Purification and Identification of Paenibacillus sp., Isolated from Diseased Larvae of Allomyrina dichotoma (Linnaeus, 1771) (Coleoptera: Scarabaeidae) in Insect Farms

Abstract

In reared populations of Allomyrina dichotoma, commercial insects, 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 their intestinal bacterial florae 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 clear identification, a whole guts was extracted from each larva showing the sign of the disease and incubated at 70 o C for 15 min to isolate spore forming bacteria. After then, each content of guts was cultured on MYPGP NAL agar medium(12.5 µg/ml of nalidixic acid) at 30 o C. The 16S rRNA gene sequence analysis for the isolated bacteria showed that they were closely related to P. rigui(97.9% similarity), to P. chinjuensis(96.1% similarity), and to P. soli(95.3% 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.

Development of Seven Microsatellite Markers Using Next Generation Sequencing for the Conservation on the Korean Population of Dorcus hopei (E. Saunders, 1854) (Coleoptera, Lucanidae)

“…The market for insects as pets has increased due to growing interest in keeping exotic pet animals. The stag beetle market is worth nearly $283 million in Japan, while the market in Korea is estimated to be worth approximately $38 million [ 7 , 8 ]. Some insect collectors called D. hopei binodulosus “Black Diamond” because its body color was black and as expensive as diamonds [ 6 , 9 , 10 ].…”

Development of 12 Microsatellite Markers in Dorcus titanus castanicolor (Motschulsky, 1861) (Lucanidae, Coleoptera) from Korea Using Next-Generation Sequencing

“…Marketing campaigns from the insect industry have contributed to a rapid increase in the number of pet insects [5,6,7]. D. titanus castanicolor is a popular pet insect [7], and the rearing method of the subspecies is well documented [8].…”