• 한국어병학회지
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• 제23권3호
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• pp.343-355
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• 2010

This study was conducted to investigate the dietary effects of lactic acid bacteria (LAB) supplementation on growth, hematological and immune responses of grey mullet, Mugil cephalus. Three replicate groups of grey mullet (body length, $62.8{\pm}1.04\;mm$ ; body weight, $3.2{\pm}0.13\;mg$) were fed the experimental diets with 0 (control), 1, 2.5 and 5,0 % of LAB for 3 months. Total body length growth rate was increased in 2.5 % supplementation group compared to control (P<0.05). No differences were observed in hematological parameters (hematocrit, hemoglobin, RBCs) and serum chemistry (calcium, magnesium, total protein, glucose, GOT and GPT). The antioxidant activity of grey mullet fed the 5.0 % LAB diets was significantly higher than that of other groups (P<0.05). Both intracellular superoxide anion production and lysozyme activity of kidney were higher in the 2.5 % LAB diet than in the control (P<0.05).

• Preventive Nutrition and Food Science
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• 제2권2호
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• pp.101-108
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• 1997

A lactic acid bacterium LAB3113, isolated from traditionally fermented Kimchi was found to produce bacteriocin whose activity was very specific toward lactobacilli and not effective against any other bacteria. Lactobacilli affected by the inhibitory substance included Lactobacillus delbrueckii-lactis, L. johnsonii, L. gsseri, and L. curvatus. Based upon biochemical and physiological characteristics, LAB3113 was classified as Lactococcus lactis, and its bacteriocin was named as lactococcin K3113. Lactococcus lactis. LAB3113 produced bacteriocin at th early stage of growth and the concentration of the bacteriocin did not decrease even after alt stationalry phase. Optimal temperature of bacteriocin production was $25^{\circ}C$ at the initial pH 7.0. Partially purified lactococcin K3113 was completely inactivated by protease, but not affected by lipase, lysozyme and RNase. The bacteriocin was very heat-stable even after autoclaving for 20 min. It was also stable in pH changes, an was not affected by th presence of solvents. lacotococcin K3113 appeared to act in bactericidal mode against L. delbrueckii-lactis ATCC4797. Molecular weight of lactococcin K3113 was calibrated as 10,500 dal by SDS-PAGE an activity staining. Lactococcus lactis LAB3113 had four residential plasmids of 3.7kb, 11.2kb, 15.5kb, and 48kh in molecular sizes. Plasmid profile analysis of mutant strain revealed that 15.5 kb plasmid was re-sponsible for the production of lactococcin K3113 and its immunity to the bacteriocin.

• Fisheries and Aquatic Sciences
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• 제26권1호
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• pp.58-68
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• 2023

This study was conducted to determine dietary threonine (Thr) requirement for juvenile olive flounder (Paralichthys olivaceus). A total of 450 juvenile fish (23.2 ± 0.4 g) were randomly distributed to 18 tanks (215 L) with 25 fish per tank. Experimental diets included with graded levels of Thr at 0.0%, 0.4%, 0.8%, 1.2%, 1.6%, and 2.0% were assigned for triplicate groups of fish and fed two times daily to apparent satiation for 12 weeks. Weight gain, specific growth rate, feed intake, feed utilization and survival were significantly (p < 0.05) increased in fish fed with dietary Thr levels over 0.8%, and no significant differences were observed between 0.8% to 2.3% levels. Non-specific immune parameters of serum lysozyme, myeloperoxidase activity, antiprotease activity, and total immunoglobulin were significantly increased by dietary Thr over 0.8%. Based on the broken-line regression analysis, the Thr requirement for the optimum growth and immune response in olive flounder is likely to be 1.03% in the diet.

