Variation in Pesticide Toxicity in the Western Honey Bee (Apis mellifera) Associated with Consuming Phytochemically Different Monofloral Honeys.

Insecticide toxicity to insect herbivores has long been known to vary across different host plants; this phenomenon has been widely documented in both foliage-feeders and sap-feeders. Species-specific phytochemical content of hostplant tissues is assumed to determine the pattern of induction of insect enzymes that detoxify insecticides, but specific phytochemicals have rarely been linked to host plant-associated variation in pesticide toxicity. Moreover, no studies to date have examined the effects of nectar source identity and phytochemical composition on the toxicity of insecticides to pollinators. In this study, we compared LD50 values for the insecticide bifenthrin, a frequent contaminant of nectar and pollen in agroecosystems, in the western honey bee, Apis mellifera, consuming three phytochemically different monofloral honeys: Nyssa ogeche (tupelo), Robinia pseudoacacia (black locust), and Fagopyrum esculentum (buckwheat). We found that bifenthrin toxicity (LD50) values for honey bees across different honey diets is linked to their species-specific phytochemical content. The profiles of phenolic acids and flavonoids of buckwheat and locust honeys are richer than is the profile of tupelo honey, with buckwheat honey containing the highest total content of phytochemicals and associated with the highest bifenthrin LD50 in honey bees. The vector fitting in the ordination analysis revealed positive correlations between LD50 values and two honey phytochemical richness estimates, Chao1 and Abundance-based Coverage Estimator (ACE). These findings suggest unequal effects among different phytochemicals, consistent with the interpretation that certain compounds, including ones that are rare, may have a more pronounced effect in mitigating pesticide toxicity.

Effects of pesticide-adjuvant combinations used in almond orchards on olfactory responses to social signals in honey bees (Apis mellifera).

Exposure to agrochemical sprays containing pesticides and tank-mix adjuvants has been implicated in post-bloom mortality, particularly of brood, in honey bee colonies brought into California almond orchards for pollination. Although adjuvants are generally considered to be biologically inert, some adjuvants have exhibited toxicity and sublethal effects, including decreasing survival rates of next-generation queens. Honey bees have a highly developed olfactory system to detect and discriminate among social signals. To investigate the impact of pesticide-adjuvant combinations on honey bee signal perception, we performed electroantennography assays to assess alterations in their olfactory responsiveness to the brood ester pheromone (BEP), the volatile larval pheromone ß-ocimene, and the alarm pheromone 2-heptanone. These assays aimed to uncover potential mechanisms underlying changes in social behaviors and reduced brood survival after pesticide exposure. We found that combining the adjuvant Dyne-Amic with the fungicide Tilt (propiconazole) and the insecticide Altacor (chlorantraniliprole) synergistically enhanced olfactory responses to three concentrations of BEP and as well exerted dampening and compensatory effects on responses to 2-heptanone and ß-ocimene, respectively. In contrast, exposure to adjuvant alone or the combination of fungicide and insecticide had no effect on olfactory responses to BEP at most concentrations but altered responses to ß-ocimene and 2-heptanone. Exposure to Dyne-Amic, Altacor, and Tilt increased BEP signal amplitude, indicating potential changes in olfactory receptor sensitivity or sensilla permeability to odorants. Given that, in a previous study, next-generation queens raised by nurses exposed to the same treated pollen experienced reduced survival, these new findings highlight the potential disruption of social signaling in honey bees and its implications for colony reproductive success.

Degree of Dietary Specialization on Furanocoumarin-Containing Hostplants in a Newly Invasive Web Building Caterpillar.

