Apis Mellifera Ligustica Research Articles

Design, Synthesis, and Antifungal Evaluation of Novel Pyrazole-5-sulfonamide Derivatives for Plant Protection.

To develop further novel environmentally friendly antifungal agents with high efficacy, a series of pyrazole-5-sulfonamide derivatives were designed and synthesized by using the active molecules synthesized in previous works as lead compounds. Their antifungal activities were evaluated in vitro against ten highly destructive plant pathogenic fungi. The bioassay results indicated that more than half of the target compounds displayed potent antifungal activities (inhibition rate ≥85%) against Valsa mali and Sclerotinia sclerotiorum at 20 mg/L. Among them, compound C22 exhibited significant broad-spectrum antifungal activities against V. mali, S. sclerotiorum, Rhizoctonia solani, Botrytis cinerea, and Trichoderma viride, with EC50 values of 0.45, 0.49, 3.06, 0.57, and 1.43 mg/L, respectively. Moreover, compounds C21 and C22 exhibited remarkable protective effects on apple Valsa canker similar to tebuconazole (89.5%) at 50 mg/L. Preliminary antifungal mechanism investigations demonstrated that compound C22 may have inhibited V. mali mycelial growth by inducing oxidative damage to the mycelium and compromising the integrity of the cell membrane. Meanwhile, compounds C21 and C22 exhibited no obvious toxicity to worker bees (Apis mellifera ligustica). Taken together, these pyrazole-5-sulfonamide derivatives, particularly compound C22, possess huge potential to be developed as novel environmentally friendly fungicides with high efficacy.

Genomic diversity and population structure of Carniolan honey bee in its native habitat

BackgroundResearch into the genetic diversity of honey bee (Apis melliferaL.) populations has become increasingly significant in recent decades, primarily due to population declines attributed to human activities and climate change. As a species of great importance, breeding programs that leverage understanding of genomic diversity could offer solutions to mitigate these challenges. The objective of this study was to examine the genomic diversity and population structure of Carniolan honey bees (Apis mellifera carnica) using the Illumina SNP chip on a large honey bee sample collected from Central and South-Eastern European countries. The study also aims to offer recommendations for future breeding programs.ResultsOur analysis involved Discriminant Analysis of Principal Components (DAPC), heterozygosity, admixture analysis, fixation indices (FST), Neighbour-Joining tree, gene flow and Isolation-by-distance analysis. DAPC indicated distinct separation between the Carniolan and Italian honey bee (Apis mellifera ligustica) populations, whereas the admixture analysis revealed varying levels of gene flow and genetic admixture within the Carniolan honey bee populations, demonstrating closer relationships between specific geographic regions (confirmed by Isolation-by-distance analysis). Furthermore, the research of heterozygosity, genomic inbreeding, pairwise FST values, and Neighbour-Joining tree provided insights into the patterns of genetic differentiation and similarity among the populations of Carniolan honey bee within its natural habitat. We have observed genetic homogeneity of the Carniolan honey bee population when considered in a broader genetic/geographical context. However, the Carniolan honey bee has sufficient genetic diversity in its geographical home range that needs to be carefully monitored and maintained.ConclusionsThis study provides important insights into the genetic composition, differentiation, and relationships among Carniolan honey bee populations in Central and South-Eastern European countries. The findings are crucial for conservation efforts, breeding programs, and sustainable beekeeping practices. They emphasise the importance of considering genetic factors and population structure in the breeding and management of honey bees. By understanding these genetic relationships, we can develop strategies to preserve genetic diversity, improve breeding outcomes, and ensure the resilience of honey bee populations in the face of environmental changes and challenges. This knowledge can also inform policy makers and stakeholders on best practices to maintain healthy bee populations, which are vital for ecosystem services and agricultural productivity.

