• Korean Journal of Veterinary Service
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• v.16 no.1
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• pp.76-81
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• 1993

To evaluate the pathogenesis of live infectious laryngotracheitis vaccine, virus, virus and E.coli were inoculated in 4-weeks old intraorbitally and intraorally. Their pathology and serology were studied. The results were as follows. 1. thicks Inoculated with ILT vaccine alone showed no clinical sign. 2. Some of chicks inoculated with ILT vaccine followed by E.coli after 1day showed nasal discharge and cough. 3. Some of chicks inoculated with E. coli followed by ILT vaccine after 1 day and inoculated ILT vaccine and E.coli concurrently, showed nasal discharge. 4. Serum neutralization titers of chicks inoculated with ILT vaccine increased 3 weeks after Inoculation, peaked at 7 to 8 weeks and decreased. 5. Antibiotic sensitivity of E.coli were higher in Enrofloxacin (Baytril) and chloramphenical.

• Journal of the Korean Society of Food Science and Nutrition
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• v.30 no.5
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• pp.802-805
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• 2001

The dapD gene of Brevibacterium lactofermentum encoding tetrahydrodipicolinate N-succinyl transferase, one of the enzymes involved in lysine biosynthesis, was cloned by complementation of Escherichia coli dapD mutnat. The recombinant plasmid pLS1 was found to contain a 3.6 kb DNA fragment. Southern hybridization analysis confirmed that the cloned DNA fragment originated from B. lactofermentum. The data of L-lysine production showed that the B. lactofermentum dapD gene was expressed in E. coli.

• Korean journal of food and cookery science
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• v.14 no.1
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• pp.9-15
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• 1998

The inhibitory effect of clove (Eugenia caryophyllata Thumb) on the growth of Escherichia coli 0157:H7 was determined. Tryptic soy broth (TSB) containing 0∼0.5% (w/v) of clove was inoculated with 10/sup/5∼10/sup/7 CFU/ml of E. coli and incubated at 5 different temperature (35, 5, -20, 50 and $55^{\circ}C$). The growth of E. coli was not inhibited at 0.1% clove and growth occured at 0.3% after a prolonged lag period while viable cells of E. coli decreased at 0.5% clove during storage at $35^{\circ}C$. During 32 days of refrigerated storage at $5^{\circ}C$, survivors of E. coli were decreased with the progress of time and increasing clove concentration. At the presence of 0.3 or 0.4% clove, bacterial cells were dead at the end of refrigerated storage. During frozen storage at -$20^{\circ}C$, survivors of E. coli at the presence of 0∼0.4% clove were decreased 2.9∼4.07 log cycles for 4 days of early period and then decreased 1.0∼2.1 log cycles through the frozen storage. There were small changes in populations of E. coli in TSB between different concentrations of clove during frozen storage. The D-values for E. coli at $50^{\circ}C$ were 105.26, 22.47, 13.76, 11.14 and 10.17 min at clove 0, 0.1, 0.2, 0.3, 0.4%, respectively. The D-values for E. coli at $55^{\circ}C$ were 10.75, 8.95, 7.40, 5.96 and 4.96 min at clove 0, 0.1, 0.2, 0.3, 0.4%, respectively. Antibacterial activity of clove against E. coli was more effective at $50^{\circ}C$ than at $55^{\circ}C$.

• Asian-Australasian Journal of Animal Sciences
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• v.27 no.7
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• pp.1013-1018
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• 2014

In this study, we developed kinetic models to predict the growth of pathogenic Escherichia coli on cheeses during storage at constant and changing temperatures. A five-strain mixture of pathogenic E. coli was inoculated onto natural cheeses (Brie and Camembert) and processed cheeses (sliced Mozzarella and sliced Cheddar) at 3 to 4 log CFU/g. The inoculated cheeses were stored at 4, 10, 15, 25, and $30^{\circ}C$ for 1 to 320 h, with a different storage time being used for each temperature. Total bacteria and E. coli cells were enumerated on tryptic soy agar and MacConkey sorbitol agar, respectively. E. coli growth data were fitted to the Baranyi model to calculate the maximum specific growth rate (${\mu}_{max}$; log CFU/g/h), lag phase duration (LPD; h), lower asymptote (log CFU/g), and upper asymptote (log CFU/g). The kinetic parameters were then analyzed as a function of storage temperature, using the square root model, polynomial equation, and linear equation. A dynamic model was also developed for varying temperature. The model performance was evaluated against observed data, and the root mean square error (RMSE) was calculated. At $4^{\circ}C$, E. coli cell growth was not observed on any cheese. However, E. coli growth was observed at $10{\circ}C$ to $30^{\circ}C$C with a ${\mu}_{max}$ of 0.01 to 1.03 log CFU/g/h, depending on the cheese. The ${\mu}_{max}$ values increased as temperature increased, while LPD values decreased, and ${\mu}_{max}$ and LPD values were different among the four types of cheese. The developed models showed adequate performance (RMSE = 0.176-0.337), indicating that these models should be useful for describing the growth kinetics of E. coli on various cheeses.

