It is essential to rapidly and precisely diagnose rabies. In this study, we evaluated four diagnostic methods, indirect fluorescent antibody test (FAT), virus isolation (VI), reverse transcriptase polymerase chain reaction (RT-PCR), and rapid immunodiagnostic assay (RIDA), to detect rabies in animal brain homogenates. Out of the 110 animal brain samples tested, 20 (18.2%) were positive for rabies according to the FAT. Compared to the FAT, the sensitivities of VI, RT-PCR, and RIDA were 100, 100, and 95%, respectively. The specificities of VI, RT-PCR and RIDA were found to be 100, 100, and 98.9%, respectively. Rabies viruses circulating in Korea were isolated and propagated in murine neuroblastoma (NG108-15) cells with titers ranging from 101.5 to 104.5 TCID50/mL. Although the RIDA findings did not completely coincide with results obtained from FAT, VI, and RT-PCR, RIDA appears to be a fast and reliable assay that can be used to analyze brain samples. In summary, the results from our study showed that VI, RT-PCR, and RIDA can be used as supplementary diagnostic tools for detecting rabies viruses in both laboratory and field settings.
Animals ; Antigens, Viral/blood ; Brain/virology ; Fluorescent Antibody Technique, Indirect/*veterinary ; Immunoassay/*veterinary ; RNA, Viral/genetics/isolation & purification ; Rabies/diagnosis/*veterinary/virology ; Rabies virus/genetics/*isolation & purification ; Republic of Korea ; Reverse Transcriptase Polymerase Chain Reaction/*veterinary ; Sensitivity and Specificity

Black rockfish (Sebastes schlegeli) is an important mariculture species in Korea. The production of this fish is drastically declined due to bacterial diseases, particularly streptococcosis caused by Lactococcus garvieae. The bacterial surface characteristics of SJ7 and TY6 were found to have capsule but not NB13 and YS18. The experiential evaluation of L. garvieae pathogenicity, the capsular isolates showed high cumulative mortality i.e. SJ7 (100%) and TY6 (60%) compared to non-capsular isolates. Based on this result the capsular isolates L. garvieae were highly suspected as the causative agent of streptococcosis in rockfish.
Agglutination Tests/veterinary ; Animals ; Bacterial Capsules ; DNA, Bacterial/genetics/isolation&purification ; Fish Diseases/*microbiology/mortality ; Fishes ; Fluorescent Antibody Technique, Indirect/veterinary ; Gram-Positive Bacterial Infections/microbiology/mortality/*veterinary ; Lactococcus/*pathogenicity ; Polymerase Chain Reaction/veterinary

OBJECTIVEFor clarifying the situation of the natural infection of rodents having hemorrhagic fever with renal syndrome (HFRS) virus and to type Hantavirus (HV) using molecular technique in Shenzhen city in 2005, and offering guidance for prevention and control of HFRS.

METHODSData on the host animals was collected from the city of Shenzhen. ELISA and indirect immunofluorscent antibody(IFA) test were applied to the specific antibodies against HV in the sera of captured rats. Direct immunofluorscece assay was adopted to determine HFRS antigens and the lung tissues of the HV infected rats were inoculated into Meriones unguiculata to isolate HV. The whole viral RNA was extracted from the lung tissues of the HV infected rats and amplified the partial M fragments with RT-nested-PCR, using the HV genotype specific primers. The amplified genes were then sequenced, and subjected to genotyping and homology analysis.

RESULTS472 rodents were captured from Shenzhen in 2005. Surveillance on rats demonstrated 9.96% rats carrying HV (with a density of 8.25%) and the main host was Rattus norvegicus. In the blood samples of rats, anti-HV IgG antibodies were detectable in 56 cases by IFA, and proved to be positive in 76 cases by ELISA. We successfully isolated a HV strain designated as SZ2083 from Rattus norvegicus for the first time in Shenzhen and was identified to SEO type by RT-nested-PCR. Compared with the coding region of the M gene of HV L99 virus strain, the homologies of nucleotide among them were 97%, but the homology was 76% of the SZ2083 with HTN 76-118 virus strain.

CONCLUSIONResults showed the existence of natural epidemic areas of HFRS in Shenzhen city. Based on the results of sequencing, it is possible that the Seoul strain of HV might be the predominant serotype of virus harbored.

