The Larvicidal and Adulticidal Effects of Selected Plant Essential Oil Constituents on Greater Wax Moths.

Some plant essential oil constituents, such as monoterpenoids and phenylpropanoids, are promising insecticides in some situations and for certain insect pests. They vary in their toxicity, depending on the target insect. Moths (Lepidoptera) appear susceptible to these compounds, making them of promise for use against greater wax moths (Galleria mellonella Fabricius, (Lepidoptera: Pyrallidae), GWM), an important pest of western honey bee (Apis mellifera Linnaeus. (Hymenoptera: Apidae)) colonies. We determined the LC50 for GWM larvae or LD50 for GWM adults of select compounds (thymol, carvacrol, (S)-(+)-carvone, estragole, citral, linalool, (S)-(-)-limonene, and γ-terpinene). Concentrations between 8 and 2,266 µg/cm3 were mixed into the diets of GWM larvae and doses ranging between 0.08 and 70.3 mg per adult were applied topically to the abdomens of GWM adults. Lethal concentrations and doses were calculated after of 72 h of exposure. All eight compounds showed insecticidal activity against all tested stages of GWMs. Thymol (LC50 µg/cm3 (95% CI) = 21 [9-56], carvacrol = 46 [26-79], citral = 63 [30-134], and carvone = 76 [33-201]) had the highest toxicity toward GWM larvae. The hydrocarbons limonene (296 [231-377]), estragole (466 [354-611]), and γ-terpinene (729 [630-857]) had the lowest toxicity toward GWM larvae. Thymol had the highest toxicity towards GWM adults (LD50 (CI) = 0.5 [0.4-0.8] mg/adult). However, carvacrol (11.6 [10.1-13.6]), linalool (12.9 [9.3-17.8]), and limonene (15.8 [13.1-19.2]) were less toxic to GWM larvae. Our data show that select plant essential oil constituents are promising controls of GWM.

A combination of Tropilaelaps mercedesae and imidacloprid negatively affects survival, pollen consumption and midgut bacterial composition of honey bee.

Tropilaelaps mercedesae is not only a major threat to honey bees in Asia but also a potential risk to global apiculture due to trade. Imidacloprid is a systemic insecticide that negatively affects individual bees. Moreover, the health of honey bees may be threatened by imidacloprid exposure and T. mercedesae infestation. We studied the effects of T. mercedesae and imidacloprid on the survival, food consumption and midgut bacterial diversity of Apis mellifera in the laboratory. Illumina 16S rRNA gene sequencing was used to determine the bacterial composition in the honey bee midgut. T. mercedesae decreased survival in parasitized honey bees compared with nonparasitized honey bees, but there was no significant difference in food consumption. The imidacloprid 50 µg/L diet significantly decreased syrup consumption of A. mellifera compared with the control diet. The combination of T. mercedesae infestation and imidacloprid 50 µg/L exposure reduced survival and increased pollen consumption of A. mellifera. T. mercedesae infestation or a combination of T. mercedesae infestation and exposure to 25 µg/L imidacloprid affected the midgut bacterial composition of honey bees. T. mercedesae infestation and imidacloprid exposure may reduce the survival and affect honey bee health.

The effect of Bt Cry9Ee toxin on honey bee brood and adults reared in vitro, Apis mellifera (Hymenoptera: Apidae).

The effects of Bt Cry9Ee toxin on honey bee, Apis mellifera L., survival, developmental rate, larval weight, pollen consumption, and midgut bacterial diversity were tested in the laboratory. Honey bee larvae and adults were reared in vitro and fed a diet that contained Cry9Ee toxin at 0.01, 0.1, 1, and 10 mg/L. Cry9Ee toxin 0.01, 0.1, and 1 mg/L in diet used in this study may represent a value closer to field relevance and the highest concentration is unlikely to be encountered in the field and thus represent a worst case scenario. The dependent variables were compared for groups of honey bees feeding on treated diet and those feeding on negative control (no addition of a test substance), solvent control (0.01 mM Na2CO3), and positive control diet (dimethoate 45 mg/L). Bt Cry9Ee toxin did not affect survival or larval weight, and the result was great confidence in accepting the null hypothesis by power analysis. The effect on development rates and pollen consumption were the inconclusive results because the post-hoc power was less than 0.8. Furthermore, the midgut bacterial structure and compositions were determined using high-throughput sequencing targeting the V3-V4 regions of the 16S rDNA. All core honey bee intestinal bacterial class such as γ-Proteobacteria, Actinobacteria, α-Proteobacteria, Bacilli, ß-Proteobacteria, and Bacteroidia were detected, and no significant changes were found in the species diversity and richness between Cry9Ee treatments and laboratory control.

The impacts of chlorothalonil and diflubenzuron on Apis mellifera L. larvae reared in vitro.

