• Korean Journal of Veterinary Service
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• v.15 no.1
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• pp.26-31
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• 1992

The oil emulsion and alhydrogel vaccines were prepared from a strain of enterotoxigenic Escherichia coli isolated from calves with diarrhea and their protective efficacy and immunogenicity were tested in Guinea-pigs. Enterotoxigenic Escherichia coli, isolated from calves with diarrhea, has K99 and F4l antigen as 46.2％ and 50.9％ with 48 and 53 strains respectively out of 104 strains. The protective efficacy of the gel and oil vaccines were 60% and 80% respectively. Agglutinin titers to sera of Guinea-pigs vaccinated with experimental gel and oil vaccines peaked at 5 and 6 weeks after vaccination.

• Korean Journal of Veterinary Research
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• v.26 no.1
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• pp.103-108
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• 1986

The oil emulsion and alhydrogel pilli vaccines were prepared from a strain(O9: K35, K99, F41) of enterotoxigenic Escherichia coli isolated from calves with diarrhea and their immunogenicity was tested in guinea-pigs, pregnant goats and cows. Serum antibody responses to K99 and F41 antigens in guinea-pigs given experimental oil and gel vaccines peaked at 4 and 6 weeks after vaccinations. At that time, the mean hemagglutination inhibition titers to K99 and F41 antigens in guinea-pigs given oil vaccine were 1:25 and 1:1, 218 and those given gel vaccine were 1:54 and 1:724 respectively. Agglutinin titers in pregnant goats given the oil vaccine were significantly higher(mean 1:2,347) compared to those of control group(mean 1:160). Less than 12.5% of goatlings from vaccinated goats developed scours compared to nearly 100% in control group after oral challenge with enterotoxigenic Escherichia coil within 24 hours after birth. The highest agglutinin titers of cow serum and colostrum and of the serum of calves 48 hours after birth from cows given oil vaccine were 1:256, 1:512 and 1:64 respectively. On the other hand, those titers of serum and colostrum and of the serum of nursing calves from nonvaccinated cows were 1:8, 1:16 and 1:20 respectively. The protective efficacy of the oil emulsion vaccine was 72.1% under field conditions. These results strongly indicated that the vaccine could be applied for protection of diarrhea caused by enterotoxigenic Escherichia coli in calves.

• Asian-Australasian Journal of Animal Sciences
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• v.31 no.8
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• pp.1308-1314
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• 2018

Objective: Gut health improvements were monitored with respect to growth performance, diarrhea incidence, fecal bacterial population and intestinal morphology of suckling pigs orally supplemented with live Lactobacillus salivarius (L. salivarius) oral suspensions and challenged with $F4^+$ enterotoxigenic Escherichia coli (ETEC). Methods: Two groups of newborn pigs from 18 multiparous sows were randomly designated as non-supplemented (control: n = 114 piglets) and L. salivarius supplemented groups (treatment: n = 87 piglets). Treatment pigs were orally administered with 2 mL of $10^9$ colony-forming unit (CFU)/mL L. salivarius on days 1 to 3, then they were orally administered with 5 mL of $10^9CFU/mL$ L. salivarius on days 4 to 10, while those in control group received an equal amount of phosphate buffered saline solution. On day 24 (2 weeks post supplementation), one pig per replicate of both groups was orally administered with $10^8CFU/mL$ $F4^+$ ETEC, then they were euthanized on day 29 of experiment. Results: Results revealed that pigs in treatment group had a statistically significant increase in average daily gain, body weight and weight gain, and tended to lower diarrhea throughout the study. Numbers of Lactobacillus population in feces of treatment pigs were higher than control pigs, especially on day 10 of study. Numbers of total bacteria in intestinal contents of control pigs were also increased, but not Coliform and Lactobacillus populations. Histological examination revealed statistically significant improvements of villous height and villous/crypt ratio of duodenum, proximal jejunum and distal jejunum parts of treatment pigs compared with controls. Duodenal pH of treatment group was significantly decreased. Conclusion: Oral supplementation of live L. salivarius during the first 10 days of suckling pig promoted growth performance and gut health, reduced diarrhea incidence, increased fecal Lactobacillus populations and improved intestinal morphology.