Distribution of honey bee mitochondrial DNA haplotypes in an Italian region where a legislative act is protecting the Apis mellifera ligustica subspecies

The conservation of the genetic integrity of Apis mellifera subspecies has emerged as an important objective. In 2019, the Emilia-Romagna region became the first Italian regional authority to issue a law specifically addressing the protection of the native Apis mellifera ligustica subspecies. In this study we analysed a highly informative portion of the mitochondrial DNA (mtDNA), widely used for assessing genetic diversity of honey bee populations. By analysing 1143 honey bees sampled after the introduction of this law, we provided a snapshot of the distribution of mtDNA haplotypes in this region. The two most frequent mtDNA haplotypes were C1 (characteristic of A. m. ligustica) and C2 (characteristic of A. m. carnica), reported in 86.5% and 11.0% of the analysed bees, respectively. About 1.3% and 1.1% of the analysed bees carried mtDNA haplotypes of the A and M lineages (haplotypes A1a, A1e, A4, A26, A65 and two novel ones, A2w and A6a; M3, M3a, M4 and M79). Continued genetic monitoring will be important to assess the impact of this regional law over the coming years. Based on the obtained results, we recommend a more stringent policy to prevent the erosion of the genetic integrity of the native subspecies A. m. ligustica.

Seasonal Occurrence of Honey Bees in Coriander

A multi-location trail was carried out at the Beekeeping Unit, TCA, Dholi (Muzaffarpur District, India) and University Apiary, RPCAU, Pusa (Samastipur District, India) during Rabi 2021-22 on Rajendra Dhania-2 variety of coriander. Spatial distribution of bee species in Pusa and Dholi showed that occurance of Italian bee Apis mellifera (F) was comparatively higher at Pusa (19.03± 6.29/ m2 / 10 min) as compared to Dholi (16.56± 5.47/ m2 / 10 min). Other species of honey bees viz. Indian hive bee, Apis cerana (F), rock bee Apis dorsata (F) and little bee Apis florea (F) were more abundant in Dholi as compared to Pusa. Maximum (number of bee/ m2 / 10 min) of Apis mellifera (24.02, 27.60), Apis dorsata (16.88, 14.89), Apis cerana (14.55, 11.64) and Apis florea (10.47, 8.63) were observed at 13:00, 15:00, 11:00 and 09:00 at Dholi and Pusa, respectively. The underlying factors of this distribution were identified as body size and dietary overlap. This study highlights the complexity of interactions between Apis spp. genus and shows how factors like body size and diet influence their distribution patterns.

Effects of a supplemented diet containing 7 probiotic strains (Honeybeeotic) on honeybee physiology and immune response: analysis of hemolymph cytology, phenoloxidase activity, and gut microbiome

BackgroundIn this study, a probiotic mixture (Honeybeeotic) consisting of seven bacterial strains isolated from a unique population of honeybees (Apis mellifera ligustica) was used. That honeybee population was located in the Roti Abbey locality of the Marche Region in Italy, an area isolated from human activities, and genetic contamination from other honeybee populations. The aim was to investigate the effects of this probiotic mixture on the innate immunity and intestinal microbiome of healthy common honeybees in two hives of the same apiary. Hive A received a diet of 50% glucose syrup, while hive B received the same syrup supplemented with the probiotics, both administered daily for 1 month. To determine whether the probiotic altered the immune response, phenoloxidase activity and hemolymph cellular subtype count were investigated. Additionally, metagenomic approaches were used to analyze the effects on gut microbiota composition and function, considering the critical role the gut microbiota plays in modulating host physiology.ResultsThe results revealed differences in hemocyte populations between the two hives, as hive A exhibited higher counts of oenocytoids and granulocytes. These findings indicated that the dietary supplementation with the probiotic mixture was safe and well-tolerated. Furthermore, phenoloxidase activity significantly decreased in hive B (1.75 ± 0.19 U/mg) compared to hive A (3.62 ± 0.44 U/mg, p < 0.005), suggesting an improved state of well-being in the honeybees, as they did not require activation of immune defense mechanisms. Regarding the microbiome composition, the probiotic modulated the gut microbiota in hive B compared to the control, retaining core microbiota components while causing both positive and negative variations. Notably, several genes, particularly KEGG genes involved in amino acid metabolism, carbohydrate metabolism, and branched-chain amino acid (BCAA) transport, were more abundant in the probiotic-fed group, suggesting an effective nutritional supplement for the host.ConclusionsThis study advocated that feeding with this probiotic mixture induces beneficial immunological effects and promoted a balanced gut microbiota with enhanced metabolic activities related to digestion. The use of highly selected probiotics was shown to contribute to the overall well-being of the honeybees, improving their immune response and gut health.