• KSBB Journal
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• 제26권3호
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• pp.199-205
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• 2011

Using tetrameric ${\beta}$-galactosidase as a model protein, anchoring motives were screened in Bacillus subtilis spore display system. Eleven spore coat proteins were selected considering their expression levels and the location in the spore coat layer. After chromosomal single-copy homologous integration in the amyE site of Bacillus subtilis chromosome, cotE and cotG were chosen as possible spore surface anchoring motives with their higher whole cell ${\beta}$-galactosidase activity. PAGE and Wester blot of extracted fraction of outer layer of purified spore, which express CotE-LacZ or CotG-LacZ fusion verified the existence of exact size of fusion protein and its location in outer coat layer of purified spore. ${\beta}$-galactosidase activity of spore with CotE-LacZ or CotG-LacZ fusion reached its highest value around 16~20 h of culture time in terms of whole cell and purified spore. After intensive spore purification with lysozyme treatment and renografin treatment, spore of BJH135, which expresses CotE-LacZ, retained only 1~2% of its whole cell ${\beta}$-galactosidase activity. Whereas spore of BJH136, which has cotG-lacZ cassette in the chromosome, retained 10~15% of its whole cell ${\beta}$-galactosidase activity, proving minor perturbation of CotG-LacZ, when incorporated in the spore coat layer of Bacillus subtilis compared to CotE-LacZ. Usage of Bacillus subtilis WB700, of which 7 proteases are knocked-out and thereby resulting in 99.7% decrease in protease activity of the host, did not prevent the proteolytic degradation of spore surface expressed CotG-LacZ fusion protein.

• Fisheries and Aquatic Sciences
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• 제24권2호
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• pp.89-98
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• 2021

The accumulation of ammonia, a consequence of intensive aquaculture activity, can damage cultured animals. We analyzed the survival rates, growth, antioxidant responses, and immune responses of abalones, Haliotis discus hannai (mean shell length 70.2 ± 4.9 mm; mean body weight 36.9 ± 3.6 g), biweekly for four weeks, to determine the accumulated concentrations of ammonia (0, 0.1, 0.2, 0.4, 0.8, and 1.6 mg/L). In our biweekly measurements, a survival rate of ≥ 95% was observed for all concentrations of ammonia. The specific growth rate (SGR) decreased at ≥ 0.8 mg/L (p < 0.05), but the daily increment in shell length (DISL) showed no significant change (p < 0.05). Regarding antioxidant responses, the superoxide dismutase (SOD) activity of the hepatopancreas was increased at ammonia concentrations over 0.8 and 0.2 mg/L (p < 0.05) at two and four weeks, respectively, and the SOD activity of the gills increased at concentrations over 0.4 and 1.6 mg/L (p < 0.05) at four weeks. The catalase (CAT) activity of the hepatopancreas and gills increased at ammonia concentrations > 0.8 mg/L (p < 0.05). Phenoloxidase (PO) activity increased at ammonia concentrations over 0.8 mg/L (p < 0.05), and Lysozyme (LZM) increased at concentrations over 0.8 and 0.4 mg/L (p < 0.05). Overall, our findings indicated that ammonia concentrations over 0.8 mg/L in seawater might damage the SGR, antioxidant responses, and immune responses in H. discus hannai. Our findings suggest the necessity of improvement or reinstallation for abalone aquaculture systems and can be used to assess the toxic effects of ammonia on H. hannai.

• 한국수산과학회지
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• 제56권1호
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• pp.89-98
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• 2023

We evaluated the effects of prebiotic (mannan oligosaccharides, Mos) and probiotic diet supplements on growth performance, innate immunity, antioxidant activity, and intestinal changes in the microbial flora of red seabream Pagrus major. A basal diet (Con) was formulated to meet the nutrient requirement of red seabream. The dietary starch in Con was replaced with 0.6% Mos, Lactobacillus plantarum, Bacillus subtilis, B. licheniformis and probiotic mixture (labeled as Mos, Pro-LP, Pro-BS, Pro-BL and Pro-Mix, respectively). We stocked 450 fish in 18 polypropylene tanks (400 L) in triplicate groups per dietary treatment. The fish were fed one of the diets twice (08:30, 18:30 h) a day for 63 days. Lysozyme activity was significantly higher in all the supplemented groups than that of the Con group. The immunoglobulin level of Pro-Mix, anti-protease activity of Pro-BL, and glutathione peroxidase and superoxide dismutase activity of Pro-BS, Pro-BL and Pro-Mix groups were significantly higher than those of the Con group. The ratio of total Vibrio/heterotrophic marine bacteria counts was significantly lower in Pro-LP, Pro-BL and Pro-Mix groups than that of the Con group. Therefore, dietary supplementation of Mos and probiotics to improves immune response and antioxidant enzyme activity and inhibits Vibrio bacteria in the intestine.

