• Journal of Veterinary Science
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• 제25권1호
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• pp.4.1-4.14
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• 2024

Background: Lawsonia intracellularis is the causative agent of proliferative enteropathy and is associated with several outbreaks, causing substantial economic loss to the porcine industry. Objectives: In this study, we focused on demonstrating the protective effect in the mouse model through the immunological bases of two vaccine strains against porcine proliferative enteritis. Methods: We used live-attenuated Salmonella Typhimurium (ST) secreting two selected immunogenic LI antigens (Lawsonia autotransporter A epitopes and flagellin [FliC]-peptidoglycan-associated lipoprotein-FliC) as the vaccine carrier. The constructs were cloned into a Salmonella expression vector (pJHL65) and transformed into the ST strain (JOL912). The expression of immunogenic proteins within Salmonella was evaluated via immunoblotting. Results: Immunizing BALB/c mice orally and subcutaneously induced high levels of LI-specific systemic immunoglobulin G and mucosal secretory immunoglobulin A. In immunized mice, there was significant upregulation of interferon-γ and interleukin-4 cytokine mRNA and an increase in the subpopulations of cluster of differentiation (CD) 4⁺ and CD 8⁺ T lymphocytes upon splenocytes re-stimulation with LI antigens. We observed significant protection in C57BL/6 mice against challenge with 10^6.9 times the median tissue culture infectious dose of LI or 2 × 10⁹ colony-forming units of the virulent ST strain. Immunizing mice with either individual vaccine strains or co-mixture inhibited bacterial proliferation, with a marked reduction in the percentage of mice shedding Lawsonia in their feces. Conclusions: Salmonella-mediated LI gene delivery induces robust humoral and cellular immune reactions, leading to significant protection against LI and salmonellosis.

• 대한수의학회지
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• 제57권1호
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• pp.9-15
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• 2017

To construct a novel vaccine candidate against bovine enterotoxigenic Escherichia coli (ETEC), FanC, the major subunit of K99 fimbriae adhesion, was inserted into secretion plasmid pYA3560 containing a ${\beta}-lactamase$ secretion system. This was then transformed into ${\Delta}asd$ ${\Delta}crp$ Salmonella (S.) Typhimurium and designated as JOL950. Secretion of recombinant fanC fimbrial antigens was confirmed by immunoblot analysis. Groups of mice were inoculated with single or double doses of JOL950. Another group was used as a negative control. Compared to control mice, all immunized mice had significantly higher levels (p < 0.05) of serum immunoglobulin (Ig)G, and secretory IgA against FanC. The IgG2a and IgG1 titer assays revealed that immunization highly induced IgG2a compared to that of IgG1, indicating that T helper-1- related cell-mediated immune responses may be elicited by JOL950. The results show that both systemic and mucosal immunities against selected fimbrial antigens of bovine ETEC expressed by a live attenuated S. Typhimurium strain are prominently produced in mice immunized with JOL950 via an oral route.

• 한국환경보건학회지
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• 제23권4호
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• pp.50-56
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• 1997

Applied for Drug Delivery System, polymer drug was prepared with chitosan and phthalylsulfathiazole. In spite of various application of chitin derivatives, commercial use of chitin has been limited due to highly resistance to chemicals and the absense of proper solvents. In this study, Chitosan were prepared from chitin which were deacetylated under various condition. The synthetic procedures of polymer drug were performed by acid chloride methods. The antibiotic activities of polymer drug exhibited growth-inhibitory activity against Staphylococcus aureus, Staphylococcus epidermidis, E. coli, Salmonella typhimurium, Klebsiella pneumoniae at the concentration of 471-514 $\mu$g/ml in general. Comparatively, Polymer drug exhibited broad antibacterial activity on MICs 897-1280 $\mu$g/ml against Gram-positive and Gram-negative bacteria including Staphylococcus aureus, Staphylococcus epidermidis and E. coli.

• Food Science and Biotechnology
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• 제18권2호
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• pp.425-431
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• 2009

Antimicrobial coating on chicken carcasses may reduce the effects of cross-contamination and improve product shelf-life and safety. Thyme oil was mixed at 0.5%(v/v) with a pre-gelatinized pea starch coating solution. The coating solution was spread on chicken breast meat after inoculation with selected spoilage and pathogenic bacteria. After inoculation, the chicken meats were packaged in plastic bags and stored at $4^{\circ}C$. During 12 day storage, total aerobic bacteria, lactic acid bacteria, and inoculated organisms were counted at 4 day intervals. Thyme oil treatments reduced the viability of Salmonella as well as the growth of Listeria and Pseudomonas by 2 log CFU/g, and appeared to eliminate inoculated Campylobacter during storage. The addition of thyme oil increased the viscosity of the pre-gelatinized pea starch solution. The results suggested that thyme oil inclusion in an edible starch coating may be a satisfactory delivery system to enhance the safety of processed fresh meat.

