The major immunogenic protein capsid (Cap) of porcine circovirus type 2 (PCV2) is critical to induce neutralizing antibodies and protective immune response against PCV2 infection. This study was conducted to investigate the immune response of recombinant adenovirus expressing PCV2b Cap and C-terminal domain of Yersinia pseudotuberculosis invasin (Cap-InvC) fusion protein in pigs. The recombinant adenovirus rAd-Cap-InvC, rAd-Cap and rAd were generated and used to immunize pigs. The phosphate-buffered saline was used as negative control. The specific antibodies levels in rAd-Cap-InvC and ZJ/C-strain vaccine groups were higher than that of rAd-Cap group (p < 0.05), and the neutralization antibody titer in rAd-Cap-InvC group was significantly higher than those of other groups during 21–42 days post-immunization (DPI). Moreover, lymphocyte proliferative level, interferon-γ and interleukin-13 levels in rAd-Cap-InvC group were increased compared to rAd-Cap group (p < 0.05). After virulent challenge, viruses were not detected from the blood samples in rAd-Cap-InvC and ZJ/C-strain vaccine groups after 49 DPI. And the respiratory symptom, rectal temperature, lung lesion and lymph node lesion were minimal and similar in the ZJ/C-strain and rAd-Cap-InVC groups. In conclusion, our results demonstrated that rAd-Cap-InvC was more efficiently to stimulate the production of antibody and protect pigs from PCV2 infection. We inferred that InvC is a good candidate gene for further development and application of PCV2 genetic engineering vaccine.
Adenoviridae ; Adenovirus Vaccines ; Antibodies ; Antibodies, Neutralizing ; Capsid ; Capsid Proteins ; Circovirus ; Genetic Engineering ; Immunization ; Interleukin-13 ; Lung ; Lymph Nodes ; Lymphocytes ; Swine ; Yersinia pseudotuberculosis

BACKGROUND: Yersinia pseudotuberculosis is known to cause fever, gastroenteritis, or acute kidney injury (AKI). There have been several Y. pseudotuberculosis infection outbreaks to date associated with ingestion of contaminated food or unsterile water. While this disease was considered to have practically been eradicated with the improvement in public health, we encountered several cases of AKI associated with Yersinia infection. METHODS: We retrospectively collected data from medical records of patients with suspected Y. pseudotuberculosis infection who visited Seoul National University Children’s Hospital in 2017. RESULTS: There were nine suspected cases of Yersinia infection (six males and three females; age range 2.99–12.18 years). Among them, five cases occurred in May, and seven patients were residing in the metropolitan Seoul area. Three patients had history of drinking mountain water. Every patient first presented with fever for a median of 13 days, followed by gastrointestinal symptoms and oliguria. Imaging studies revealed mesenteric lymphadenitis, terminal ileum wall thickening, and increased renal parenchymal echogenicity. Creatinine levels increased to 5.72 ± 2.18 mg/dL. Urinalysis revealed sterile pyuria, proteinuria, and glycosuria. Oliguria continued for 4 to 17 days, and two patients required dialysis; however, all of them recovered from AKI. Mucocutaneous manifestations developed later. In the diagnostic work-up, Yersinia was isolated from the stool culture in one patient. Anti-Yersinia immunoglobulin (Ig) A and IgG were positive in 6 patients. CONCLUSION: Y. pseudotuberculosis infection is an infrequent cause of interstitial nephritis presenting with AKI. When a patient presents with fever, gastroenteritis, and AKI not resolving despite hydration, the clinician should suspect Y. pseudotuberculosis infection.
Acute Kidney Injury ; Creatinine ; Dialysis ; Disease Outbreaks ; Drinking ; Eating ; Female ; Fever ; Gastroenteritis ; Glycosuria ; Humans ; Ileum ; Immunoglobulin G ; Immunoglobulins ; Male ; Medical Records ; Mesenteric Lymphadenitis ; Nephritis, Interstitial ; Oliguria ; Proteinuria ; Public Health ; Pyuria ; Retrospective Studies ; Seoul ; Urinalysis ; Water ; Yersinia Infections ; Yersinia pseudotuberculosis ; Yersinia

