Circular RNAs (circRNAs) are a new class of non-coding RNAs, which have been confirmed to regulate insect gene expression and immune response through multiple manners such as competing endogenous RNA (ceRNA) regulatory network. Currently, function of circRNA in honey bee immune response remains unclear. In this study, PCR and Sanger sequencing were performed to validate the back splicing (BS) site of ame_circ_000115 (in short ac115). RT-qPCR was used to detect the expression profile of ac115 in larval guts of Apis mellifera ligustica stressed by Ascosphaera apis. Dual-luciferase reporter gene assay was conducted to verify the binding relationship between ac115 and ame-miR-13b. Interference of ac115 in larval guts was carried out by feeding specific siRNA, followed by determination of the effect of ac115 interference on expression of six genes relevant to host immune response. The results confirmed the existence of BS site within ac115. Compared with the un-inoculated group, the expression of ac115 in 4-day-old larval gut of the A. apis-inoculated group was up-regulated with extreme significance (P < 0.000 1), while that in 5- and 6-day-old larval guts were significantly up-regulated (P < 0.05). The brightness of specific band for ac115 in 4-, 5- and 6-day-old larval guts of the siRNA-circ_000115-fed group gradually became weak, whereas that of the siRNA-scrambl-fed group was pretty high without obvious variation. Compared with that of the siRNA-scramble-fed group, the expression of ac115 in 4-day-old larval gut of the siRNA-circ_000115-fed group was significantly down-regulated (P < 0.05), whereas that of the 5- and 6-day-old larval guts were down-regulated with extreme significance (P < 0.001). Ame-miR-13b was truly existed and expressed in A. m. ligustica larval guts, and there was true binding relationship between ac115 and ame-miR-13b. Compared with that of the siRNA-scramble-fed group, the expression of antimicrobial peptide genes hymenoptaecin and abaecin in 6-day-old larval gut of the siRNA-circ_000115-fed group was significantly up-regulated (P < 0.05), while that of ecdysone receptor (Ecr) was down-regulated with extreme significance (P < 0.01). These results indicate that ac115 is truly expressed in A. m. ligustica larval guts, BS site truly exists within ac115, and effective interference of ac115 in A. m. ligustica larval guts can be achieved via feeding siRNA. Moreover, ac115 potentially regulates Ecr expression through adsorption of ame-miR-13b and expression of hymenoptaecin and abaecin using a non-ceRNA manner, further participating in host stress-response.
Bees/genetics* ; Animals ; Larva/metabolism* ; RNA, Circular/genetics* ; RNA, Small Interfering/genetics* ; MicroRNAs/genetics*

As the most efficient perception system in nature, the perception mechanism of the insect (such as honeybee) antennae is the key to imitating the high-performance sensor technology. An automated experimental device suitable for collecting electrical signals (including antenna reaction time information) of antennae was developed, in response to the problems of the non-standardized experimental process, interference of manual operation, and low efficiency in the study of antenna perception mechanism. Firstly, aiming at the automatic identification and location of insect heads in experiments, the image templates of insect head contour features were established. Insect heads were template-matched based on the Hausdorff method. Then, for the angle deviation of the insect heads relative to the standard detection position, a method that calculates the angle of the insect head mid-axis based on the minimum external rectangle of the long axis was proposed. Eventually, the electrical signals generated by the antennae in contact with the reagents were collected by the electrical signal acquisition device. Honeybees were used as the research object in this study. The experimental results showed that the accuracy of template matching could reach 95.3% to locate the bee head quickly, and the deviation angle of the bee head was less than 1°. The distance between antennae and experimental reagents could meet the requirements of antennae perception experiments. The parameters, such as the contact reaction time of honeybee antennae to sucrose solution, were consistent with the results of the manual experiment. The system collects effectively antenna contact signals in an undisturbed state and realizes the standardization of experiments on antenna perception mechanisms, which provides an experimental method and device for studying and analyzing the reaction time of the antenna involved in biological antenna perception mechanisms.
Animals ; Arthropod Antennae ; Bees

