Honey, a natural healthy liquid bee product, is rich in amino acids, vitamins, and other essential nutrients. Different origin honeys also varied in organic acids. The objective of this study is to establish an efficient solid-phase extraction-gas chromatography–mass spectrometry (SPE-GC–MS) method to eliminate interference of sugar and other impurities for accurately estimating 22 organic acids in honey by optimizing extraction, purification, derivatization, and gas chromatography–mass spectrometry (GC–MS) analytical conditions. After being extracted, purified and derivatized, organic acids in honey were qualified and quantified by GC–MS. This method was evaluated experimentally, and the results showed that, within a certain concentration range, the standard curve linear relationship was satisfactory (R² > 0.9942), and the target organic acid recovery rate was 86.74% ~ 118.68%. Besides, precision (relative standard deviation, RSD = 2.98% ~ 13.42%), detection limit (LOD = 0.002 ~ 0.2 mg kg⁻¹), and quantification limit (LOQ = 0.008 ~ 0.5 mg kg⁻¹) met the target requirements. Also, based on this analytical method, the organic acids in five types of honey (acacia, jujube, vitex, canola and linden honey) were estimated. Notably, they all contained 22 different kinds of organic acids, and significant differences (p < 0.05) in the organic acid content and composition among different honey varieties were observed. PCA analysis showed that the five honeys could be differentiated based on the content of 22 organic acids.

Temperature is a pivotal ecological factor in the regulation of insect survival and reproduction

Honeybees are vital pollinators with functional differentiation as a key survival strategy. The Chinese honeybee (Apis cerana cerana) exhibits exceptional nectar foraging in complex terrains, yet how alternative splicing (AS) and polyadenylation (APA) regulate its labor division remains unclear. Here, we applied PacBio full-length transcriptome sequencing to annotate worker bee transcriptomes across three developmental stages (Ac3d, Ac10d, Ac21d), calibrating the third-generation sequencing data with second-generation sequencing to enhance transcriptome annotation accuracy. We identified 17,961 isoforms and 1922 APA genes, finding that alternative first exon was the major type of AS, while APA enhances transcriptomic diversity via dual polyadenylation sites in most genes. Functional analyses revealed AS enrichment in growth signaling (Vg6, CYP15A1) and immune pathways (PTPRR), whereas APA regulated growth signaling (Vg6), energy metabolism (Acsl1, AcceFE), and oxidative stress (PTPRR, PPO2). Validation by PCR and 3′RACE confirmed stage-specific AS/APA events in key genes. These findings significantly enhance the A. cerana cerana reference genome annotation and provide valuable insights into the mechanisms of AS and APA regulation underlying honeybee development and functional transitions.

Honey bees are important pollinators that enhance food safety and promote biodiversity. However, honey bees are increasingly threatened by the use of insecticides. Matrine, a plant-derived insecticide, has been used in plants such as pear trees, citrus, and cotton, whose flowers serve as a honey source. Despite matrine's economic significance, its impact on bees is rarely reported. In this study, newly emerged honey bee workers (Apis cerana) were randomly divided into four groups and reared to adulthood within a simulated colony environment with suitable temperatures, humidity, and food sources. The adult workers were exposed to four concentrations (0, 1.2, 10, and 40 mg/L) of matrine. Continuous matrine intake for more than 9 days caused the rapid death of honey bees but had no significant effect on food consumption and body weight of the honey bees. Oral exposure to matrine decreased the abundance of beneficial Gilliamella, Dorea, and Lachnoclostridium in gut microbiota. It also led to the up-regulation of argininosuccinic acid and spermidine, and the down-regulation of arachidonic acid and LOC107998471 (Aldh), which control the decomposition of spermidine. This finding underscored the role of these microbes, metabolites, and genes in honey bees under matrine stress. This study provides a comprehensive understanding of matrine toxicity to honey bees, revealing the crucial roles of microbiota, metabolites, and genes. These findings provide important references for honey bee poisoning and the associated detoxification mechanism.

Declining wild honeybee (Apis) resources could cause the extinction of symbiotic relationships between plants and honeybees, reducing food production. Therefore, the protection of genetic resources and the diversity of wild honeybees are crucial. This study examined the honeybees in a plant nature reserve (Leigongshan) to better understand their genetic status in a purely natural state and explored the potential role of this plant nature reserve on pollinators. Nuclear (15 microsatellites) and mitochondrial markers (tRNA^leu ~ COII and COI) were used to investigate the genetic resources and diversity of eastern honeybees (Apis cerana). Here, 31 new honeybee haplotypes were discovered from the inside and outside regions of the reserve. The honeybees showed a high genetic diversity level (He: 0.6564 ± 0.1594, Na: 3–18, PIC: 0.6191 ± 0.1718, H_d in tRNA^leu ~ COII: 0.6797, H_d in COI: 0.8103), and no apparent genetic differentiation was observed between the two regions. Thus, we speculate that Leigongshan Nature Reserve protects honeybees from human disturbance and prevents a decline in their population size, preserving the honeybee genetic resources and high genetic diversity level. Furthermore, our results have revealed the practical function of the plant nature reserve on honeybee genetic resources, which explains the decreasing honeybee resources.

Nature reserves regulate the genetic conservation of endangered wildlife, plants, and ecosystems. Since honey bees are severely threatened, understanding the role of nature reserves in protecting their genetic variation is necessary. This study used two robust mitochondrial genetic markers (tRNA^leu ~ COII and COI) to analyze the genetic structure of the eastern honey bee Apis cerana Fabricius in and around Fanjingshan National Nature Reserve, China. Private and new reported haplotypes were identified in 197 honey bee colonies in the study area. Based on the network, phylogenetic tree, structure analysis, low F-statistics (0.01251 in tRNA^leu ~ COII, 0.00738 in COI), and great gene flow (35.17 in tRNA^leu ~ COII, 129.82 in COI), all the honey bees inside and outside Fanjingshan Nature Reserve revealed private haplotypes without genetic differentiation between them. We speculated that the nature reserve played a limited positive role in protecting eastern bee genetic resources since no specific population was formed until now. However, new private haplotypes were discovered inside the reserve. This work demonstrates the impact of nature reserves on honey bee genetic resources.