A multiplex polymerase chain reaction (PCR) was developed for the simultaneous detection and differentiation among Nosema apis and Nosema ceranae in honeybee. Three sets of primers were selected from different genomic sequences to specifically amplify a 831 bp amplicon within the SSU rRNA gene, specific for both N. apis and N. ceranae (MSSR primer); a 375 bp amplicon within the SSU rRNA gene, specific for N. apis (NA primer); and a 1,131 bp amplicon with in SSU rRNA gene, specific for N. ceranae (NC primer). Using the primers in conjunction (multiplex PCR) we were able to N. apis and N. ceranae and to differentiate between them. The sensitivity of this PCR assay was approximately 102spores per milliliter. We proposed that the multiplex PCR was sensitive, specific and rapid tool that can serve as a useful differential diagnostic tool for detecting N. apis and N. ceranae in honeybee.
This study was conducted to investigated the distribution, ecological character and life cycle of Black Soldier Fly(BSF), Hermetia illucens. The distribution of BSF was defined in all parts of the country in Korea. Its main habitat was found to be areas near cattle sheds, manure sheds, living waste dump grounds, and food waste dump grounds. Observed characteristics of BSF by developmental stage may be summarized as follows: eggs were a long oval shape of 886.9±19.7 ㎛ in major axis and 190.1±9.7 ㎛ in minor axis; they were 24.0±1.6 ㎍ in weight. One adult insect laid 1001±247 eggs in quantity; days to hatch from eggs (27℃, 60% R.H.) were 81.3±12.5 days. Last instar larva ranged from 20.7±1.1 mm in size, the length of larval stage was approximately 15~20 days. Pupae exhibited red brown, 19.2±1.1 mm in size; pupal state lasted 15.5±1.4 days for female, 14.7±1.4 days for male, exhibiting the tendency of males having shorter period than females. Adult insects were sized about 13~20 mm and colored black. Mating started 2 days after emergence and was most active during the 3rd day. Mating mostly occurred between 10:00 and 16:00 during which light intensity is high. Laying eggs started 3 days after emergence and was most frequent during days 4~6. Time of laying eggs during the day was similar to copulation time, showing the highest laying rate between 10:00 and 16:00.