• Korean Journal of Veterinary Research
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• v.38 no.4
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• pp.745-750
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• 1998

A sensitive and rapid enzyme immunoassay(EIA) to detect Escherichia coli O157 in ground beef was developed by using a sandwich type assay with polyclonal antibodies to E coli O157. E coli O157 in ground beef could be detected within 15hr, including incubation for 12hr in enrichment broth and 3hr in immunoassay. The EIA could detect $1.3{\times}10^5$ cells of E coli O157/g of ground beef without enrichment. The lowest limit of detection was 0.23 E coli O157 per g of meat after enrichment. Confirmation was required in the positive specimens in the EIA by culture method even though the negative specimens were not. These results suggested that the immunoassay could be a very efficient method for the screening E coli O157 in meat.

• Food Science of Animal Resources
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• v.38 no.4
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• pp.829-834
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• 2018

The objective of this study was to determine the minimum enrichment time for different types of food matrix (pork, beef, and fresh-cut lettuce) in an effort to improve Escherichia coli detection efficiency. Fresh pork (20 g), beef (20 g), and fresh-cut lettuce (20 g) were inoculated at 1, 2, and 3 Log CFU/g of Escherichia coli. Samples were enriched in filter bags for 3 or 5 h at $44.5^{\circ}C$, depending on sample type. E. coli cell counts in the samples were enriched in E. coli (EC) broth at 3 or 5 h. One milliliter of the enriched culture medium was used for DNA extraction, and PCR assays were performed using primers specific for uidA gene. To detect E. coli (uidA) in the samples, a 3-4 Log CFU/mL cell concentration was required. However, E. coli was detected at 1 Log CFU/g in fresh pork, beef, and fresh-cut lettuce after 5, 5, and 3-h enrichment, respectively. In conclusion, 5-h enrichment for fresh meats and 3-h enrichment for fresh-cut lettuce in EC broth at $44.5^{\circ}C$, and PCR analysis using uidA gene-specific primers were appropriate to detect E. coli rapidly in food samples.

• Korean Journal of Soil Science and Fertilizer
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• v.47 no.6
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• pp.432-436
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• 2014

While searching for healthier diets, people became more attentive to agricultural organic products. However, organic foods may be more susceptible to microbiological contamination because of the use of livestock manure compost and liquid manure, potential sources of pathogenic bacteria. This study was undertaken to investigate the persistence of Salmonella enterica, Escherichia coli O157:H7, and Listeria monocytogenes in soil, liquid manure amended soil, and liquid manure. Loamy soil, liquid manure amended soil, and liquid manure were inoculated with S. enterica, E. coli O157:H7, and L. monocytogenes. Samples were incubated in consistent moisture content at $25^{\circ}C$. Samples had been periodically collected during 120 days depending on the given conditions. S. enterica and E. coli O157:H7 survived over 120 days in loamy soil and over 60 days in liquid manure amended soil, respectively. L. monocytogenes decreased faster than other pathogens in soil. S. enterica, E. coli O157:H7, and L. monocytogenes survived for up to 5 days in liquid manure. S. enterica and E. coli O157:H7 in soil decreased by 2 to $2.5log\;CFU\;g^{-1}$ for 120 days. S. enterica and E. coli O157:H7 in liquid manure amended soil decreased slowly for 21 days. However, S. enterica, E. coli O157:H7, and L. monocytogenes sharply decreased after 21 days. S. enterica, E. coli O157:H7, and L. monocytogenes in soil increased by 0.5 to $1.0log\;CFU\;g^{-1}$ for 7 days. Foodborne pathogens in soil and liquid manure amended soil gradually decreased over time.

