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Imidacloprid is a global health threat that severely poisons the economically and ecologically important honeybee pollinator, Apis mellifera . However, its effects on developing bee larvae remain largely unexplored. Our pilot study showed that imidacloprid causes developmental delay in bee larvae, but the underlying toxicological mechanisms remain incompletely understood. In this study, we exposed bee larvae to imidacloprid at environmentally relevant concentrations of 0.7, 1.2, 3.1, and 377 ppb. There was a marked dose-dependent delay in larval development, characterized by reductions in body mass, width, and growth index. However, imidacloprid did not affect on larval survival and food consumption. The primary toxicological effects induced by elevated concentrations of imidacloprid (377 ppb) included inhibition of neural transmission gene expression, induction of oxidative stress, gut structural damage, and apoptosis, inhibition of developmental regulatory hormones and genes, suppression of gene expression levels involved in proteolysis, amino acid transport, protein synthesis, carbohydrate catabolism, oxidative phosphorylation, and glycolysis energy production. In addition, we found that the larvae may use antioxidant defenses and P450 detoxification mechanisms to mitigate the effects of imidacloprid. Ultimately, this study provides the first evidence that environmentally exposed imidacloprid can affect the growth and development of bee larvae by disrupting molting regulation and limiting the metabolism and utilization of dietary nutrients and energy. These findings have broader implications for studies assessing pesticide hazards in other juvenile animals.

Nosema ceranae is the pathogen of nosemosis in the honey bee, which can bring great economic loss to apiculture. Chitin acts as a major component of the endospore of microsporidia and plays an essential role to form the bridges across the endospore. Here, Chitin Spore Coats (CSCs) of N. ceranae were successfully extracted by optimized hot alkaline treatment. SDS-PAGE and Calcofluor White Stain (CWS) staining indicated that the obtained CSCs were protein-free and the transmission electron microscopy analysis showed that CSCs performed the intact and loose chitin spore coats. Western blotting and indirect immunofluorescence analysis (IFA) demonstrated that CSCs could interact with three spore wall proteins (rNcSWP7, rNcSWP8, and rNcSWP12). Our method was effective to extract CSCs of N. ceranae and this could be very useful for screening spore wall proteins involved in endospore composition, which could be helpful to uncover the biological structure and pathogenesis of microsporidia.

Worker honey bees are crucial for colony stability and ecosystem pollination. However, the cross-scale aging features and underlying mechanisms in the Eastern honey bee (Apis cerana) remain poorly understood. This study systematically investigated age-related changes in A. cerana workers across youth (1~5 days post-emergence, dpe), middle age (29 dpe), and old age (50 dpe) through integrated morphological, ultrastructural, and transcriptomic analyses. With increasing age, the phenotypic deterioration in the old bees (OBs) was significant: the body color brightness decreased by 16.7% compared to the young bees (YBs) (p < 0.001), and the hair density of the head, thorax, and abdomen declined by 63.5%, 97.2%, and 91.5%, respectively (p < 0.0001). The wing wear index (WWI) increased to 96.7% (p < 0.0001). The locomotor performance declined sharply, with only 6.7% of the OBs successfully reaching the feeding platform within 15 s (p < 0.0001). Ultrastructural analysis revealed sensory organ abrasion, flattened thoracic bristles, thickened cuticle, and 90.4% increased mitochondrial damage (p < 0.0001). The autophagosomes showed dynamic changes, with 81.8% reduction versus those of mid-aged bees (MBs) (p < 0.001), which suggests that mitochondrial dysfunction and autophagy dysregulation may be the core driving factors behind aging. Transcriptomics identified 67 differentially expressed genes enriched in lifespan regulation, glutathione metabolism, and lysosomal pathways. Fifteen key aging-related genes were identified, such as major royal jelly protein 3 (MRJP3), synaptic vesicle glycoprotein 2A (SV2A), and apidermin 3 (APD3), whose expression dynamics have been shown to be closely related to nutritional metabolism, behavioral perception, and the decline of epidermal barrier function. This work establishes the first multidimensional aging evaluation system for A. cerana, providing critical insights into bee senescence mechanisms and colony health optimization.

