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Western honey bees (Apis mellifera), a cornerstone to crop pollination in the U.S., are faced with an onslaught of challenges from diseases caused by parasites, pathogens, and pests that affect this economically valuable pollinator. Natural products (NPs), produced by living organisms, including plants and microorganisms, can support health and combat disease in animals. NPs include both native extracts and individual compounds that can reduce disease impacts by supporting immunity or directly inhibiting pathogens, pests, and parasites. Herein, we describe the screening of NPs in laboratory cage studies for their effects on honey bee disease prevention and control. Depending on the expected activity of compounds, we measured varied responses, including viral levels, honey bee immune responses, and symbiotic bacteria loads. Of the NPs screened, several compounds demonstrated beneficial activities in honey bees by reducing levels of the critical honey bee virus deformed wing virus (DWV-A and-B), positively impacting the gut microbiome or stimulating honey bee immune responses. Investigations of the medicinal properties of NPs in honey bees will contribute to a better understanding of their potential to support honey bee immunity to fight off pests and pathogens and promote increased overall honey bee health. These investigations will also shed light on the ecological interactions between pollinators and specific floral food sources.

RNA viruses that contain single-stranded RNA genomes of positive sense make up the largest group of pathogens infecting honey bees. Sacbrood virus (SBV) is one of the most widely distributed honey bee viruses and infects the larvae of honey bees, resulting in failure to pupate and death. Among all of the viruses infecting honey bees, SBV has the greatest number of complete genomes isolated from both European honey bees Apis mellifera and Asian honey bees A. cerana worldwide. To enhance our understanding of the evolution and pathogenicity of SBV, in this study, we present the first report of whole genome sequences of two U.S. strains of SBV. The complete genome sequences of the two U.S. SBV strains were deposited in GenBank under accession numbers: MG545286.1 and MG545287.1. Both SBV strains show the typical genomic features of the Iflaviridae family. The phylogenetic analysis of the single polyprotein coding region of the U.S. strains, and other GenBank SBV submissions revealed that SBV strains split into two distinct lineages, possibly reflecting host affiliation. The phylogenetic analysis based on the 5&prime, UTR revealed a monophyletic clade with the deep parts of the tree occupied by SBV strains from both A. cerane and A. mellifera, and the tips of branches of the tree occupied by SBV strains from A. mellifera. The study of the cold stress on the pathogenesis of the SBV infection showed that cold stress could have profound effects on sacbrood disease severity manifested by increased mortality of infected larvae. This result suggests that the high prevalence of sacbrood disease in early spring may be due to the fluctuating temperatures during the season. This study will contribute to a better understanding of the evolution and pathogenesis of SBV infection in honey bees, and have important epidemiological relevance.

China is the largest producer and exporter of royal jelly (RJ) in the world, supplying >90% of the global market. The high production of RJ in China is principally owing to the high RJ-producing lineage of honeybees (Apis mellifera ligustica Spinola, 1806) established by beekeepers in the 1980s. We describe the development of high royal jelly-producing honeybees and the management of this lineage today. Previous research and recent advances in the genetic characterization of this lineage, and the molecular markers and mechanisms associated with high RJ production are summarized. The gaps in our knowledge and prospects for future research are also highlighted.

China is a country with high honeybee diversity, long history of beekeeping and large amount of managed honeybee colonies. Based on the development of a high royal jelly-producing lineage of honeybees and related techniques and tools, China is producing nearly all of royal jelly in the world market. Over the past decade, great development has occurred in Chinese beekeeping industry, e.g., the development of honeybee pollination market, the promotion of beekeeping with Apis cerana, and the mechanization of beekeeping. However, there are also challenges facing the industry, e.g., honey adulteration and colony losses caused by biotic and/or non-biotic factors. The exploitation of native bumble bees and stingless bees is still in the very early initial stage. The role of honeybee pollination in agriculture has been recognized by government and farmers. We believe that Chinese beekeeping is entering an era full of opportunities.