• Microbiology and Biotechnology Letters
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• v.32 no.3
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• pp.212-217
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• 2004

The gene coding for mannanase from Bacillus subtilis WL-7, a number of glycosyl hydrolase family 26, was hyperexpressed in Escherichia coli. Two recombinant plasmids, pE7MAN and pENS7, were constructed by introducing the complete mannanase gene and the mature mannanase gene lacking N-terminal signal peptide region into a expression vector pET24a(＋), respectively. The level of mannanase produced by E. coli BL21 (DE3) carrying pENS7, which included the mature mannanase gene, was considerably higher than that by E. coli BL21 (DE3)/pE7MAN. Almost mannanase produced by the recombinant E. coli carrying pENS7 at growth temperature of $37^{\circ}C$ existed as inactive enzyme of insoluble form. Growth at temperature below $31^{\circ}C$ increased the soluble fraction of mannanase having catalytic activity in the recombinant E. coli cells. The highest productivity of active mannanase was observed in cell-free extract of the recombinant E. coli grown at growth temperature ranging from $25^{\circ}C$ to $28^{\circ}C$, while mannanase activity per soluble protein of the cell-free extract was highest in the cells grown at $^31{\circ}C$.

• Microbiology and Biotechnology Letters
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• v.14 no.2
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• pp.155-160
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• 1986

$\alpha$-Amylase gene of B. amyloliquefaciens was cloned to E. coli-yeast shuttle vector YEp-13 and expressed in E. coli. Chromosomal DNA of B. amyloliquefaciens was partially digested with Sau3Al and YEp13 plasmid was cleaved with BamH1. The hybrid plasmid, pHA28, was constructed by shotgun method and transformed to E. coli C600 and HB101. The amount of $\alpha$-amylase produced by transformants of E. coli was about 20% to 30% of that produced by B. amyloli-quefaciens. About 65% of $\alpha$-amylase produced by transformant was secreted into periplasm and the others were located in cytoplasm. $\alpha$-Amylase production was maximal when transformants were cultivated for 15hr to 20hr. As the result of agarose gel electrophoresis, pHA28 plasmid was found to be various in its size. This result suggested that pHA28 plasmid was segregated.

• Journal of Microbiology and Biotechnology
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• v.24 no.6
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• pp.862-868
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• 2014

Certain Escherichia coli (E. coli) strains have the ability to cause diarrheal disease. Five types of diarrheagenic E. coli have been identified, including EHEC, ETEC, EPEC, EAEC, and EIEC. To detect these five diarrheagenic types rapidly, we developed a one-step multiplex PCR (MP-PCR) assay using nine primer pairs to amplify nine virulence genes specific to the different virotypes, with each group being represented (i.e., stx1 and stx2 for EHEC, lt, sth, and stp for ETEC, eaeA and bfpA for EPEC, aggR for EAEC, and ipaH for EIEC). The PCR primers were constructed using MultAlin. The sensitivity and specificity of the constructed multiplex PCR primers were measured using DNA isolated from diarrheagenic E. coli strains representing each group. The limits of detection were as follows: $5{\times}10^1CFU/ml$ for EHEC, $5{\times}10^3CFU/ml$ for ETEC expressing lt and sth, $5{\times}10^4CFU/ml$ for ETEC expressing stp, $5{\times}10^2CFU/ml$ for EPEC, $5{\times}10^4CFU/ml$ for EAEC, and $5{\times}10^2CFU/ml$ for EIEC. To confirm the specificity, C. jejuni, C. perfringens, S. Typhimurium, V. parahaemolyticus, L. monocytogenes, Y. enterocolitica, B. cereus, and S. aureus were used as negative controls, and no amplification was obtained for these. Moreover, this kit was validated using 100 fecal samples from patients with diarrhea and 150 diarrheagenic E. coli strains isolated in Korea. In conclusion, the multiplex PCR assay developed in this study is very useful for the rapid and specific detection of five diarrheagenic E. coli types. This single-step assay will be useful as a rapid and economical method, as it reduces the cost and time required for the identification of diarrheagenic E. coli.