The oriental fruit fly is a major polyphagous insect pest with a worldwide distribution. We investigated the development time, survivorship, longevity and fecundity of B. dorsalis at six different temperatures. We applied the computer programs – TWOSEX-MSChart and TIMING-MSChart - to analyze our data. We obtained the population parameters – adult preoviposition period (APOP), total preoviposition period (TPOP), oviposition days, eggs per reproductive female, first age of survival rate <50%, proportion of male and female individuals, propoortion of N-type individual, and life table parameters (net reproductive rate, intrinsic rate of increase, finite rate of increase, and mean generation time) - and population projection using two computer programs.

The oriental fruit fly is a major polyphagous insect pest with a worldwide distribution. The effects of temperature on stage-specific development were investigated at eight constant temperatures (13.0, 14.4, 16.2, 19.5, 23.8, 27.7, 31.8 and 34.8℃). Stage-specific lower developmental thresholds and thermal constants were determined using linear regression. The lower and higher temperature threshold (TL and TH) were estimated using the Sharpe-Schoolfield-Ikemoto (SSI) model. The daily adult emergence frequency of B. dorsalis was estimated in relation to adult age and temperature. Thermal performance was compared among B. dorsalis populations from different locations in Taiwan.

The dynastid beetle Allomyrina dichotoma has been used as a herbal medicine. Recently, we performed de novo RNAsequencing of Allomyrina dichotoma and identified several antimicrobial peptide candidates based on in silico analysis.Among them, cationic antimicrobial peptide, Allomyrinasin, was selected and we assessed the anti-inflammatory activitiesof Allomyrinasin against mouse macrophage Raw264.7 cells. The results showed that Allomyrinasin decreased the nitricoxide production of the lipopolysaccharide-induced Raw264.7 cells. In addition, quantitative RT-PCR, ELISA and Westernblot analysis revealed that Allomyrinasin reduced cytokine expression levels in the Raw264.7 cells. Taken together, thesedata indicated that Allomyrinasin had anti-inflammatory activity in the lipopolysaccharide-induced Raw264.7 cells.

Previously, we have performed de novo RNA sequencing of Scolpendra subpinipes mutilans using next generation sequencing technology and identified several AMP candidates. Among them, a synthetic peptide (scolopendrasin I) was designed based on SVM algorithm. In this study, we reported that the synthetic peptide scolopendrasin I had an antimicrobial and anticancer activity. As a result, scolopendrasin I showed antibacterial activities against Gram positive and Gram negative bacteria strains in radial diffusion assay and colony count assay without hemolytic activity. In addition, we confirmed that scolopendrasin I bound to the surface of bacteria via a specific interaction with lipoteichoic acid and lipopolysaccharide, which is one of bacteria cell membrane components. In addition, we found that scolopendrasin I had anticancer activities in the human leukemic T lymphocyte cell line Jurkat using MTS assay. In conclusion, our results suggested that scolopendrasin I could be useful for developing peptide antibiotics and anticancer agents.

This study describes the developmental process of gonads in chameleon goby, Tridentiger trigonocephalus from the stage of hatching to 100 days after hatching (DAH). Based on histological observation, the primordial germ cells were observed in mesentery between mesonephric duct and gut at 15 DAH (total length, TL: 6.8±0.2 mm). At 20 DAH (TL: 7.9±0.1 mm), the primordial gonad began to protrude into peritoneal cavity and developed between mesonephric duct and gut. Initial ovarian differentiation was identified by the presence of ovarian cavity and oogonia in the gonads at 55 DAH (TL: 21.1±1.3 mm). Testicular differentiation started at 65 DAH (TL: 23.7±0.9 mm) with appearance of spermatogonial cells in the gonads. These findings indicate that sex differentiation in T. trigonocephalus occurs earlier in females than males, suggesting that this species can be classified as an undifferentiated gonochorist.

The objective of this study was to characterize the reproductive cycle of the chameleon goby, T. trigonocephalus. Gonadal development was investigated using a histological method. Specimens were collected monthly, from April 2009 to March 2010. The gonadosomatic index (GSI) of females began to increase in April, reaching the maximum in May, and declined sharply in August. In males, the GSI began to increase in April and reaching the maximum in July. The annual reproductive cycle of T. trigonocephalus can be divided into four successive stages in females: the growing (November- March), maturing (April-May), ripe and spawning (June-July), and recovery (August-October) stages. Males passed through growing (November-March), maturing (April-June), ripe and spermiation (July-August), and recovery (September-October) stages. These results indicate the spawning season is from June to July. The relationship between fecundity (Fc) and body length (BL) was Fc=86.1511BL2.6506. Fecundity was ranged from 3,448-9,654 eggs in a BL of 4.8-7.2 cm and it was increased as BL increased.

