Feasibility and Stability of Liver Biopsy before Treatment for Preclinical Nonalcoholic Fatty Liver Studies

채연지,전대원,Waqar Khalid Saeed,강현태,오주희,이승민,장기석 대한의학회 2019 Journal of Korean medical science Vol.34 No.2

Background: The heterogeneity of histological findings in preclinical diet-induced nonalcoholic fatty liver disease (NAFLD) animal models is highly challenging. Here, we aimed to evaluate the feasibility and stability of repeated liver biopsy in NAFLD animal models. Methods: Heterogeneity of diet-induced NAFLD was evaluated at different time points in 52 high-fat diet (HFD), 35 methionine choline-deficiency diet (MCD), and 166 western diet (WD) induced NAFLD mice. Serial liver biopsies (left lateral, right medial, and left medial lobes) were performed monthly for up to 3 months. Mortality rates and changes in food intake, body weight, and liver enzymes were assessed. Results: At 12 weeks, of the HFD animals, 14% and 30% did not develop steatosis and lobular inflammation, respectively; of the MCD animals, 7% did not develop lobular inflammation; and of the WD animals, 14% and 51% did not develop steatosis and lobular inflammation, respectively. The mortality rate of repeated liver biopsy was 1.62% (2/123 mice died). Repeated liver biopsy can be used to trace disease progression. Although body weight, food intake, and liver enzymes slightly changed after biopsy, all recovered within a week. Repeated liver biopsy did not affect the degrees of inflammation and steatosis of the other liver lobes. Conclusion: The diet-induced NAFLD models were quite heterogeneous. Our results suggest that the repeated liver biopsy before treatment was applicable and stable in this NAFLD animal study.

기수역 요각류 위내용물 유전자 분석: 소화기관 내외부 유전자의 선택적 처리방법

채연지,오혜지,김용재,장광현,조현빈 한국하천호수학회 2021 생태와 환경 Vol.54 No.3

Understanding the selective feeding behavior of zooplankton on phytoplankton is essential for evaluating the nutrient cycle and energy flow in the food web. Although many studies have been conducted regarding the feeding behaviors of zooplankton through gut content analyses, there are limitations in the visual identification of digested contents using a microscope. DNA techniques have been applied to overcome these limitations since they can detect and amplify small amounts of prey DNA remaining in the gut contents. We designed a method to extract prey DNA from the gut contents of the whole body of the copepod specimen and tested the resolution of DNA identification for the prey phytoplankton. The common brackish species, Sinocalanus tenellus, were collected from Saemangeum Reservoir in different sites and seasons, and gut content DNA was extracted using 2.5% bleach treatment for 2 min for removal of potential contamination sources existing in preserved specimens without dissolution of the body. The sequences of the extracted gut contents were confirmed using BLASTn suite based on the NCBI database. The phytoplankton species detected in the gut showed temporal and spatial differences. Although DNA analysis of small copepod gut contents has been suggested as an effective method to examine the dynamics of primary prey sources at the genus or species level, uncertainties such as misidentification and limitations in the detailed information of the composition still exist.

정수생태계 동물플랑크톤 채집 시 네트 인양 유형 및 수심에 따른 군집 정보 비교

오혜지,채연지,구도영,김유진,왕정현,최보형,지창우,곽인실,박영석,남귀숙,김용재,장광현,OH, Hye-Ji,Chae, Yeon-Ji,Ku, Doyeong,Kim, Yu-Jin,Wang, Jeong-Hyeon,Choi, Bohyung,Ji, Chang Woo,Kwak, Ihn-Sil,Park, Young-Seuk,Nam, Gui-Sook,Kim, Yong-Ja 한국하천호수학회 2020 생태와 환경 Vol.53 No.4

