This study focuses on developing diagnostic compositions, kits, and information provision methods for identifying species-specific genes in domestically residing Reticulitermes speratus and Reticulitermes kanmonensis, as well as the recently introduced Cryptotermes domesticus. The core innovation of this invention lies in the utilization of species-specific genetic markers to facilitate rapid and accurate species identification using a PCR (polymerase chain reaction)-based diagnostic technique. This approach enables swift identification of termites at quarantine stages, contributing to efficient management of imported goods and minimizing ecological and economic damages caused by termites. Through genome analysis of termites, this research has identified candidate species-specific genetic markers, developed diagnostic compositions and kits based on these markers, and proposed a rapid diagnostic method capable of determining termite species within a day, optimally within three hours. This invention provides a groundbreaking tool for termite management and research, significantly contributing to pest control and biodiversity conservation efforts.

Radish is an important root vegetable in the world, and many cultivars have been developed with various molecular marker systems to identify these cultivars. Recently developed markers for radish cultivar identification require only 11 primer pairs, but they still use conventional PCR with different annealing temperatures and time-consuming gel electrophoresis. To improve the genotyping method, we applied touchdown PCR with 11 primers with M13 tails among 105 radish cultivars. Touchdown PCR successfully generated amplicons in all 11 M13-tailed primers with a condition of annealing temperature starting from 55℃, decreased by 1°C and 33 cycles at 53°C. The 11 M13-tailed primers followed by fragment analysis produced 71 amplicons, which produced more amplicons than gel electrophoresis that produced 23 amplicons. Especially, simple sequence repeats produced more amplicons, 12 on average, than the other marker types. The present study requires less effort and provides more accurate results compared to genotyping using gel electrophoresis. Besides, a database can be established using digitized genotyping results among radish cultivars.

Parrots have been threatened by global trade to meet their high demand as pets. Controlling parrot trade is essential because parrots play a vital role in the ecosystem. Accurate species identification is crucial for controlling parrot trade. Parrots have been traded as eggs due to their advantages of lower mortality rates and more accessible transport than live parrots. A molecular method is required to identify parrot eggs because it is difficult to perform identification using morphological features. In this study, DNAs were obtained from 43 unidentified parrot eggs using a non-destructive sampling method. Partial cytochrome b (CYTB ) gene was then successfully amplified using polymerase chain reaction (PCR) and sequenced. Sequences newly obtained in the present study were compared to those available in the GenBank by database searching. In addition, phylogenetic analysis was conducted to identify species using available sequences in GenBank along with sequences reported in previous studies. Finally, the 43 parrot eggs were successfully identified as seven species belonging to two families and seven genera. This non-destructive sampling method for obtaining DNA and molecular identification might help control the trade of parrot eggs and prevent their illegal trade.

Recently, extreme terrorist attacks have frequently occurred around the world and are threatening the international community. It is no longer a safe zone for terrorism in our country. Therefore, domestic nuclear facilities as the highest level of national security facilities have established a physical protection system to protect facilities and lives against terrorist attacks. In addition, security search and access control are conducted for controlled items and unauthorized person. However, with the development of science and technology, disguised weapons or homemade explosives used in terrorism are becoming very sophisticated. Therefore, nuclear facilities need to strengthen security search of weapons or homemade explosives. Since these disguised weapons or homemade explosives are difficult to find only through security search, it is also necessary to actively identify unspecified people who possess disguised weapons or do abnormal behavior. For this reason, the “Abnormal Behavior Detection Method”, which is very effective in preemptive response to potential terrorist risks, has been introduced and operated in aviation security field. Korea Institute of Nuclear Nonproliferation and Control (KINAC) has established a “Practice Environment for Identifying Disguised Weapons” in 2020 for trainees to recognize the dangers of controlled items and to use for physical protection education. This Practice environment has not only the basic explanation of the controlled items of nuclear facilities, but also various actual disguised weapons were displayed. It also introduces actual terrorist incidents using homemade explosives such as attempted bombing of a cargo plane bound for Chicago and the Boston Marathon bombing. And then a model of the disguised explosives actually used is displayed and used for education. In addition, in 2022, the “Abnormal behavior detection method” education module was developed and used for physical protection education. In this module, the outline and introduction of the “Abnormal Behavior Detection Method” and “Behavior Detection Officer (BDOs)” are explained. In this way, the access control and security search system of nuclear facilities require the overall monitoring system, not only for dangerous goods but also for identification of persons possess and carrying them. This study describes the development of the Curriculum for “Disguised Weapon Identification” and “Abnormal Behavior Detection Method” to enhance the effectiveness of physical protection education.