• Journal of Microbiology and Biotechnology
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• 제32권6호
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• pp.816-823
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• 2022

The rapid spread of superbugs leads to the escalation of infectious diseases, which threatens public health. Endolysins derived from bacteriophages are spotlighted as promising alternative antibiotics against multi-drug resistant bacteria. In this study, we isolated and characterized the novel Salmonella typhimurium phage PBST08. Bioinformatics analysis of the PBST08 genome revealed putative endolysin ST01 with a lysozyme-like domain. Since the lytic activity of the purified ST01 was minor, probably owing to the outer membrane, which blocks accessibility to peptidoglycan, antimicrobial peptide cecropin A (CecA) was fused to the N-terminus of ST01 to disrupt the outer membrane. The resulting CecA::ST01 has been shown to have increased bactericidal activity against gram-negative pathogens including Pseudomonas aeruginosa, Klebsiella pneumoniae, Acinetobacter baumannii, Escherichia coli, and Enterobacter cloacae and the most affected target was A. baumannii. In the presence of 0.25 µM CecA::ST01, A. baumannii ATCC 17978 strain was completely killed and CCARM 12026 strain was wiped out by 0.5 µM CecA::ST01, which is a clinical isolate of A. baumannii and resistant to multiple drugs including carbapenem. Moreover, the larvae of Galleria mellonella could be rescued up to 58% or 49% by the administration of CecA::ST01 upon infection by A. baumannii 17978 or CCARM 12026 strain. Finally, the antibacterial activity of CecA::ST01 was verified using 31 strains of five gram-negative pathogens by evaluation of minimal inhibitory concentration. Thus, the results indicate that a fusion of antimicrobial peptide to endolysin can enhance antibacterial activity and the spectrum of endolysin where multi-drug resistant gram-negative pathogens can be efficiently controlled.

• 한국양식학회:학술대회논문집
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• 한국양식학회 2003년도 추계학술발표대회 논문요약집
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• pp.68-69
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• 2003

The present study was conducted to investigate the effects of dietary supplementation of $\beta$-1,3 glucan and feed stimulants(BAISM) as a feed additive for juvenile olive flounder (Paralichthys olivaceus). Eight experimental diets supplemented with $\beta$-1,3 glucan and feed stimulants at 0%, $\beta$-1,3 glucan 0.05% ＋ Baism 0.45%, $\beta$-1,3 glucan 0.05% ＋ Baism 0.95%, $\beta$-1,3 glucan 0.10% ＋ Baism 0.90%, $\beta$-1,3 glucan 0.10% ＋ Baism 1.90%, $\beta$-1,3 glucan 0.15% ＋ Baism 1.35%, $\beta$-1,3 glucan 0.15% ＋ Baism 2.85% and $\beta$-1,3 glucan 0.30% ＋ Baism 2.70% of diets as a dry-matter(DM) basis were prepared. Three replicate groups of fish averaging 9.2 $\pm$ 0.1g (Mean $\pm$ SD) were randomly distributed in each aquarium as a group of 15 fish and fed one of eight experimental diets for seven weeks. After the feeding trial, $\beta$-1,3 glucan 0.10% ＋ Baism 0.90%, $\beta$-1,3 glucan 0.10% ＋ Baism 1.90% diets had a higher weight gain (WG), feed efficiency(FE), specific growth rate(SGR) and protein efficiency ratio(PER) than did fish fed 0%, $\beta$-1,3 glucan 0.05% ＋ Baism 0.45%, $\beta$-1,3 glucan 0.05% ＋ Baism 0.95%, $\beta$-1,3 glucan 0.15% ＋ Baism 2.85% and $\beta$-1,3 glucan 0.30% ＋ Baism 2.70% (P<0.05). however, there was no significant difference among fish fed $\beta$-1,3 glucan 0.05% ＋ Baism 0.45%, $\beta$-1,3 glucan 0.05% ＋ Baism 0.95%, $\beta$-1,3 glucan 0.15% ＋ Baism 2.85% and $\beta$-1,3 glucan 0.30% ＋ Baism 2.70%(P>0.05). and $\beta$-1,3 glucan 0.10% ＋ Baism 0.90% diets had a higher peak value of CL(Chemiluminescence) and lysozyme activity, than did fish fed the other diets (P<0.05). These results indicated that dietary sipplementation of $\beta$-1, 3 glucan and Baism affected growth, feed efficiency, specific growth rate, protein efficiency ratio, Peak value of CL and Lysozyme activity, and the optimum dietary supplementation level of $\beta$-1, 3 glucan and Baism as a feed additive could be approximately $\beta$-1, 3 glucan 0.10% ＋ Baism 0.90% of diet in juvenile olive flounder (Paralichthys olivaceus).

