial assessment was performed in the laboratory arena and showed that the TARI-LED trap with 373-nm wavelength for R. dominica rapidly increased in the first 30 min, reaching the highest trapping rate (68.5%) after 3 h. In addition, no significant difference was observed between the suction fan turned on or off. The field tests showed that the 373-nm wavelength had the highest effectiveness for trapping R. dominica in the two paddy rice storehouses, and no significant difference was observed in the number of R. dominica trapped by the 373-nm TARI-LED trap or the CDC-UV light trap. In conclusion, our TARI-LED trap 373 nm exhibited high efficiency in trapping R. dominica in paddy rice storehouses. Moreover, a suction fan-free design should benefit long-term and safe use in paddy rice storehouses trapping R. dominica.

Chronic Effects of Imidacloprid on Honey Bee Worker Development-Molecular Pathway Perspectives.

Sublethal dosages of imidacloprid cause long-term destructive effects on honey bees at the individual and colony levels. In this review, the molecular effects of sublethal imidacloprid were integrated and reported. Several general effects have been observed among different reports using different approaches. Quantitative PCR approaches revealed that imidacloprid treatments during the adult stage are expressed as changes in immuneresponse, detoxification, and oxidation-reduction response in both workers and queens. In addition, transcriptomic approaches suggested that phototransduction, behavior, and somatic muscle development also were affected. Although worker larvae show a higher tolerance to imidacloprid than adults, molecular evidence reveals its potential impacts. Sublethal imidacloprid treatment during the larval stage causes gene expression changes in larvae, pupae, and adults. Transcriptome profiles suggest that the population and functions of affected differentially expressed genes, DEGs, vary among different worker ages. Furthermore, an early transcriptomic switch from nurse bees to foragers was observed, suggesting that precocious foraging activity may occur. This report comprehensively describes the molecular effects of sublethal dosages of imidacloprid on the honey bee Apis mellifera. The corresponding molecular pathways for physiological and neurological responses in imidacloprid-exposed honey bees were validated. Transcriptomic evidence suggests a global and sustained sublethal impact of imidacloprid on honey bee development.

Do I stay or do I go? Shifts in perch use by lizards during morning twilight suggest anticipatory behaviour.

Anticipatory behaviour is the expectation of a near-future event based on information processed in the past and influences an animal's tactical decisions, particularly when there are significant fitness consequences. The grass lizard (Takydromus viridipunctatus) perches on blades of grass at night which likely reduces the probability of predation by terrestrial predators such as snakes, rodents and shrews. During twilight (starting 30 min before sunrise), they move from above the grass to within grass clumps and this is thought to afford the lizard protection while reducing detection by avian predators. Here, we examined how lizards shift their behaviour as a function of visual detectability to their primary predator, the cattle egret (Bubulcus ibis). We show that the lizards shift from their perch site during twilight at the earliest time at which egrets depart communal roosts. At the same time, visual modelling shows a dramatic increase in the detectability of the lizards to the visual system of egrets. Therefore, anticipatory behaviour in response to environmental cues acts to reduce predation risk as lizards become more conspicuous and predators become more active. Grass lizard anticipatory behaviour appears to be finely tuned by natural selection to adjust to temporal changes in predation risk.

Missing Nurse Bees-Early Transcriptomic Switch From Nurse Bee to Forager Induced by Sublethal Imidacloprid.

The environmental residue/sublethal doses of neonicotinoid insecticides are believed to generate a negative impact on pollinators, including honey bees. Here we report our recent investigation on how imidacloprid, one of the major neonicotinoids, affects worker bees by profiling the transcriptomes of various ages of bees exposed to different doses of imidacloprid during the larval stage. The results show that imidacloprid treatments during the larval stage severely altered the gene expression profiles and may induce precocious foraging. Differential expression of foraging regulators was found in 14-day-old treated adults. A high transcriptome similarity between larvae-treated 14-day-old adults and 20-day-old controls was also observed, and the similarity was positively correlated with the dose of imidacloprid. One parts per billion (ppb) of imidacloprid was sufficient to generate a long-term impact on the bee's gene expression as severe as with 50 ppb imidacloprid. The disappearance of nurse bees may be driven not only by the hive member constitution but also by the neonicotinoid-induced precocious foraging behavior.

Honey Bee Colony Population Daily Loss Rate Forecasting and an Early Warning Method Using Temporal Convolutional Networks.

