The recent increase in the occurrence of common bed bug and tropical bed bug in shared areas highlights the need for rapid species identification at infestation sites, which is crucial for implementing targeted control measures due to differences in genetic and physiological traits. In this study, molecular diagnostic methods were developed using species-specific ITS2 sequences. Both multiplex PCR and loop-mediated isothermal amplification (LAMP) protocols with a DNA release method successfully distinguished between the two bed bug species regardless of developmental stages in 0.5~2.5 hours, even with dead specimens. Especially, LAMP's simplicity and speed make it applicable for rapid and accurate bed bug diagnosis at infestation sites.

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.

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.

A single postlarva (9.72 mm in standard length) specimen of Crossorhombus azureus (Alcock, 1889) belonging to the family Bothidae, was collected using a bongo net from the southern sea of Korea on December 10, 2019. This species is characterized by having spines on post basipterygial processes. It has two eyes located at the opposite side in head, which accordingly belongs to pre-metamorphosis stage. Melanophores are distributed on the dorsal and anal fin base on the right side (blind side), which is regarded as a useful identification key distinguishing C. azureus from congeneric species in their postlarval stage. A molecular analysis based on mitochondrial DNA COI sequences showed that our specimen was closely matched to adult C. azureus (K2P distance = 0.017). As there is no Korean name for the genus Crossorhombus in spite of presence of Crossorhombus kobensis and its Korean name “Go-be-dung-geul-neob-chi” in Korea, we proposed a new Korean name “Dung-geul-neob-chi-sog” for the genus Crossorhombus and “Pa-lang-dung-geul-neob-chi” for the species C. azureus.

Doublesex and mab-3 related transcription factor (dmrt) play crucial roles in sex determination and sex differentiation in vertebrates and invertebrates. Although dmrt genes have been identified in vertebrates, little is known about aquatic invertebrates. In this study, two dmrt genes, namely, Dc_dmrt93B and Dc_dmrt99B, were identified from brackish water flea, Diaphanosoma celebensis. Transcriptional changes were observed in the dmrt genes when the flea was exposed to bisphenol (BP), an endocrine disruptor. Sequence and phylogenetic analyses showed that both dmrt genes contained two conserved domains, namely, DM and DMA, closely clustered with those of Daphnia spp. Additionally, a significant increase in the Dc_dmrt99B mRNA expression level was observed upon exposure to intermediate concentrations of BP (bisphenol A>bisphenol S=bisphenol F, p<0.05), while the expression of Dc_dmrt93B mRNA was slightly modulated. These findings imply that the two dmrt genes may be involved in sex differentiation of D. celebensis. Furthermore, it was found that the ability of BP to modulate dmrt genes could affect development and reproduction. This study provides a basis for understanding the function of the dmrt genes and the molecular mode of action of BP in small crustaceans.

During the ichthyoplankton survey around the Chilsan Island (previously known as the biggest spawning ground of the Sciaenidae) in the southwestern sea of Korea from April to June in 2019 using ring nets, we collected a total of 12 individuals belonging to the family Sciaenidae. Using molecular methods, we identified five sciaenid species (Collichthys lucidus, Collichthys niveatus, Johnius grypotus, Nibea albiflora, and Pennahia argentata), and described and compared them on the basis of the preflexion larval stage. C. lucidus was well distinguished by the presence of occipital crests at preflexion stage. Although there were no occipital crests, preflexion larva of C. niveatus was distinguished by the absence of melanophore except for the upper part of the abdominal cavity. J. grypotus and N. albiflora were very similar morphologically, but were distinguished by myomere height (15.22-15.53% in J. grypotus vs. 11.66-12.78% in N. albiflora) in the percentage of notochord length, and eye diameter (32.58-33.37% in J. grypotus vs. 40.32-42.53% in N. albiflora) in the percentage of head length between specimens of similar size (J. grypotus: 3.22-3.23 mm, N. albiflora: 3.04-3.13 mm). P. argentata were distinguished by distribution of ventral caudal melanophore (one row of small spot in P. argentata vs. irregular patches on the central part of caudal in J. grypotus and N. albiflora). Comparative morphological studies using more diverse species must be conducted for more comprehensive understanding of the morphogenesis of Sciaenidae.

이취미 물질인 2-MIB를 합성하는 원인종에 대한 정보는 담수생태계에서 이와 관련된 환경 및 경제적 문제를 해결하는 데 필수적이다. 본 연구는 북한강 수계에서 출현하는 Pseudanabaena strain을 분리·배양하고, 16S rDNA 염기서열을 이용하여 종 수준의 동정과 2-MIB 합성 유전자 탐색을 통해 이취미 물질 발생 잠재성을 분석하였다. 북한강 본류 지역 (삼봉리, 조암면, 의암호 지역)에서 분리한 Pseudanabaena strain은 총 11개로서 NHUA201911과 NHPD201909 strain을 제외하고 단일세포의 크기는 서로 유사하였다. 그러나 16S rDNA 계통분석을 통한 유전자 염기서 열의 유연관계를 분석한 결과, 분리된 strain들은 총 5개 종 (P. cinerea, P. yagii, P. mucicola, P. galeata, P. redekei)으 로 분류되었다 (40~55% 유사도). 2-MIB를 합성하는 mibC 유전자는 P. cinerea 07 strain (NHUA202007-07)와 P. yagii (NHUA202007-08), P. redekei (NHUA201911)에서만 발견 되었으며, 가스크로마토그래피 분석에 따라 실질적인 2-MIB 합성은 P. cinerea와 P. redekei 종에서 확인되었다. 본 연구 결과는 분자생물학적 수준에서 북한강 수역에서 발생하는 Pseudanabaena속 남조류의 다양도에 대한 증거를 제공하는 연구로서 북한강 수계에서 2-MIB 생산 원인종에 대한 중요한 정보를 제공한다.

A single juvenile malacanthid specimen (Hoplolatilus chlupatyi) was collected from the South Sea of Korea and identified by DNA barcoding. This species is readily distinguished from other malacanthid species in having well developed rostral spine and elongated spines in the posttemporal and preopercle. A molecular analysis based on mitochondrial DNA COI sequences showed that this species is matched to adult H. chlupatyi (genetic distance = 0.005). Therefore, this is the first record of H. chlupatyi in Korea. We propose new Korean names, “Mu-ji-gae-og-dom-sog” for the genus Hoplolatilus and “Mu-ji-gae-og-dom” for the species H. chlupatyi.