Aims: Symbiotic bee fungus associated with the stingless bee larval food has been revealed to play a major role in performing a mutual relationship with the host. The fungus is believed capable to produce crucial nutrients that are required for larval pupation. However, detailed information on the fungus identification isolated from the larval food of our native Indo-Malaya stingless bee, Heterotrigona itama (Hymenoptera: Apidae), is poorly understood. Hence, this study was conducted to determine the effect of fungus isolated from the stingless bee larval food on the survival of H. itama larvae and identify the isolated fungus using both morphological and molecular analyses. Methodology and results: Elisa plates designed with F-bottom type were used as artificial brood cells in a controlled condition. The eggs transferred to Elisa plates were kept in the incubator with 75%-100% humidity and 25 ± 2.5 °C of temperature. This study carried out in three different treatments: (1) microbes-free larval food, (2) fresh larval food and (3) microbes-free larval food with fungus supplement. Results showed that the survival of H. itama larvae depends on the presence of beneficial fungus with the highest survival rate (Treatment 2 = 85.71%). The absence of the beneficial fungus in the fluid food of H. itama showed the lowest survival rate (Treatment 1 = 37.14%). Microscopy analysis showed that the fungus had various forms, including unconjugated asci, round-shaped ascospore and pseudohyphae. The molecular characterisation of the isolated fungi was performed using 23S rRNA gene sequencing using universal primers ITS1 and ITS4. DNA barcoding of three isolated fungi confirmed all isolated fungi matched Panus lecomtei (Basidiomycetes; Poriales; Polyporaceae) with 99.70% of similarity. Conclusion, significance and impact of study This study provides information on the importance of whitish fungus in appearance that existed in the brood cell, which is proposed to be the crucial component of in-vitro stingless bee queen rearing protocol.
Bees--microbiology

bees (Apis mellifera), and then, within 10 min, amplified the target cDNA by ultra-rapid qPCR. The PCR products were hybridized to a DNA-chip for confirmation of target gene specificity. The results of this study suggest that our method might be a useful tool for detecting DWV, as well as for the diagnosis of RNA virus-mediated diseases on-site.]]>
Beekeeping ; Bees ; Diagnosis ; DNA, Complementary ; Methods ; Polymerase Chain Reaction ; RNA ; RNA, Viral ; Sensitivity and Specificity

BACKGROUND: Understanding the epidemiology of anaphylaxis is imperative for appropriate diagnosis and treatment, but the prevalence reportedly varies and only a few studies have compared the clinical features of anaphylaxis with the underlying causes in Korea. OBJECTIVE: This study aimed to investigate the etiology and clinical features of anaphylaxis. METHODS: We retrospectively reviewed the medical records of 319 anaphylaxis patients who visited our emergency room and extracted information on the causes, clinical characteristics, and subsequent outpatient visits. RESULTS: Food, drugs, and environmental factors were common causes of anaphylaxis. Statistically significant differences (p<0.001) were observed between children (<18 years of age) and adults (≥18 years of age), with food and drugs identified as the most common causes, respectively. Clinical characteristics of the patients were compared according to the common causes of anaphylaxis. Cutaneous symptoms were observed more frequently in food-induced cases (n=137, 95.1%) compared to drug-induced (n=73, 77.7%) and bee sting-induced (n=18, 78.3%) cases (p<0.001), whereas neurological symptoms were observed more frequently in drug-induced (n=37, 39.4%) and bee sting-induced (n=11, 47.8%) cases than in food-induced (n=18, 12.5%) cases (p<0.01). Drug-induced (n=44, 46.8%) and bee sting-induced (n=11, 47.8%) cases were more severe than food-induced cases (n=32, 22.2%). Out of the 319 assessed patients, only 25, 14, and 14 patients were referred to the pediatrics, allergy, and dermatology departments, respectively, after visiting the emergency room. CONCLUSION: Clinical characteristics of anaphylactic patients differed according to the underlying cause, but these findings are presumably influenced by factors determining the severity of anaphylaxis. We found that subsequent follow-up care in other departments to identify the cause of anaphylaxis was inadequate for most patients.
Adult ; Anaphylaxis ; Bees ; Child ; Dermatology ; Diagnosis ; Emergencies ; Emergency Service, Hospital ; Epidemiology ; Follow-Up Studies ; Humans ; Hypersensitivity ; Korea ; Medical Records ; Outpatients ; Pediatrics ; Prevalence ; Retrospective Studies ; Tertiary Care Centers