The genus Depressaria (Lepidoptera: Depressariidae) mostly comprises specialist herbivores with varying capacity for detoxification of defensive phytochemistry. Depressaria depressana, a Eurasian moth recently introduced into North America, is a family-level specialist of the Apiaceae, whose hosts include more than a dozen species in multiple tribes; Depressaria radiella is a super-specialist of Eurasian origin that feeds exclusively on species in the genera Pastinaca and Heracleum throughout its native and introduced range. In eastern North America, it feeds upon Pastinaca sativa, an invasive European species, and Heracleum maximum, a native species. We determined whether differences in furanocoumarin metabolism exist between D. depressana and two isolated populations of D. radiella, feeding exclusively on either P. sativa or H. maximum. We also compared gravimetric estimates of feeding efficiency to assess D. depressana larval performance on different diets. Both populations of D. radiella metabolized furanocoumarins at a greater rate than D. depressana. Although there was no difference in rates of metabolism of linear furanocoumarins in the two populations of D. radiella, individuals collected from H. maximum metabolized angular furanocoumarins more rapidly. The gravimetric assessments of feeding efficiencies revealed that D. depressana exhibited highest efficiencies consuming Daucus carota; moreover, this species survived to pupation consuming fruits of Zizia aurea, an apiaceous species native to North America. Our preliminary phylogenetic analysis, building on an earlier morphological analysis, incorporates mitochondrial cytochrome oxidase subunit 1 data from the BOLD database and revealed that the presence or absence of furanocoumarins is not a strong predictor of species-level evolution in Depressaria.

Increase in longevity and amelioration of pesticide toxicity by natural levels of dietary phytochemicals in the honey bee, Apis mellifera.

For the past decade, migratory beekeepers who provide honey bees for pollination services have experienced substantial colony losses on a recurring basis that have been attributed in part to exposure to insecticides, fungicides, or their combinations applied to crops. The phytochemicals p-coumaric acid and quercetin, which occur naturally in a wide variety of bee foods, including beebread and many types of honey, can enhance adult bee longevity and reduce the toxicity of certain pesticides. How variation in concentrations of natural dietary constituents affects interactions with xenobiotics, including synthetic pesticides, encountered in agroecosystems remains an open question. We tested the effects of these two phytochemicals at a range of natural concentrations on impacts of consuming propiconazole and chlorantraniliprole, a triazole fungicide and an insecticide frequently applied as a tank mix to almond trees during bloom in California's Central Valley. Propiconazole, even at low field concentrations, significantly reduced survival and longevity when consumed by adult bees in a sugar-based diet. The effects of propiconazole in combination with chlorantraniliprole enhanced mortality risk. The detrimental effects of the two pesticides were for the most part reduced when either or both of the phytochemicals were present in the diet. These findings suggest that honey bees may depend on non-nutritive but physiologically active phytochemical components of their natural foods for ameliorating xenobiotic stress, although only over a certain range of concentrations; particularly at the high end of the natural range, certain combinations can incur additive toxicity. Thus, efforts to develop nectar or pollen substitutes with phytochemicals to boost insecticide tolerance or immunity or to evaluate toxicity of pesticides to pollinators should take concentration-dependent effects of phytochemicals into consideration.

Fungicide suppression of flight performance in the honeybee (Apis mellifera) and its amelioration by quercetin.

As a managed agricultural pollinator, the western honeybee Apis mellifera frequently encounters agrochemicals as contaminants of nectar and pollen. One such contaminant, the fungicide boscalid, is applied at bloom in orchards for fungal floral pathogen control. As an inhibitor of complex II in the mitochondrial electron transport chain of fungi, boscalid can potentially interfere with high energy-demanding activities of bees, including flight. We designed an indoor flight treadmill to evaluate impacts of ingesting boscalid and/or quercetin, a ubiquitous phytochemical in bee food that also affects mitochondrial respiration. Boscalid reduced the wingbeat frequencies of foragers during flight but did not alter the duration of flight. At the colony level, boscalid ingestion may thereby affect overall health by reducing forager efficiency. The consumption of quercetin, by contrast, led to higher adenosine triphosphate levels in flight muscles and a higher wingbeat frequency. Consuming the two compounds together increased wingbeat frequency, demonstrating a hitherto unrecognized mechanism by which dietary phytochemicals may act to ameliorate toxic effects of pesticides to promote honeybee health. In carrying out this work, we also introduce two methodological improvements for use in testing for pesticide effects on flight capacity-a 'force-feeding' to standardize flight fuel supply and a novel indoor flight treadmill.

Behavioral responses of honey bees (Apis mellifera) to natural and synthetic xenobiotics in food.