Hydroxy Fatty Acid Synthesis-Related mRNA as the Biomarker for Detecting Mislabeling of Honey Entomological Origin.

The authentication of the entomological origin of honey is a widespread concern, necessitating the prompt establishment of an effective approach for distinguishing between Apis cerana cerana honey (ACH) and Apis mellifera ligustica honey (AMH). Hydroxy fatty acids (HFAs) found in honey are bee-derived components synthesized by the mandibular glands of worker bees. We previously discovered significant variations in the hydroxy fatty acid composition between ACH and AMH, suggesting their potential as indicators for identifying the authenticity of the entomological origin of honey. Herein, we identified differentially expressed genes associated with HFA synthesis by conducting transcriptome sequencing of the mandibular glands of AC and AM honeybees. Subsequently, we proposed a method for the relative quantitative analysis of bee-derived RNA components using real-time fluorescence quantitative polymerase chain reaction, which was supplemented by multivariate statistical analysis to further discern differences in HFA synthesis-related mRNA between ACH and AMH. The results showed that the mRNAs of FAXDC2 (fatty acid hydroxylase domain-containing protein 2) and FAS (fatty acid synthase) may serve as indicators to discern the entomological origin of honey. This study presents two novel biomarkers for detecting mislabeling of the entomological origin in ACH and AMH based on variations in bee-derived components.

The black honey bee genome: insights on specific structural elements and a first step towards pangenomes

BackgroundThe honey bee reference genome, HAv3.1, was produced from a commercial line sample that was thought to have a largely dominant Apis mellifera ligustica genetic background. Apis mellifera mellifera, often referred to as the black bee, has a separate evolutionary history and is the original type in western and northern Europe. Growing interest in this subspecies for conservation and non-professional apicultural practices, together with the necessity of deciphering genome backgrounds in hybrids, triggered the necessity for a specific genome assembly. Moreover, having several high-quality genomes is becoming key for taking structural variations into account in pangenome analyses.ResultsPacific Bioscience technology long reads were produced from a single haploid black bee drone. Scaffolding contigs into chromosomes was done using a high-density genetic map. This allowed for re-estimation of the recombination rate, which was over-estimated in some previous studies due to mis-assemblies, which resulted in spurious inversions in the older reference genomes. The sequence continuity obtained was very high and the only limit towards continuous chromosome-wide sequences seemed to be due to tandem repeat arrays that were usually longer than 10 kb and that belonged to two main families, the 371 and 91 bp repeats, causing problems in the assembly process due to high internal sequence similarity. Our assembly was used together with the reference genome to genotype two structural variants by a pangenome graph approach with Graphtyper2. Genotypes obtained were either correct or missing, when compared to an approach based on sequencing depth analysis, and genotyping rates were 89 and 76% for the two variants.ConclusionsOur new assembly for the Apis mellifera mellifera honey bee subspecies demonstrates the utility of multiple high-quality genomes for the genotyping of structural variants, with a test case on two insertions and deletions. It will therefore be an invaluable resource for future studies, for instance by including structural variants in GWAS. Having used a single haploid drone for sequencing allowed a refined analysis of very large tandem repeat arrays, raising the question of their function in the genome. High quality genome assemblies for multiple subspecies such as presented here, are crucial for emerging projects using pangenomes.

The Nutritional Year-Cycle of Italian Honey Bees (Apis mellifera ligustica) in a Southern Temperate Climate

Nutrition is a key aspect influencing honey bee health and overwintering. Since honey bee seasonality in southern temperate climates represents a significant research gap, this study conducted long-term monitoring of honey bees in the Mediterranean (Sassari, Italy). Specifically, individual weight, fat body, and size measurements (head, thorax, abdomen, and total body) were recorded monthly so to detect changes in the nutrient storage of worker bees during an annual cycle. Data were analysed according to sampling date, climate (temperature, precipitation, and daylength), and flower diversity and were conducted for nurse and forager bees separately. The nutritional honey bee year-cycle generally followed the nectar flow and showed two critical timepoints: summer and winter dearth. A short cessation of activities in late fall/early winter coupled with an increase in nutrient storage indicated the presence of winter bees. Precipitation was found to play an important role in honey bee nutrition in the study area through its impacts on colony demography and plants in particular illustrating how climate change could pose a threat to European honey bee populations in the future. These results provide valuable insights into the nutritional dynamics of Apis mellifera ligustica in the Mediterranean that could support management decisions to improve overwintering success and prevent unnecessary colony losses.

