To obtain information on the sanitary of indoor environment in greenhouses used for shiitake cultivation, bacteria associated with larvae and imagoes of Sciaridae flies, pest to shiitake mushroom, were isolated and identified. A total of 1,048 bacterial colonies were isolated from the flies’ larvae and 984 bacterial colonies were isolated from the flies’ imagoes. Based on molecular analysis of the 16S rDNA sequence, Achromobacter xylosoxidans and Tetrathiobacter kashmirensis of β-proteobacteria, Enterobacter asburiae and Raoultella ornithinolytica of γ -proteobacteria, Curtobacterium sp. and Microbacterium thalassium of Actinobacteria, and Penibacillus taichungensis of Firmicutes were identified from the colonies of the flies’ larvae. While, Bacillus megaterium, B. thuringiensis, Lysinbacillus sphaericus and L. fusifomis of Firmicutes, Microbacterium thalassium and Citricoccus parietis of Actinobacteria, and Enterobacter asburiae of γ-proteobacteria were identified from the flies’ imagoes. Some of the isolated bacterial species were known be human pathogens. Overall, the results of this study suggested that mushroom fly carrying human pathogenic bacteria is one of sources impact on the sanitary of indoor environment of greenhouses used for shiitake cultivation.

The 304 stainless steel powders were prepared by high energy ball milling and subsequently sintered byspark plasma sintering, and the microstructural characteristics and micro-hardness were investigated. The initial size ofthe irregular shaped 304 stainless steel powders was approximately 42 µm. After high energy ball milling at 800 rpmfor 5h, the powders became spherical with a size of approximately 2 µm, and without formation of reaction compounds.From TEM analysis, it was confirmed that the as-milled powders consisted of the aggregates of the nano-sized particles.As the sintering temperature increased from 1073K to 1573K, the relative density and micro-hardness of sintered sampleincreased. The sample sintered at 1573K showed the highest relative density of approximately 95% and a micro-hard-ness of 550 Hv.

Mosquito-borne viral pathogens infect millions of people worldwide, often resulting in fatal infections. Our research interests in mosquito vector biology focuses on understanding of the molecular and genetic basis of mosquito resistance to arbovirus infections. Unlike mammalian cells including humans, for instance, mosquito cells do not show pathologic symptoms when infected by arboviruses. This observation led us to embarking a microarray study to investigate mosquito-virus interactions using Anopheles gambiae and o’nyong-nyong virus (ONNV) as a model system. As a result, we found that transcription of the hsc70B gene is increased about 2.6-fold in ONNV-infected An. gambiae compared to non-infected controls. Subsequently, in vivo RNAi silencing of the hsc70B transcript caused enhanced ONNV replication in female mosquitoes. Therefore, these results suggest that the hsc70B protein has an inhibitory effect on ONNV replication. A promoter analysis of the hsc70B locus further demonstrated that the hsc70B promoter is able to induce transcription of hsc70B in response to ONNV infection. In addition, hsc70B transcription was also induced by West Nile or La Crosse virus infection. Collectively, our findings indicate that hsc70B plays a role in suppressing virus replication as a general antiviral mechanism. Implications of hsc70B research and our other research endeavors toward the control of mosquito-borne infectious diseases will be discussed.

14-3-3 proteins are known to play a pivotal role in a diverse array of cellular events such as cell survival, apoptosis, and signal transduction. Numerous 14-3-3 ζ have been cloned and characterized from a host of eukaryotic organisms including human, plants, yeast, fruit fly and silkworm. However, no study on Spodoptera exigua 14-3-3ζ in conjunction with virus infection has so far been reported in insects. It appears that expression of Se14-3-3ζ was decreased starting 24 h post-SeNPV infection as SeNPV titers seemed to increase as evidenced by intense bands of SeNPV IAP3. Interestingly, confocal microscopic analysis revealed that Se14-3-3ζ is expressed at the apical side of the NPV-uninfected gut cells, whereas it was detected mainly in the nucleus of the NPV-infected cells. Thus, despite the biological significance of Se14-3-3ζ in S. exigua in conjunction with molecular interactions between SeNPV and S. exigua is unclear now, our data suggest that Se14-3-3 ζ protein plays a role to protect S. exigua from the infection or inhibit replication of SeNPV.

Janus kinase-signal transducer and activator of transcription (JAK-STAT) signaling pathway is known to play a pivotal role in various cellular events and antiviral responses in both vertebrates and insects. In an attempt to elucidate the potential involvement of STAT on S. exigua-SeNPV interactions, the full length cDNA of SeSTAT was cloned from S. exigua. Analysis of temporal expression patterns shows that SeSTAT is expressed in all stages of life cycle such as larvae, pupae, and adult. Spatial expression analysis shows that it is highly expressed in fat body and Malpighian tubule. Interestingly, SeSTAT is induced at 24 h in response to either laminarin or LTA injection in larvae. Electrophoretic mobility shift assay (EMSA) shows that the binding of nuclear extracts from fat body cells immune-challenged with LTA to STAT5 probe was observed. In addition, SeSTAT was nuclear-translocalized in both fat body and gut cells that were challenged with LTA and laminarin, respectively. Finally, gene silencing of SeSTAT shows that SeNPV number appears to be increased. It suggests that SeSTAT may act as a negative regulator against SeNPV in midgut.

The techniques of IVM, IVF and IVC of canine oocytes may provide useful information for gamete salvage programs and the conservation of endangered canidae. This investigation has been made to determine the efficiency of in vitro maturation (IVM) as a basic experiment to study the development of canine oocytes after in vitro fertilization (IVF). The rate of oocytes developing to the MII stage was higher in the hormone treated group (10 IU/ml hCG+eCG, 14.7%, p<0.05) than in the control group (0 IU/ml hCG+eCG, 10.0%). The monospermy and pronuclear rates of canine oocytes were investigated after caffeine treatment on IVF. Canine oocytes were fertilized in the Fert‐TALP medium supplemented with 0, 10, 20 or 30 mM caffeine (Fert I, Fert II, Fert III or Fert IV, respectively). The highest pronuclear formation rate was obtained in the Fert I for 24 h IVF (6.7%, 6/89). Therefore, it is believed that unlike in other mammals, caffeine in canine IVF does not increase the efficiency of fertilization rate, and is not an important factor.