OBJECTIVE: Primary dysmenorrhea typically occurs with no associated pelvic pathology and is common in adolescents and young women. This study evaluated the effect of bee propolis on relief of primary dysmenorrhea. METHODS: The study was performed in 2018 in Hamadan, in western Iran, among female students with primary dysmenorrhea. Participants were randomly divided into two groups, using balanced block randomization, and were administered a placebo or bee propolis capsules for 5 days during two menstruation cycles. The number of participants required was estimated to be 86 in total, with 43 students in each group according to the inclusion criteria. We used the visual analog scale to assess pain severity. The independent t-test was conducted for comparing between two groups, using SPSS 16.0. RESULTS: A significant change was found in the mean pain scores during the first (P<0.001) and second (P<0.001) months after using bee propolis in comparison with placebo. The means of the pain scores in the bee propolis group were 5.32±2.28 and 4.74±2.40 in first and second months after the intervention, respectively, whereas the means of the pain scores in the placebo group were 7.40±1.21 and 7.17±1.24 in first and second months after the intervention, respectively. CONCLUSION: Our study showed that the use of bee propolis for two months compared with placebo reduced primary dysmenorrhea during the first and second months after use, with no adverse effects. Therefore, it could be used as an alternative to nonsteroidal anti-inflammatory drugs for relief of primary dysmenorrhea.
Adolescent ; Bees ; Capsules ; Dysmenorrhea ; Female ; Humans ; Iran ; Menstruation ; Pathology ; Propolis ; Random Allocation ; Visual Analog Scale

PURPOSE: To report a patient stung by a bee, who was diagnosed with sterile endopthalmitis and another patient diagnosed with optic neuritis, with decreasing visual acuity, after refined bee venom injection around the orbital tissue. CASE SUMMARY: A 82-year-old female visited our hospital for decreased visual acuity in the right eye and ocular pain due to a bee sting. The bee sting penetrated the sclera into the vitreous. In the anterior segment, severe cornea edema and anterior chamber cells were seen. Using ultrasonography, inflammation was seen around the intravitreal area. After 3 months, intravitreal inflammation regressed but the patient's visual acuity was light perception negative, and corneal opacity, neovascularization, and phthisis bulbi were detected. A 55-year-old male visited our hospital for ocular pain in the right eye and decreasing visual acuity after refined bee venom injection around the orbital tissue. The best-corrected visual acuity in the right eye was 15/100, there was moderate injection on the conjunctiva. A relative afferent pupillary defect, abnormal color vision test results, and a defect in the visual field test were observed. There was no pain during external ocular movement, and other general blood tests, and a brain MRI were normal. Based on these symptoms, methylprednisolone megatherapy was started for treatment of optic neuritis. After treatment, visual acuity of the right eye was 9/10 and all other clinical optic neuritis symptoms regressed. CONCLUSIONS: Based on these two cases, ocular toxicity from bee venom could result from both direct and indirect courses. Treatment using refined bee venom might be harmful, and caution is recommended in its use.
Aged, 80 and over ; Anterior Chamber ; Bee Venoms ; Bees ; Bites and Stings ; Brain ; Color Vision ; Conjunctiva ; Cornea ; Corneal Opacity ; Edema ; Female ; Hematologic Tests ; Humans ; Inflammation ; Magnetic Resonance Imaging ; Male ; Methylprednisolone ; Middle Aged ; Optic Neuritis ; Orbit ; Pupil Disorders ; Sclera ; Ultrasonography ; Visual Acuity ; Visual Field Tests