Effects of tefluthrin and guadipyr on the midgut bacteria of adult Apis mellifera

The objective of this study is to assess the potential impact of tefluthrin and guadipyr on the gut microbial composition and metabolism in adult Apis mellifera ligustica, thereby elucidating the underlying mechanisms of insecticide action and its practical implications for bee protection. In this investigation, A. mellifera were subjected to one of three dietary conditions: (1) control sugar water, (2) tefluthrin-infused sugar water, or (3) guadipyr-infused sugar water. After a 10-day exposure period, genomic DNA from the gut bacteria was extracted. High-throughput sequencing was employed to evaluate the potential influence of tefluthrin and guadipyr treatments on the diversity and abundance of gut bacteria. Among the A. mellifera specimens, a total of twenty species of gut bacteria were identified, spanning across five phyla, six classes, eleven orders, eleven families, and fifteen genera. The dominant phyla within the gut bacterial community were Proteobacteria and Bacteroidetes. In comparison to the control group, both the tefluthrin-treated and deltamethrin-treated groups exhibited alterations in the composition of their gut bacterial flora. At the phylum level, there was a significant decrease in the relative abundance of Cyanobacteria (P < 0.05). On the genus level, the tefluthrin group displayed a significant increase in the relative abundance of Bartonella and Serratia (P < 0.05). In the guadipyr-treated group, the relative abundance of Gilliamella and Frischella increased significantly (P < 0.05), while the relative abundance of norank_o_Chloroplast and Enterobacter decreased significantly (P < 0.05). Further analysis of cluster of orthologous genes predicted functional changes in gut microbial metabolism following tefluthrin exposure but no significant changes after guadipyr exposure. Consequently, exposure to tefluthrin and guadipyr can induce shifts in both the composition and metabolic activity of the gut bacteria in A. mellifera. Notably, the impact of tefluthrin on the gut bacteria of A. mellifera appears to be more pronounced compared to that of guadipyr.

Comparative assessment of food consumption, longevity, thermoregulation, and molecular health markers in mite-resistant and Italian honey bee stocks

Identifying traits for adaptation to different management and environmental regimes is key to maintaining robust honey bee populations under global climate change. We compared mite-resistant (Pol-line and Russian) and Italian honey bee stocks in variable-temperature cage experiments (200 bees per cage) with respect to food consumption, thermoregulation, gene expression, and lifespan, in 3 experiments over 2 years. The Italian stock bees consumed more syrup and pollen on average than the mite-resistant stocks, but the mite-resistant stocks maintained higher cluster temperatures and had median lifespans 8 days longer, consistent with the increased expression of vitellogenin relative to Italian stock. Model results indicated that, to maintain the same colony size as the mite-resistant stocks, Italian stock colonies would need about 13% more sealed brood to offset reduced worker lifespans. These differences among bee stocks likely influence colony-level productivity and health, and showed the importance of experimental replication.

The Effects of Artificial Diets on the Expression of Molecular Marker Genes Related to Honey Bee Health.

Honey bees are commonly used to study metabolic processes, yet the molecular mechanisms underlying nutrient transformation, particularly proteins and their effects on development, health, and diseases, still evoke varying opinions among researchers. To address this gap, we investigated the digestibility and transformation of water-soluble proteins from four artificial diets in long-lived honey bee populations (Apis mellifera ligustica), alongside their impact on metabolism and DWV relative expression ratio, using transcriptomic and protein quantification methods. Diet 2, characterized by its high protein content and digestibility, was selected for further analysis from the other studied diets. Subsequently, machine learning was employed to identify six diet-related molecular markers: SOD1, Trxr1, defensin2, JHAMT, TOR1, and vg. The expression levels of these markers were found to resemble those of honey bees who were fed with Diet 2 and bee bread, renowned as the best natural food. Notably, honey bees exhibiting chalkbrood symptoms (Control-N) responded differently to the diet, underscoring the unique nutritional effects on health-deficient bees. Additionally, we proposed a molecular model to elucidate the transition of long-lived honey bees from diapause to development, induced by nutrition. These findings carry implications for nutritional research and beekeeping, underscoring the vital role of honey bees in agriculture.

