The dynastid beetle Allomyrina dichotoma has been used as a herbal medicine. Recently, we performed de novo RNAsequencing of Allomyrina dichotoma and identified several antimicrobial peptide candidates based on in silico analysis.Among them, cationic antimicrobial peptide, Allomyrinasin, was selected and we assessed the anti-inflammatory activitiesof Allomyrinasin against mouse macrophage Raw264.7 cells. The results showed that Allomyrinasin decreased the nitricoxide production of the lipopolysaccharide-induced Raw264.7 cells. In addition, quantitative RT-PCR, ELISA and Westernblot analysis revealed that Allomyrinasin reduced cytokine expression levels in the Raw264.7 cells. Taken together, thesedata indicated that Allomyrinasin had anti-inflammatory activity in the lipopolysaccharide-induced Raw264.7 cells.

The objective of this study was to evaluate the effect of the Allomyrina dichotoma larva (AD) on allergy and inflammation.We examined inhibitory effect of AD on allergic reactions in mast cells (RBL-2H3) activated by Compound 48 / 80and inflammatory response in macrophages (Raw 264.7) activated by LPS. Anti-allergy and anti-inflammatory actions ofAD water extract had no cytotoxicity. At these concentrations AD inhibited ẞ-hexosaminidase, tumor necrosis factor- α(TNF- α), interleukin-4 (IL-4), interleukin-6(IL-6) and cyclooxygenase-2(COX-2). AD also inhibited the expression of inducibleNO synthase (iNOS). AD reduced the release of inflammatory cytokines including IL-4, IL-6, TNF-α, and ẞ-hexosaminidase.These results suggest that AD may be potential anti-allergy and anti-inflammatory therapeutic agent.

In 2015, we reported a viral disease extremely fatal to Allomyrina dichotoma larvae spread in the majority of the larva-rearing farms in Korea. Currently, the virus-infected larva is diagnosed by PCR-based amplification but this requires laboratory equipment and agarose gel electrophoresis. Loop-mediated isothermal amplification (LAMP), a highly sensitive DNA amplification method, uses DNA polymerase isolated from Bacillus stearothemophilus and a set of six primers. It has great potential for field use because DNA is amplified under single temperature and the overall reaction completes in 30 min without laboratory equipments. Here, we report the development of on-site diagnosis method for Korean horn beetle larva infected by Allomyrina dichotoma Nudivirus.

Since we reported a viral disease extremely fatal to A. dichotoma, had landed on Korea in 2015, we have investigated this disease nationwide and it was turned out the viral disease has spread in the majority of the larva-rearing farms all over the country. The transmission of this disease is most severe when the larvae are harvested and stored for overwintering. During this period, even a few virus-infected larvae can easily transmit the virus and about two hundreds of larvae stored in a container can be killed. In a peroral infection experiment, 80% of infected larvae were killed but survived larvae became adults fully functional physiologically. These survived beetles were confirmed as live virus carrier. Also, about 75% of larvae hatched from the eggs laid by the infected beetles were infected and killed by the virus. This vertical transmission is due to the feeding and secretion activity of the infected parents.

The current market size of insect industry in Korea is estimated at 300 million dollars and more than 500 local farms are related to many insect industry. One of the strong candidates for insect industry is Korean horn beetle, Allomyrina dichotoma. Early this year, we reported a viral disease extremely fatal to A. dichotoma larvae. While we were proceeding a nationwide investigation of this disease, several farmers answered that similar disease symptom has been occurred occasionally during past 10 years. The symptom can be easily confused with early stage of bacterial infection or physiological damage including temperature and moisture. A peroral infection with the purified virus to healthy larvae produced a result that only 21% of larvae survived and became pupae. Although some of the survived adult beetle were deformational, many of them had no abnormal appearance and even succeeded in mating. Later, these beetles were examined if they were carrying the virus, and all except one were confirmed as virus carrier. This implies that these beetle may fly out and spread the disease throughout the country. We found the evidence by collecting wild A. dichotoma larvae which were virus infected, near two local farms rearing the horn beetle. So far, transovarial transmission of this virus to the eggs, or horizontal transmission to other commercially reared insects are not known yet.

In Korea, the value of insect resources has been increased immensely and the insect industry is expected to increase up to 40 billion dollars by 2020. Among them, Korean horn beetle, Allomyrina dichotoma, is one of the strongest candidates for insect industry as medicinal purpose for various liver diseases and diabetes. In many local farms rearing larvae, a few bacterial and fungal pathogens causing disease of A. dichotoma larvae have been identified including Bacillus thuringiensis and Metarhizium anisopliae. For past few years, a disease seemingly due to viral infection has been reported for causing death of larvae en masse. The pathogen of this disease has not been identified so far. In Southeastern Asia, Oryctes rhinoceros nudivirus (OrNV) is known to cause severe disease in coconut palm rhinoceros beetle, Oryctes rhinoceros, and is used as a biological control agent to reduce the pest population. Here, we report for the first time that OrNV may have landed on Korea and may be the major pathogen for the diseased larvae of A. dichotoma. After peroral inoculation with the purified virus extracted from the hemolymph of diseased larvae, we confirmed that over 60% of infected larvae perished in six weeks. This viral disease spreads very fast in many farms nationwide throughout Korea. This threat not only makes economic loss of local farms rearing larvae, but also may disturb the ecosystem by transmitting to wild A. dichotoma.

