Beekeeping and Managed Bee Diversity in Indonesia: Perspective and Preference of Beekeepers

There is a high diversity of bees in the tropics, including honey bees and stingless bees, which are the main sources for honey and other ecosystem services. In Indonesia, beekeeping practices have been developed for centuries, and they have been part of many cultural practices in many traditional communities. The objective of this research was to study the beekeeping status and managed bee diversity in Indonesia and to investigate beekeepers’ perspectives on the factors and obstacles related to beekeeping. Direct interview and online interview were conducted to gain data on bees and beekeepers. In total, 272 beekeepers were interviewed across 25 provinces. Samplings of honey bees and stingless bees were also done during direct interviews for further identification and, when possible, pollen identification. All data and specimens were then sent to IPB Bogor for compilation and identification. We recorded 22 species of bees, including 3 species of honey bees and 19 species of stingless bees, that are reared by Indonesian beekeepers, with Apis cerana and Tetragonula laeviceps as the most common species. Our research also found that the majority of beekeepers fall into the category of the younger generation (30–39 years old) with educational background mostly from senior high school. Based on the beekeepers’ perspectives, there are several obstacles to beekeeping, especially the occurrence of death of bee foragers attributed to climate, food source, and pesticides. In conclusion, there is a need to develop a strategy for beekeeping and bee conservation in Indonesia, especially for adaptation and mitigation from environmental changes with a particular focus on climate and land-use change.

Comparative transcriptome investigation of Nosema ceranae infecting eastern honeybee workers

Apis cerana is the original host for Nosema ceranae, a widespread fungal parasite resulting in bee nosemosis, which leads to severe losses for apiculture industry throughout the world. However, knowledge of N. ceranae infecting eastern honeybees is extremely limited. Currently, the mechanism underlying N. ceranae infection is still largely unknown. Based on our previously gained high-quality transcriptome datasets, comparative transcriptomic investigation was conducted in this work, with a focus on virulence factor-associated differentially expressed genes (DEGs). Microscopic observation showed that A. c. cerana workers midguts were effectively infected after inoculation with clean spores of N. ceranae. Totally, 1411, 604, and 38 DEGs were identified from NcCK vs. NcT1, NcCK vs. NcT2 and NcT1 vs. NcT2 comparison groups. Venn analysis showed that ten up-regulated genes and nine down-regulated ones were shared by aforementioned comparison groups. GO category indicated these DEGs were involved in a series of functional terms relevant to biological process, cellular component, and molecular function, such as metabolic process, cell part, and catalytic activity. Additionally, KEGG pathway analysis suggested that the DEGs were engaged in an array of pathways of great importance, such as metabolic pathway, glycolysis, and biosynthesis of secondary metabolites. Further, expression clustering analysis demonstrated that majority of genes encoding virulence factors such as ricin B lectins and polar tube proteins displayed apparent up-regulation, whereas a few virulence factor-associated genes such as hexokinase gene and 6-phosphofructokinase gene presented down-regulation during the fungal infection. Finally, the expression trend of 14 DEGs was confirmed by RT-qPCR, validating the reliability of our transcriptome datasets. These results together demonstrated that an overall alteration of the transcriptome of N. ceranae occurred during the infection of A. c. ceranae workers, and most of virulence factor-related genes were induced to activation to promote the fungal invasion. Our findings not only lay a foundation for clarifying the molecular mechanism underlying N. ceranae infection of eastern honeybee workers, but also shed light on developing novel targets for microsporidiosis control.

Diversity and abundance of some insect fauna in Bhawal and Madhupur Sal forests of Bangladesh