Animals ; Bacterial Proteins ; genetics ; metabolism ; Biofilms ; growth & development ; Gene Expression Regulation, Bacterial ; Lipopolysaccharides ; chemistry ; metabolism ; Operon ; Promoter Regions, Genetic ; Protein Binding ; Siphonaptera ; microbiology ; Species Specificity ; Transcription, Genetic ; Transferases ; genetics ; metabolism ; Virulence ; Yersinia pestis ; genetics ; metabolism ; pathogenicity ; Yersinia pseudotuberculosis ; genetics ; metabolism

Two trials of external quality assessment for clinical microbiology laboratories were performed in 2008. A total of 16 specimens were distributed. Eight specimens were distributed to 330 laboratories with 319 (96.7%) returns in Trial I, and 8 specimens to 335 laboratories with 319 returns (95.2%) in Trial II. Two slide specimens for mycobacterium stain (AFB) were distributed in Trial I and II. The acceptable percentages of Gram stain were relatively good for both stainability and morphology except for Acinetobacter baumannii. The acceptable percentages of bacterial identification (correct answers to species level) on Klebsiella pneumoniae, Staphylococcus aureus, Neisseria meningitidis, Serratia marcescens, Erysipelothrix rhusiopathiae and Candida albicans (Trial I) were 97.4%, 99.2%, 55.6%, 97.0%, 79.2%, and 92.0%, respectively. The acceptable percentages of bacterial identification on A. baumannii, Enterococcus faecalis, Streptococcus pyogenes, Haemophilus parainfluenzae, Elizabethkingia meningoseptica, and Yersinia pseudotuberculosis (Trial II) were 92.0%, 90.8%, 4.5%, 53.1%, 74.8% and 94.3%, respectively. The acceptable percentages for antimicrobial susceptibility tests on K. pneumoniae and S. aureus (Trial I), and A. baumannii and E. faecalis, (Trial II) were relatively good compared to data of the last year. The acceptable percentages for AFB stain in Trial I and II were relatively high. In summary, the acceptable percentages of bacterial stain and identification were relatively good except some cases with poor specimen quality. However, it is still necessary that the quality assurance of the individual laboratories should be improved for antimicrobial susceptibility tests, and the selection of the most appropriate antimicrobial agents to test should be also considered.
Acinetobacter baumannii ; Anti-Infective Agents ; Candida albicans ; Enterococcus faecalis ; Erysipelothrix ; Haemophilus parainfluenzae ; Klebsiella pneumoniae ; Korea ; Mycobacterium ; Neisseria meningitidis ; Pneumonia ; Serratia marcescens ; Staphylococcus aureus ; Streptococcus pyogenes ; Yersinia pseudotuberculosis

BACKGROUND: Yersinia pseudotuberculosis is recognized throughout the world as a cause of water-or food born infections in human and animals. Although many attempts have been made to define optimal conditions for the isolation of the organism from water, their isolation yields remain low; therefore, we tried to find an effective method for the recovery of Y. pseudotuberculosis from water. METHODS: Water samples were deliberately contaminated with Y. pseudotuberculosis at various levels and then processed by the following three isolation METHODS: centrifugation, direct filtration, and intracellular culture. For the centrifugation method, the water samples were centrifuged at 5000 rpm for 1 hr and the final precipitates were inoculated in cefsulodin-irgasan-novobiocin(CIN) media. For the filtration method, the water samples were filtered by negative pressure and the filter papers were put directly on CIN media. For the intracellular culture method, the organisms were extracted from the HeLa cells that had been infected with Y. pseudotuberculosis and inoculated on CIN media. We also examined the efficacy of the filtration method after cold enrichment with a mixture of Y. pseudotuberculosis, Escherichia coli, and Citrobacter freundii. RESULTS: With the concentration of 3x10(2)/100 mL, Y. pseudotuberculosis was isolated only by the filtration method; however, none of the culture methods were good enough to recover the organism from the water sample when the concentration was 3x10/100 mL. With cold enrichment, however, the recovery was much more efficient; the organism grew after direct inoculation or after filter inoculation when the starting concentrations were 3x10(2)/100 mL or 3x10/100 mL, respectively. CONCLUSION: A combined use of direct filtration and filter inoculation after cold enrichment is the most effective method to yield Y. pseudotuberculosis isolation. The introduction of effective methods for the isolation of Y. pseudotuberculosis from untreated drinking water would increase the awareness by the public of the health hazard of spring water.
Animals ; Centrifugation ; Citrobacter freundii ; Drinking Water ; Escherichia coli ; Filtration ; HeLa Cells ; Humans ; Water* ; Yersinia pseudotuberculosis* ; Yersinia*