Chlorothalonil is a broad-spectrum fungicide and diflubenzuron is an insect growth regulator used to control many insect larvae feeding on agricultural, forest and ornamental plants. Honey bee larvae may be exposed to both via contaminated pollen, in the form of beebread, added to their diet by their adult nurse sisters. In this study, we determined how single (acute: 72 h mortality) and repeated (chronic: mortality until emergence as adults) exposure to chlorothalonil and diflubenzuron in their diet affected honey bee larvae reared in vitro. The tested doses of chlorothalonil (20, 100, or 200 mg/L) did not impact 72 h larval mortality acutely relative to that of the solvent control. The 72 h mortality of larvae exposed to 1.6 mg/L and higher doses of diflubenzuron acutely in their diet (47.2-63.9% mortality) was significantly higher than that of larvae fed the solvent control, with no predictable dose dependent pattern observed. In the chronic toxicity tests, consuming an artificial diet with 30 or 100 mg/L chlorothalonil and 0.8, 1.3 or 2 mg/L diflubenzuron significantly lowered the survival of honey bee larvae over that of larvae feeding on the solvent control diet. We calculated risk quotients (RQs) for both compounds using the data we generated in our experiments. Collectively, the RQs suggest that neither compound is likely to affect larval mortality directly at field relevant doses given that pollen composes only a fraction of the total larval diet. Nevertheless, our data do not preclude any sublethal effects that chronic exposure to either compound may cause.

Ingestion of Bt rice pollen does not reduce the survival or hypopharyngeal gland development of Apis mellifera adults.

Because of its ecological and economic importance, the honey bee Apis mellifera is commonly used to assess the environmental risk of insect-resistant, genetically modified plants. In the present study, feeding-exposure experiments were used to determine whether pollen from transgenic rice harms A. mellifera worker bees. In 1 experiment, the survival and mean acinus diameter of hypopharyngeal glands of adult bees were similar when bees were fed on pollen from Bt rice lines or from a non-Bt rice line, but bee survival was significantly reduced when they received pollen that was mixed with potassium arsenate as a positive control. In a second experiment, bee survival and hypopharyngeal gland development were not reduced when adult bees were fed on non-Bt pollen and a sucrose solution supplemented with Cry2A at 400 µg/g, Cry1C at 50 µg/g, or bovine serum albumin (BSA) at 400 µg/g, but bee survival and hypopharyngeal gland development were reduced when the diet was supplemented with soybean trypsin inhibitor as a positive control. In both experiments, the uptake of Cry proteins by adult bees was confirmed. Overall, the results indicate that the planting of Bt rice lines expressing Cry2A or Cry1C protein poses a negligible risk to A. mellifera worker bees. Environ Toxicol Chem 2017;36:1243-1248. © 2016 SETAC.

Bt Cry1Ie Toxin Does Not Impact the Survival and Pollen Consumption of Chinese Honey Bees, Apis cerana cerana (Hymenoptera, Apidae).

The cry1Ie gene may be a good candidate for the development of Bt maize because over-expression of Cry1Ie is highly toxic to Lepidopteran pests such as Heliothis armigera Hübner and Ostrinia furnacalis Guenée. The Bt cry1Ie gene also has no cross resistance with other insecticidal proteins such as Cry1Ab, Cry1Ac, Cry1Ah, or Cry1F. Chinese honey bees (Apis cerana cerana) are potentially exposed to insect-resistant genetically modified (IRGM) crops expressing Cry1Ie toxin via the collection of IRGM crop pollen. In this study, we tested whether Chinese honey bee workers are negatively affected by sugar syrup containing 20, 200, or 20,000 ng/ml Cry1Ie toxin and 48 ng/ml imidacloprid under controlled laboratory conditions. Our results demonstrated that the Cry1Ie toxin does not adversely impact survival and pollen consumption of Chinese honey bees. However, imidacloprid decreases Chinese honey bee survival and the total pollen consumption on the 5th, 6th, and 18th d of exposure. The described bioassay is suitable to assess the effects of GM expressed toxins against honey bee.

Field assessment of Bt cry1Ah corn pollen on the survival, development and behavior of Apis mellifera ligustica.

Honeybees may be exposed to insecticidal proteins from transgenic plants via pollen. An assessment of the impact of such exposures on the honeybee is an essential part of the risk assessment process for transgenic Bacillus thuringiensis corn. A field trial was conducted to evaluate the effect of transgenic Bt cry1Ah corn on the honeybee Apis mellifera ligustica. Colonies of honeybees were moved to Bt or non-Bt corn fields during anthesis and then sampled to record their survival, development and behavior. No differences in immature stages, worker survival, bee body weight, hypopharyngeal gland weight, colony performance, foraging activity or olfactory learning abilities were detected between colonies that were placed in non-Bt corn fields and those placed in Bt corn fields. We conclude that cry1Ah corn carries no risk for the survival, development, colony performance or behavior of the honeybee A. mellifera ligustica.