Transcriptional dynamics and regulatory function of milRNAs in Ascosphaera apis invading Apis mellifera larvae.

In the present study, small RNA (sRNA) data from Ascosphaera apis were filtered from sRNA-seq datasets from the gut tissues of A. apis-infected Apis mellifera ligustica worker larvae, which were combined with the previously gained sRNA-seq data from A. apis spores to screen differentially expressed milRNAs (DEmilRNAs), followed by trend analysis and investigation of the DEmilRNAs in relation to significant trends. Additionally, the interactions between the DEmilRNAs and their target mRNAs were verified using a dual-luciferase reporter assay. In total, 974 A. apis milRNAs were identified. The first base of these milRNAs was biased toward U. The expression of six milRNAs was confirmed by stem-loop RT-PCR, and the sequences of milR-3245-y and milR-10285-y were validated using Sanger sequencing. These miRNAs grouped into four significant trends, with the target mRNAs of DEmilRNAs involving 42 GO terms and 120 KEGG pathways, such as the fungal-type cell wall and biosynthesis of secondary metabolites. Further investigation demonstrated that 299 DEmilRNAs (novel-m0011-3p, milR-10048-y, bantam-y, etc.) potentially targeted nine genes encoding secondary metabolite-associated enzymes, while 258 (milR-25-y, milR-14-y, milR-932-x, etc.) and 419 (milR-4561-y, milR-10125-y, let-7-x, etc.) DEmilRNAs putatively targeted virulence factor-encoded genes and nine genes involved in the MAPK signaling pathway, respectively. Additionally, the interaction between ADM-B and milR-6882-x, as well as between PKIA and milR-7009-x were verified. Together, these results not only offer a basis for clarifying the mechanisms underlying DEmilRNA-regulated pathogenesis of A. apis and a novel insight into the interaction between A. apis and honey bee larvae, but also provide candidate DEmilRNA-gene axis for further investigation.

Exploring the hidden riches: Recent remarkable faunistic records and range extensions in the bee fauna of Italy (Hymenoptera, Apoidea, Anthophila).

The area sourrounding the Mediterranean basin is recognised as a major biodiversity hotspot for bees, and Italy is amongst the European countries with the highest bee species richness. Detailed knowledge of bee distribution is crucial for understanding bee biology and designing tailored conservation strategies, but is still insufficient in southern European countries, especially in Italy. We report recent finds of 48 bee species that yield significant novelties for the Italian bee fauna. Eight species, namely Andrenaconfinis Stöckhert, Anthidiellumbreviusculum Pérez, Coelioxysalatus Foerster, Lasioglossumalgericolellum Strand, Megachilelapponica Thomson, Megachileopacifrons Pérez, Megachilesemicircularis auct. nec Zanden and Trachusaintegra Eversmann are reported as new for Italy. In addition, Andrenabinominata Smith, Andrenacompta Lepeletier, Colletesacutus Pérez, Lasioglossumstrictifrons Vachal, Rhodanthidiumsiculum Spinola and Rhodanthidiumsticticum Fabricius are newly recorded from mainland Italy, Osmiaheteracantha Pérez from Sardegna and Nomadaflavopicta Kirby from Sicilia. We also report significant range extensions for other bee species and recent records of species that had long gone unrecorded in Italy. The combination of morphology and DNA barcoding provided reliable identifications even for the most challenging specimens. As several of our records come from areas neglected by bee experts in the past, this study stands out as a key indicator of a bee faunistic richness still awaiting discovery and hopefully it will stimulate the interest of taxonomists and stakeholders in pursuing bee research in Italy in the near future.

Increased Mass-Rearing of Queens in High Royal-Jelly-Producing Honey Bee Colonies (Apis mellifera ligustica) Generates Smaller Queens with Comparable Fecundity

The mass rearing of high-quality queen bees is an essential beekeeping practice for producing new queens to maintain colony productivity. A strain of high royal-jelly-producing bees (RJBs; Apis mellifera ligustica) in China exhibits high potential for the rapid mass rearing of queens. To explore the potential changes in the quality of mass-reared queens, a set of morphometric traits and the sealed brood area were compared between the queens reared from 64 and 320 queen cells in RJB colonies. The increase in the queen cell number was found to induce a slightly but significantly reduced body weight and smaller wing length and thorax width in the reared queens at emergence. However, the ovariole number and sealed brood area, an indicator of the queen fecundity, were not observed to be significantly influenced. With respect to body weight and ovariole number, all the reared queens satisfied the current criteria for high-quality queens. Our findings provide evidence for the efficient mass production of high-quality queens using RJB colonies.