호소 생태계에 대한 생물측정망 조사 및 평가지침은 동물플랑크톤 조사의 편의성을 고려하여 출현 종 수와 개체군 밀도, 군집 지수 등을 상대적으로 비교하기 위해 수심 5m 이하의 얕은 호소-수변부에서 사선끌기, 수심 20 m 이상 호소-전수심 수직끌기, 그 이상의 깊은 호소-20 m까지의 수심을 대상으로 한 수직끌기를 제시하고 있다. 본 연구에서는 지침에서 제시하는 방법 중 사선끌기법과 20 m 수직 끌기법을 각각 전수심을 대상으로 한 조사 방법과 비교하여 동물플랑크톤 군집 정보의 차이 및 특성에 대해 분석하였다. 군집 지수의 경우 수심이 얕은 호수에서의 사선끌기법/수직끌기법 비교에서는 차이를 보이지 않은 반면, 수심이 깊은 호소에서는 끌기 수심을 20 m로 제한할 경우보다 전수심 수직끌기를 적용했을 때 다양도 및 풍부도 지수가 상승하는 것으로 나타났다. 또한, 사선끌기 및 20 m 수직끌기를 통해 채집한 동물플랑크톤 시료로부터 표면~저층 상층부까지의 전수심을 채집 대상으로 설정한 경우보다 약 3배 정도 높은 개체군 밀도가 계산되어, 동물플랑크톤 총 개체 밀도가 크게 과대 평가되는 것으로 나타났다. 이러한 차이는 동물플랑크톤 수직 분포 특성상 발생하는 수층별 동물플랑크톤 개체 밀도 차이 및 여과된 원수량의 차이에서 비롯된 결과로 판단되며, 이에 따라 호소를 대변하여 수체 간 또는 수체 내 변동을 보다 정확히 파악하고 호 내 2차 생산과 관련한 기능적 정량 지표로서 동물플랑크톤 군집 정보의 활용을 고려할 경우, 수심에 따른 개체군 밀도 분포와 전체 개체수 환산 시 수층별 기여율을 고려한 전수심 수직끌기 방법의 적용이 보다 적절할 것으로 분석되었다. Biomonitoring Survey and Assessment Manual for lake ecosystem suggest zooplankton collection methods to compare relatively the number of species, population density, and community indices, taking into account the convenience of the field sampling according to the sites' water depth. In this study, the oblique towing and 20 m vertical towing methods presented in the manual were respectively compared with the whole water column-vertical towing and we analyzed the differences and characteristics of zooplankton community information gathered by each collection method. For community indices, there was no difference in the comparison of oblique/vertical towing methods in the shallow lake, but in the deep lake, the diversity and richness indices increased when vertically towing through whole water column rather than when limiting the towing depth to 20 m. In addition, the total zooplankton density collected by the oblique/20 m vertical towing methods was about three times higher than the whole water column-vertical towing method, which means that the density of zooplankton community can be overestimated depending on the collection methods. It appears to be results of differences in the zooplankton density by water layer arising from their vertical distribution and in filtered raw water quantity according to the towing depth/distance. Hence, for zooplankton community information to be used as a functional quantitative indicator representing the entire lake, it would be more appropriate to apply the whole water column-vertical towing method with considering the distribution of zooplankton density by depth and contribution rate of each water layer when converting total zooplankton density.

동물플랑크톤 연구에 있어 DNA 분석 기법의 활용 방법과 과제: 개체 동정에서 군집 분석, 생물학적 상호작용 분석까지

오혜지,채연지,최예림,구도영,허유지,곽인실,조현빈,박영석,장광현,김현우 한국하천호수학회 2021 생태와 환경 Vol.54 No.3

Traditional morphological identification difficulties, such as phenotypic plasticity, misidentification of cryptic species, and larval stage species, can be compensated for by using DNA analysis techniques, such as DNA barcoding, in surveying zooplankton populations, including species identification. Recently, the rapid development of DNA sequencing techniques has allowed DNA-based community analysis not only for zooplankton assemblages in various aquatic ecosystems but also for the gut contents of zooplankton that are limited by conventional methods such as visual and microscopic identification. Therefore, the application of DNA sequencing can help understand biological interactions through the analysis of zooplankton food sources. The present paper introduces the major DNA-based approaches in zooplankton research topics, including taxonomic approaches by DNA barcoding, community-level approaches by metabarcoding, and gut content analyses, summarizes the analysis methods, and finally suggests the methodological topics that need to be considered for future applications.

영주댐 유사조절지 상류의 용존유기물 (Dissolved Organic Matter) 특성과 물리 . 화학 및 생물학적 환경 요인과의 연관성 연구

오혜지,김도균,최지수,채연지,오종민,신경훈,최광순,김동균,장광현 한국하천호수학회 2021 생태와 환경 Vol.54 No.4

A sediment control dam is an artificial structure built to prolong sedimentation in the main dam by reducing the inflow of suspended solids. These dams can affect changes in dissolved organic matter (DOM) in the water body by changing the river flow regime. The main DOM component for Yeongju Dam sediment control of the Naeseongcheon River was analyzed through 3D excitation-emission matrix (EEM) and parallel factor (PARAFAC) analyses. As a result, four humic-like components (C1~C3, C5), and three proteins, tryptophan-like components (C2, C6~C7) were detected. Among DOM components, humic-like components (autochthonous: C1, allochthonous: C2~C3) were found to be dominant during the sampling period. The total amount of DOM components and the composition ratio of each component did not show a difference for each depth according to the amount of available light (100%, 12%, and 1%). Throughout the study period, the allochthonous organic matter was continuously decomposing and converting into autochthonous organic matter; the DOM indices (fluorescence index, humification index, and freshness index) indicated the dominance of autochthonous organic matter in the river. Considering the relative abundance of cyanobacteria and that the number of bacteria cells and rotifers increased as autochthonous organic matter increased, it was suggested that the algal bloom and consequent activation of the microbial food web was affected by the composition of DOM in the water body. Research on DOM characteristics is important not only for water quality management but also for understanding the cycling of matter through microbial food web activity.