Two tree frogs, Dryophytes suweonensis and Dryophytes japonicus, inhabiting Korea, are morphologically similar and share the same habitats. Therefore, they are identified mainly through their calls, especially for males. Dryophytes suweonensis is registered as an endangered (IUCN: EN grade) and protected species in South Korea. Thus, it is necessary to develop a method to rapidly identify and discriminate the two species and establish efficient protection and restoration plans. We identified significant genetic variation between them by sequencing a maternallyinherited mitochondrial 12S ribosomal DNA region. Based on the sequence data, we designed a pair of primers containing 7 bp differences for high resolution melting (HRM) analysis to rapidly and accurately characterize their genotypes. The HRM analysis using genomic DNA showed that the melting peak for D. suweonensis was 76.4±0.06°C, whereas that of D. japonicus was 75.0±0.05°C. The differential melt curve plot further showed a distinct difference between them. We also carried out a pilot test for the application of HRM analysis based on immersing D. suweonensis in distilled water for 30 min to generate artificial environmental DNA (eDNA). The results showed 1.10-1.31°C differences in the melting peaks between the two tree frog samples. Therefore, this HRM analysis is rapid and accurate in identifying two tree frogs not only using their genomic DNA but also using highly non-invasive eDNA.

Known for its effectiveness in weight loss and diabetes prevention, Gymnema sylvestre products can be found in the US, Japanese, and Indian markets. However, the recommended dosage or safety of these products has not yet been proven. Therefore, development of an analytical method for detecting the content of Gymnema sylvestre in food products is required. Accordingly, this study proposes an analysis method that can examine Gymnema sylvestre in food using LC-ESI-MS/MS and KASP (Kompetitive Allele-Specific PCR) markers. In LC-ESI-MS/MS, a simultaneous analysis method for gymnemic acid and deacylgymnemic acid was optimized using negative ionization mode, and its validation test was completed for solid and liquid samples. In addition, KASP markers were prepared by finding the specific SNP of G. sylvestre in ITS2 and matK through DNA barcodes. The two KASP markers returned positive FAM fluorescence result when combined with G. sylvestre, and this aspect was confirmed in raw G. sylvestre as well. The applicability of the method was tested on 21 different food and healthy functional products containing G. sylvestre purchased on the internet. As a result, although there was a difference in the ratios of gymnemic acid and deacylgymnemic acid in LC-ESI-MS/MS, the index component was detected in all 21 products samples. In the KASP analysis, 9 products returned positive FAM result, and the rest of the products were found to be containing G. sylvestre extract. This study is the first study to use the dual system of LC-ESI-MS/MS and KASP for the analysis of G. sylvestre. The study has confirmed that these two methods are applicable to the examine G. sylvestre content in food products.

동물플랑크톤이 식물플랑크톤을 선택적으로 섭식하는 특성에 대한 이해는 수생태계 먹이사슬 내의 물질 이동에 중요하다. 하지만 해부를 통한 위내용물 추출 방법은 소형 요각류를 대상으로 적용하기에는 적절하지 않고, 유전자가 유실되거나 위내용물이 아닌 개체 외부의 유전자로 인해 오염될 가능성이 존재한다. 본 연구에서 호내 식물 플랑크톤 조성 및 기타 환경이 상이한 두 지점을 선정하여 모든 지점에서 지속적으로 출현하는 기수성 요각류인 Sinocalanus tenellus를 대상으로 위내용물의 유전자 분석을 수행하였다. 요각류 개체 외부의 DNA를 제거하는 데 2.5%로 희석한 시판용 표백제(차아염소산나트륨 5.4%) 에 2분간 처리하여 증류수로 2회 세척한 뒤 유전자를 추출하였다. 추출된 유전자는 23S rRNA을 증폭하여 서열분석을 실시하였다. Capillary sequencing 분석 결과, 원수와 처리수 및 요각류 위내용물에서 다양한 분류군 (규조강, 녹조 강, 남조강, 와편모조강, 은편모조강, 황갈조강)에 속하는 식물플랑크톤이 검출되었으며, 새만금호 내 시공간 차이에 따라 상이한 경향을 보였다. 현미경을 이용하여 동정한 식물플랑크톤 군집 조성의 경우 규조강이 우점한 반면, 동일한 원수의 유전자 분석 (capillary sequencing) 결과에서는 주로 녹조강, 남조강 및 와편모조강이 우점하여 다소 상반된 경향을 나타냈다. 본 연구에서 적용한 위내용물 분석에 특화된 외부 유전자 제거 전처리 방법은 농도와 처리시간 조절 등의 응용방법에 따라 다양한 동물플랑크톤 분류군에 적용이 가능할 것으로 사료된다.