While the natural foods of the western honey bee (Apis mellifera) contain diverse phytochemicals, in contemporary agroecosystems honey bees also encounter pesticides as floral tissue contaminants. Whereas some ubiquitous phytochemicals in bee foods up-regulate detoxification and immunity genes, thereby benefiting nestmates, many agrochemical pesticides adversely affect bee health even at sublethal levels. How honey bees assess xenobiotic risk to nestmates as they forage is poorly understood. Accordingly, we tested nine phytochemicals ubiquitous in nectar, pollen, or propolis, as well as five synthetic xenobiotics that frequently contaminate hives-two herbicides (atrazine and glyphosate) and three fungicides (boscalid, chlorothalonil, and prochloraz). In semi-field free-flight experiments, bees were offered a choice between paired sugar water feeders amended with either a xenobiotic or solvent only (control). Among the phytochemicals, foragers consistently preferred quercetin at all five concentrations tested, as evidenced by both visitation frequency and consumption rates. This preference may reflect the long evolutionary association between honey bees and floral tissues. Of pesticides eliciting a response, bees displayed a preference at specific concentrations for glyphosate and chlorothalonil. This paradoxical preference may account for the frequency with which these pesticides occur as hive contaminants and suggests that they present a greater risk factor for honey bee health than previously suspected.

Impacts of Dietary Phytochemicals in the Presence and Absence of Pesticides on Longevity of Honey Bees (Apis mellifera).

Because certain flavonols and phenolic acids are found in pollen and nectar of most angiosperms, they are routinely ingested by Apis mellifera, the western honey bee. The flavonol quercetin and the phenolic acid p-coumaric acid are known to upregulate detoxification enzymes in adult bees; their presence or absence in the diet may thus affect the toxicity of ingested pesticides. We conducted a series of longevity assays with one-day-old adult workers to test if dietary phytochemicals enhance longevity and pesticide tolerance. One-day-old bees were maintained on sugar syrup with or without casein (a phytochemical-free protein source) in the presence or absence of quercetin and p-coumaric acid as well as in the presence or absence of two pyrethroid insecticides, bifenthrin and ß-cyfluthrin. Dietary quercetin (hazard ratio, HR = 0.82), p-coumaric acid (HR = 0.91) and casein (HR = 0.74) were associated with extended lifespan and the two pyrethroid insecticides, 4 ppm bifenthrin (HR = 9.17) and 0.5 ppm ß-cyfluthrin (HR = 1.34), reduced lifespan. Dietary quercetin enhanced tolerance of both pyrethroids; p-coumaric acid had a similar effect trend, although of reduced magnitude. Casein in the diet appears to eliminate the life-prolonging effect of p-coumaric acid in the absence of quercetin. Collectively, these assays demonstrate that dietary phytochemicals influence honey bee longevity and pesticide stress; substituting sugar syrups for honey or yeast/soy flour patties may thus have hitherto unrecognized impacts on adult bee health.

Structural and Mechanical Properties of Cocoons of Antherina suraka (Saturniidae, Lepidoptera), an Endemic Species Used for Silk Production in Madagascar.

Antherina suraka Boisduval (Saturniidae, Lepidoptera) produces a silken cocoon that has been the focus of efforts to create a commercial wild silk industry in Madagascar. In this study, structural and mechanical properties of the cocoon of A. suraka from two sites were measured and compared to the cocoon of Bombyx mori L. (Bombycidae, Lepidoptera) the world's most common source for silk. Results of environmental scanning electron microscopy and mechanical testing showed that the silk sheet of A. suraka cocoon is less compact, with greater thickness and lower tensile strength and stiffness than that of B. mori Confirming these results, stiffness and cell and thread density were found to be negatively correlated with thickness, and the cell and thread volumes were positively correlated with thickness. Antherina suraka showed no major differences between silk sheets from Kirindy and Isalo sites in either structural or mechanical properties, except for mean cell volume, which was greater in cocoons from Kirindy. Comparison between the two layers forming the cocoon showed that the inner layer has greater elastic modulus, denser silk distribution and lower porosity. Cocoons from both Kirindy and Isalo are suitable for sericulture. Although the inner layer of cocoon silk is of higher quality than the outer layer, the fact that both layers are of great but lower tensile strength than B. mori silk suggests that the current practice of sewing the two layers together for making one single layer fabric should be continued in efforts to produce a commercially viable product.

Firefly light flashing: oxygen supply mechanism.