The Varroa paradox: infestation levels and hygienic behavior in feral scutellata-hybrid and managed Apis mellifera ligustica honey bees

The Varroa destructor mite is a parasitic threat to managed and feral honey bee colonies around the world. Beekeepers use miticides to eliminate Varroa in commercial hives, but these chemicals can diminish bee health and increase miticide resistance. In contrast, feral honey bees have developed multiple ways to counteract mites without chemical treatment. We compared mite levels, grooming habits, and mite-biting behavior between feral Africanized honey bees (genomically verified Apis mellifera scutellata hybrids) and managed Italian honey bees (A. mellifera ligustica). Surprisingly, there was no difference in mite infestation levels between scutellata-hybrids and managed bees over one year despite the regular use of miticides in managed colonies. We also found no differences in the social immunity responses of the two groups, as measured by their hygienic habits (through worker brood pin-kill assays), self-grooming, and mite-biting behavior. However, we provide the first report that both scutellata-hybrids and managed honey bees bite off mite chemosensory forelegs, which the mites use to locate brood cells for reproduction, to a significantly greater degree than other legs (a twofold greater reduction in foreleg length relative to the most anterior legs). Such biting may impair mite reproduction.

First Description of the Occurrence of Slow Bee Paralysis Virus-1 and Deformed Wing Virus B in Apis mellifera ligustica Honeybee in Italy

Among the causes of bee colony death, viruses are among the number of pathogens that can contribute to the ill health of a colony. This study focuses on two of the several honeybee viruses, Slow Bee Paralysis Virus-1 (SBPV-1) and Deformed Wing Virus B (DWV-B), both belonging to the Iflavirus genus. To date, there is limited information available on the presence of these viruses in Italy. For this research project (IZS LT 04/19 RC), funded by the Ministry of Health, the presence and positivity of several honeybee viruses were evaluated in Italy using molecular methods. Convenience sampling was used, and these samples were analyzed for the two viruses and/or other main honeybee viruses, using specific PCR protocols and Sanger sequencing when necessary. A statistical analysis was conducted to study the independence between the viruses. Our data demonstrate for the first time the presence of SBPV-1 in Italian territory with a rather low number in most of the regions investigated, except for Emilia-Romagna and Lazio where it was detected more frequently, while DWV-B was detected at a higher level in all the regions investigated.

Analyzing the evolution of insect TMED gene and the expression pattern of silkworm TMED gene

Transmembrane emp24 domain (TMED) gene is closely related to immune response, signal transduction, growth and disease development in mammals. However, only the Drosophila TMED gene has been reported on insects. We identified the TMED family genes of silkworm, Tribolium castaneum, tobacco moth and Italian bee from their genomes, and found that the TMED family gene composition patterns of one α-class, one β-class, one δ-class and several γ-classes arose in the common ancestor of pre-divergent Hymenoptera insects, while the composition of Drosophila TMED family members has evolved in a unique pattern. Insect TMED family γ-class genes have evolved rapidly, diverging into three separate subclasses, TMED6-like, TMED5-like and TMED3-like. The TMED5-like gene was lost in Hymenoptera, duplicated in the ancestors of Lepidoptera and duplicated in Drosophila. Insect TMED protein not only has typical structural characteristics of TMED, but also has obvious signal peptide. There are seven TMED genes in silkworm, distributed in six chromosomes. One of seven is single exon and others are multi-exons. The complete open reading frame (ORF) sequences of seven TMED genes of silkworm were cloned from larval tissues and registered in GenBank database. BmTMED1, BmTMED2 and BmTMED6 were expressed in all stages and tissues of the silkworm, and all genes were expressed in the 4th and 5th instar and silk gland of the silkworm. The present study revealed the composition pattern of TMED family members, their γ class differentiation and their evolutionary history, providing a basis for further studies on TMED genes in silkworm and other insects.

Monitoring Immune Modulation in Season Population: Identifying Effects and Markers Related to Apis mellifera ligustica Honey Bee Health.

