• Journal of Veterinary Science
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• v.25 no.1
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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.

• Korean Journal of Veterinary Research
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• v.57 no.1
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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.

• Journal of Environmental Health Sciences
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• v.23 no.4
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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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• v.18 no.2
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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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• v.25 no.4
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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.

• Journal of Food Hygiene and Safety
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• v.12 no.3
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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.

• Proceedings of the Korean Vacuum Society Conference
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• 2014.02a
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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.

• Journal of Food Hygiene and Safety
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• v.28 no.1
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• pp.69-74
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• 2013

This study has been performed for 150 days from February 1 - June 31, 2012 aiming at analyzing biologically hazardous factors in order to develop HACCP system for the vinegared pickle radishes. A process chart was prepared as shown on Fig. 1 by referring to manufacturing process of manufacturer of general vinegared pickle radishes regarding process of raw agricultural products of vinegared pickle radishes, used water, warehousing of additives and packing material, storage, careful selection, washing, peeling off, cutting, sorting out, stuffing (filling), internal packing, metal detection, external packing, storage and consignment (delivery). As a result of measuring Coliform group, Staphylococcus aureus, Salmonella spp., Bacillus cereus, Listeria Monocytogenes, E. coli O157:H7, Clostridium perfringens, Yeast and Mold before and after washing raw radishes, Bacillus cereus was $5.00{\times}10$ CFU/g before washing but it was not detected after washing and Yeast and Mold was $3.80{\times}10^2$ CFU/g before washing but it was reduced to 10 CFU/g after washing and other pathogenic bacteria was not detected. As a result of testing microorganism variation depending on pH (2-5) of seasoning fluid (condiment), pH 3-4 was determined as pH of seasoning fluid as all the bacteria was not detected in pH3-4. As a result of testing air-borne bacteria (number of general bacteria, colon bacillus, fungus) depending on each workplace, number of microorganism of internal packing room, seasoning fluid processing room, washing room and storage room was detected to be 10 CFU/Plate, 2 CFU/Plate, 60 CFU/Plate and 20 CFU/Plate, respectively. As a result of testing palm condition of workers, as number of general bacteria and colon bacillus was represented to be high as 346 $CFU/Cm^2$ and 23 $CFU/Cm^2$, respectively, an education and training for individual sanitation control was considered to be required. As a result of inspecting surface pollution level of manufacturing facility and devices, colon bacillus was not detected in all the specimen but general bacteria was most dominantly detected in PP Packing machine and Siuping machine (PE Bulk) as $4.2{\times}10^3CFU/Cm^2$, $2.6{\times}10^3CFU/Cm^2$, respectively. As a result of analyzing above hazardous factors, processing process of seasoning fluid where pathogenic bacteria may be prevented, reduced or removed is required to be controlled by CCP-B (Biological) and threshold level (critical control point) was set at pH 3-4. Therefore, it is considered that thorough HACCP control plan including control criteria (point) of seasoning fluid processing process, countermeasures in case of its deviation, its verification method, education/training and record control would be required.

• Journal of Food Hygiene and Safety
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• v.29 no.2
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• pp.79-84
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• 2014

This study has been performed for about 270 days at analyzing biologically hazardous factors in order to develop HACCP system for the non heat-frozen carrot juice. A process chart was prepared by manufacturing process of raw agricultural products of non heat-frozen carrot juice, which was contained water and packing material, storage, washing, cutting, extraction of the juice, internal packing, metal detection, external packing, storage and consignment (delivery). As a result of measuring Coliform group, Staphylococcus aureus, Salmonella spp., Bacillus cereus, Listeria Monocytogenes, Enterohemorrhagic E. coli before and after washing raw carrot, Standard plate count was $4.7{\times}10^4CFU/g$ before washing but it was $1.2{\times}10^2CFU/g$ detected after washing. As a result of testing airborne bacteria (Standard plate count, Coliform group, Yeast and Fungal) depending on each workplace, number of microorganism of in packaging room, shower room and juice extraction room was detected to be 10 CFU/Plate, 60 CFU/Plate, 20 CFU/Plate, respectively. As a result of testing palm condition of workers, as number of Standard plate count, Coliform group and Staphylococcus aureus was represented to be high as $6{\times}10^4CFU/cm^2$, $0CFU/cm^2$ and $0CFU/cm^2$, respectively, an education and training for individual sanitation control was considered to be required. As a result of inspecting surface pollution level of manufacturing facility and devices, Coliform group was not detected in all the specimen but Standard plate count was most dominantly detected in scouring kier, scouring kier tray, cooling tank, grinding extractor, storage tank and packaging machine-nozzle as $8.00{\times}10CFU/cm^2$, $3.0{\times}10CFU/cm^2$, $4.3{\times}10^2CFU/cm^2$, $7.5{\times}10^2CFU/cm^2$, $6.0{\times}10CFU/cm^2$, $8.5{\times}10^2CFU/cm^2$ respectively. As a result of analyzing above hazardous factors, processing process of ultraviolet ray sterilizing where pathogenic bacteria may be prevented, reduced or removed is required to be controlled by CCP-B (Biological) and critical level (critical control point) was set at flow speed is 4L/min. Therefore, it is considered that thorough HACCP control plan including control criteria (point) of seasoning fluid processing process, countermeasures in case of its deviation, its verification method, education/training and record control would be required.