Neonicotinoid insecticides are now the most widely used insecticides in the world. Previous studies have indicated that sublethal doses of neonicotinoids impair learning, memory capacity, foraging, and immunocompetence in honey bees (Apis mellifera, Linnaeus) (Hymenoptera: Apidae). Despite these, few studies have been carried out on the molecular effects of neonicotinoids. In this study, we focus on the second-generation neonicotinoid thiamethoxam, which is currently widely used in agriculture to protect crops. Using high-throughput RNA-Seq, we investigated the transcriptome profile of honey bees after subchronic exposure to 10 ppb thiamethoxam over 10 d. In total, 609 differentially expressed genes (DEGs) were identified, of which 225 were upregulated and 384 were downregulated. Several genes, including vitellogenin, CSP3, defensin-1, Mrjp1, and Cyp6as5 were selected and further validated using real-time quantitative polymerase chain reaction assays. The functions of some DEGs were identified, and Gene Ontology-enrichment analysis showed that the enriched DEGs were mainly linked to metabolism, biosynthesis, and translation. Kyoto Encyclopedia of Genes and Genomes pathway analysis showed that thiamethoxam affected biological processes including ribosomes, the oxidative phosphorylation pathway, tyrosine metabolism pathway, pentose and glucuronate interconversions, and drug metabolism. Overall, our results provide a basis for understanding the molecular mechanisms of the complex interactions between neonicotinoid insecticides and honey bees.
The Earth has undergone a significant climate switch from greenhouse to icehouse during the Plio-Pleistocene transition (PPT) around 2.7-2.4 million years ago (Ma), marked by the intensification of the Northern Hemisphere glaciation (NHG) ~2.7 Ma. Evidence based on oceanic CO2 [(CO2)aq], supposed to be in close equilibrium with the atmospheric CO2 [(CO2)atm], suggests that the CO2 decline might drive such climate cooling. However, the rarity of direct evidence from [CO2]atm during the interval prevents determination of the atmospheric CO2 level and further assessment on the impact of its fluctuation. Here, we reconstruct the [CO2]atm level during 2.77-2.52 Ma based on a new developed proxy of stomatal index on Typha orientalis leaves from Shanxi, North China, and depict the first [CO2]atm curve over the past 5 Ma by using stomata-based [CO2]atm data. Comparisons of the terrestrial-based [CO2]atm and the existed marine-based [CO2]aq curves show a similar general trend but with different intensity of fluctuations. Our data reveal that the high peak of [CO2]atm occurred at 2.77-2.52 Ma with a lower [CO2]aq background. The subsequent sharp fall in [CO2]atm level might be responsible for the intensification of the NHG based on their general temporal synchronism. These findings shed a significant light for our understanding toward the [CO2]atm changes and its ecological impact since 5 Ma.
Bacteria utilize a quorum sensing (QS) system to coordinate gene expression by monitoring the concentration of molecules known as autoinducers (AI). In the present study, we confirmed the presence of a LuxS/AI-2 dependent QS system in vancomycin-resistant Enterococcus faecalis V583. Then, the cellular targets controlled by AI-2 were identified by comparative proteomics analysis in order to elucidate the possible role of AI-2 in E. faecalis. Results demonstrated 15 proteins that are differentially expressed upon the addition of AI-2, including proteins involved in metabolism, translation, energy production and/or conversion, and cell wall biogenesis. Glyceraldehyde-3-phosphate dehydrogenase and malate dehydrogenase associated with carbohydrate metabolism and energy production were up-regulated upon inducing by AI-2. In addition, externally added AI-2 could down-regulate acetyl-coenzyme A carboxylase and ornithine carbamoyltransferase, two key enzyme involved in metabolism. All these data suggest that AI-2 signaling may play a role in the regulation of a number of important metabolic properties of E. faecali. We further investigated the role of AI-2 in biofilm formation by E. faecalis, showing the addition of AI-2 to E. faecalis V583 cultures resulted in increased biofilm formation. Our results provide important clues to the role of a LuxS/AI-2 dependent QS system in vancomycin-resistant E. faecalis.
As one of the most widely used insecticides, thiamethoxam (Th) plays an important role in agriculture pest control but is hazardous to the health of honeybees (Apis mellifera ligustica ). In agricultural practice, several types of insecticides are often applied simultaneously, but their synergistic effects on honeybees remain elusive. Here, we treated honeybees with various binary mixtures of insecticides in which a sublethal dose of Th was mixed with three other common insecticides: λ-cyhalothrin (λ-cy), β-cypermethrin (β-cy), and abamectin (Ab). The mortality rates of the groups treated with the insecticide mixtures were higher than that obtained with the pesticides alone. Specifically, the Th + λ-cy and Th + Ab combinations exerted a synergistic effect (P < 0.05) on bees and resulted in significantly higher mortality than that obtained with the single treatments. However, the Th + β-cy mixture had no synergistic effect (P > 0.05) on bees, and the mortality rate of this group was almost identical to those of the groups administered these pesticides alone. Furthermore, the Th + λ-cy and Th + Ab mixtures significantly suppressed the activity and expression of glutathione S-transferases (GSTs), which are key detoxification enzymes in insects. This result suggests that the Th + λ-cy and Th + Ab combinations at a given concentration adversely affect the survival of honeybees. Our study provides novel information for the selection of pesticide combinations based on protecting honeybee populations.
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