Honey bees play a significant role in ecology, producing biologically active substances used to promote human health. However, unlike humans, the molecular markers indicating honey bee health remain unknown. Unfortunately, numerous reports of honey bee collapse have been documented. To identify health markers, we analyzed ten defense system genes in Apis mellifera ligustica honey bees from winter (Owb) and spring (Fb for foragers and Nb for newly emerged) populations sampled in February and late April 2023, respectively. We focused on colonies free from SBV and DWV viruses. Molecular profiling revealed five molecular markers of honey bee health. Of these, two seasonal molecular markers-domeless and spz genes-were significantly downregulated in Owb compared to Nb and Fb honey bees. One task-related marker gene, apid-1, was identified as being downregulated in Owb and Nb compared to Fb honey bees. Two recommended general health markers, SOD and defensin-2, were upregulated in honey bees. These markers require further testing across various honey bee subspecies in different climatic regions. They can diagnose bee health without colony intervention, especially during low-temperature months like winter. Beekeepers can use this information to make timely adjustments to nutrients or heating to prevent seasonal losses.

Expression of Cry1Ab/2Aj Protein in Genetically Engineered Maize Plants and Its Transfer in the Arthropod Food Web

While transgenic Bacillus thuringiensis (Bt) maize provides pest resistance and a reduced application of chemical pesticides, a comprehensive environmental risk assessment is mandatory before its field release. This research determined the concentrations of Bt protein in plant tissue and in arthropods under field conditions in Gongzhuling City, northeastern China, to provide guidance for the selection of indicator species for non-target risk assessment studies. Bt maize expressing Cry1Ab/2Aj and non-transformed near-isoline were grown under identical environmental and agricultural conditions. Cry1Ab/2Aj was detected in plant tissues and arthropods collected from Bt maize plots during pre-flowering, flowering, and post-flowering. The expression of Cry1Ab/2Aj varied across growth stages and maize tissues, as well as in the collected arthropods at the three growth stages. Therefore, representative species should be chosen to cover the whole growing season and to represent different habitats and ecological functions. Dalbulus maidis (Hemiptera: Cicadellidae), Rhopalosiphum padi (Hemiptera: Aphididae), Heteronychus arator (Coleoptera: Scarabaeidae), and Somaticus angulatus (Coleoptera: Tenebrionidae) are suitable non-target herbivores. Propylea japonica (Coleoptera: Coccinellidae), Paederus fuscipes (Coleoptera: Staphylinidae), Chrysoperla nipponensis (Neuroptera: Chrysopidae), and spiders are suggested predators. Apis cerana and Apis mellifera ligustica (both Hymenoptera: Apidae) represent pollinators and Folsomia candida (Collembola: Isotomidae) decomposers.

Exploratory comparison of flower visiting behavior and pollination ability of mason bees, bumble bees, and honey bees.

This study explored the flower visiting behaviors and pollination abilities of mason bees (Osmia excavata Alfken (Hymenoptera: Megachilidae)), bumble bees (Bombus terrestris (Linnaeus, 1758) (Hymenoptera: Apidae)), and Italian honey bees (Apis mellifera ligustica Spinola (Hymenoptera: Apidae)) in apple orchards in early spring in Jinan (located in the central region of Shandong) and Yantai (located in the Peninsula of Shandong). We compared the pollen collection patterns, flower visiting behavior, flying speed, and effects on apple pollination of the 3 types of bees. The frequencies of flower visits were significantly higher for mason bees (12.89/min in Jinan and 10.63/min in Yantai) than bumble bees and Italian honey bees in the 2 regions. The single flower residence times were significantly higher for Italian honey bees (8.22 s in Jinan and 9.43 s in Yantai), but Italian honey bees were most affected by the climate. The 3 bees differed significantly in terms of the amount of apple pollen collected and their effects on the fruit setting rate in apples (mason bees > bumble bees > Italian honey bees). The results showed that the mason bee was the most suitable pollinating species for spring apple orchards; Bumble bees were more suitable as alternative pollinators during cloudy and low temperatures; Italian honey bees were able to take advantage of their large number of worker bees in sunny and warm weather. Compared to individual bee species, a combination of 2 or 3 species of bees might be more advantageous in dealing with complex and variable weather conditions.