The population loss rate of a honey bee colony is a critical index to verify its health condition. Forecasting models for the population loss rate of a honey bee colony can be an essential tool in honey bee health management and pave a way to early warning methods in the understanding of potential abnormalities affecting a honey bee colony. This work presents a forecasting and early warning algorithm for the population daily loss rate of honey bee colonies and determining warning levels based on the predictions. Honey bee colony population daily loss rate data were obtained through embedded image systems to automatically monitor in real-time the in-and-out activity of honey bees at hive entrances. A forecasting model was trained based on temporal convolutional neural networks (TCN) to predict the following day's population loss rate. The forecasting model was optimized by conducting feature importance analysis, feature selection, and hyperparameter optimization. A warning level determination method using an isolation forest algorithm was applied to classify the population daily loss rate as normal or abnormal. The integrated algorithm was tested on two population loss rate datasets collected from multiple honey bee colonies in a honey bee farm. The test results show that the forecasting model can achieve a weighted mean average percentage error (WMAPE) of 17.1 ± 1.6%, while the warning level determination method reached 90.0 ± 8.5% accuracy. The forecasting model developed through this study can be used to facilitate efficient management of honey bee colonies and prevent colony collapse.

Effects of artificial light at night (ALAN) on gene expression of Aquatica ficta firefly larvae.

Artificial light at night (ALAN) is a major driver of firefly population declines, but its physiological effects are not well understood. To investigate the impact of ALAN on firefly development, we exposed larval Aquatica ficta fireflies to ALAN for two weeks. High larval mortality was observed in the periods of 1-68 days and 106-134 days post-treatment, which may represent the short- and long-term impacts of ALAN. We then profiled the transcriptome of larval Aquatica ficta fireflies following two weeks of ALAN exposure. A total of 1262 (1.67% out of 75777 unigenes) were differentially expressed in the treatment group: 1157 were down-regulated, and 105 were up-regulated. Up-regulated unigenes were related to regulation of hormone levels, ecdysteroid metabolic process, and response to stimulus; down-regulated unigenes were related to negative regulation of insulin receptor signaling, germ cell development, oogenesis, spermatid development, and regulation of neuron differentiation. Transcriptome results suggest that the endocrine, reproductive, and neural development of firefly larvae could be impaired by even relatively brief period of ALAN exposure. This report contributes a much-needed molecular perspective to the growing body of research documenting the fitness impacts of ALAN on bioluminescent fireflies.

Social interactions upregulate hemolymph tryptophan and tyrosine levels in the male lobster cockroach.

In the present study, we found that tryptophan (TRP) and tyrosine (TYR) levels are increased in hemolymph of male Nauphoeta cinerea after social contact with either male or female conspecifics. Hemolymph was collected from individual males before and after the social interactions, and samples were analyzed by HPLC-ECD; analyte identities were confirmed by UPLC/MS. After a male-male first encounter fight, hemolymph TRP and TYR levels were significantly increased in dominants compared with the levels before the encounter. Conversely, TRP and TYR in subordinates were maintained at levels similar to those before the encounter. While after-fight TRP and TYR levels were significantly higher in dominants than subordinates, no significant differences were found in the contestants before the fight. Moreover, contact with an isolated male antenna was sufficient to stimulate attack behavior and increase hemolymph TRP and TYR titers to levels similar to those seen in dominants. After a male-female interaction, two distinct outcomes could be observed. Either hemolymph TRP and TYR levels were increased in successfully mated males, or TRP and TYR levels were unchanged in males that only exhibited premating wing-raising behavior but failed in mating. After contacting the antenna of a socially naïve male with an isolated female antenna, three patterns of behavior and related amino acid response were observed: 1) only premating wing-raising behavior with significant increase of TRP and TYR levels, 2) only attack behavior with significant increase of TRP and TYR levels, and 3) mixed wing-raising and attack behaviors with no significant changes in TRP and TYR levels. The present results show a robust response of hemolymph TRP and TYR to social contact. In light of previously characterized responses in pheromone and juvenile hormone levels, these amine responses suggest that the physiological response of N. cinerea to social contact is multi-dimensional.

An update of the Worldwide Integrated Assessment (WIA) on systemic insecticides. Part 2: impacts on organisms and ecosystems.

