The H9N2 strains of avian influenza viruses (AIVs) circulate worldwide in poultry and cause sporadic infection in humans. To better understand the evolution of these viruses while circulating in poultry, an H9N2 chicken isolate was passaged 19 times in chickens via aerosol inoculation. Whole-genome sequencing showed that the viruses from the initial stock and those after the 8th and 19th passages (P0, P8, and P19) all had the same monobasic cleavage site in the hemagglutinin (HA), typical for viruses of low pathogenicity. However, at position 226 of the HA protein the ratio of glutamine (which favors avian-type receptor binding) to leucine (which favors mammalian-type receptor binding) decreased from 54:46 in P0, to 87:13 in P8, and then 0:100 in P19. In chickens exposed to aerosols of P0, P8, or P19, replication of the viruses was similar and mainly limited to the respiratory tract. None of the infected chickens showed any clinical signs. Over the 19 passages the viruses maintained relatively stable infectivity but gradually lost lethality to chicken embryos. According to the hemagglutination inactivation assay, P8 was slightly and P19 significantly (P < 0.05) less thermostable than P0. Collectively, after 19 passages in chickens the H9N2 AIVs retained low pathogenicity with a positive selection of L226 in the HA. These findings suggest that H9N2 viruses might acquire mammalian specificity after asymptomatic circulation in avian species.

Colibacillosis is a disease caused by avian pathogenic E. coli (APEC) and is one of the principle cause of morbidity and mortality in poultry worldwide which is represented by a complex syndrome characterized by multiple organ lesions. This study was carried out to determine phylogenetic grouping and virulenceassociated genes contained by E. coli isolates which is related in causing disease in chicken. E. coli isolates obtained from clinical cases of Veterinary ResearchInstitute were re-identified by conventional methods. Phylogenetic grouping of the isolates was determined by triplex polymerase chain reaction (PCR), and the presence of eight virulence genes were identified by multiplex PCR. A total of 125 E. coli isolates were subjected toanalysis of phylogenetic background and virulence associated genes profiling. Phylogenetic analysis demonstrated that most of the E. coli isolated from chicken in this study belonged to group B1 (36.0%),group D (28.0%), group A (27.2%) and group B2 (8.8%). Multiplex PCR analysis demonstrated that 96 (78.6%) of the E. coli isolates harbored at least one virulencegene, while 29 (23.3%) did not contain any virulence genes tested. The most prevalent virulence genes identified were iss (51.2%), followed by iucD (36.0%),tsh (32.8%), vat (16.0%), astA (13.6%), irp2 (11.2%), papC (9.6%) and the least is cva/cvi gene (0%). None of the isolates harbored more than four virulence genes.Each of phylogenetic groups presented with different combinations of virulence genes, with no specific combinations of virulence genes found to correlate withE. coli phylogroups. None of the E. coli isolates harbored more than four virulence genes, suggesting that E. coli isolates from chicken in this study appear to bederived from commensal strains and may relate to environmental predispose factors especially stress factors in the host to establish infection.

The prevalence of thermophilic Campylobacter (C.) spp. in stray, breeding, and household dogs was 25.2, 12.0, and 8.8%, respectively. C. jejuni and C. upsaliensis were the predominant Campylobacter spp. from household dogs. cdtA, cdtB, and cdtC were detected by PCR in all isolates. Despite the high cytolethal distending toxin (CDT) gene prevalence, only 26 (31%) C. jejuni strains and one (15.3%) C. coli strain showed evidence of CDT production in HEp-2 cell cytotoxicity assays. Virulence-associated genes detected in the C. jejuni and C. coli isolates were cadF, dnaJ, flaA, racR, ciaB, iamA, pldA, virB11, ceuE, and docC. cadF, dnaJ, flaA, and ceuE were found in all C. jejuni and C. coli isolates. When detecting Guillain-Barre syndrome-associated genes (galE, cgtB, and wlaN), galE was identified in all isolates. However, cgtB and wlaN were more prevalent in C. jejuni isolates from humans than those from dogs. Adherence and invasion abilities of the C. jejuni and C. coli strains were tested in INT-407 cells. A considerable correlation (adjusted R©�= 0.678) existed between adherence and invasion activities of the Campylobacter spp. isolates.

A total of 18 Newcastle disease virus (NDV) isolates that were recovered from 1949 through 1997 were characterized and pathotyped. All viruses were highly virulent as determined by intracerebral pathogenicity indices greater-than or equal to 1.81 in day-old. These pathotypes are typical for viscerotropic velogenic NDV (VVNDV) pathotype viruses. Some differences were observed for the chicken red blood cell elution rate and thermostability of the hemagglutinin at 56℃. Three antigenic groups were identified by a hemagglutination-inhibition assay using NDV monoclonal antibodies. And the predominant gross lesions were as follows: discharge from the nasal cavity, tracheal mucus, petechial hemorrhage in the heart fat, kidney urates and hemorrhage with or without necrosis in the gastrointestinal tract. Severe hemorrhagic or necrotic lesions were also noted in the lymphoid organs and were localized primarily in the spleen and cecal tonsil. However, differences in the occurrence and frequency of the gross lesions were observed between the virus strains. Among them, NDV strains that induced neurological symptoms belonged only to genotype Ⅵ. This strain had spread throughout Korea during the late 1980s to the 1990s, which suggests that specific VVNDVs genotypes might result in neurological symptoms.