Firefly luminescence is an intriguing phenomenon with potential technological applications, whose biochemistry background was only recently established. The physics side of this phenomenon, however, was still unclear, specifically as far as the oxygen supply mechanism for light flashing is concerned. This uncertainty is due to the complex microscopic structure of the tracheal system: without fully knowing its geometry, one cannot reliably test the proposed mechanisms. We solved this problem using synchrotron phase contrast microtomography and transmission x-ray microscopy, finding that the oxygen consumption corresponding to mitochondria functions exceeds the maximum rate of oxygen diffusion from the tracheal system to the photocytes. Furthermore, the flashing mechanism uses a large portion of this maximum rate. Thus, the flashing control requires passivation of the mitochondria functions, e.g., by nitric oxide, and switching of the oxygen supply from them to photoluminescence.

Impaired olfactory associative behavior of honeybee workers due to contamination of imidacloprid in the larval stage.

The residue of imidacloprid in the nectar and pollens of the plants is toxic not only to adult honeybees but also the larvae. Our understanding of the risk of imidacloprid to larvae of the honeybees is still in a very early stage. In this study, the capped-brood, pupation and eclosion rates of the honeybee larvae were recorded after treating them directly in the hive with different dosages of imidacloprid. The brood-capped rates of the larvae decreased significantly when the dosages increased from 24 to 8000 ng/larva. However, there were no significant effects of DMSO or 0.4 ng of imidacloprid per larva on the brood-capped, pupation and eclosion rates. Although the sublethal dosage of imidacloprid had no effect on the eclosion rate, we found that the olfactory associative behavior of the adult bees was impaired if they had been treated with 0.04 ng/larva imidacloprid in the larval stage. These results demonstrate that a sublethal dosage of imidacloprid given to the larvae affects the subsequent associative ability of the adult honeybee workers. Thus, a low dose of imidacloprid may affect the survival condition of the entire colony, even though the larvae survive to adulthood.

Visualization of the spatial and spectral signals of orb-weaving spiders, Nephila pilipes, through the eyes of a honeybee.

It is well known that the honeybee has good color vision. However, the spectral range in which the bee can see is different from that of the human eye. To study how bees view their world of colors, one has to see through the eyes of the bee, not the eyes of a human. A conventional way to examine the color signals that animals can detect is to measure the surface reflectance spectra and compute the quantum catches of each photoreceptor type based on its known spectral sensitivity. Color signal and color contrast are then determined from the loci of these quantum catches in the color space. While the point-by-point measurements of the reflectance spectra using a standard spectrometer have yielded a significant amount of data for analyzing color signals, the lack of spatial information and low sampling efficiency constrain their applications. Using a special filter coating technique, a set of filters with transmission spectra that were closely matched to the bee's sensitivity spectra of three photoreceptor types (UV, blue, and green) was custom made. By placing these filters in front of a UV/VIS-sensitive CCD camera and acquiring images sequentially, we could collect images of a bee's receptor with only three shots. This allowed a direct visualization of how bees view their world in a pseudo-color RGB display. With this imaging system, spatial and spectral signals of the orb-weaving spider, Nephila pilipes, were recorded, and color contrast images corresponding to the bee's spatial resolution were constructed and analyzed. The result not only confirmed that the color markings of N. pilipes are of high chromatic contrast to the eyes of a bee, but it also indicated that the spatial arrangement of these markings resemble flower patterns which may attract bees to visit them. Thus, it is likely that the orb-weaving spider (N. pilipes) deploys a similar strategy to that of the Australian crab spider (Thomisus spectabilis) to exploit the bee's pre-existing preference for flowers with color patterning.

Chromatic cues to trap the oriental fruit fly, Bactrocera dorsalis.

Various colors have been used as visual cues to trap insect pests. For example, yellow traps for monitoring and control of the oriental fruit fly (Bactrocera dorsalis) have been in use for a very long time. However, the chromatic cue of using color traps has never been meticulously investigated. In this study, the spectral sensitivities of the photoreceptors in the compound eyes of B. dorsalis were measured intracellularly, and the theory of receptor quantum catch was applied to study the chromatic cue of fly attracting. Responses to five wavelength categories with peak wavelengths of 370, 380, 490, and 510 nm, and one with dual peaks at 350 and 490 nm were recorded. Based on spectral sensitivities, six colored papers were chosen to test the color preference of the fly, and an additional UV preference test was done to confirm the effect of the UV stimuli. It was concluded that UV and green stimuli (spectra: 300-380 nm and 500-570 nm) would enhance the attractiveness of a colored paper to the oriental fruit fly, and blue stimuli (380-500 nm) would diminish the attractiveness.