Spermatogonial stem cells (SCCs) is foundation for spermatogenesis throughout male adult life because they have ability of self-renewal and differentiation into spermatozoa. Storage of such SSCs is very important to study on male reproduction, which would contribute human male infertility to be treated. However, during cryopreservation, the most cells are damaged by cryoinjury such as apoptosis, necrosis, osmotic stress, oxidative stress and so on. For the reason, in cryopreservation technique, targeting purpose is what cells are stored stably without cryoinjury. The purpose of this study was to develop the cryoprotectant for decrease in cryoinjury of SSCs by using melatonin and necrostatin-1 as additive cryoprotectant. The SSCs with melatonin or necrostatin-1 was frozen for 1 month, and then thawed to evaluate survival, recovery and proliferation rate. The result showed that necrostatin-1 50 mM was significantly greater than DMSO control. Furthermore, we conducted the characterization of cryo-thawed SSCs with necrostatin-1 50 mM to confirm whether the SSCs could maintain the undifferentiated state. As a result, the normal expression of each marker, which is PLZF, GFRa1 and VASA, was observed except for C-kit, meaning that the cells could maintain the undifferentiated state regardless of cryopreservation. Therefore, the result indicates that the cryo-thawed SSCs have ability of proliferation and self-renewal. In conclusion, our finding verifies that cryopreservation of SSC with necrostatin-1 50 mM could be helpful to preserve the SSCs stably, contributing to various studies on male reproduction and infertility treatment

The effects of some essential oils and Sopophora extract on Frankliniella occidentalis adults, Myzus persicae adults, Plutella xylostella larvae, and Spodoptera exigua larvae were investigated. Insecticidal and phytotoxicity activities of emulsion in water formulations containing the active essential oils as active ingredients under laboratory and pot conditions were also tested. Based on these results, NRS-13 and NRS-24 formulations were selected to further assess as follows; the GC and GC-MS analysis on the active essential oils, the quantity analysis of the major components contained in the formulations, their insecticidal activities under pot and field conditions, and their oral, dermal, skin and eye irritation, and fish toxicities. Thus, the NRS-13 (BaechooSaferTM) and NRS-24 (JindiOutTM) formulations could be considered as insecticides for pest control.

The habitat of the cockroach varies by species. The German cockroach (Blattella germanica) lives in human dwellings, while the Japanese field roach (Blattella nipponica) lives in a mountainous region. Based on phylogenetic analysis of mtCO I, the two species are closely related to each other and B. germanica is divergent from wild species such as B. nipponica. Their habitats and walking speed differ even though the two species have similar morphology. We hypothesized that habitats might influence walking speed via changes to appendage morphology and enzyme-based physiological differences. We found that phenotype such as appendage length and esterase isozyme expression were clearly different between the two species. These differences might be responsible for the observed difference in walking speed.

Even though klotho deficiency in mice exhibits multiple aging-like phenotypes, studies using large animal models such as pigs, which have many similarities to humans, have been limited due to the absence of cell lines or animal models. The objective of this study was to generate homozygous klotho knockout porcine cell lines and cloned embryos. A CRISPR sgRNA specific for the klotho gene was designed and sgRNA (targeting exon 3 of klotho) and Cas9 RNPs were transfected into porcine fibroblasts. The transfected fibroblasts were then used for single cell colony formation and 9 single cell–derived colonies were established. In a T7 endonuclease I mutation assay, 5 colonies (#3, #4, #5, #7 and #9) were confirmed as mutated. These 5 colonies were subsequently analyzed by deep sequencing for determination of homozygous mutated colonies and 4 (#3, #4, #5 and #9) from 5 colonies contained homozygous modifications. Somatic cell nuclear transfer was performed to generate homozygous klotho knockout cloned embryos by using one homozygous mutation colony (#9); the cleavage and blastocyst formation rates were 72.0% and 8.3%, respectively. Two cloned embryos derived from a homozygous klotho knockout cell line (#9) were subjected to deep sequencing and they showed the same mutation pattern as the donor cell line. In conclusion, we produced homozygous klotho knockout porcine embryos cloned from genome-edited porcine fibroblasts.

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.

This study was conducted to evaluate a visual surveillance system. The advancement of recording technology and network service make it easy to record and transfer the videos. Moreover, progressed recognition technology help to make a distinction each other. Cows show distinguishing behaviors during their estrus period. The mounting is one of the behaviors. The result was different depending on the breed of the cows and the size of the farm. In the case of Korean native cattle, the estrus detection rate was 71.15%, however, dairy cows, the estrus detection rate was 39.38%. At the farms having below 6 modules, the estrus detection rate was 87.41%. On the other hand, at the farms having over 6 modules, the estrus detection rate was 77.78%. With the proper progress, the visual surveillance system can be used to detect heat detection.