메탄가스는 주요 온실가스 중 하나로 반추동물의 장내발효를 통해 발생하며, 이러한 경로의 메탄가스는 대한민국 축산부문 총 온실가스 발생량의 40% 이상을 차지한다. 이런 이유로 많은 연구자들은 반추동물에서 발생하는 메탄생성량을 줄이기 위한 시도를 계속해 왔다. 본 연구는 반추동물의 메탄발생량을 측정하기 위해 호흡대사챔버를 개발하고 호흡대사챔버의 정확성을 검증하기 위해 실시하였다. 호흡대사챔버는 25.4 m3 크기로 스테인리 스 플레이트로 내부를 완전히 밀폐하였다. 직경 Φ100의 공기 유입관과 배출관을 설치하였고, 공기 배출관에 에어모터를 설치하여 내부공기를 제거함과 동시에 유입관을 통해 배출된 만큼의 공기가 외부에서 유입되도록 하였다. 챔버 내 메탄가스 회수기능을 검증하기 위해 메탄표준가스 5L를 각 챔버에 주입하여 균일하게 확산시킨 후, 챔버 내부의 공기를 900 L/min의 속도로 제거하였다. 제거된 공기의 메탄가스 농도를 연속적으로 측정하여 주입된 메탄가스와 비교함으로써 회수율을 평가하였다. 챔버 내 표준가스는 평균 100분에 완전히 제거되었으며, 메탄가스의 회수율은 평균 109 ± 6.7%로 측정되었다. 호흡대사챔버의 실제 이용성을 평가하기 위해, 평균체중 581.9 ± 33.8 kg 의 한우 거세우 4두에게 비육후기 배합사료 9 kg과 볏짚 1 kg을 급여하며 메탄발생량을 측정하였다. 한우의 장내발효에 의한 메탄발생량은 평균 236.4 ± 105.44 L/day로 측정되었다. 본 연구에서 개발·검증한 호흡대사챔버는 국내 한우의 장내발효 메탄가스발생량 측정 시험에 적용 가능할 것이다.

Pine wood nematode (Bursaphelenchus xylophilus), the pathogen of pine wilt disease (PWD), is transmitted to new host trees through Monochamus spp., vector insects. In Korea, aerial application has been allowed conventionally to eliminate the vectors in their flying season. In this study, thiacloprid was applied to four-year-old Pinus densiflora, which were fed to adult M. alternatus emerged from PWD infected logs for 72 h in the condition of 1 DAT (Days after treatment of thiacloprid) and 15 DAT, respectively. We confirmed the presence of nematodes in each beetle applied to experiment by Baermann funnel method and PCR with species-specific ITS primers. After 72 h of 1 DAT and 15 DAT, thiacloprid residues in pine trees were 0.59-16.32 and 0.043-0.131 mg/kg and the mortalities of the beetle were 100 and 99.2%, respectively. Detection rate of B. xylophilus was 51.3%. The incidence rate of PWD in 1 DAT and 15 DAT showed no mortality and 5.55% compared with 55.5% of control, respectively.

Pine wilt disease (PWD) induced by the pinewood nematode (PWN), Bursaphelenchus xylophilus, is a great threat to pine forests in Europe and East Asia. Identification of interrelation between Monochamus spp., the major vectors of PWD and PWN is the key factor for effective control of PWD because PWN is trasmitted to healthy pine trees only by Monochamus vectors. In this study, direct injection system is designed to apply PWN artificially to M. alternatus just before emergence to adults. After injection of PWN (300~500 nematodes in 10 μl of 1X PBS) using adapted glass pipets, the survival rate of M. alternatus and detection rate of live PWN were 80.0% and 68.8% at 168 h respectively. As a result, we demonstrated the artificial injection methods of nematode to Monochamus vectors for further study of coeffect between vectors and specific stage of PWN including 3rd stage dispersal juvenile.

Hertwig's epithelial root sheath (HERS) consists of bilayered cells derived from the inner and outer dental epithelia and plays important roles in tooth root formation as well as in the maintenance and regeneration of periodontal tissues. With regards to the fate of HERS, and although previous reports have suggested that this entails the formation of epithelial rests of Malassez, apoptosis or an epithelialmesenchymal transformation (EMT), it is unclear what changes occur in the epithelial cells in this structure. This study examined whether HERS cells undergo EMT using a keratin-14 (K14) cre:ROSA 26 transgenic reporter mouse. The K14 transgene is expressed by many epithelial tissues, including the oral epithelium and the enamel organ. A distinct K14 expression pattern was found in the continuous HERS bi-layer and the epithelial diaphragm were visualized by detecting the β-galactosidase (lacZ) activity in 1 week postnatal mice. The 2 and 4 week old mice showed a fragmented HERS with cell aggregation along the root surface. However, some of the lacZ-positive dissociated cells along the root surface were not positive for pan-cytokeratin. These results suggest that the K14 transgene is a valuable marker of HERS. In addition, the current data suggest that some of the HERS cells may lose their epithelial properties after fragmentation and subsequently undergo EMT.

The working mechanism of bisphosphonate on bone cells is unclear despite its powerful inhibitory activity on bone resorption. The differentiation and activation of osteoclasts are essential for bone resorption and are controlled by the stimulatory RANKL and inhibitory OPG molecules. Teeth exhibit a range of movement patterns during their eruption to establish their form and function, which inevitably accompanies peripheral bone resorption. Hence, the mandible, which contains the teeth during their eruption processes, is a good model for revealing the inhibitory mechanism of bisphosphonate upon bone resorption. In the present study, RANKL and OPG expression were examined immunohistochemically in the mandible of rats with developing teeth after alendronate administration (2.5 mg/kg). The preeruptive mandibular first molars at postnatal days 3 to 10 showed the developing stages from bell to crown. No morphological changes in tooth formation were observed after alendronate administration. The number of osteoclasts in the alveolar bone around the developing teeth decreased markedly at postnatal days 3, 7 and 10 compared with the control group. RANKL induced strong positive immunohistochemical reactions in the dental follicles and stromal cells around the mandibular first molar. In particular, many osteoclasts with strongly positive reactions to RANKL appeared above the developing mandibular first molars at postnatal days 3 and 10. Immunohistochemical reactions with RANKL after alendronate administration were weaker than the control groups. However, the immunohistochemical reactivity to OPG was stronger after alendronate administration, at postnatal days 3 and 10. These results suggest that alendronate may decrease bone resorption by regulating the RANKL/OPG pathway in the process of osteoclast formation, resulting in a delay in tooth eruption.

Teeth develop via a reciprocal induction between the ectomesenchyme originating from the neural crest and the ectodermal epithelium. During complete formation of the tooth morphology and structure, many cells proliferate, differentiate, and can be replaced with other structures. Apoptosis is a type of genetically-controlled cell death and a biological process arising at the cellular level during development. To determine if apoptosis is an effective mechanism for eliminating cells during tooth development, this process was examined in the rat mandible including the developing molar teeth using the transferase-mediated dUTP-biotin nick labeling (TUNEL) method. The tooth germ of the mandibular first molar in the postnatal rat showed a variety of morphological appearances from the bell stage to the crown stage. Strong TUNEL-positive reactivity was observed in the ameloblasts and cells of the stellate reticulum. Odontoblasts near the prospective cusp area also showed a TUNEL positive reaction and several cells in the dental papilla, which are the forming pulp, were also stained intensively in this assay. Our results thus show that apoptosis may take place not only in epithelial-derived dental organs but also in the mesenchyme-derived dental papilla. Hence, apoptosis may be an essential biological process in tooth development.