• Journal of Microbiology and Biotechnology
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• v.27 no.1
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• pp.112-121
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• 2017

Edwardsiella tarda is a gram-negative pathogenic bacterium in aquaculture that can cause hemorrhagic septicemia in fish. Many secreted proteins have already been identified as virulent factors of E. tarda. Moreover, since virulent phenotypes are based on the expression regulation of virulent genes, understanding the expression profile of virulent genes is important. A quantitative RT-PCR is one of the preferred methods for determining different gene expressions. However, this requires the selection of a stable reference gene in E. tarda, which has not yet been systematically studied. Accordingly, this study evaluated nine candidate reference genes (recA, uup, rpoB, rho, topA, gyrA, groEL, rpoD, and 16S rRNA) using the Excel-based programs BestKeeper, GeNorm, and NormFinder under different culture conditions. The results showed that 16S rRNA was more stable than the other genes at different culture growth phases. However, at the same culture time, topA was identified as the reference gene under the conditions of different strains, different culture media, and infection, whereas gyrA was identified under the condition of different temperatures. Thus, in experiments, the expression of gapA and fbaA in E. tarda was analyzed by RT-qPCR using 16S rRNA, recA, and uup as the reference genes. The results showed that 16S rRNA was the most suitable reference gene in this analysis, and that using unsuitable reference genes resulted in inaccurate results.

• Proceedings of the Korea Society of Environmental Toocicology Conference
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• 2003.05a
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• pp.181-182
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• 2003

Exposure to some synthetic environmental chemicals and their metabolites cause reproductive problems in a variety of vertebrate via endocrine mechanisms. However, in most cases, the link between these compounds and adverse effects on humans, fish, and wildlife has not been established, which necessitates a closer look at the molecular, functional, and clinical implications of these chemicals in the environment. Calbindin-D9k (CaBP-9k) is a member of a largo family of intracellular calcium binding proteins that have high affinities for calcium. It was reported that the estrogen level of uterus affected the expression of the CaBP-9k gene in rat uterus. We examined the dose-dependent CaBP-9K gene expression in the uterus for three-days injection of methoxychlor (HC) in the overectomized immature rats and the relation with uterotrophic response of the compoundsand compared the responses induced by MC according to the route of administration.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.48 no.4
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• pp.417-425
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• 2015

Semi-dried conger eel Conger myriaster is considered as a health food in Korea due to its richness in vitamins, minerals, proteins and omega-3 fatty acids. This study characterize the sanitary quality of commercial semi-dried conger eel for developing high quality semi-dried products and suggested the guideline for controlling the quality. Moisture content ranged from 41.2% to 73.4% (mean: 61.5%), volatile basic nitrogen ranged from 16.1 to 93.6 mg/100 g (mean: 55.1 mg/100 g), and peroxide value ranged from 15.2 to 69.8 meq/kg. Viable cell counts ranged between 6.51 and 8.53 log CFU/g, while the Escherichia coli count ranged from undetectable to 4.6 log CFU/g. Based on these chemical and microbial findings, we suggest that provisions be established for development of high quality semi-dried conger eel as follows: 50-68% for moisture content, < 50 mg/100 g for volatile basic nitrogen content, < 60 meq/kg for peroxide value, and negative for E. coli. Among the 16 commercial semi-dried conger eel products used in this study, the standard-passed product was only GS-L.

• Environmental Analysis Health and Toxicology
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• v.28
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• pp.16.1-16.8
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• 2013