• 대한수의학회지
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• 제57권2호
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• pp.117-126
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• 2017

Bovine mastitis is an important microbial disease in the dairy industry. We investigated the frequencies of bacterial pathogens in 62 farms and pathogen antibiotic resistance from mastitis samples (n = 748). We tested the antimicrobial activity of chicken and duck egg white and lysozyme purified from chicken egg white. Moreover, we compared the microbiomes of normal and mastitic raw milk obtained by 16S rRNA gene pyrosequencing and culture methods. The results showed that the frequencies of Gram-positive pathogens (Enterococcus faecalis 37% and Staphylococcus aureus 36%) were higher than that of a Gram-negative pathogen (Escherichia coli 15%). Resistance frequencies to ampicillin and norfloxacin were lowest in Staphylococcus aureus (21%), Enterococcus faecalis (23%), and Escherichia coli (33%), and the antimicrobial activity of chicken egg white was higher than those of lysozyme and duck egg white. Pyrosequencing results revealed clear differences between the microbiomes of mastitic and normal raw milk samples and revealed a slightly similar, but clearly different, composition of pathogens compared to that from the culture method. Thus, pyrosequencing may be useful for elucidating changes in microbiomes during mastitis progression and treatment. A chicken egg white and antibiotic combination may help with mastitis treatment; however, further studies are needed.

• 한국어병학회지
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• 제18권3호
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• pp.277-285
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• 2005

홍삼추출물을 다양한 농도 (1-50 mg/kg)로 어체의 복강내에 주입하여 3일 후 어체를 희생시켜서 Ht, lysozyme, TIC 및 NBT reduction을 측정한 바, 10 ㎍/㎎ 이상의 처치군에 유의성 있는 증가를 보였다. 한편, in vitro에서 홍삼 추출물을 처리시 림프구의 증식능에 미치는 영향은 PHA 혹은 Con A와 혼합처리 경우에는 10 ${\mu}g/m{\ell}$농도에서 가장 높은 증가를 보였으나, 고농도 (100 ${\mu}g/m{\ell}$)에서는 억제되었다. 그렇지만, RGE 단독 혹은 LPS혼합 처리군에서는 증가효과가 유의성 있게 나타나지 않았다. 한편, 백혈구의 유주능 및 발생기산소 (ROI)의 생성은 10 ${\mu}g/m{\ell}$이상의 처리군에서 유의성 있는 증가를 보였다. 이러한 결과는 홍삼추출물을 복강내로 처리시 림프구의 증식 및 탐식세포의 기능을 증가시키는 것을 알 수 있었으며, 향후 면역보조제로 사용하기 위해서는 경구투여에 따른 실행이 수행되어져야 할 것으로 사료된다.