• Journal of Microbiology
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• v.44 no.5
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• pp.548-555
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• 2006

FaeG is the key factor in the infection process of K88ad enterotoxigenic Escherichia coli (ETEC) fimbrial adhesin. In an attempt to determine the possibility of expressing recombinant FaeG with immunogenicity for a new safe and high-production vaccine in E. coli, we constructed the recombinant strain, BL21 (DE3+K88), which harbors an expression vector with a DNA fragment of faeG, without a signal peptide. Results of 15% SDS-polyacrylamide slab gel analysis showed that FaeG can be stably over-expressed in BL21 (DE3+K88) as inclusion bodies without FaeE. Immunoglobulin G (IgG) and M (IgM) responses in pregnant pigs, with boost injections of the purified recombinant FaeG, were detected 4 weeks later in the sera and colostrum. An in vitro villius-adhesion assay verified that the elicited antibodies in the sera of vaccinated pigs were capable of preventing the adhesion of K88ad ETEC to porcine intestinal receptors. The protective effect on the mortality rates of suckling piglets born to vaccinated mothers was also observed one week after oral challenge with the virulent ETEC strain, $C_{83907}$ (K88ad, $CT^+,\;ST^+$). The results of this study proved that the adhesin of proteinaceous bacterial fimbriae or pili could be overexpressed in engineered E. coli strains, with protective immune responses to the pathogen.

• Journal of fish pathology
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• v.32 no.2
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• pp.105-111
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• 2019

Citurs unshiu Markovich is a medicinal product of dried tangerine peel (DTP). It is effective on antioxidation, and getting fame as a medicine and functional food. By utilizing DTP as a feed additive, we aim to enhance the growth rate, innate immunity, and bacterial infection resistance to Tilapia. The DTP extract was added to the feed weight by 0.1, 0.5, 1, 5% and then fed to tilapia for 7 days to evaluate the innate immunity parameter, growth rate and anti-bacterial activity. Innate immunity parameter results showed that the ROI was significantly higher in the 5% group added at high concentration, while showing decrease or no differences in other experimental groups. In other parameters, all the experimental groups showed no significant difference or decreased compared to the control group. The challenge test showed a high survival rate of 71% in the 0.5% group and the lowest in the control group (36%). For the growth rate, the feed efficiency was improved in all groups except for the 0.1% group compared to the control group. In conclusion, DTP extract has bacterial resistant effect in while not affecting innate immune system of fish. Also, it has shown the potential as a possible feed additive as it has brought the improvement on feed efficiency ratio.

• Journal of fish pathology
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• v.10 no.1
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• pp.45-52
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• 1997

A protein-bound polysaccharide preparation, PS-K, isolated from Coriolus versicolor was evaluated for its ability to enhance resistance against Edwardsiella tarda, causal agent of classical edwardsiellosis in tilapia, Oreochromis niloticus. Fish treated with a dose rate of 0.1 mg PS-K $g^{-1}$ body weight were challenged by intraperitoneal injection of E. tarda with different concentrations. Against low burdens of injections($1.2{\times}10^7$ and $1.2{\times}10^8$ cfu/fish), PS-K treated fish did not show any mortality compared to 50% and 100% mortality of control groups, respectively. However there was no increase of resistance against challenge with high concentrations of E. tarda ($1.2{\times}10^9$ and $1.2{\times}10^{10}$cfu /fish) except few days delaying of death. Tilapia injected with PS-K one day after intraperitoneal inoculation of E tarda($1.2{\times}10^7$ cfu/fish) showed 100% survival rate compared to control group of 50% survival rate. The result indicates the potential of PS-K as a prophylactic agent in aquaculture. The increased antibody response in fish received PS-K one week prior to FKC administration suggested the influence of PS-K on the specific humoral immunity and increased resistance against bacterial infection.

• Journal of fish pathology
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• v.10 no.2
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• pp.143-152
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• 1997

The effect of $\beta$-glucan as an immunostimulant to increase resistance to bacterial diseases by enhancing non-specific defense mechanism in rockfish (Sebastes schlegeli) was examined by oral and bath administration. After oral or bath administration with $\beta$-glucan, the injection challenges with Vibro ordalii, Staphylococcus epidermidis and Edwardsiella tarda were performed to assess $\beta$-glucan efficacy. After injection of V. ordalii, oral administration for 30 days with 1% $\beta$-glucan showed 25% of survival rate. But all control fish died within 3 days after the injection. After injection of S, epidermidis, oral administration group for 20 and 30 days showed a remarkably increased survival rate of 95%. But oral administration of $\beta$-glucan to rockfish did not induce protection against experimental E. tarda infection. $\beta$-Glucan bath administration with or without formalin-killed V. ordalii showed that no protection was observed at 10 days after challenge. The results show that $\beta$-glucan to rockfish was effective to increase survival rate of bacterial infections of S. epidermidis and V. ordalii but not against E. tarda.