Microsporidia have attracted much attention because they infect a variety of species ranging from protists to mammals, including immunocompromised patients with AIDS or cancer. Aside from the study on Nosema ceranae, few works have focused on elucidating the mechanism in host response to microsporidia infection. Nosema bombycis is a pathogen of silkworm pébrine that causes great economic losses to the silkworm industry. Detailed understanding of the host (Bombyx mori) response to infection by N. bombycis is helpful for prevention of this disease. A genome-wide survey of the gene expression profile at 2, 4, 6 and 8 days post-infection by N. bombycis was performed and results showed that 64, 244, 1,328, 1,887 genes were induced, respectively. Up to 124 genes, which are involved in basal metabolism pathways, were modulated. Notably, B. mori genes that play a role in juvenile hormone synthesis and metabolism pathways were induced, suggesting that the host may accumulate JH as a response to infection. Interestingly, N. bombycis can inhibit the silkworm serine protease cascade melanization pathway in hemolymph, which may be due to the secretion of serpins in the microsporidia. N. bombycis also induced up-regulation of several cellular immune factors, in which CTL11 has been suggested to be involved in both spore recognition and immune signal transduction. Microarray and real-time PCR analysis indicated the activation of silkworm Toll and JAK/STAT pathways. The notable up-regulation of antimicrobial peptides, including gloverins, lebocins and moricins, strongly indicated that antimicrobial peptide defense mechanisms were triggered to resist the invasive microsporidia. An analysis of N. bombycis-specific response factors suggested their important roles in anti-microsporidia defense. Overall, this study primarily provides insight into the potential molecular mechanisms for the host-parasite interaction between B. mori and N. bombycis and may provide a foundation for further work on host-parasite interaction between insects and microsporidia.

Vairimorpha (Nosema) ceranae is a pathogen that affects Apis mellifera and Apis ceranae Fabricius, capable of spreading within and between honeybee colonies. The spore wall of microsporidia is the initial structure to contact the host cell directly, which may play a crucial role in the infection process. Currently, several spore wall proteins have been identified in microsporidia, but only two spore wall proteins from V. ceranae have been characterized. Here, we report the expression and identification of a novel spore wall protein, NcSWP8, with a molecular mass of 21.37 kDa in V. ceranae. Subcellular localization analysis revealed that NcSWP8 was localized on the spore wall of V. ceranae. Co-immunoprecipitation and Far-Western blotting experiments demonstrated that NcSWP8 could stably interact with polar tube proteins, NcPTP2 and NcPTP3. The antibody blocking assay significantly decreased their infection rate, indicating that NcSWP8 played a significant role in the process of V. ceranae infection. These results together suggested that NcSWP8 was a new spore wall protein localized to the spore wall and interacted with the polar tube proteins, playing a crucial role in supporting the formation of the spore wall and potentially affecting the process of infection of V. ceranae.

Chinese sacbrood disease (CSD), which is caused by Chinese sacbrood virus (CSBV), is a major viral disease in Apis cerana cerana larvae. Analysis of lipid composition is critical to the study of CSBV replication. The host lipidome profiling during CSBV infection has not been conducted. This paper identified the lipidome of the CSBV–larvae interaction through high-resolution mass spectrometry. A total of 2164 lipids were detected and divided into 20 categories. Comparison of lipidome between healthy and CSBV infected-larvae showed that 266 lipid species were altered by CSBV infection. Furthermore, qRT-PCR showed that various sphingolipid enzymes and the contents of sphingolipids in the larvae were increased, indicating that sphingolipids may be important for CSBV infection. Importantly, Cer (d14:1 + hO/21:0 + O), DG (41:0e), PE (18:0e/18:3), SM (d20:0/19:1), SM (d37:1), TG (16:0/18:1/18:3), TG (18:1/20:4/21:0) and TG (43:7) were significantly altered in both CSBV_24 h vs. CK_24 h and CSBV_48 h vs. CK_48 h. Moreover, TG (39:6), which was increased by more than 10-fold, could be used as a biomarker for the early detection of CSD. This study provides evidence that global lipidome homeostasis in A. c. cerana larvae is remodeled after CSBV infection. Detailed studies in the future may improve the understanding of the relationship between the sphingolipid pathway and CSBV replication.

Climate change affects public health, and improving eco-efficiency means reducing the various pollutants that are the result of economic activities. This study provided empirical evidence of the quantitative impact analysis of climate change on the health conditions of residents across China due to improvements that have been made to eco-efficiency. First, the indicators that were collected present adequate graphical trends and regional differences with a priori evidence about their relationships to each other; second, the present study applied Sensitivity Evaluation with Support Vector Machines (SE-SVM) to Chinese provincial panel data, taking the Visits to Hospitals, Outpatients with Emergency Treatment, and Number of Inpatients as proxy variables for the health conditions of the residents in each area and temperature, humidity, precipitation, and sunshine as the climate change variables, simultaneously incorporating the calculated eco-efficiency with six controlling indicators; third, we compared in-sample forecasting to acquire the optimal model in order to conduct elasticity analysis. The results showed that (1) temperature, humidity, precipitation, and sunshine performed well in forecasting the health conditions of the residents and that climate change was a good forecaster for resident health conditions; (2) from the national perspective, climate change had a positive relationship with Visits to Hospitals and Outpatients with Emergency Treatment but a negative relationship with the Number of Inpatients; (3) An increase in regional eco-efficiency of 1% increase the need for Visits to Hospitals and Outpatients with Emergency Treatment by 0.2242% and 0.2688%, respectively, but decreased the Number of Inpatients by 0.6272%; (4) increasing the regional eco-efficiency did not show any positive effects for any individual region because a variety of local activities, resource endowment, and the level of medical technology available in each region played different roles. The main findings of the present study are helpful for decision makers who are trying to optimize policy formulation and implementation measures in the cross-domains of economic, environmental, and public health.