• Korean Journal of Veterinary Service
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• v.40 no.4
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• pp.235-244
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• 2017

Enterotoxigenic Escherichia coli (ETEC) strains producing each F4, F5, F6 and F41 fimbriae were lysed by GI24 peptide. The lysate cells were used as ETEC vaccine candidate. This study was carried out to examine whether intramuscular (im) immunization of pregnant sows with the novel vaccine candidate could effectively protect their neonatal piglets against ETEC colibacillosis. All pregnant sows were primed at 11 weeks and were boosted at 14 weeks of pregnancy. Group A sows were im inoculated with PBS. Group B sows were im immunized with $2{\times}10^9$ the mixture. Seral IgG, colostral IgA and IgG titers from group B sows, and seral IgG and IgA levels in group B piglets were significantly higher than those of group A sows and piglets, respectively. After challenge with wild-type ETEC, diarrhea and mortality was not observed in group B piglets. However, diarrhea was observed in 66.7% of group A piglets, and 33.3% mortality was observed. These findings indicate that im immunization of sows with the mixture of the novel vaccine candidate can effectively protect their offspring from ETEC colibacillosis.

• Journal of Veterinary Science
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• v.23 no.1
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• pp.7.1-7.14
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• 2022

Background: Enterotoxigenic Escherichia coli (ETEC) infection is a primary cause of livestock diarrhea. Therefore, effective vaccines are needed to reduce the incidence of ETEC infection. Objectives: Our study aimed to develop a multivalent ETEC vaccine targeting major virulence factors of ETEC, including enterotoxins and fimbriae. Methods: SLS (STa-LTB-STb) recombinant enterotoxin and fimbriae proteins (F4, F5, F6, F18, and F41) were prepared to develop a multivalent vaccine. A total of 65 mice were immunized subcutaneously by vaccines and phosphate-buffered saline (PBS). The levels of specific immunoglobulin G (IgG) and pro-inflammatory cytokines were determined at 0, 7, 14 and 21 days post-vaccination (dpv). A challenge test with a lethal dose of ETEC was performed, and the survival rate of the mice in each group was recorded. Feces and intestine washes were collected to measure the concentrations of secretory immunoglobulin A (sIgA). Results: Anti-SLS and anti-fimbriae-specific IgG in serums of antigen-vaccinated mice were significantly higher than those of the control group. Immunization with the SLS enterotoxin and multivalent vaccine increased interleukin-1β (IL-1β) and tumor necrosis factor-α (TNF-α) concentrations. Compared to diarrheal symptoms and 100% death of mice in the control group, mice inoculated with the multivalent vaccine showed an 80% survival rate without any symptom of diarrhea, while SLS and fimbriae vaccinated groups showed 60 and 70% survival rates, respectively. Conclusions: Both SLS and fimbriae proteins can serve as vaccine antigens, and the combination of these two antigens can elicit stronger immune responses. The results suggest that the multivalent vaccine can be successfully used for preventing ETEC in important livestock.

• Journal of Animal Science and Technology
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• v.62 no.4
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• pp.543-552
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• 2020