The abundance and diversity of insect fauna were studied from two deciduous sal forests of Bhawal and Madhupur located at central part of Bangladesh. A total of 544 individuals of insects of 61 species belonging to 54 genera, 33 families and 11 orders have been identified with Hymenoptera (31%) as the dominant order in species richness followed by Coleoptera (13%), Orthoptera (11%), Diptera (10%), Hemiptera (8%), Lepidoptera (8%), Odonata (8%), Homoptera (3%), Isoptera (3%), Neuroptera (3%) and Dictyoptera (2%). Bhawal scores higher Shannon-Weaver diversity index (Hʹ=3.725) compared to Madhupur (Hʹ=3.340). The Bhawal Sal Forest with the collected 341 (63%) insects and identified 53 (59%) species belonging to 10 orders was found more diverse in species richness than the Madhupur Sal Forest with 37(41%) species belonging to 11 orders identified from the collected 203 (37%) insect samples. Insects of the order Neuroptera were not recorded from Bhawal. Off the 61 species, 29(48%) species were common in both the forests, 24(39%) species were exclusive to Bhawal and eight (13%) species were exclusive to the Madhupur Sal Forest. Apis cerana of Hymenoptera was identified as the dominant species having 9% of the identified samples followed by dipteran species Musca domestica with 6% of the samples. Among the insect species 30 (49%) species were found playing beneficial role as biological control agents, predators, pollinators, honey producers and also organic debris recycler. On the other hand, 31(51%) species were found to be harmful causing damage to forest vegetation as well as human and wildlife at variable degrees. J. Biodivers. Conserv. Bioresour. Manag. 2021, 7(1): 11-24

Impact of Nosema ceranae invasion on sucrose solution consumption, midgut epithelial cell structure, and lifespan of Apis cerana cerana workers

Nosema ceranae is an intracellular fungal parasite for honeybees, leading to chronic disease named bee nosemosis with worldwide distribution. Asian honeybee (Apis cerana) is the original host for N. ceranae, but the impact of N. ceranae infection on A. cerana physiology is largely unknown. In this current work, workers of Apis cerana cerana, a subspecies of Asian honeybee, were artificially inoculated with N. ceranae spores and reared under lab conditions, followed by detection of fungal spore load as well as host sucrose solution consumption, midgut epithelial cell structure, and lifespan. The result of spore counting suggested that the spore load in the host midgut decreased significantly during 1 dpi-2 dpi, whereas that displayed an elevated trend among 2 dpi-13 dpi. The sucrose solution consumption of workers in N. ceranae-inoculated groups among 1 dpi-20 dpi was always higher than that of workers in un-inoculated groups; additionally, the difference of sucrose solution consumption between these two groups at 4 dpi, 5 dpi, and 13 dpi was of significance. Based on microscopic observation of paraffin sections, darkly stained parasites were clearly detected in the midgut epithelial cells of N. ceranae-inoculated workers at 7 dpi-10 dpi, whereas no parasite was observed in those of un-inoculated workers. In addition, the boundaries of un-inoculated host epithelial cells were intact and the darkly stained nucleus were clear, while the boundaries of midgut epithelial cells of N. ceranae-inoculated workers were blurred, the nucleus were almost disappeared, and the nucleic acid substances were diffused. Moreover, the survival rates of workers in both N. ceranae-inoculated groups and un-inoculated groups at 1 dpi-5 dpi were pretty high and then started to decrease at 5 dpi; the survival rate of workers in N. ceranae-inoculated groups was always lower than that in un-inoculated groups, with significant difference between these two groups during 11 dpi-20 dpi. These results together indicate that the quantity of fungal spores continuously elevated with the microsporidian multiplication, causing energetic stress for workers and host cell structure damage, which further negatively affected the host lifespan. Our findings offer a solid basis not only for exploring the molecular mechanism underlying N. ceranae infection but also for investigating the interaction between N. ceranae and eastern honeybee.

Efficacy of Nyagrodh Twak Lepa in Honey bee sting: In-vivo study

Introduction- As Acharya Charaka has explained the local application of Kshirivruksha Twak to cure all types of keeta visha, hence Nyagrodh (Ficus benghalensis L.) Twak Lepa with water as base is selected as Trial drug on Apis Cerana Indica bee sting poisoning. Material and Methods- An in-vivo study on albino mice to know the efficacy of trial drug has been planned after animal ethical clearance. 18 albino mice were prorated into three groups with 6 animals in each group viz. Control group, Trial drug (Nyagrodh Twak Lepa Churna) group and Standard drug (Beclomethasone Dipropionate 0.025% w/w) group. 6 stings were given to each mice and 3 stings were removed after sting operation. All mice were observed for allergic reactions viz. erythema, scaling, fissures, oedema and mortality for a period of 7 days. Histo-pathological changes were also noted after completion of study. Statistical analysis was done using Paired t test. Results- Results revealed that Trial drug had worked more efficiently on Erythema and Oedema while Standard drug worked more efficiently on Scaling and Fissure. Histo-pathology showed that wound healed with Nyagrodh twak lepa and Standard drug have shown almost similar changes while wound in control group showed extensive areas of necrosis. Conclusion- Present study suggests that both Nyagrodh and Beclomethasone can be used in Honey bee sting poisoning but as Nyagrodh being a religious tree can be easily identified by a common man, it can be employed as preliminary treatment for the same before reaching hospital.