Regional eco-efficiency affects local public health through intermediaries such as economic and environmental impacts. Considering multiple factors, the implicit and uncertain relationship with regional characteristics, and the limited data availability, this paper investigated the forecasting of changes in local public health—including the number of visits to hospitals (VTH), outpatients with emergency treatment (OWET), number of inpatients (NI), number of health examinations (NOHE), and patients discharged (PD)—using calculated regional eco-efficiency with the Least Square-Support Vector Machine-Forecasting Model and acquired empirical evidence, utilizing the province-level data in China. Results: (1) regional eco-efficiency is a good predictor in both a single and multi-factor situation; (2) the prediction accuracy for five dimensions of the changes in local public health was relatively high, and the volatility was lower and more stable throughout the whole forecasting period; and (3) regional heterogeneity, denoted by three economic and demographic factors and three medical supply and technical level factors, improved the forecasting performance. The findings are meaningful for provincial-level decision-makers in China in order for them to know the current status or trends of medical needs, optimize the allocation of medical resources in advance, and enable ample time to tackle urgent emergencies, and, finally, the findings can serve to evaluate the social effects of improving regional eco-efficiency via local enterprises or individuals and adopting sustainable development strategies.

Imidacloprid is a global health threat that severely poisons the economically and ecologically important honeybee pollinator, Apis mellifera . However, its effects on developing bee larvae remain largely unexplored. Our pilot study showed that imidacloprid causes developmental delay in bee larvae, but the underlying toxicological mechanisms remain incompletely understood. In this study, we exposed bee larvae to imidacloprid at environmentally relevant concentrations of 0.7, 1.2, 3.1, and 377 ppb. There was a marked dose-dependent delay in larval development, characterized by reductions in body mass, width, and growth index. However, imidacloprid did not affect on larval survival and food consumption. The primary toxicological effects induced by elevated concentrations of imidacloprid (377 ppb) included inhibition of neural transmission gene expression, induction of oxidative stress, gut structural damage, and apoptosis, inhibition of developmental regulatory hormones and genes, suppression of gene expression levels involved in proteolysis, amino acid transport, protein synthesis, carbohydrate catabolism, oxidative phosphorylation, and glycolysis energy production. In addition, we found that the larvae may use antioxidant defenses and P450 detoxification mechanisms to mitigate the effects of imidacloprid. Ultimately, this study provides the first evidence that environmentally exposed imidacloprid can affect the growth and development of bee larvae by disrupting molting regulation and limiting the metabolism and utilization of dietary nutrients and energy. These findings have broader implications for studies assessing pesticide hazards in other juvenile animals.

Background Microsporidia is an obligate intracellular eukaryote, which is capable of parasitizing vertebrates and invertebrates . Nosema ceranae, which can infect both Apis mellifera and Apis cerana, poses a serious threat and causes heavy losses to the worldwide apiculture. During infection, polar tube, a highly specialized invasion structure, ejected from the spore to deliver the sporoplasm into host cells to cause infection. Although seven different polar tube proteins (PTP1 ~ 7) have been reported from various microsporidia and showed key functions associated with spore invasion and proliferation, no systematic analysis on identification and characterization of polar tube proteins from N. ceranae was found. Methods and results The polar tube proteins 2 (NcPTP2) was identified from the total polar tube proteins of N. ceranae by LC_MS/MS and the transcriptional profile was performed by RT-PCR. Sequence characterization analysis revealed that NcPTP2 was rich in lysine and had a signal peptide at the N-terminal. It had 3 potential O-glycosylation sites and 6 potential N-glycosylation sites. 25 phosphorylation sites were found on serine, tyrosine and threonine sites. Sequence alignment analysis revealed that NcPTP2 was homologous and had conserved cysteine residues with PTP2 proteins from other microsporidia. Indirect immunofuorescence analysis (IFA) and Immunoelectron Microscopy analysis (IEM) confirmed that NcPTP2 was localized on the polar tube of the germinated spores. The interaction between NcPTP2 and spore wall protein in N. ceranae indicated its potential function in anchoring and coiling of polar tube in spore. Conclusion NcPTP2 was the first subcellular localized polar tube protein in N. ceranae and this work could provide an important basis for further analyzing the biological functions of polar tube proteins and uncovering the infection mechanism of N. ceranae to the host cells.