For efficient prevention and treatment of enteric colibacillosis, understanding about latest virulence factors and antimicrobial resistance of Escherichia coli is essentially needed. The aim of this study was to survey antimicrobial resistance and determine the prevalence of fimbriae and enterotoxin genes among 118 pathogenic E. coli isolates obtained from Korean pigs with diarrhea between 2016 and 2017. The genes for the toxins and adhesins were amplified by polymerase chain reaction (PCR). The susceptibility of the E. coli isolates to antimicrobials were tested using the standard Kirby-Bauer disk diffusion method. The most prevalent fimbrial antigen was F18 (40.7%), followed by F4 (16.9%), and the most prevalent combinations of toxin genes were Stx2e (21.2%), STb:EAST-1 (19.5%), and STa:STb (16.9%), respectively. Among the pathotypes, enterotoxigenic E. coli (ETEC) was the most predominant (67.8%), followed by Shiga-toxin producing E. coli (STEC, 23.7%). We confirmed high resistance rates to chloramphenicol (88.1%), tetracycline (86.4%), streptomycin (86.4%), and ampicillin (86.4%). And the majorities of isolates (90.7%) showed multi-drug resistance which means having resistance to 3 or more subclasses of antimicrobials. Results of this study can be a source of valuable data for investigating the epidemiology of and control measures for enteric colibacillosis in Korean piggeries.

• Korean Journal of Agricultural Science
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• v.41 no.2
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• pp.135-139
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• 2014

Pathogenic E. coli associated post-weaning diarrhea (PWD) and edema disease are common diseases in commercially-housed weanling pigs. An enterotoxigenic E. coli (ETEC) oral challenge model has been used to mimic the physiological responses observed in commercial conditions. However, an oral challenge procedure has two major limitations: (1) the ETEC cell density is unknown at the point of oral inoculation, and (2) blending ETEC with traditional TSB (trypticase soy broth) is not palatable and hence decreases acceptability by piglets. Therefore, the purposes of this study were to (1) establish a regression equation that can be used for estimation of ETEC concentration in dilution media using the spectrophotometric measurement of cell density; and (2) examine survival of ETEC after blending either with TSB, sweetener or dextrose. A strain of ETEC (serogroup beta-hemolytic E. coli O149; K91; F4; toxins LT, STa, STb) was grown in TSB for 3.5 hours, centrifuged, the supernatant was discarded, and the ETEC pellet was then blended either with TSB (100 mL), sweetener (60 mL TSB + 40 mL fruit flavored concentrate), or dextrose (50 mL TSB + 50 mL dextrose; 0.5g/mL dextrose). Cell density was measured using the colorimetric method and also plated on a 5% sheep blood agar for counting of ETEC colony forming units at 0, 5, 35, 65 and 125 min after blending. The optical density at 600 nm explained 83% of ETEC colony forming units, indicating that the established linear equation (y= 6E+08x - 4E+07, P<0.004) can be used for robust quantification of ETEC cell density in TSB, sweetener and dextrose media. When ETEC was blended with sweetener and dextrose, survival of ETEC was decreased by 45% and 72% within 5 min post-blending. Therefore, further research is required to find out the suitable medium that has potential to improve palatability without compromising survival of ETEC.

• Asian-Australasian Journal of Animal Sciences
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• v.21 no.4
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• pp.489-493
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• 2008

In 2004, Jorgensen and coworkers proposed the MUC4 gene as a candidate gene of enterotoxigenic Escherichia coli (ETEC) F4ab/ac receptor in piglets and a mutation of $G{\rightarrow}C$ in intron 7 of MUC4 was identified to be associated with the ETEC F4ab/ac adhesion phenotypes. In this study, we used 310 piglets of three breeds (Landrace, Large White and Chinese Songliao Black) to analyze the relationship between this mutation and the F4ab/ac adhesion phenotype. The results show that the genotypes at this site and the ETEC F4ab/ac adhesion phenotypes were not completely consistent, although they are very strongly associated. Among the individuals with genotype CC, which was identified as a resistant genotype to F4ab/ac adhesion, only 72.1% (124/172) were non-adhesive to ETEC F4ab and 77.9% (134/172) were non-adhesive to ETEC F4ac infections. This suggests that this mutation may not be the causative mutation for ETEC F4ab/ac adhesion, rather, the actual causative mutation may be in another gene closely linked to MUC4, or at aother site within the MUC4 gene. Our results also suggest that the receptors of F4ab and F4ac may be determined by two different but closely linked loci. In order to screen other genes related to F4ab/ac adhesion in piglets, the mRNA profiles from six full sib piglets, of which three were adhesive to ETEC F4ab/ac and three non-adhesive, were analyzed by suppression subtractive hybridization (SSH). One up-regulated gene, Ep-CAM, was selected for further analysis based on its role in the intestinal epithelial cells adhesion. Using real-time RT-PCR, we found that the Ep-CAM gene was significantly up-regulated in the piglets adhesive to F4ab/ac. It was mapped to SSC3q11-q14 by radiation hybrid mapping.