• Toxicological Research
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• v.34 no.1
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• pp.65-73
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• 2018

Pretreatment of low-dose lipopolysaccharide (LPS) induces a hyporesponsive state to subsequent secondary challenge with high-dose LPS in innate immune cells, whereas super-low-dose LPS results in augmented expression of pro-inflammatory cytokines. However, little is known about the difference between super-low-dose and low-dose LPS pretreatments on immune cell-mediated inflammatory and hepatic acute-phase responses to secondary LPS. In the present study, RAW 264.7 cells, EL4 cells, and Hepa-1c1c7 cells were pretreated with super-low-dose LPS (SL-LPS: 50 pg/mL) or low-dose LPS (L-LPS: 50 ng/mL) in fresh complete medium once a day for 2~3 days and then cultured in fresh complete medium for 24 hr or 48 hr in the presence or absence of LPS ($1{\sim}10{\mu}g/mL$) or concanavalin A (Con A). SL-LPS pretreatment strongly enhanced the LPS-induced production of tumor necrosis factor (TNF)-${\alpha}$, interleukin (IL)-6, TNF-${\alpha}$/IL-10, prostaglandin E2 ($PGE_2$), and nitric oxide (NO) by RAW 264.7 cells compared to the control, whereas L-LPS increased IL-6 and NO production only. SL-LPS strongly augmented the Con A-induced ratios of interferon (IFN)-${\gamma}$/IL-10 in EL4 cells but decreased the LPS-induced ratios of IFN-${\gamma}$/IL-10 compared to the control, while L-LPS decreased the Con A- and LPS-induced ratios of IFN-${\gamma}$/IL-10. SL-LPS enhanced the LPS-induced production of IL-6 by Hepa1c1c-7 cells compared to the control, while L-LPS increased IL-6 but decreased IL-$1{\beta}$ and C reactive protein (CRP) levels. SL-LPS pretreatment strongly enhanced the LPS-induced production of TNF-${\alpha}$, IL-6, IL-10, $PGE_2$, and NO in RAW 264.7 cells, and the IL-6, IL-$1{\beta}$, and CRP levels in Hepa1c1c-7 cells, as well as the ratios of IFN-${\gamma}$/IL-10 in LPS- and Con A-stimulated EL4 cells compared to L-LPS. These findings suggest that pre-conditioning of SL-LPS may contribute to the mortality to secondary infection in sepsis rather than pre-conditioning of L-LPS.

• Asian-Australasian Journal of Animal Sciences
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• v.21 no.12
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• pp.1753-1759
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• 2008

Forty weanling piglets ($5.6{\pm}0.5kg$ and 26 to 30 d of age) were used in a 28-d experiment to determine the effects of ${\beta}$-glucan from Paenibacillus polymyxa and L-theanine on growth performance. Piglets were randomly allotted to four groups (n = 10, 2 animals per pen) provided with the basal feed (control), ${\beta}$-glucan 400 mg/kg feed, L-theanine 80 mg/kg feed or ${\beta}$-glucan plus l-theanine (combination of the above-mentioned concentrations). Body weight and feed consumption were recorded during four weeks. Subsequently, the immunomodulatory effects of ${\beta}$-glucan and L-theanine were investigated for lipopolysaccharide (LPS)-induced cytokine production in vitro and in vivo on day 28. Although there were no significant differences in the growth performances among the treatment groups, ${\beta}$-glucan plus L-theanine had 5.6% greater ADG (p = 0.074) on day 21 to 28. ${\beta}$-Glucan alone or plus L-theanine increased interleukin (IL)-10 levels and decreased interferon (IFN)-$\gamma$ and tumor necrosis factor (TNF)-${\alpha}$ levels in cultured medium by LPS treatment (p<0.05). Plasma IL-10 levels were also increased in the piglets fed with ${\beta}$-glucan alone or plus L-theanine after LPS challenge ($25{\mu}g/kg$, i.p.), whereas plasma IFN-$\gamma$ and TNF-${\alpha}$ levels were decreased (p<0.05). The levels of IFN$\gamma$ in piglets fed with ${\beta}$-glucan plus L-theanine showed the greatest inhibition after LPS challenges. In conclusion, treatment of ${\beta}$-glucan alone or plus L-theanine might lessen inflammatory responses against Gram-negative bacterial infection via the inhibition of pro-inflammatory cytokine production and enhancement of anti-inflammatory cytokine production. Further studies are needed to determine an optimal concentration of ${\beta}$-glucan and L-theanine for improved growth performance.