The Asian honeybee Apis cerana is one of two bee species that have been commercially kept with immense economic value. Here we present the analysis of genomic sequence and transcriptomic exploration for A. cerana as well as the comparative genomic analysis of the Asian honeybee and the European honeybee A. mellifera. The genome and RNA-seq data yield new insights into the behavioral and physiological resistance to the parasitic mite Varroa the evolution of antimicrobial peptides, and the genetic basis for labor division in A. cerana. Comparison of genes between the two sister species revealed genes specific to A. cerana, 54.5% of which have no homology to any known proteins. The observation that A. cerana displayed significantly more vigilant grooming behaviors to the presence of Varroa than A. mellifera in conjunction with gene expression analysis suggests that parasite-defensive grooming in A. cerana is likely triggered not only by exogenous stimuli through visual and olfactory detection of the parasite, but also by genetically endogenous processes that periodically activates a bout of grooming to remove the ectoparasite. This information provides a valuable platform to facilitate the traits unique to A. cerana as well as those shared with other social bees for health improvement.

The content of 10-hydroxy-trans-2-decenoic acid (10-HDA), a marker compound in royal jelly (RJ), is the most important criterion in grading RJ for commercial trade and varies with its origin. To identify the effect of geographical origin on 10-HDA content in RJ, 138 samples were collected from 19 provinces of China (divided into three groups) produced by either Apis mellifera ligustica Spinola, 1806 or a hybrid of A. m. ligustica and Apis mellifera carnica Pollman, 1879 and analyzed for moisture, sugar, crude protein, ash, acid, and 10-HDA concentration. The results show that RJ from western China has a significantly higher 10-HDA level (2.01 +/- 0.05%) than those from northeastern (1.87 +/- 0.05%) and eastern (1.75 +/- 0.03%) China. RJ secreted by hybrid bees contained more 10-HDA (1.89 +/- 0.03%) than that secreted by A. m. ligustica (1.78 +/- 0.03%). The 10-HDA content of RJ produced during flowering of rape (Brassica campestris L.), lime (Tilia amurensis Ruprecht), and vitex (Vitex negundo L. variety heterophylla (Franch.) Rehder) was 1.92, 1.80, and 1.68%, respectively. The results would be helpful during the process of price determination of RJ by providing some basis of geographical, bee strain, and botanical information for commercial trade.

International audience; AbstractIn honey bee colonies, workers, in particular of “anarchistic” lineages, can activate their ovaries and lay eggs, even in the presence of the queen. We identified three queenright colonies showing typical signs of worker reproduction. To characterize this new lineage, we extracted the mandibular gland and analyzed it using gas chromatography. The total amounts of the five main components of the mandibular gland, namely methyl p-hydroxyben-zoate (HOB), 9-oxo-2(E)-decenoic acid (9-ODA), (S)-9-hydroxy-(E)-2-decenoic acid (9-HDA), 10-HDA, and 10-hydroxyde-canoic acid (10-HDAA) were significantly higher in the mandibular gland profiles of workers with activated ovaries (AWs, 8.88 ± 1.71 μg) compared to workers with inactivated ovaries (IAWs, 4.00 ± 2.09 μg). Furthermore, the chemical profiles of IAWs were dominated by the “worker substances” 10-HDA (34.64 ± 8.19 %) and its precursor 10-HDAA (22.88 ± 4.95 %), while the chemical profiles in AWs were dominated by the precursor of the queen substance 9-HDA (40.04 ± 7.55 %). The ratios of two precursor substances 10-HDAA/9-HDA of IAWs were more worker like (>1.0) whereas AWs were more queen like (≤1.0). These results suggest that the mandibular pheromones of anarchistic workers resemble a more queen-like reproductive active profile and that these workers may represent a reversion to a more basal reproductive phenotype.

The sequences of mitochondrial ND2, CO2, and 16S rRNA genes and nuclear ITPR gene were obtained from 22 samples of 5 Apis species from China. The characteristics of the sequences and the pairwise distances among species were analyzed. Phylogenetic trees were reconstructed for Apis species using maximum parsimony, neighbor-joining and maximum likelihood methods together with the sequences of the other 4 Apis species downloaded from GenBank. Results supported that Apis species were divided into three major clusters: dwarf bees (A. florea and A. andreniformis), giant bees (A. dorsata and A. laboriosa), and cavity-nesting bees (A. mellifera, A. cerana, A. koschevnikovi, A. nigrocinta, and A. nuluensis). The dwarf honey bees were confirmed as basal lineage. Our study also revealed a high level of genetic divergence between A. dorsata from Hainan Island and China mainland.