TCR subunits are members of membrane-bound receptors which allow the fast and efficient elimination of the specific fish pathogens have regulated function in adaptive immunity. Sequence structure of TCR subunits have been reported for various teleosts, but the information of each TCR subunit functional characterization through expression analysis in fish was unknown. In this study, we examined the gene expression of TCR subunits in the early developmental stages and observed transcript levels in various tissues from healthy adult olive flounder by RT-PCR. The mRNA expression of alpha subunit was already detected in the previous hatching step. But the transcripts of another TCR subunit were not observed during embryo development and increased after hatching and maintained until metamorphosis at the same level. It was found that all TCR subunits mRNAs are commonly expressed in the immune-related organ such as spleen, kidney and gill, also weak expressed in fin and eye. TCR alpha and beta subunit were expressed in brain, whereas gamma and delta were not expressed same tissue. The sequence alignment analysis shows that there are more than 80% sequence homology between TCR subunits. Because it has a high similarity of amino acid sequence to expect similar in function, but expression analysis show that will have may functional diversity due to different time and place of expression.

The innate immune system is the only defense weapon that invertebrates have, and it is the fundamental defense mechanism for fish. The innate immune response is important in newly hatched flounders because it is closely involved in the initial feeding phase, which is why it is essential for survival during the juvenile period. The expression analysis of genes involved in the innate immune response in the olive flounder (Paralichthys olivaceus) in the days after hatching is incomplete. Therefore, we have begun to examine the expression patterns of genes specifically induced during the development of the innate immune system in newly hatched flounders. Microscopic observation showed that pronephron formation corresponded with the expression of perforin-encoding gene. These results suggest that perforin plays a vital role in the innate immunity of the kidney during developmental stages. Perforin expression was strong at the start of the development of the innate immune response, and continued throughout all the development stages. Our findings have important implications with respect to perforin’s biological role and the evolution of the first defense mechanisms in olive flounder. Further studies are required to elucidate the perforin-mediated innate immunity response and to decipher the functional role of perforin in developmental stages.

Fish larvae are immediately exposed to microbes from hatching to maturation of their lymphoid organs, therefore effective innate mechanisms is very important for survival in such an environment. The key component of innate immune system, C3 is central protein of all activation pathways of the complement system, leading to inflammatory reactions, such as opsonisation, chemotaxis, and cell lysis of pathogens. Although, innate mechanisms is essential for survival in the early stage of development, little is known about defence mechanisms. In this study, the alignment analysis showed that amino acid sequence of C3 from olive flounder liver EST homologous to other known C3 sequences with 73-99% identity. Also, we examined the tissue distribution of olive flounder C3 and analyzed expression pattern from the fertilized egg until 28 days post hatching. As a result, olive flounder C3 mRNA was expressed only in the liver and the mRNA level more increased as developmental proceed during the early stage. These results may suggest that olive flounder C3 plays an important function in the early immune response of olive flounder larvae.

Olive flounder (Paralichthys olivaceus) is one of the commercial important flatfish species in Korea. The ocular signal transduction pathway is important in newly hatched flounders because it is closely involved in the initial feeding phase thus essential for survival during the juvenile period. However, the study of gene expression during ocular development is incomplete in olive flounder. Therefore we examined the expression analysis of specifically induced genes during the development of the visual system in newly hatched flounders. We searched ocular development-involved gene in the database of expressed sequence tags (ESTs) from olive flounder eye and this gene similar to arrestin with a partial sequence homology. Microscopic observation of retinal formation corresponded with the time of expression of the arrestin gene in the developmental stage. These results suggest that arrestin plays a vital role in the visual signal transduction pathway of the retina during ocular development. The expression of arrestin was strong in the ocular system during the entirety of the development stages. Our findings regarding arrestin have important implications with respect to its biological role and evolution of G-protein coupled receptor (GPCR) signaling in olive flounder. Further studies are required on the GPCR-mediated signaling pathway and to decipher the functional role of arrestin.

Cathepsins are members of the multigene family of lysosomal cysteine proteinases and have regulated function in several life processes. The potential role of cathepsin F cysteine gene was expected as protease in the yolk processing mechanism during early developmental stage, but expression analysis was unknown after fertilization. The alignment analysis showed that amino acid sequence of cathepsin F from olive flounder liver expressed sequence tag (EST) homologous to cathepsin F of other known cathepsin F sequences with 87－98% identity. In this study, we examined the gene expression analysis of cathepsin F in various tissues at variety age flounder. Tissue distribution of the cathepsin F mRNA has been shown to be ubiquitous and constitutive pattern regardless of age in each group, although derived from cDNA library using liver sample. The mRNA level of cathepsin F more increased as developmental proceed during embryogenesis and early developmental stage, especially increased in the blastula, hatching stage and 3 days post hatching (dph). As a result, it may suggest that the proteolysis of yolk proteins (YPs) has been implicated as a mechanism for nutrient supply during early larval stages in olive flounder.