Objectives Fish vitellogenin (VTG) is produced in the female liver during oogenesis through the estradiol cycle and produced in the male liver by endocrine disrupting chemicals (EDCs) such as alkylphenols. In this study, we propose that the VTG concentration in the pale chub could be detected using monoclonal antibodies and polyclonal antibodies against vitellin (Vn) in a VTG enzyme-linked immunosorbent assay (ELISA) system. Methods Monoclonal antibodies and polyclonal antibodies were produced using the Vn extracted from the matured ovum of the ovary. The VTG was extracted from the plasma of the male pale chub. The Vn and VTG were confirmed by measuring the molecular weight of their proteins using sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and the specificity of the antibodies was checked through western blotting methods. The assay system was validated with respect to optimal assay concentrations, specificity, recovery, and intra- and inter-assay variations. Results The Vn consisted of two protein bands with apparent molecular weights of 64 and 37 kDa. The SDS-PAGE indicated protein weights of 146 and 77 kDa in the VTG. The assay range was 15.6 ng/mL to 2,000 ng/mL, and the value of the intra- and inter-assay variations were within 10.0% and 14.7%, respectively. The recovery rate was $99.5{\pm}5.5%$. Conclusions A sandwich ELISA was developed that could be used to qualify the VTG of pale chub in screening for EDCs. Pale chub is an ideal species for observing estrogen activity in the environment because of its extensive habitat and extensive food chain. The ELISA developed here would be more favorable than those for other species for determining the effect of long-term food chain accumulation of EDCs in aquatic environments.

• Journal of Nutrition and Health
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• v.21 no.1
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• pp.36-46
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• 1988

This study was carried out to investigate the effects of different protein and fat levels on the growth and on immune response in rats. In experiment 1, Sprague-Dawly male rats were fed diets containing 6%, 15%, or 30% casein with 2 levels of fat(2% and 30%) at each protein level. In experiment 2 and 3, rats were devided into 8 diet groups ; 4 different sources of proteins(casein, meat protein, fish protein, and gluten) were used at 15% level of the diet with 2% or 30% of dietary fat. The results show as follows 1) The rats in 6% casein group showed lower body weight gain and organ weight than those in 15% and 30% casein groups. There was no significant difference between 15% and 30% casein groups. In experiment 2, the gluten diet group showed the lowest growth rate and epididymal fat pad weight among 4 different dietary protein groups regardless the level of dietary fat. 2) There was no significant difference in immune response according to the sources and levels of dietary protein. However, the rats fed high fat diet showed the lower plaque-forming cell response than those fed low fat diet regardless dietary protein.

• Journal of Aquaculture
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• v.9 no.1
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• pp.93-100
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• 1996

Previously, as an effort to make an autonomously replicating expression vector in fish, an ARS (autonomously replicating sequence) was cloned from MAR (matrix attachment region) of Misgurnus mizolepis. The DNA fragment composed of 443 base pairs contains ARS core consensus sequences, topoisomerase II consensus sequences, and A or T box sequences which are homologous to the known consensus sequences originated from other organisms. The clond ARS, as other DNA replication origins, contains inverted repeat sequences and several potential hairpin loop structures. These consensus sequences and hairpin structures may serve as recognition signals for regulatory proteins of DNA replication initiation.

• Asian-Australasian Journal of Animal Sciences
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• v.13 no.8
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• pp.1178-1187
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• 2000

Many immunostimulating substances have been developed to improve immunity of domestic animals, although their exact mode of action and effects are not clearly defined, and they are now widely used in feed industry. Bacterial lipopolysaccharides, called endotoxin, in particular may have a profound effect not only on the immune system but also on macrophages of the reticuloendothelial system. Glucans from a variety of yeast cell wall have been shown to stimulate both specific and non-specific immune responses and to increase growth performance in pigs. Recently, there has been great interest in the role of complex carbohydrates in disease prevention and treatment. Mannanoligosaccharide is a glucomannoprotein complex derived from the cell wall of yeast. Generally, it was also known that the deficiencies of some major vitamins (vitamin A, E and C) and minerals (chromium and selenium) lead to impaired immune system and, as a result, immune function is depressed and recovery delayed. On the other hand, many researchers suggested that one possible reason for the superior performance observed in pigs fed plasma protein may be because of the presence of biologically active plasma proteins (e.g., immunoglobulins) which are known to contribute to the health of the starter pig. And, immunoglobulins present in plasma protein have been implicated as contributing to the overall immunocompetence of the newborn pig. Other immunostimulants, lactoferrin and lysozyme, mainly found in milk and egg white, have been known as having bacteriocidal and bacteriolytic effect. When considering practical use of immunostimulants, the concept of using immunostimulants is new to many people and, in most cases, it is poorly understood how and why such compounds act, and how they should be used in practice. Therefore, in order to clarify the reason for discrepancies in results, special attention should be paid to the dose/response relationship of immunostimulants and the duration of the effect.