In this study, we isolated 12 of Brucella (B.) spp. from cattle, which have been positive in Rose Bangal test and tube agglutination test in Gyeongbuk province in 2009. According to AMOS PCR analysis, isolated 12 strains were identified as B. abortus. Murine derived macrophage, RAW 264.7 cells, were infected with isolated 12 strains or reference strain (B. abortus 544), and bacterial internalization were characterized. According to these results, we divided the isolated strains into the following three groups: class Ⅰ, lower internalization than that of B. abortus 544; class Ⅱ, similar internalization to that of that of B. abortus 544; class Ⅲ, higher internalization than that of B. abortus 544 within RAW 264.7 cells. Furthermore, intracellular growth, bacterial adherent assay, LAMP-1 colocalization, virulence in mice and surface protein pattern were characterized. From these results, representative strains of class Ⅲ showed lower LAMP-1 colocalization, higher adherent efficiency, higher virulence in mice than those of B. abortus 544, and showed different pattern of surface proteins. These results suggest that B. abortus field strains, isolated from cattle in Korea, possess various virulence properties and higher internalization ability of field strain may have an important role for its virulence expression.

The pathogenicity and acute toxicity of Lactobacillus (L.) pentosus PL11 from eel (Anguilla japonica) were investigated using male and female Sprague-Dawley rats. The pathogenicity of L. pentosus PL11 was examined after treating the rats with 10∨11 CFU/mL, 10∨9 CFU/mL or 10∨7 CFU/mL doses of L. pentosus PL11 culture or 0.85% NaCl (Control) intragastrically. For acute toxicity studies, rats were treated with dried culture broth of L. pentosus PL11 at doses of 5,000 mg/mL, 2,500 mg/mL, 1,250 mg/mL or 625 mg/mL or Lactobacilli MRS broth (Control), and clinical signs or mortalities were monitored for two weeks. The results of the present investigation revealed no mortalities or obvious clinical signs in rats administered with the live bacterial cultures or dried culture broth at any investigated dose level. Also, no significant differences were observed in net body weight gain, gross pathological findings, feed and water consumption and body temperature among the different treatment groups and between the treated and control rats. It can be concluded from the above findings that L. pentosus PL11 is a safe probiotic strain with potential as feed additive to increase the feed efficiency or health of fish.

A stable and reliable Helicobacter pylori (H. pylori) infection animal model would be necessary for evaluating vaccine efficacy and helpful for understanding the pathological mechanism of the organism. The aim of the present study is to investigate the effect of ethanol treatment prior to H. pylori inoculation on associated gastric mucosal injury and to establish ethanol-pretreating animal model to study H. pylori infection. Male Mongolian gerbils were used for the study. H. pylori was orally inoculated after 12 h fasting.

The shigellae are common etiological agents of bacillary dysentery in humans and primates. During four years from 2002 to 2005, 22 strains of Shigella spp. were isolated from the diarrheic patients in Seoul region. All of them were identified as S. flexneri by biochemical tests and serotyping. The prevalence of serotypes were variable by year, but the major serotypes were 2a and 3a. In an antimicrobial susceptibility test, all of the isolates were resistant to streptomycin and tetracycline, and susceptible to amikacin, kanamycin, cefoxitin, and gentamicin.

Objective-To test horses for serologic evidence of an association between vesiviral antibodies and abortion. Sample Population-Sera from 141 horses. Procedures-2 experiments were conducted. Experiment 1 comprised sera obtained in 2001 and 2002 from 3 groups of horses (58 mares from farms with a history of abortion problems, 25 mares between 3 and 13 years of age with unknown reproductive histories that were sold at auction breeding-age control mares, and 29 mixed-age males and yearling females sold at auction negative control population). Experiment 2 comprised sera from 3 groups of pregnant mares (10 pregnant mares fed Eastern tent caterpillars ETCs, 9 pregnant mares fed ETC frass only, and 10 pregnant control mares). Sera were analyzed for antibodies against vesivirus by use of a validated recombinant vesivirus-specific peptide antigen in an indirect ELISA. Results-For experiment 1, 37 of 58 (63.8%) mares from farms with abortion problems were seropositive for vesivirus antibodies, whereas 10 of 25 (40%) breeding-age control mares were seropositive. All 29 mixed-age males and yearling females were seronegative for vesivirus antibodies. For experiment 2, 17 of 29 mares aborted (some from each group). Seropositive status for vesivirus antibodies increased from 47.1% (8/17) to 88.2% (15/17) for the pregnant mares that aborted during the experiment. Conclusion and Clinical Relevance-Significant association was detected between seropositive status for vesivirus and abortion in mares; consequently, vesivirus appears to be a pathogenic virus associated with abortion in mares. These data support adding vesivirus antibody testing into diagnostic screening to determine the cause for abortion in mares.

The early interaction of Lawsonia intracellularis with host cells was examined with the use of porcine ileum models. Two conventional swine were anesthetized, and ligated ileum loops were prepared during abdominal surgery. The loops were inoculated with 10⁸ L. intracellularis or saline. After 60 min, samples of each loop were processed for routine histologic and electron microscopic study. Histologic and ultrathin sections of all the loops appeared normal, with no apposition of bacteria and host cells or bacterial entry events in any loop. Portions of ileum from a single gnotobiotic piglet were introduced as xenografts into the subcutis of each flank of 5 weaned mice with severe combined immunodeficiency disease. After 4 wk, 10⁸ L. intracellularis were inoculated into each of 4 viable xenografts with a sterile needle; the other 3 viable xenografts received saline. Histologic and ultrathin sections of all the xenografts 3 wk after inoculation showed relatively normal porcine intestinal architecture, with normal crypts, crypt cell differentiation, and low villous structures; the xenografts treated with the bacteria also showed intracytoplasmic L. intracellularis within crypt and villous epithelial cells. Thus, entry of L. intracellularis into target epithelial cells and multiplication may not be sufficient alone to directly cause cell proliferation. A proliferative response may require active division of crypt cells and differentiation in conjunction with L. intracellularis growth.