In order to investigate pathogenic genes and genetic relationships of Y. pseudotuberculosis strains, We isolated 9 strains of Y. pseudotuberculosis from 380 spring water sites in Seoul from 2000 to 2003. All isolates were distributed to the northeast area in Seoul. The isloates were analyzed for chromosomal virulence gene (inv) and plasmid-borne genes (yadA and lcrF) using PCR to assume pathogenicity. As a result, all isolates were positive for the inv gene, but only five isolates (55.6%) were positive for the yadA and lcrF genes. RAPD and PCR-ribotyping were tested and all isolates were grouped with 90% similarity. RAPD revealed 4 clusters and PCR-ribotyping revealed 2 clusters. The result of this experiments confirmed the view that RAPD had better powerful discrimination than PCR-ribotyping and RAPD typing was effective to distinguish between various strains of Y. pseudotuberculosis from spring water.
Discrimination (Psychology) ; Molecular Typing* ; Polymerase Chain Reaction ; Seoul* ; Virulence ; Yersinia pseudotuberculosis* ; Yersinia*

A 40-yr-old buddhist monk was admitted to the hospital with abdominal pain, fever, and confusion. He had a history of drinking untreated mountain spring water in his temple, and experienced the above symptoms for several days before admission. In past medical history, he had suffered from hepatic cirrhosis. Yersinia pseudotuberculosis was isolated from his blood and ascitic fluid. The mountain spring water that he had ingested was cultivated and Y. pseudotuberculosis was also isolated. For identification of pathogenic Y. pseudotuberculosis, each isolate from the three sources (blood, ascitic fluid, and drinking water) was also analysed for the inv gene for Y. pseudotuberculosis and the virF gene for virulent plasmid by PCR. All strains were positive for both the virF and the inv genes and also positive for autoagglutination test. For relationship study, each isolate from the three sources was also analysed with serotyping and restriction endonuclease analysis of virulence plasmid DNA (REAP) using BamHI. All belonged to the serotype 4b and REAP pattern D. Thus, all these findings supported that the mountain spring water was the source of the Y. pseudotuberculosis infection in this case.
Adhesins, Bacterial/genetics ; Adult ; Agglutination Tests ; Bacterial Proteins/genetics ; DNA, Bacterial/analysis ; Feces/microbiology ; Food ; Human ; Male ; Plasmids ; Restriction Mapping ; Septicemia/diagnosis/microbiology ; Serotyping ; Virulence Factors/genetics ; *Water Supply ; Yersinia pseudotuberculosis/classification/*genetics/isolation & purification ; Yersinia pseudotuberculosis Infections/*diagnosis/*transmission