Respiratory inflammation is a frequent and fatal pathologic state encountered in veterinary medicine. Although diluted bee venom (dBV) has potent anti-inflammatory effects, the clinical use of dBV is limited to several chronic inflammatory diseases. The present study was designed to propose an acupoint dBV treatment as a novel therapeutic strategy for respiratory inflammatory disease. Experimental pleurisy was induced by injection of carrageenan into the left pleural space in mouse. The dBV was injected into a specific lung meridian acupoint (LU-5) or into an arbitrary non-acupoint located near the midline of the back in mouse. The inflammatory responses were evaluated by analyzing inflammatory indicators in pleural exudate. The dBV injection into the LU-5 acupoint significantly suppressed the carrageenan-induced increase of pleural exudate volume, leukocyte accumulation, and myeloperoxidase activity. Moreover, dBV acupoint treatment effectively inhibited the production of interleukin 1 beta, but not tumor necrosis factor alpha in the pleural exudate. On the other hand, dBV treatment at non-acupoint did not inhibit the inflammatory responses in carrageenan-induced pleurisy. The present results demonstrate that dBV stimulation in the LU-5 lung meridian acupoint can produce significant anti-inflammatory effects on carrageenan-induced pleurisy suggesting that dBV acupuncture may be a promising alternative medicine therapy for respiratory inflammatory diseases.
Acupuncture ; Acupuncture Points ; Animals ; Bee Venoms ; Bees ; Carrageenan ; Complementary Therapies ; Exudates and Transudates ; Hand ; Inflammation ; Interleukin-1beta ; Leukocytes ; Lung ; Mice ; Peroxidase ; Pleurisy ; Tumor Necrosis Factor-alpha ; Veterinary Medicine

Royal jelly (RJ) from honeybee has been widely used as a health promotion supplement. The major royal jelly proteins (MRJPs) have been identified as the functional component of RJ. However, the question of whether MRJPs have anti-senescence activity for human cells remains. Human embryonic lung fibroblast (HFL-I) cells were cultured in media containing no MRJPs (A), MRJPs at 0.1 mg/ml (B), 0.2 mg/ml (C), or 0.3 mg/ml (D), or bovine serum albumin (BSA) at 0.2 mg/ml (E). The mean population doubling levels of cells in media B, C, D, and E were increased by 12.4%, 31.2%, 24.0%, and 10.4%, respectively, compared with that in medium A. The cells in medium C also exhibited the highest relative proliferation activity, the lowest senescence, and the longest telomeres. Moreover, MRJPs up-regulated the expression of superoxide dismutase-1 (SOD1) and down-regulated the expression of mammalian target of rapamycin (MTOR), catenin beta like-1 (CTNNB1), and tumor protein p53 (TP53). Raman spectra analysis showed that there were two unique bands related to DNA synthesis materials, amide carbonyl group vibrations and aromatic hydrogens. These results suggest that MRJPs possess anti-senescence activity for the HFL-I cell line, and provide new knowledge illustrating the molecular mechanism of MRJPs as anti-senescence factors.
Animals ; Bees ; Cattle ; Cell Line ; Cell Proliferation ; Cellular Senescence/drug effects* ; Culture Media ; Dose-Response Relationship, Drug ; Fatty Acids/chemistry* ; Fibroblasts/drug effects* ; Humans ; Insect Proteins/chemistry* ; Lung/drug effects* ; Serum Albumin/metabolism* ; Spectrum Analysis, Raman ; Superoxide Dismutase-1/metabolism* ; TOR Serine-Threonine Kinases/metabolism* ; Tumor Suppressor Protein p53/metabolism* ; beta Catenin/metabolism*

Bee stings result in diverse clinical manifestations from localized pain, rash to life-threatening systemic allergic reactions or toxic reactions. Toxic reactions include skin necrosis, pancreatitis, acute renal failure, hemolysis or coagulopathy, while systemic allergic reactions present with IgE-mediated anaphylaxis. We experienced a 63-year-old woman who developed rhabdomyolysis and diabetic ketoacidosis after bee sting. The patient was accompanied by pulmonary edema due to acute kidney injury, which was recovered by intensive hemodialysis treatment. Here, we report a rare and serious case induced by bee sting with a review of the literature.
Acute Kidney Injury ; Anaphylaxis ; Bees* ; Bites and Stings* ; Diabetic Ketoacidosis* ; Exanthema ; Female ; Hemolysis ; Humans ; Hypersensitivity ; Middle Aged ; Necrosis ; Pancreatitis ; Pulmonary Edema ; Renal Dialysis ; Rhabdomyolysis* ; Skin