본 연구에서 multiplex PCR과 real-time PCR을 이용하여 창난젓의 원료를 감별할 수 있는 새로운 판별법을 개발하였다. 명태와 가이양의 종 특이 프라이머를 디자인하고, 명태와 가이양의 genomic DNA를 template로 single PCR과 multiplex PCR을 실시하였다. PCR을 실시한 결과, single PCR에서 명태(297 bp)와 가이양(132 bp)에 해당하는 PCR 밴드를 확인하였으며 교차 반응이 일어나지 않는 것을 확인하였다. Multiplex PCR에서 명태와 가이양 사이에 교차 반응없이 증폭이 일어나는 것을 확인하였다. Real-time PCR 결과, 명태 종 판별 프라이머에서 명태의 Ct 평균값은 20.765±0.691, 가이양 시료에서 Ct 평균값은 35.719±1.828이 었으며, 가이양 종 판별 프라이머에서 명태 시료의 Ct 평균 값은 35.996±1.423, 가이양 시료의 Ct 평균값은 20.096±0.793 으로 프라이머의 효율성, 특이성 및 교차 반응성에서 유의한 차이가 나타났다. 이러한 결과를 바탕으로 시중에서 판매되는 7개 제품을 multiplex PCR 및 real-time PCR로 확인 하였으며, 모든 시료에서 유효한 결과를 확인하였다. 본 연구에서 제작된 명태와 가이양에 대한 종 특이적 프라이머는 가공된 젓갈 시료의 원료의 판별 가능하며, 이러한 결과는 식품안전관리에 기여할 수 있을 것으로 기대된다.

참돔은 등쪽에 푸른 반점이 흩어져 있고, 홍민어는 꼬리 쪽에 검은 점이 있어 원물 형태는 쉽게 구분할 수 있다. 그러나 횟감이나 필렛으로 이용 시 참돔과 홍민어를 구분하기 어려워, 참돔의 종 특이 primer를 개발 및 검증하고 모니터링을 통해 참돔의 위변조 사례를 조사하고자 하였다. 시료에서 gDNA를 추출하여 PCR을 진행하였으며 참돔 primer의 product size는 468 bp, 홍민어 primer의 product size는 181 bp으로 설계하였다. Multiplex PCR 결과 참돔과 홍민어에 대한 종 특이적 증폭이 확인되었다. 또한, 참돔과 홍민어 PCR 민감도 실험 결과 참돔 primer는 1 ng, 홍민어 primer는 0.1 ng 까지 밴드가 확인되었다. 모니터링 결과 참돔으로 구매한 시료 19건 모두 참돔으로 판정되었다. 따라서 본 연구에서 제작된 참돔과 홍민어 종에 대한 종 특이적 primer는 횟감 등 수산물에도 적용할 수 있어 현재 유통 및 판매되고 있는 참돔 및 홍민어 판 별에 적합성을 확인하였다.