• Journal of Microbiology and Biotechnology
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• 제25권4호
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• pp.554-558
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• 2015

Drug delivery systems (DDSs) incorporating bacterial minicells have been evaluated as a very powerful tool in view of biocompatibility. However, limited studies have been carried out on these systems, mainly using minicells from Salmonella sp. and Escherichia coli. Thus, we generated a new minicell-producing strain from an endotoxin-free Corynebacterium glutamicum by the inactivation of genes related to cell division. The two knockout strains, ${\Delta}parA$ and ${\Delta}ncgl1366$, showed distinct abilities to produce minicells. The resulting minicells were purified via sequential antibiotic treatments and centrifugations, which resulted in reproducible yields.

• 한국식품위생안전성학회지
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• 제12권3호
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• pp.228-233
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• 1997

Drug Dclivcry system (DDS) purpose to getting better remedial result by improving medication from ordinary methods. Applied for DDS, to improve selectivity and comtinuity during absorbing and delivery step, polmer drug (prodrug) was prepared by the esterification with dextran in such of biodegradable polymer and phthalylsulfathiazole with is efficient for entilitis. The polymer durg was prepared with dextran and phthalylsulfathiazole by the esterification. The synthetic procedures of polymer drug was performed by acid chloride and DCC methods. Polymer drug was synthesized in high yield by acid chloride method than DCC method. The antibiotic activities of polymer drug exhibited growth-inhibitory activity against Staphylococcus aureus, Staphylococcus epidermidis, E. coli, Salmonella typhimurium, Klebsiella pneumoniae at the concentration of 500 *g/m* in general through in vitro. As a result of test, polymer drug has 1/2 MIC than phthalylsulfathiazole. Also, it has high level MIC as much as phthalylsulfathiazole with Proteus, Pseudomonas. We conducted possibility of DDS as an applied for medicine with synthesized polymer drug by using natrural polymer. We consider that clinical research must be followed to verify safety and efficacy for controlled release, activity and toxicity.

• 한국진공학회:학술대회논문집
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• 한국진공학회 2014년도 제46회 동계 정기학술대회 초록집
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• pp.411.1-411.1
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• 2014

This study evaluates the utility of an antibacterial microneedle composed of green tea extract (GT) and hyaluronic acid (HA), for the efficient delivery of GT. These microneedles have the potential to be a patient-friendly method for the conventional sustained release of drugs. In this study, a fabrication method using a mold-based technique to produce GT/HA microneedles with a maximum area of ${\sim}60mm^2$ with antibacterial properties was used to manufacture transdermal drug delivery systems. Fourier transform infrared (FTIR) spectrometry was carried out to observe the potential modifications in the microneedles, when incorporated with GT. The degradation rate of GT in GT/HA microneedles was controlled simply by adjusting the HA composition. The effects of different ratios of GT in the HA microneedles were determined by measuring the release properties. In HA microneedles loaded with 70% GT (GT70), a continuous higher release rate were sustained for 72 h. The in vitro cytotoxicity assays demonstrated that GT/HA microneedles are not generally cytotoxic to chinese hamster ovary cells (CHO-K1), human embryonic kidney cells (293T), and mouse muscle cells (C2C12), which were treated for 12 and 24 h. Antimicrobial activity of the GT/HA microneedles was demonstrated by ~95% growth reduction of gram negative [Escherichia coli (E. coli), Pseudomonas putida (P. putida) and Salmonella typhimurium (S. typhimurium)] and gram positive bacteria [Staphylococcus aureus (S. Aureus) and Bacillus subtilis (B. subtilis)], with GT70. Furthermore, GT/HA microneedles reduced bacterial growth in the infected skin wound sites and improved skin wound healing process in rat model.

• 한국식품위생안전성학회지
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• 제28권1호
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• pp.69-74
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• 2013

