The bumblebee, Bombus terrestris, has played an important role as one of the alternative pollinators since the outbreak of honeybee collapse disorder. Recently, pathogens and parasites such as viruses, bacteria and mites, which affect the life span and fecundity of their host, have been discovered in B. terristris. In order to detect the microsporidian pathogen, Nosema Spp. in the field populations of B. terristris, we collected adults and isolated their genomic DNA for diagnostic PCR. The PCR primers specific for Nosema Spp. were newly designed and applied to gene amplification for cloning. Only small subunit ribosomal RNA(SSU rRNA) gene of N. ceranae was successfully amplified and sequenced among examined genes, which indicates that N. ceranae mainly infects the examined field population of B. terristris. To detect of SSU rRNA gene, two regions of SSU rRNA gene were selected by primary PCR analysis and further analyzed in quantitative real-time PCR(qRT-PCR). The qRT-PCR analysis demonstrated that SSU rRNA of N. ceranae was detected at concentrations as low as 0.85 ng/μl genomic DNA. This result suggests that the detection via qRT-PCR can be applied for the rapid and sensitive diagnosis of N. ceranae infection in the field population as well as risk assessment of B. terristris.

This study was conducted to evaluate the feeding value of the spent mushroom (Pleurotus eryngii) substrates (SMS) in laying hens (Hy-Line Brown). The fresh spent mushroom (Pleurotus eryngii) substrates collected from the DOJUN farm were fermented with Bacillus subtilis EJ3 for 2 weeks. A total of twenty-four laying hens were fed corn-soy based experimental diets containing 0% (control), 5% (T1), 10% (T2) and 15%(T3) fermented SMS for 7 weeks. There were no significant differences among the treatments in egg production, egg weight, egg mass, feed conversion and viability during the experimental period. Feed intake was significantly lowered in control (118.3 g) than T1 (121.9 g), T2 (120.3 g) and T3 (122.4 g). There were no significant differences among the treatments eggshell breaking strength, thickness and haugh unit, whereas the yolk color of T1, T2 and T3 were significantly heavy than T0. The palatability of boiled meat was significantly better in the T3 laying hens than in the T0 laying hens. In conclusion, fermented SMS can be used as resource of feed in laying hen feed at 5.0-15% level without effect on performance and egg qualify.