New information on the lethal and sublethal effects of neonicotinoids and fipronil on organisms is presented in this review, complementing the previous Worldwide Integrated Assessment (WIA) in 2015. The high toxicity of these systemic insecticides to invertebrates has been confirmed and expanded to include more species and compounds. Most of the recent research has focused on bees and the sublethal and ecological impacts these insecticides have on pollinators. Toxic effects on other invertebrate taxa also covered predatory and parasitoid natural enemies and aquatic arthropods. Little new information has been gathered on soil organisms. The impact on marine and coastal ecosystems is still largely uncharted. The chronic lethality of neonicotinoids to insects and crustaceans, and the strengthened evidence that these chemicals also impair the immune system and reproduction, highlights the dangers of this particular insecticidal class (neonicotinoids and fipronil), with the potential to greatly decrease populations of arthropods in both terrestrial and aquatic environments. Sublethal effects on fish, reptiles, frogs, birds, and mammals are also reported, showing a better understanding of the mechanisms of toxicity of these insecticides in vertebrates and their deleterious impacts on growth, reproduction, and neurobehaviour of most of the species tested. This review concludes with a summary of impacts on the ecosystem services and functioning, particularly on pollination, soil biota, and aquatic invertebrate communities, thus reinforcing the previous WIA conclusions (van der Sluijs et al. 2015).

Floral Color Diversity: How Are Signals Shaped by Elevational Gradient on the Tropical-Subtropical Mountainous Island of Taiwan?

Pollinators with different vision are a key driver of flower coloration. Islands provide important insights into evolutionary processes, and previous work suggests islands may have restricted flower colors. Due to both species richness with high endemism in tropical-subtropical environments, and potentially changing pollinator distributions with altitude, we evaluated flower color diversity across the mountainous island of Taiwan in a comparative framework to understand the cause of color diversity. We sampled flower color signaling on the tropical-subtropical island of Taiwan considering altitudes from sea level to 3300 m to inform how over-dispersion, random processes or clustering may influence flower signaling. We employed a model of bee color space to plot loci from 727 species to enable direct comparisons to data sets from continental studies representing Northern and Southern Hemispheres, and also a continental mountain region. We observed that flower color diversity was similar to flowers that exist in mainland continental studies, and also showed evidence that flowers predominantly had evolved color signals that closely matched bee color preferences. At high altitudes floras tend to be phylogenetically clustered rather than over-dispersed, and their floral colors exhibited weak phylogenetic signal which is consistent with character displacement that facilitated the co-existence of related species. Overall flower color signaling on a tropical-subtropical island is mainly influenced by color preferences of key bee pollinators, a pattern consistent with continental studies.

Lamelloplasts and minichloroplasts in Begoniaceae: iridescence and photosynthetic functioning.

Iridoplasts (modified plastids in adaxial epidermal cells) reported from Begonia were originally hypothesized to cause iridescence, which was broadly accepted for decades. However, several species of Begonia with iridoplasts are not iridescent causing confusion. Here chloroplast ultrastructure was observed in 40 taxa of Begoniaceae to explore the phenomenon of iridescence. However, 22 Begonias and Hillebrandia were found to have iridoplasts, but only nine display visually iridescent blue to blue-green leaves. Unexpectedly, a new type of plastid, a 'minichloroplast,' was found in the abaxial epidermal cells of all taxa, but was present in adaxial epidermal cells only if iridoplasts were absent. Comparative ultrastructural study of iridoplasts and a shading experiment of selected taxa show that a taxon with iridoplasts does not inevitably have visual iridescence, but iridescence is greatly affected by the spacing between thylakoid lamellae (stoma spacing). Thus, we propose instead the name 'lamelloplast' for plastids filled entirely with regular lamellae to avoid prejudging their function. To evaluate photosynthetic performance, chlorophyll fluorescence (F v /F m ) was measured separately from the chloroplasts in the adaxial epidermis and lower leaf tissues by using leaf dermal peels. Lamelloplasts and minichloroplasts have much lower photosynthetic efficiency than mesophyll chloroplasts. Nevertheless, photosynthetic proteins (psbA protein of PSII, RuBisCo and ATPase) were detected in both plastids as well as mesophyll chloroplasts in an immunogold labeling. Spectrometry revealed additional blue to blue-green peaks in visually iridescent leaves. Micro-spectrometry detected a blue peak from single blue spots in adaxial epidermal cells confirming that the color is derived from lamelloplasts. Presence of lamelloplasts or minichloroplasts is species specific and exclusive. High prevalence of lamelloplasts in Begoniaceae, including the basal clade Hillebrandia, highlights a unique evolutionary development. These new findings clarify the association between iridescence and lamelloplasts, and with implications for new directions in the study of plastid morphogenesis.

Short- and mid-wavelength artificial light influences the flash signals of Aquatica ficta fireflies (Coleoptera: Lampyridae).