• Journal of Food Hygiene and Safety
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• v.35 no.3
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• pp.271-278
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• 2020

Enterotoxigenic Escherichia coli is one of the major causative infectious agents of diarrhea in newborn and post-weaning pigs and leads to a large economic loss worldwide. However, there is limited information on the distribution and characterization of virulence genes in E. coli isolated from diarrheic piglets, which also applies to the current status of pig farms in Korea. To investigate the prevalence and characterization of virulence genes in E. coli related to diarrhea in piglets, the rectal swab samples of diarrheic piglets (aged 2 d to 6 w) were collected from 163 farms between 2013 and 2016. Five to 10 individual swab samples from the same farm were pooled and cultured on MacConkey agar plates, and E. coli were identified using the API 32E system. Three sets of multiplex PCRs were used to detect 13 E. coli virulence genes. As a result, a total of 172 E. coli isolates encoding one or more of the virulence genes were identified. Among them, the prevalence of individual virulence gene was as follows, (1) fimbrial adhesins (43.0%): F4 (16.9%), F5 (4.1%), F6 (1.7%), F18 (21.5%), and F41 (3.5%); (2) toxins (90.1%): LT (19.2%), STa (20.9%), STb (25.6%), Stx2e (15.1%), EAST1 (48.3%); and (3) non-fimbrial adhesin (19.6%): EAE (14.0%), AIDA-1 (11.6%) and PAA (8.7%), respectively. Taken together, various pathotypes and virotypes of E. coli were identified in diarrheic piglets. These results suggest a broad array of virulence genes is associated with coliform diarrhea in piglets in Korea.

• Korean Journal of Veterinary Service
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• v.44 no.4
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• pp.271-281
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• 2021

Pathogenic Escherichia coli (E. coli) is one among the most important agents of diarrhea in calves. From January to December 2021, 108 isolates from feces of calves with diarrhea were investigated for enterotoxigenic E. coli (ETEC), enteropathogenic E. coli (EPEC), shiga toxin-producing E. coli (STEC), enteroaggregative E. coli (EAEC), and enteroinvasive E. coli (EIEC) using real-time PCR. In addition, the genes for F5, F17 and F41 fimbriae were detected by PCR. The most frequently isolated pathotypes were EPEC/STEC (29 isolates), and ETEC/EPEC/STEC (29 isolates). ETEC/EPEC, and ETEC/STEC were also found in 10 isolates. EPEC, STEC, and ETEC were detected in 13, 11, and 6 respectively. EAEC, and EIEC was not detected. Antimicrobial resistance test was carried out by agar disc diffusion method with 14 antimicrobials. Among 108 pathogenic E. coli isolates, 107 isolates were resistant to at least one of 14 antibiotics used in this study, 99 (91.7%) were resistant to two or more antimicrobials, and a single remarkable isolate was resistant to 14 antimicrobials. The isolates were primarily resistant to penicillins, streptomycin, tetracycline, ceftiofur, Trimethoprim/sulfamethoxazole, Kanamycin, and Ciprofloxacin. The high rate of resistance in pathogenic E. coli, sometimes to multiple drugs, may complicate future options for treating human infections. These results may bu used for diagnosis and therpeitic purposes in calves with diarrhea.