To evaluate the effects of benzo[a]pyrene (B[a]P), one of polycyclic aromatic hydrocarbons (PAHs), on in vitro oocyte maturation (GVBD) and sex steroid hormone production, maturing oocytes (oocyte diameters=0.74, 0.88 and 0.93 mm) of the longchin goby, Chasmichthys dolichognathus were incubated with B[a]P (1, 10 and 100 ng/mL) for 24 hours. After incubation, the oocytes were fixed with clearing solution (ethanol:formalin:glacial acrtic acid=6:3:1). The location of the germinal vesicle was observed under low-power magnification using a dissecting microscope. Steroids in aliquots of the incubation media were extracted twice using five volumes of ethylacetate:cyclohexane (1:1). Then, the T, E2 and 17α20βP levels were measured by RIA. In oocytes 0.74 mm diameter (vitellogenic oocytes), B[a]P had no significant effect on GVBD at the concentrations tested. In oocytes 0.88 mm diameter (fully vitellogenic oocytes), B[a]P inhibited GVBD significantly at 1 and 100 ng/mL. T production was decreased and the ratio of E2/T was increased significantly at 1 and 10 ng/mL compared with control. In 0.93 mm diameter oocytes (germinal vesicle located near the center of oocytes), B[a]P induced GVBD significantly at 10 and 100 ng/mL and decreased the ratio of E2/T significantly at 1 and 10 ng/mL compared with control. These findings suggest that B[a]P has different sensitivity to the oocyte maturation according to the oocyte diameters.

We investigated the changes in plasma sex steroid hormones, testosterone (T), estradiol-17β (E2), 17,20 β-dihydroxy-4-pregnen-3-one (17α20βOHP), 11-ketotestosterone (11KT) and cortisol levels from ribbed gunnel, Dictyosoma burgeri in associated with annual reproductive cycle. The fish were collected at the coastal waters of Dadaepo, Busan, Korea from August 2002 to March 2003. Body weight, liver weight and gonad weight were measured for calculation of hepatosomatic index (HSI) and gonadosomatic index (GSI). Plasma steroid and cortisol levels were measured by radioimmunoassay and data were analyzed using the Kruskal-Wallis test and the Spearman rank correlation. The GSI of females increased from November, peaked in February (13.21±2.27) and decreased rapidly from March (1.05±0.23). The GSI of males also increased from November, peaked in December (0.36±0.12) and remained high to February. In females, E2 levels increased and remained high from December to February (19.81±7.46～22.85±6.18 ng/ ㎖). The levels of T showed a similar tendency and correlated (rs=0.813, p=0.0001) significantly with E2 levels. The levels of 17α20βOHP increased rapidly in February (4.78±1.01 ng/㎖) and peaked in July (5.08±0.65 ng/㎖). Cortisol level was peaked in March (2081.07±1140.02 ng/㎖) and correlated significantly with 17α20βOHP levels (rs=0.790, p=0.0001). In males, the levels of T was peaked in January (2.43±0.17 ng/㎖) and then decreased rapidly. The levels of 11KT were remained high from October to January (3.27±1.55～4.28±1.53 ng/㎖) and correlated significantly with T levels (rs=0.513, p=0.002). On the other hand, the levels of 17α20βOHP fluctuated during reproductive cycle and the variation pattern of cortisol levels were similar to females. These results suggest that plasma sex steroids in ribbed gunnels have annual periodicity, and that cortisol may involve in maturation of females.

본 연구에서는 해산어를 이용하여 bisphenol A(BPA)와 nonylphenol(NP)이 난모세포 성숙 과정에 어떤 영향을 미치는지 조사하기 위해 성숙단계에 있는 노래미(Hexagrammos agrammus) 난모세포(난경 약 1.88 ㎜)를 대상으로 in vitro에서 BPA와 NP 처리에 의한 난모세포의 성스테로이드 생성농도를 조사하였다. 난모세포에 BPA와 NP를 농도구별(0.1, 1, 10, 100, 1,000 ng/㎖)로 첨가하고, 50 IU의 human chorionic gonadotropin(HCG)를 농도구별 BPA 또는 NP와 함께 첨가하거나 하지 않고 48시간 동안 배양하였다. 배양 후 배양액 내의 17α,20β-dihydroxy-4-pregnen-3-one(17α20βOHP), estradiol-17β(E2) 그리고 testosterone(T)의 농도를 방사면역측정법(RIA)을 통해 정량하였다. BPA 처리구에서는 100 ng/㎖의 농도구에서 HCG 처리 유무에 상관없이 E2 생성이 촉진되었다. HCG 처리하에서 0.1 ng/㎖의 농도구에서 T 생성은 촉진되었으나, HCG를 처리하지 않은 실험구의 모든 농도구에서 T 생성은 저해되었다. NP 처리구에서는 HCG를 처리하지 않은 실험구의 10 ng/㎖의 농도구에서 17α20βOHP와 T 생성이 촉진되었고, 1 ng/㎖의 농도구에서는 E2 생성이 억제되었다. 이상의 결과를 종합하면, 노래미의 성숙단계의 난모세포에서 BPA는 약한 estrogen-agonistic 효과를, NP는 estrogen- antagonistic 효과를 지니는 것으로 사료된다.