International audience; We investigated the genetic structure of Chinese Apis dorsata population based on DNA microsatellites. The results show that populations from different geographical regions are significantly differentiated. Furthermore, the A. dorsata population from Hainan Island very probably diverged from that of the China mainland and also exhibits a lower level of genetic diversity, which is probably the result of founder effects and genetic drift. The results provide useful information for effective management and conservation of A. dorsata in China, particularly with regard to the population of Hainan Island.

Although the phylogenetic relationships of Apis species (Hymenoptera: Apidae) have been generally accepted, genetic variation within most species and some new possible species' status have not been studied in detail. In this study, we surveyed the genetic variation of mitochondrial DNA sequences in Apis species from China and investigated the phylogeography of Apis dorsata F. in China and neighboring Asian areas. The phylogeny of Chinese Apis species was consistent with the generally accepted Apis phylogeny, and a substructure within A. dorsata was suggested. Furthermore, results of analysis of molecular variance revealed significant genetic structuring of A. dorsata between Hainan Island and China mainland. When compared with neighboring Asian areas, Chinese A. dorsata grouped with one A. dorsata sample from Palawan Island of Philippines and two samples from Malaysia. Another two samples from Malaysia clustered with one Nepalese sample and the cluster from Thailand also was supported. These results support the hypothesis that glaciations and deglaciations during the Pleistocene could have greatly influenced the distribution and divergence of A. dorsata in China and Southeast Asia. In addition, the species status of Apis dorsata breviligula Maa and Apis dorsata binghami Cockerell were supported in our study and genetic variation may exist in Apis laboriosa Smith despite of limited natural distribution.

SummaryMultivariate morphometric analyses were performed on the giant honey bees Apis dorsata F. and Apis laboriosa F. of China. The two giant honey bee species were readily separated by principal components, cluster and discriminant analyses. Apis laboriosa showed higher values for the variables of size and cubital index than A. dorsata (ranging respectively as the width of the wax mirror 2.83~2.99 mm and 2.36~2.56 mm, width of sternum 6 3.61~3.86 mm and 3.03~3.26 mm, cubital index 10.36~14.56 and 6.38~9.61). Most of the wing venation angles were also significantly different in the two giant honey bees (angle J10 were 38.2°~40.7° and 34.7°~37.7° respectively). A. dorsata is distributed widely across China, and cluster and discrimination analyses indicated that they could be subdivided into two morphometric subclusters: one from the Chinese mainland and another from Hainan Island which reflects their relative geographic isolation. Few characters differed in the two A. dorsata morphometric subclusters, how...

The content and composition of fatty acids in the seed oil from 4 blackberry varieties were compared in this paper to provide further guidelines for use of blackberry seeds. Blackberry seed oil contains rich α-linolenic and linoleic acid making it an excellent dietary source of 18:3n-3 and essential fatty acids. The oil is a good dietary source of natural antioxidants for health promotion and disease prevention. The presence of n-3 fatty acid and antioxidants also suggests potential value-added utilization of blackberry seeds for preparing functional foods or supplemental products.

In order to understand the nutritional characteristics of blueberries (Vaccinium spp.), provide the fundamental basis for its processing and utilization, 13 blueberry varieties were studied on their major nutrient compositions in fruit. The results showed that blueberry fruit contained 10.31 ± 0.98% of soluble solids, of which 77.13% was soluble sugar and 89.02% of soluble sugar was reducing sugar. Average content of organic acids was 0.79 ± 0.23% and sugar-acid ratio was 10.91 ± 3.38. Total polyphenol content was 4.921 ± 0.738 mg·g-1, of which anthocyanin was 1.441 ± 0.435 mg·g-1 and ellagic acid 0.921 ± 0.098 mg·g-1. Nutrient content and their components had difference between blueberries varieties.