Yersinia pseudotuberculosis is a relatively infrequent cause of human infections, mostly as intestinal yersinosis. A septicemic form of Y. pseudotuberculosis infection has been reported only rarely. It is usually seen in patients with underlying disorders such as diabetes, hepatic cirrhosis or iron overload. A 63-year-old man with diabetes mellitus and liver fibrosis was admitted to Asan Medical Center via emergency department because of epigastric pain, fever and watery diarrhea; he was septic. The stool culture did not grow Salmonella, Shigella, or Yersinia. But, in the blood culture Y. pseudotuberculosis grew from one anaerobic vial among two sets of aerobic and anaerobic blood cultures. Serotype of Y. pseudotuberculosis strain was could not be determined because it was a rough type. The isolate was positive in the autoagglutination test and polymerase chain reaction for the virF gene. The serum levels of iron, TIBC and ferritin were within normal range. The patient received ceftriaxone therapy for 3 days and was discharged with a clinical improvement.
Ceftriaxone ; Chungcheongnam-do ; Diabetes Mellitus ; Diarrhea ; Emergency Service, Hospital ; Ferritins ; Fever ; Humans ; Iron ; Iron Overload ; Liver Cirrhosis ; Middle Aged ; Polymerase Chain Reaction ; Reference Values ; Salmonella ; Sepsis* ; Shigella ; Yersinia pseudotuberculosis* ; Yersinia*

A 40-year-old man with systemic lupus erythematosus (SLE) and diabetes was found to have sepsis with multiple small hepatic abscesses secondary to Yersinia pseudotuberculosis which were detected by computed tomography (CT) scan and blood cultures. Sepsis with Y. pseudotuberculosis is uncommon but usually seen in patients with underlying liver diseases or diabetes. A few of those patients are accompanied by liver abscesses. Those patients with liver abscesses invariably have multiple small abscesses. CT scan of the liver was important in demonstrating the multiple small liver abscesses. Identification of the pathogen on blaod culture and elevated serum antibody titer to Y. pseudotu-berculosis are useful for diagnosis. Although rare, Y. pseudotuberculosis should be also considered as a possible cause in febrile patient with immunocompromised state such as SLE, diabetes or hemochromatosis.
Abscess ; Adult ; Diagnosis ; Hemochromatosis ; Humans ; Liver Abscess* ; Liver Diseases ; Liver* ; Lupus Erythematosus, Systemic* ; Sepsis* ; Tomography, X-Ray Computed ; Yersinia pseudotuberculosis* ; Yersinia*

BACKGROUND: Yersinia pseudotuberculosis, a member of genus Enterobactericeae, is a main etiologic organism of diarrhea in childhood. Because a mouse and a unchlorinated spring water are main reservoirs of Y. pseudotuberculosis, the strains from a contaminated spring water and mouse could be involved in human epidemic. The purpose of this study was to investigate a clonality between the strains from patients and those from an unchlorinated spring water and a mouse by restriction enzyme analysis of plasmid DNA and pulsed-field gel electrophoresis (PFGE). METHOD: We isolated 15 Y. pseudotuberculosis strains including 8 isolates from patients (S1-S8), 6 isolates from mountain water (W1-W6), 1 isolate from a mouse (M1) in northeast area of Seoul. Plasmid and chromosomal DNA of all strains were analyzed by REAP with Bam H1 restriction and by PFGE with Xba I restriction , respectively. RESULTS: Restriction enzyme analysis of plasmid DNA was classified into type B and type D. All 7 strains of serotype 15 were classified as type B and 8 strains of serotype 4b were classified as type D. PFGE were classified into 6 different types. Among them, strains of PFGE type I, II, III, IV belong to Y. pseudotuberculosis serotype 15 and Y. pseudotuberculosis 4b strains were classified into PFGE type V, VI. S1 and W1 were classified into PFGE type I . S8, W6 and M1 were classfied into PFGE type VI. CONCLUSIONS: PFGE revealed clonality among strains from patients, a water and a mouse. PFGE was more discriminative than REAP to characterize the Y. pseudotuberculosis outbreaks in Korea.
Animals ; Diarrhea ; Disease Outbreaks ; DNA* ; Electrophoresis, Gel, Pulsed-Field ; Humans ; Korea* ; Mice ; Plasmids* ; Restriction Mapping* ; Seoul ; Yersinia pseudotuberculosis* ; Yersinia*