• Journal of Microbiology and Biotechnology
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• v.23 no.6
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• pp.811-817
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• 2013

The present study dealt with the isolation, identification and enzyme characterization of potential luminous bacteria from water, sediment, squid, and cuttle fish samples of the Karaikal coast, Bay of Bengal, India during the study period September 2007 - August 2008. Bioluminescent strains were screened in SWC agar and identified using biochemical tests. As Shewanella henadai was found to be the most common and abundant species with maximum light emission [69,702,240 photons per second (pps)], the optimum ranges of various physicochemical parameters that enhance the luciferase activity in Shewanella hanedai were worked out. The maximum luciferase activity was observed at the temperature of $25^{\circ}C$ (69,674,387 pps), pH of 8.0 (70,523,671 pps), salinity of 20 ppt (71,674,387 pps), incubation period of 16 h (69,895,714 pps), 4% peptone (70,895,152 pps) as nitrogen source, 0.9% glycerol (71,625,196 pps), and the ionic supplements of 0.3% $CaCO_3$ (73,991,591 pps), 0.3% $K_2HPO_4$ (73,919,915 pps), and 0.2% $MgSO_4$ (72,161,155 pps). Shewanella hanedai was cultured at optimum ranges for luciferase enzyme characterization. From the centrifuged supernatant, the proteins were precipitated with 60% ammonium sulfate, dialyzed, and purified using anion-exchange chromatography, and then luciferase was eluted with 500 mM phosphate of pH 7.0. The purified luciferase enzyme was subjected to SDS-PAGE and the molecular mass was determined as 78 kDa.

• Molecules and Cells
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• v.37 no.8
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• pp.592-597
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• 2014

The cell membrane provides critical cellular functions that rely on its elaborate structure and organization. The structure of turtle membranes is an important part of an ongoing study of erythrocyte membranes. Using a combination of atomic force microscopy and single-molecule force spectroscopy, we characterized the turtle erythrocyte membrane structure with molecular resolution in a quasi-native state. High-resolution images both leaflets of turtle erythrocyte membranes revealed a smooth outer membrane leaflet and a protein covered inner membrane leaflet. This asymmetry was verified by single-molecule force spectroscopy, which detects numerous exposed amino groups of membrane proteins in the inner membrane leaflet but much fewer in the outer leaflet. The asymmetric membrane structure of turtle erythrocytes is consistent with the semi-mosaic model of human, chicken and fish erythrocyte membrane structure, making the semi-mosaic model more widely applicable. From the perspective of biological evolution, this result may support the universality of the semi-mosaic model.

• The Korean Journal of Food And Nutrition
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• v.26 no.3
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• pp.329-338
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• 2013

This study is conducted to compare the problems affecting dietary habits and health status between a long-term exercise group and non-exercise group of males older than 50 years. Most subjects of the two groups consider regular exercise to be the most important factor for maintaining health, and recognize hypertension as the number one concern. The most common nutritional supplement among subjects is multiple vitamins, and sleeping time range from 6~8 hours. Breakfast fasting rate was 15.9% among non-exercise group (NEG), where overeating and eating out rates were higher among exercise group (EG) when comparing to NEG. Fasting rates of breakfast, and consumption rates of milk and dairy products, vegetables such as kimchi, and fruits and fruit juices are higher among the EG. Overeating and eating out rates are lower among NEG, whereas eating rates of proteins like meat, fish, eggs, beans, fried foods, high-fat meats carbonated drinks, ice cream, and salt intakes are higher. Rates of health, dietary habits, and nutrient intakes are higher among EG. Energy intakes among the two groups were below the standard level, Vitamin $B_2$ intake is low in EG, and folic acid and calcium intakes were higher in NEG. The exercising group care more about health, eating habits and nutrient intakes, and low smoking rates are being observed. Thus, specific dietary improvement programs for adult men, are required, and it is necessary to consider the practice of nutrition education.