본 연구는 PCR 기반 RFLP (Restriction Fragment Length Polymorphism; 제한절편 길이 다형성) 분자기법을 활용하여 난 및 치어 대상 납자루아과 어류 3종의 동정을 좀 더 빠르고 정확하게 파악하고 납자루아과 어류의 종별 산란양상 및 번식생태 이해에 대한 기여가 목적이다. 본 연구를 위해 기존 선행된 문헌자료를 확인하고 납자루아과 어류가 2종 이상 동서하고 있는 지역을 확인하여 현지조사를 수행하였다. 현지조사 결과 확인된 납자루아과 어류는 묵납자루 (Acheilognathus signifer), 줄납자루 (A. yamatsutae) 및 각시붕어 (Rhodeus uyekii)로 총 3종이 확인되었으며, 확인된 납자루아과 어류와 동서하고 있는 숙주조개 (작은말조개; Unio douglasiae sinuolatus)를 채집하여 숙주조개 속 납자루아과 어류의 난 및 치어를 확보하였다. 현지조사 결과 확인된 납자루아과 어류 3종을 대상으로 미토콘드리아 DNA COI과 cyt b 유전자 염기서열을 비교하여 각각 종별로 특이성을 지닌 부위 (단일염기변이; Single Nucleotide Variation: SNV)에 맞는 제한효소를 선정하였고, 숙주조개 속 난 및 치어를 대상으로 genomic DNA를 추출하여 PCR-RFLP 실험을 수행한 결과 현지조사 시 확인된 납자루아과 어류 3종의 독특한 제한절편 길이 양상을 전기영동을 통하여 확인하였다. 본 연구를 통해 묵납자루, 줄납자루 및 각시붕어의 종을 판별할 수 있는 RFLP 마커를 개발하였으며, 숙주조개 난 및 치어를 대상으로 정확한 종의 동정을 보다 빠르고 효과적으로 수행하여 각각 납자루아과 종별 산란양상을 보다 정확히 규명하고 향후 이들 자연개체군의 효과적인 유지, 관리 및 보전 방법 개발에 유용하게 활용될 수 있을 것으로 판단된다.

This paper validates a system identification method using mathematical optimization using sea trial measurement data as a benchmark. A fast time simulation tool, SIMOPT, and a Rheinmetall Defence mathematical model have been adopted to conduct initial hydrodynamic coefficient estimation and simulate ship modelling. Calibration for the environmental effect of sea trial measurement and sensitivity analysis have been carried out to enable a simple and efficient optimization process. The optimization process consists of three steps, and each step controls different coefficients according to the corresponding manoeuvre. Optimization result of Step 1, an optimization for coefficient on x-axis, was similar compared to values applying an empirical regression formulae by Clarke and Norrbin, which is used for SIMOPT. Results of Steps 2 and 3, which are for linear coefficients and nonlinear coefficients, respectively, was differ from the calculation results of the method by Clarke and Norrbin. A comparison for ship trajectory of simulation results from the benchmark and optimization results indicated that the suggested stepwise optimization method enables a coefficient tuning in a mathematical way.

Monochamus alternatus (M. alternatus) and Monochamus saltuarius (M. saltuarius) are major vectors for Bursaphelenchus xylophilus in South Korea. When an adult, they are easily distinguishable by several morphological classification. However, it is difficult to identification between M. alternatus and M. saltuarius when they are larvae as they have very similar morphological characters. Thus, they are not easily distinguishable without expertise about Cerambycidae taxonomy. Furthermore, during epidemiological investigation, sometimes, adults or larvae would not be founded in death pine trees. For these reasons, in this experiment, we are able to identified between M. alternatus and M. saltuarius by mitochondrial 12S rRNA gene primers that are specific to 12S rRNA gene fragment of M. alternatus using larvae tissue and frass. Moreover, we had examined whether vectors that were already escaped from dead pine tree have Bursaphelenchus xylophilus or not by multiplex PCR using larva frass that was remained in dead pine tree.

The rapid growth of complexity and scale can be witnessed in the design and development of modern systems. As such, the severity of damages in the occasional accidents has attracted great deal of attention lately. Although a variety of methods have so far been studied to overcome or reduce the disastrous results of hazards, the issues seem still persistent and even complicated due to the situation mentioned above. The concept of functional safety has been regarded as one approach to handling the matters by shifting up to the functions level from the consideration of each physical component itself. The outcomes of those efforts would be the international standards on functional safety such as IEC 61508 and its relatives including IEC 62278, EN 50128, ISO26262, and so on. In this paper, a method of how hazards can be analyzed to be coped with those standards has been studied. In the method proposed, the systems modeling language (SysML) is playing a key role to model and analyze the hazards from the viewpoint of functional safety. The approach taken has been applied in the analysis of the hazards in railroad systems. In spite of focusing on the individual components hazards, the method based on functional safety has analyzed them collectively with the added effect of identifying the cause originated from the interface between the functions.