Nisin is an antimicrobial polypeptide produced by Lactococcus lactis and is believed nontoxic to humans. The objective of this study was to evaluate a nisin-based formulation for the treatment of bovine clinical mastitis in lactating dairy cattle. A total of 92 cows with 107 clinically mastitic quarters were randomly assigned to nisin- (48 cows with 51 quarters) and gentamicin (GM)-treated (44 cows with 56 quarters) groups. In the nisin-treated group, cows received an intramammary infusion of nisin at a dose of 2,500,000 IU; in the GM-treated group, intramammary infusion of GM was administered at a dose of 0.8 g. Results indicated that nisin offered a clinical cure rate similar to GM (90.2 vs. 91.1%) and no difference in bacteriological cure rate than GM-treated group (60.8 vs. 44.6%, respectively). Proportion of the quarters with milk somatic cell counts500,000 cells/mL was not different in the nisin-treated group (50.0 and 47.8%) compared with the GM-treated group (33.3 and 37.3%) 1 and 2 wk after treatment. Of 17 Staphylococcus aureus isolates, 82.5% were resistant to penicillin, and 35.3% to GM, but none of them to nisin. Nisin therapy eliminated 54.5% (6 of 11) of S. aureus IMI, whereas GM eliminated 33.3% (2 of 6). Nisin in milk (4.5 +/- 0.8 IU/mL) was detected only at 12 h following intramammary infusion, which was much lower than the upper limit (500 mg/mL) allowed as preservative in milk by the China authority. Because of its efficacy in the treatment of bovine clinical mastitis, especially resistant Staph. aureus-caused IMI, as well as its safety in humans, nisin deserves further study to clarify its effects on mastitis caused by different mastitis pathogens on a larger scale.

In an attempt to isolate and identify the antigenic epitopes on ferritin of Schistosoma japonicum (SjFer) and to test their protective potentiality against Schistosoma japonicum (S.j), polyclonal antisera against SjFer was prepared to screen a 12-mer random peptide library. Three rounds of biopanning were performed and resulted in an enrichment. Six peptides selected randomly from the third elute were all found to be positive by evaluating the binding to anti-SjFer sera by ELISA and Western blotting. Three amino acid sequences were deduced from the six phage clones by sequencing. When they were used to vaccinate mice, the three peptides could induce significant reduction in adult worms (26.7%, 20.4%, and 25.9%) as well as in liver eggs per gram (LEPG) (40.0%, 38.2%, and 40.8%). This result showed that three mimotopes on SjFer were obtained and they could induce significant protective efficacy against S.j.

Caste differentiation in the female honey bee is one of the most intriguing polyphenism phenomena. This developmental switch depends on the differential expression of entire suites of the genes involved in the larval fate between the queens and workers. In this study, we compared the transcriptome differences between full-sister queen- (QL) and worker-destined larvae (WL) using high-throughput RNA-Seq. QL and WL at fourth (L4) and fifth instar (L5) were used to prepare four libraries and to generate 50,191,699 (QL4), 57,628,541 (WL4), 56,613,619 (QL5), and 58,626,829 (WL5) usable reads, which were assembled into groups of 7,952, 7,993, 7,971, and 8,023 genes, respectively. The transcriptome changes were investigated using the DEGs Package (DEGseq), which resulted in more than 4,500 differentially expressed genes (DEGs) between the castes. Eight of the DEGs were verified by quantitative real-time RT-PCR (qRT-PCR), and the results supported our sequencing data. All of the DEGs were analysed using Web Gene Ontology Annotation Plot (WEGO) and then mapped using the Kyoto Encyclopedia of Genes and Genomes (KEGG) database. These results suggest that over 70% of the DEGs in each instar were more highly expressed in QL than in WL, possibly suggesting that the QL genes had higher transcriptional activity than the WL genes during differentiation. The same gene set is active (but differentially expressed) in both castes, which in turn result in dimorphic females. The L4 stage is a very active gene expression period for both QL and WL before their pupal stage. The activity of the mTOR (a target of rapamycin) encoding gene in the mTOR signalling pathway is higher in QL4 than in WL4, and this difference was no longer present by the L5 feeding stage. The genes down-stream of mTOR maintained this change at the L5 stage. These results could contribute to an in-depth study of the candidate genes during honey bee caste differentiation and improve our current understanding of the polyphenism phenomenon in insects.