Allomyrina dichotoma is a typical pet beetles in Korea. From 2010, similar symptoms of milky spore disease were found collectively in grubs of the species reared in insect farms. They shared a specific symptom that the skin of last instar larvae was changed softer with opaque white and infested grubs eventually died. To clarify the cause of the symptom, we collected the larvae of A. dichotoma from five farms and examined intestinal bacterial florae of them using pyrosequencing technique. From those results, a member of Paenibacillus was found only in the larvae showing the symptom of disease. Through PCR analysis using a Paenibacillus specific primer set, we obtained the partial 16S rRNA gene sequence and confirmed the microbe as Paenibacillus sp. For detailed characterizing, a whole guts was extracted from each larva showing the sign of the disease and incubated at 70℃ for 15 min to isolate spore forming bacteria. After then, each content of guts was cultured on MYPGPNAL agar medium (12.5 μg/ml of nalidixic acid) at 30℃. The 16S rRNA gene sequence analysis for six isolate showed that they were closely related to P. rigui (97.5~98.0% similarity) and to P. chinjuensis (95.9~96.6% similarity). Additional tests including API test and cellular fatty acid composition analysis were performed, but the strain couldn’t be identified at species level, suggesting it may represent novel species of the genus Paenibacillus.

A bacterium was isolated from feces of Allomyrina dichotoma larva that feeds on well-rotted sawdust of woods. The isolate was identified as a Gram-positive and spore-forming stain. We named the isolate as Bacillus amyloliquefaciens on the basis of morphological and biochemical properties as well as 16S rRNA gene sequences. The culture of bacteria beneficially increased root and shoot growth of tomato, pepper and cucumber plants compared to the distilled water control. In addition, the bacterial culture strongly inhibited the mycelial growth of several fungal phytopathogens. The drop collapse assay with this culture showed a surfactant activity that is a major indicator for the selection of biocontrol agent. Also, a bacterium has ability of wastewater treatment. These data demonstrate the potential application of B. amyloliquefaciens as a biocontrol agent.

An extract of Allomyrina dichotoma larva (ADL), one of the insects used most frequently in traditional Chinese medicine for the treatment of liver diseases such as hepatocirrhosis and hepatofibrosis, was assessed for antioxidant bioactivity in this study.

The larvae of Asian rhinoceros beetle, Trypoxylus [Allomyrina] dichotoma (Coleoptera: Scarabaeidae) feed on decaying organic matter and can be found in compost heaps or old dead logs. Mass rearing of this insect with a sawdust-based diet has made it popular commercial pet in many countries of Asia. Recently, lignocellulose, or biomass has been focused as renewable and sustainable resources for energy and many chemical products. It is well-known that gut symbiotic microbes in consortia are inevitable for the lignocellulose digestion in many insects. The cellulase and xylanase activity of the crude extracts from each gut contents and tissues, feces, and blood as a control from Asian rhinoceros beetle were analyzed with the overlaid agar-plate assay and spectrophotometric reduced sugar determination. In the plate assay, the highest activity of cellulase and xylanase was observed from midgut tissue extracts, and next from midgut contents, and minor activity from feces. Whereas no enzyme activity was detected from the samples from blood, crop, and hind gut. Similar results were confirmed from the reducing sugar assay with substrates of Avicell and xylan from birchwood, respectively. Further research on the biochemical and genetic property of symbiotic microbes and their enzymes relating on the lignocellulose degradation is planned.

Imago"s flight activities of Protaetia brevitarsis were from mid June to early September, and those of Allomyrina dichotoma were from mid June to late August. These activities were mainly influenced by amount and duration of rain. They were more active when there was small amount and short period of rain in a year. Distribution depth of the 3<SUP>rd</SUP> larvae of P. brevitarsis were not affected to the temperature. But A. dichotoma were sensitively react to the temperature, almost larvae were near the surface when high temperature. The distribution of P. brevitarsis was not affected by humidity, but most of A. dichotoma were near the surface when high humidity. Larvae of P. brevitarsis usually ate their dead individuals. When we supplied as food the pork, chicken, fresh mackerel, bread, apple, and pear. They took all of these food but larvae of A. dichotoma took only bread. Two species were lived in similar environment but those feeding habit was very different.

우리나라에서 한약재로 활용되고 있는 흰점박이꽃무지(Protaetia brevitarsis)와 장수풍뎅이(Allomyrina dichotoma)의 산란과 발육에 관여하는 환경요인을 밝히고자 본 시험을 실시하였다. 부엽토수분함량은 두 종 모두에서 과습이나 과습은 산란을 저하시켰고 산란 시 최적의 수분함량은 로 추정되었다. 부엽토 수분함량은 흰점박이꽃무지와 장수풍뎅이의 알 및 유충의 생존율과 발육기간, 그리고 유충의 섭식활동에 크게 영향을 주는 것으로 이들의 생육에 관여하는 가장 중요한 요인이었다.

본 연구는 굼벵이를 이용하여 동충하초의 자실체를 유도하기 위하여 3종의 동충하초(Cordyceps militaris) CHO-7208, CHO-7845, CHO-7846를 사용하여 배양조건을 조사하였다. PDA배지에서 13일간 배양 결과 CHO-7208는 25 ±2mm, CHO-7845는 26±2mm와 CHO-7846은 16±2mm 생장하였으며 굼벵이를 배지로서 이용하기 위해서는 전분이나 밀가루, 쌀을 이용하여 사육을 했을 때 전분의 생존율이 높았다. 굼벵이 배지에서는 CHO-7298과 CHO-7846에서 자실체가 발생하였으며 27일간 생장량은 각각 51±5mm와 51 ±5mm로 생장하였다. 또한 굼벵이 에서의 자실체 발생은 기중균사 형성이 왕성한 것이 균주로서 적합하였으며 굼벵이 배지에 동충하초를 배양하여도 자실체의 발생은 양호하였다.