• Journal of fish pathology
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• v.24 no.2
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• pp.53-64
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• 2011

In quantitative studies of clinical signs, rock bream of adults and juveniles infected with Megalocytivirus IVS-1 isolated from rock bream (Oplegnathus fasciatus) in Korea showed average $4.49{\pm}1.13$ and $4.85{\pm}1.06$ of spleen index respectively. In challenge experiments, Megalocytivirus IVS-1 induced 100% cumulative mortality in both adult and juvenile rock bream. However we found 60% cumulative mortality in juvenile red sea bream (Pagrus major) even after 30 days of injection, which contradicted with the results of other laboratories. Interestingly, IVS-1 infected red sea bream of the same juvenile size with rock bream showed lower spleen index compared to that of rock bream. In real-time PCR, there was continuous increasing of the numbers of viral copies ($2.03{\times}10^7$ copies/mg) in the spleen of juvenile rock bream infected, which were different from those in adult rock bream showing plateau level after reaching to the peak level. Moreover, enlarged cell numbers in the infected spleen were also increased continuously in the juvenile but not in adult of rock bream, even decreased after reaching to peak level. Consequently, significant differences in clinical signs: cumulative mortality. spleen index and viral copy number were found between rock bream and red sea bream, but not between adult and juvenile rock bream. Certainly quantitative expression of clinical sign as in this study may be a way to compare the progression of megalocitiviral disease more accurately in different species or physiological conditions.

• BMB Reports
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• v.42 no.1
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• pp.59-64
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• 2009

Hepatitis B virus (HBV) infection is highly prevalent worldwide. The major challenge for current antiviral treatment is the elevated drug resistance that occurs via rapid viral mutagenesis. In this study, we developed AAV vectors to simultaneously deliver two or three shRNAs targeting different HBV-related genes. These vectors showed markedly better antiviral effects than ones that delivered a single shRNA in vitro. A dual shRNA expression vector (AAV-157i/1694i), which simultaneously expressed two shRNAs targeted the S and X genes of HBV, reduced HBsAg, HBeAg and HBV DNA levels by $87{\pm}4$, $80.3{\pm}2.6$ and $86.2{\pm}7%$ respectively, eight days post-transduction. In a mouse model of prophylactic treatment, HBsAg and HBeAg were reduced to undetectable levels and the serum HBV DNA level was reduced by at least 100 fold. These results indicate that AAV-157i/1694i generates potent anti-HBV effects and that the strategy of constructing multi-shRNA expression vectors may lead to enhanced anti-HBV efficacy and overcome the evading mechanism of the virus and thus the development of drug resistance.

• Parasites, Hosts and Diseases
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• v.26 no.3
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• pp.179-188
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• 1988

The life cycle of Fibricola seoulensis was studied in the laboratory and in the field, with special interests in the larval developments within the eggs and in the intermediate hosts. The first emergence of miracidia after incubation of eggs in 26C water began on the ninth day. The miracidia, elongate and cylindrical shape, had epidermal plates in the formula of 6, 9, 4 and 3, with two pairs of flame cells and lateral processes. A kind of fresh water snail, Hippeutis (H.) cantori, was found to shed furcocercous cercariae from the 13th day after experimental challenge with miracidia while Physa acute failed to shed. The same kind of snail collected from the field also shed the same cercariae. The cercariae were equipped with 2 pairs of penetration glands and 5 pairs of fame cells. The tadpoles of Rana nigromaculata were found susceptible to experimental infection with the cercariae. The same kind of tadpoles collected from various areas were also found naturally infected. The metacercariae in the tadpoles which were infected experimentally became infective to the definitive host in 21 days. The metacercariae were located free in the body cavity of tadpoles, and attained sexual maturity in rats in 7 days. The present study successfully followed the complete life cycle of F. seoulensis and found that it is possible to maintain the life cycle in the laboratory.

• Journal of Life Science
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• v.26 no.12
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• pp.1409-1414
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• 2016

In the larvae of the black soldier fly, Hermetia illucens, innate immunity mechanisms are activated in response to various pathogens and stimulants, resulting in the expression of antimicrobial peptides (AMPs). To induce the mass production of AMPs, H. illucens fifth instar larvae were immunized with five different kinds of bacteria. We isolated from the hemolymph of the H. illucens larvae after bacterial challenge, and their antimicrobial activities against Gram-positive bacteria (Staphylococcus aureus) and Gram-negative bacteria (Escherichia coli) were measured using the inhibition zone assay. Among these five different kinds of bacteria, the hemolymph of Bacillus subtilis-challenged H. illucens larvae showed the strongest antimicrobial activity against both Gram-positive bacteria and Gram-negative bacteria. The antimicrobial activity of the hemolymph of $1{\times}10^9cfu/ml$ B. subtilis-challenged H. illucens peaks at 24 hr at 48 hr post-infection and gradually declines with time. Moreover, the immunized hemolymph also showed strong antimicrobial activity against various poultry pathogens such as S. enteritidis, S. typhimurium, and S. pullorum. These results suggest that the expression of AMP genes in B. subtilis-challenged H. illucens is up-regulated by innate immune responses, and that B. subtilis-challenged H. illucens overexpressing AMPs may be useful as a feed additive in livestock diets to reduce the need for antibiotics.