• Asian-Australasian Journal of Animal Sciences
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• v.30 no.4
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• pp.576-584
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• 2017

Objective: To generate recombinant Bacillus subtilis (B. subtilis) engineered for expression of porcine ${\beta}-defensin-2$ (pBD-2) and cecropin P1 (CP1) fusion antimicrobial peptide and investigate their anti-bacterial activity in vitro and their growth-promoting and disease resisting activity in vivo. Methods: The pBD-2 and CP1 fused gene was synthesized using the main codons of B. subtilis and inserted into plasmid pMK4 vector to construct their expression vector. The fusion peptide-expressing B. subtilis was constructed by transformation with the vector. The expressed fusion peptide was detected with Western blot. The antimicrobial activity of the expressed fusion peptide and the recovered pBD-2 and CP1 by enterokinase digestion in vitro was analyzed by the bacterial growth-inhibitory activity assay. To analyze the engineered B. subtilis on growth promotion and disease resistance, the weaned piglets were fed with basic diet supplemented with the recombinant B. subtilis. Then the piglets were challenged by enteropathogenic Escherichia coli (E. coli). The weight gain and diarrhea incidence of piglets were measured after challenge. Results: The recombinant B. subtilis engineered for expression of pBD-2/CP1 fusion peptide was successfully constructed using the main codons of the B. subtilis. Both expressed pBD-2/CP1 fusion peptide and their individual peptides recovered from parental fusion peptide by enterokinase digestion possessed the antimicrobial activities to a variety of the bacteria, including gram-negative bacteria (E. coli, Salmonella typhimurium, and Haemophilus parasuis) and grampositive bacteria (Staphylococcus aureus). Supplementing the engineered B. subtilis to the pig feed could significantly promote the piglet growth and reduced diarrhea incidence of the piglets. Conclusion: The generated B. subtilis strain can efficiently express pBD-2/CP1 fusion antimicrobial peptide, the recovered pBD-2 and CP1 peptides possess potent antimicrobial activities to a variety of bacterial species in vitro. Supplementation of the engineered B. subtilis in pig feed obviously promote piglet growth and resistance to the colibacillosis.

• Korean Journal of Veterinary Service
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• v.34 no.4
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• pp.341-352
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• 2011

Colibacillosis in pigs remain a major swine industry bruden worldwide. Although some progress has been made in treating collibacillosis in pigs by using biosecurity and antimicrobials, it still remain a considerable problem. The use of host-specific bateriophages as a biocontrol is one possible alternative. The purpose of this study was to evaluate the effect of bacteriophage against enterotoxigenic Escherichia coli (ETEC) K88 infection in piglets. Twenty-eight piglets were randomly divided into four groups and each group was allocated with 7 pigs. Group B, C and D were inoculated with 5 ml of ETEC K88 ($1{\times}10^8$ CFU/ml) per head of piglet via oral. Group C and D were fed with bacteriophages (Group C, $1.0{\times}10^6$ PFU/g; Group D, $1.0{\times}10^8$ PFU/g; CJ CheilJedang Corp., Korea) orally as treatment. In piglets administrated bacteriophages and challenged with ETEC K88 (Group C and D), Clinical signs and the growth performance were improved and antibody titers were maintained low level compared with piglets challenged with ETEC K88 (Group B, P<0.05). Group B were shown high pH in the alimentary tracts compared with other piglets (P<0.05). In quantitative analysis by real-time PCR, the results of Group C and D were lower than those Group B in faecal and intestinal samples (P<0.05). Severe villus atrophy and crypt hyperplasia were observed in Group B consequently V/C ratio increased, compared with other piglets. These results indicate that feeding with bacteriophage has effect to prevent ETEC K88 infection in piglets and suggest that use of bacteriophage can be considered a valid antibiotic alternative.

• Korean Journal of Environmental Agriculture
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• v.32 no.1
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• pp.70-77
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• 2013

BACKGROUND: Nitroxoline is an antibiotic agent. It is used for the treatment of the second bacterial infection by the colibacillosis, salmonellosis and viral disease of the poultry. When the nitroxoline is indiscreetly used, the problem about the abuse of the antibiotics can occur. Therefore, this study presented the residue analytical method of nitroxoline in food for the safety management of animal farming products. METHODS AND RESULTS: A simple, sensitive and specific method for nitroxoline in chicken muscle by high performance liquid chromatograph with PDA was developed. Sample extraction with acetonitrile, purification with SPE cartridge (MCX) were applied, then quantitation by HPLC with C18 column under the gradient condition with 0.1 % tetrabutylammonium hydroxide-phosphoric acid and methanol was performed. Standard calibration curve presented linearity with the correlation coefficient ($r^2$) > 0.999, analysed from 0.02 to 0.5 mg/L concentration. Limit of quantitation in chicken muscle showed 0.02 mg/kg, and average recoveries ranged from 72.9 to 88.1 % in chicken muscle. The repeatability of measurements expressed as coefficient of variation (CV %) was less than 12 % in 0.02 and 0.04 mg/kg. CONCLUSION(S): Newly developed method for nitroxoline in chicken muscle was applicable to food inspection with the acceptable level of sensitivity, repeatability and reproducibility.

• Korean Journal of Poultry Science
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• v.18 no.3
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• pp.151-159
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• 1991

As a part of investigation on causes of drop in egg production in domestic chicken breeding farm, bacteriological contamination on air, feed, drinking water and artificial insemination instruments of randomly selected three farms was surveyed. Total bacterial population in the air was very high in all of the chicken houses tested and was not significantly different among these farms . However, total bacterial counts in the air of the problem house having egg drop problem and colibacillosis was higher than normal house within the problem farm. Bacterial population in the assorted feed was low before or after administration on the normal farm while it was much more increased after administration than before administration on the problem farm. Bacterial population of the drinking water in the source of water supply was very low and has no differences among farms tested. Also, bacterial population in the normal farm was not significantly different between source of water supply and after administration. However, population of total bacteria and coliform bacteria after administration was increased. Bacterial population was much higher in the artificial insemination instrument of problem farm than normal farm. However, this bacterial population in the problem farm was decreased to those of normal farm after these instruments were sanitized.