DIVERSITY OF THE CLOSED-NESTED HONEY BEES (APIDAE: APIS SPP.) AND THE TRADITIONAL HONEY COLLECTING AND BEEKEEPING IN FOUR ISLANDS OF INDONESIA

The closed-nested honey bees are an important group that has been successfully bred traditionally and in a modern way. The traditional honey beekeeping practices are still favorable by local people living near natural habitats. Many rural areas in Indonesia are well known as producers of honey from the traditional honey collecting and traditional honey beekeeping of the closed-nested honey bees. However, there is limited information on the diversity of the honey bees that had supported the honey productions and their traditional honey beekeeping. This research was to provide an overview of the diversity of the honey bee species that are used in the wild honey collecting and their traditional honey beekeeping in four selected study sites in the islands of Java, Bawean, Kalimantan, and Peleng. We recorded three species of closed-nested native honey bees in the traditional honey collecting and traditional honey beekeeping, namely Apis cerana, A. koschevnikovi, and A. nigrocincta. We observed that traditional beekeeping of A. cerana was carried out in Tasikmalaya and Bawean Island, and that of A. cerana and A. koschevnikovi were carried out in Kayan Hilir. On Peleng Island, people do not do beekeeping but collect honey directly from the forest. Honey collecting and beekeeping practices are related to changes in the seasons of the flowering period in their habitats. The knowledge of the flowering period is needed to know the seasonal movement of honey bees from forest to village and vice versa.

Development of a Kit for Rapid Immunochromatographic Detection of Sacbrood Virus Infecting Apis cerana (AcSBV) Based on Polyclonal and Monoclonal Antibodies Raised against Recombinant VP1 and VP2 Expressed in Escherichia coli

A Korean isolate of the sacbrood virus infecting Apis cerana (AcSBV-Kor) is the most destructive honeybee virus, causing serious economic damage losses in Korean apiculture. To address this, here, we attempted to develop an assay for the rapid detection of AcSBV-Kor based on immunochromatographic detection of constituent viral proteins. Genes encoding VP1 and VP2 proteins of AcSBV-Kor were cloned into an expression vector (pET-28a) and expressed in Escherichia coli BL21(DE3). During purification, recombinant VP1 (rVP1) and VP2 (rVP2) proteins were found in the insoluble fraction, with a molecular size of 26.7 and 24.9 kDa, respectively. BALB/c mice immunized with the purified rVP1 and rVP2 produced polyclonal antibodies (pAbs) such as pAb-rVP1 and pAb-rVP2. Western blot analysis showed that pAb-rVP1 strongly reacted with the homologous rVP1 but weakly reacted with heterologous rVP2. However, pAb-rVP2 strongly reacted not only with the homologous rVP2 but also with the heterologous rVP1. Spleen cells of the immunized mice fused with SP2/0-Ag14 myeloma cells produced monoclonal antibodies (mAbs) such as mAb-rVP1-1 and mAb-rVP2-13. Western blot analysis indicated that pAb-rVP1, pAb-rVP2, mAb-rVP1-1, and mAb-rVP2-13 reacted with AcSBV-infected honeybees and larvae as well as the corresponding recombinant proteins. These antibodies were then used in the development of a rapid immunochromatography (IC) strip assay kit with colloidal gold coupled to pAb-rVP1 and pAb-rVP2 at the conjugate pad and mAb-rVP1-1 and mAb-rVP2-13 at the test line. One antibody pair, pAb-rVP1/mAb-VP1-1, showed positive reactivity as low as 1.38 × 103 copies, while the other pair, pAb-rVP2/mAb-VP2-13, showed positive reactivity as low as 1.38 × 104 copies. Therefore, the antibody pair pAb-rVP1/mAb-VP1-1 was selected as a final candidate for validation. To validate the detection of AcSBV, the IC strip tests were conducted with 50 positive and 50 negative samples and compared with real-time PCR tests. The results confirm that the developed IC assay is a sufficiently sensitive and specific detection method for user-friendly and rapid detection of AcSBV.