본 연구는 식초절임 무의 HACCP (Hazard Analysis Critical Control Point)시스템 구축을 위하여 생물학적 위해요소분석을 위한 목적으로 2012년 2월 1일~6월 31일까지 약 150일 동안 수행하였다. 일반적인 식초절임 무 제조업체의 제조공정을 참고로 하여 공정도를 작성하였으며, 식초절임 무의 원료 농산물(무), 용수, 첨가물과 포장재료에 입고, 보관, 정선, 세척, 표피제거, 절단, 선별, 충진, 내포장, 금속검출, 외포장, 보관 및 출하공정에 대하여 작성하였다. 원료 무의 세척 전, 세척 후의 Coliform group, Staphylococcus aureus, Salmonella spp., Bacillus cereus, Listeria Monocytogenes, E. coli O157:H7, Clostridium perfringens, Yeast 와 Mold를 측정한 결과 Bacillus cereus는 세척 전 $5.00{\times}10$ CFU/g이었으나, 세척 후 검출되지 않았으며, Yeast 와 Mold 는 세척 전 $3.80{\times}10^2$ CFU/g, 세척 후 10 CFU/g로 감소되었으며 나머지 병원성균은 검출되지 않았다. 조미액의 pH(2~5)별 미생물의 변화를 시험한 결과 모든 균이 검출되지 않은 pH 3~4를 조미액의 pH로 결정하였다. 작업장별 공중낙하균 (일반세균수, 대장균, 진균수) 시험결과 내포장실, 조미액가공실, 세척실, 보관실의 미생물수는 10 CFU/Plate, 2 CFU/Plate, 60 CFU/Plate 그리고 20 CFU/Plate 가 검출되었다. 종사자 손바닥 시험결과 일반세균수 346 $CFU/Cm^2$, 대장균군 23 $CFU/Cm^2$로 높게 나타나 개인위생관리에 대한 교육 및 훈련이 요구 되었다. 제조설비 및 기구의 표면오염도를 검사한 결과 모든 시료에서 대장균군은 검출되지 않았고, 일반세균은 PP Packing machin과 Siuping machine (PE Bulk)에서 가장 많은 $4.2{\times}10^3CFU/Cm^2$, $2.6{\times}10^3CFU/Cm^2$ 검출되었다. 위의 위해분석 결과 병원성미생물을 예방, 감소 또는 제거할 수 있는 조미액 가공 공정이 CCP-B (Biological)로 관리되어야 하고, 한계기준은 pH 3~4로 결정하였다. 따라서 전통한과생산에의 HACCP 모델 적용을 위한 미생물학적 위해도 평가에서와 같이 조미액 가공 공정의 관리기준 및 이탈시 조치방법, 검증방법, 교육 훈련과 기록관리 등 철저한 HACCP 관리계획이 필요할 것으로 생각된다.

• 한국식품위생안전성학회지
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• 제29권2호
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• pp.79-84
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• 2014

본 연구는 비가열 섭취 냉동 당근주스의 HACCP (Hazard Analysis Critical Control Point)시스템 구축을 위하여 생물학적 위해요소분석을 위한 목적으로 2013년 6월 21일~2014년 3월 30일까지 약 270일간 제주도 제주시 구좌읍 소재에 있는 구좌농협에서 수행하였다. 일반적인 과채주스 제조업체의 제조공정을 참고로 하여 공정도를 작성하였으며, 원료 농산물(당근), 용수와 포장재료 입고, 보관, 세척, 분쇄, 착즙, 냉각, 내포장, 금속검출, 외포장, 보관 및 출하공정에 대하여 Fig. 1과 같이 작성하였다. 원료 당근의 세척 전, 세척 후의 Coliform group, Staphylococcus aureus, Salmonella spp., Bacillus cereus, Listeria Monocytogenes, 장출혈성대장균수를 측정한 결과 Bacillus cereus 는 세척 전 $4.70{\times}10^4CFU/g$이었으나, 세척 후 $1.02{\times}10^2CFU/g$ 검출되었으며, 나머지 병원성균은 검출되지 않았다. 자외선살균공정에서 당근주스의 유속를 변화시키면서 미생물의 변화를 시험한 결과 유속 4 L/min을 한계기준으로 결정하였다. 작업장별 공중낙하균(일반세균수, 대장균, 진균수) 시험결과 세척실의 미생물수는 20 CFU/Plate가 검출되었다. 작업자 손 세척 전후 시험결과 세척 전 일반세균수가 $6{\times}10^4CFU/cm^2$로 높게 나타났으나 손 세척 후에는 검출되지 않아 손 세척 및 소독에 대한 중요성을 교육하고 훈련해야 할 것이다. 제조설비 및 기구의 표면오염도를 검사한 결과 모든 시료에서 대장균군은 검출되지 않았고, 일반세균은 포장기 노즐에서 가장 많은 $8.5{\times}10^4CFU/cm^2$ 검출되었다. 위해분석 결과 병원성미생물을 예방, 감소 또는 제거할 수 있는 자외선살균 공정이 CCP-B (Biological)로 관리되어야 하고, 한계기준은 유속 4 L/min로 결정하였다. 따라서 Kwon의 유산균을 함유한 녹즙의 HACCP에 관한 연구에서와 같이 자외선살균 공정의 한계기준 및 이탈시 조치방법, 검증방법, 교육 훈련과 기록관리 등 철저한 HACCP 관리계획이 필요할 것으로 생각된다.