Urbanization can radically disrupt natural ecosystems through alteration of the sensory environment. Habitat disturbances are predicted to favor behaviorally flexible species capable of adapting to altered environments. When artificial light at night (ALAN) is introduced into urban areas, it has the potential to impede reproduction of local firefly populations by obscuring their bioluminescent courtship signals. Whether individual fireflies can brighten their signals to maintain visibility against an illuminated background remains unknown. In this study, we exposed male Aquatica ficta fireflies to diffused light of varying wavelength and intensity, and recorded their alarm flash signals. When exposed to wavelengths at or below 533 nm, males emitted brighter signals with decreased frequency. This is the first evidence of individual-level light signal plasticity in fireflies. In contrast, long wavelength ambient light (≥ 597 nm) did not affect signal morphology, likely because A. ficta cannot perceive these wavelengths. These results suggest long wavelength lighting is less likely to impact firefly courtship, and its use in place of broad spectrum white lighting could augment firefly conservation efforts. More generally, this study demonstrates benefits of bioluminescent signal plasticity in a "noisy" signaling environment, and sheds light on an important yet understudied consequence of urbanization.

Reply to 'Pitfalls of using confocal-microscopy based automated quantification of synaptic complexes in honeybee mushroom bodies (response to Peng and Yang 2016)'.

In a comment on our Article "Sublethal Dosage of Imidacloprid Reduces the Microglomerular Density of Honey Bee Mushroom Bodies", Rössler et al. assert that our reported counts are overall lower than previously reported due to the use of automated quantification. We address these issues in this reply.

Gene expression changes in honey bees induced by sublethal imidacloprid exposure during the larval stage.

Honey bee larvae exposed to sublethal doses of imidacloprid show behavioural abnormalities as adult insects. Previous studies have demonstrated that this phenomenon originates from abnormal neural development in response to imidacloprid exposure. Here, we further investigated the global gene expression changes in the heads of newly emerged adults and observed that 578 genes showed more than 2-fold changes in gene expression after imidacloprid exposure. This information might aid in understanding the effects of pesticides on the health of pollinators. For example, the genes encoding major royal jelly proteins (MRJPs), a group of multifunctional proteins with significant roles in the sustainable development of bee colonies, were strongly downregulated. These downregulation patterns were further confirmed through analyses using quantitative reverse transcription-polymerase chain reaction on the heads of 6-day-old nurse bees. To our knowledge, this study is the first to demonstrate that sublethal doses of imidacloprid affect mrjp expression and likely weaken bee colonies.

Regulation of genes related to immune signaling and detoxification in Apis mellifera by an inhibitor of histone deacetylation.

The western honeybee (Apis mellifera) is essential for the global economy due to its important role in ecosystems and agriculture as a pollinator of numerous flowering plants and crops. Pesticide abuse has greatly impacted honeybees and caused tremendous loss of honeybee colonies worldwide. The reasons for colony loss remain unclear, but involvement of pesticides and pathogen-pesticide interactions has been hypothesized. Histone deacetylase inhibitors (HDACis) inhibit the activity of histone acetylase, which causes the hyperacetylation of histone cores and influences gene expression. In this study, sodium butyrate, an HDACi, was used as a dietary supplement for honeybees; after treatment, gene expression profiles were analyzed using quantitative PCR. The results showed that sodium butyrate up-regulated genes involved in anti-pathogen and detoxification pathways. The bioassay results showed that honeybees treated with sodium butyrate were more tolerant to imidacloprid. Additionally, sodium butyrate strengthened the immune response of honeybees to invasions of Nosema ceranae and viral infections. We also performed a bioassay in which honeybees were exposed to pesticides and pathogens. Our results provide additional data regarding the mechanism by which honeybees react to stress and the potential application of HDACis in beekeeping.

Magnetic Sensing through the Abdomen of the Honey bee.

Honey bees have the ability to detect the Earth's magnetic field, and the suspected magnetoreceptors are the iron granules in the abdomens of the bees. To identify the sensing route of honey bee magnetoreception, we conducted a classical conditioning experiment in which the responses of the proboscis extension reflex (PER) were monitored. Honey bees were successfully trained to associate the magnetic stimulus with a sucrose reward after two days of training. When the neural connection of the ventral nerve cord (VNC) between the abdomen and the thorax was cut, the honey bees no longer associated the magnetic stimulus with the sucrose reward but still responded to an olfactory PER task. The neural responses elicited in response to the change of magnetic field were also recorded at the VNC. Our results suggest that the honey bee is a new model animal for the investigation of magnetite-based magnetoreception.