Animals ; China ; epidemiology ; Cities ; Data Collection ; Enzyme-Linked Immunosorbent Assay ; Fluorescent Antibody Technique, Indirect ; Genotype ; Hantavirus ; classification ; genetics ; isolation & purification ; Hantavirus Infections ; epidemiology ; veterinary ; Rats ; virology ; Rodentia ; virology

A virus strain, showing cytopathic effect in Vero cell, was isolated from plasma of a fattening pig in Gyeonggi province, Korea in October 1999. The evaluation of physicochemical/biological properties of the isolate showed that the virus, KV1899, inoculated suckling mouse showed paralysis and died within 7 days post-inoculation, the mouse brain suspension had hemagglutinating activity with goose RBC. Pathogenicity of isolate was carried out by intracranial and intraperitoneal inoculation of 3-4 weeks mice. The mice inoculated with isolate showed 10 4.5 LD50/ 0.03 ml and 10 3.0 LD50/0.5 ml according to the inoculation route. The isolate was identified as RNA and enveloped virus using IUDR and chloroform sensitivity test. The virus particles within the infected Vero cell were measured to be 40-50 nm in size by electron microscopy. The isolate was further characterized by immuno-fluorescence assay using Japanese encephalitis virus (JEV) specific monoclonal antibodies. Reverse transcription polymerase chain reaction (RT-PCR) revealed the presence of JE specific conserved sequences in this isolate. The artificially inoculated pigs had HI titer of 320 to 2,560 against JEV at 14 to 42 days post inoculation. We confirmed this isolate as Japanese encephalitis virus. It was the second isolation of JEV in pigs in Korea.
Animals ; Antibodies, Viral/analysis ; Cercopithecus aethiops ; Cytopathogenic Effect, Viral ; Encephalitis Virus, Japanese/*classification/*isolation & purification/ultrastructure ; Encephalitis, Japanese/pathology/*veterinary/virology ; Fluorescent Antibody Technique, Indirect/veterinary ; Hemagglutination Inhibition Tests/veterinary ; Hemagglutination Tests/veterinary ; Korea ; Mice ; Microscopy, Electron/veterinary ; RNA, Viral/analysis ; Reverse Transcriptase Polymerase Chain Reaction/veterinary ; Swine ; Swine Diseases/pathology/*virology ; Vero Cells/virology

During the period from January to December of 2001, a total of 3,391 swine sera were submitted to our laboratory from 256 farms for the diagnosis of porcine reproductive and respiratory syndrome (PRRS). The antibody to porcine reproductive and respiratory syndrome virus (PRRSV) was tested by the indirect immunofluorescent antibody (IFA) test. Of the 256 farms tested, 230 farms (89.8%) were positive for the PRRSV antibody. The overall seroprevalence of the PRRSV antibody was 52.1% (1765/3391). Most of the pigs seemed to be infected with PRRSV at around 50 to 60 days old. The seroprevalence of the antibody became higher with age, and peaked at around 100 days old. More than one-third of the adult pigs, including boars, gilts, and sows, was positive for the PRRSV antibody. The infection of PRRSV was chronic and confined to growers and/or finishers in most farms. However, the antibody was detected in all production phases at some farms.
Age Factors ; Animals ; Antibodies, Viral/blood ; Female ; Fluorescent Antibody Technique, Indirect/veterinary ; Korea/epidemiology ; Male ; Porcine Reproductive and Respiratory Syndrome/diagnosis/epidemiology/*virology ; Porcine respiratory and reproductive syndrome virus/immunology/*isolation &purification ; Seroepidemiologic Studies ; Sex Factors ; Swine

Amplification of the 16S rRNA gene from a blood sample obtained from a dog in southeastern Brazil was used to confirm a naturally acquired Ehrlichia (E.) canis infection. Following isolation and culturing of the new bacterial strain called Uberlandia, partial sequences of the dsb and p28 genes were obtained. The dsb partial sequence of the novel strain was 100% similar to dsb gene sequences of E. canis obtained from different geographic areas around the world. Conversely, the p28 partial sequence for the E. canis Uberlandia strain differed at several nucleotides from other sequences available in GenBank. To confirm the antigenic profile of the Uberlandia strain, an indirect immunofluorescence assay against E. canis antigens was performed using dog sera collected from two different areas in Brazil (Uberlandia and Sao Paulo). The results suggest that both antigens were able to identify animals seropositive for E. canis in Brazil since these Brazilian strains appear to be highly conserved.
Animals ; Antigens, Bacterial/blood/*diagnostic use ; Bacterial Outer Membrane Proteins/genetics/metabolism ; Bacterial Proteins/*genetics/metabolism ; Base Sequence ; Brazil ; Dog Diseases/diagnosis/*microbiology ; Dogs ; Ehrlichia canis/*genetics/*immunology/isolation & purification ; Ehrlichiosis/diagnosis/microbiology/*veterinary ; Fluorescent Antibody Technique, Indirect/veterinary ; Male ; Molecular Sequence Data ; Polymerase Chain Reaction/veterinary ; RNA, Ribosomal, 16S/genetics/metabolism ; Sequence Alignment/veterinary

To determine the immune responses in pigs to hog cholera virus after treatment with an ionized alkali mineral complex (IAMC), 40 healthy pigs (28-32 days old) from a commercial swine farm were purchased and housed into 4 groups (n=10 each). All pigs were vaccinated intramuscularly (1 ml) with an attenuated live hog cholera virus (HCV, LOM strain) at 28-32 days old and challenged with a virulent hog cholera virus at 8 weeks after vaccination. Each group was treated with PowerFeelTM sprayed diet as 0.05% (w/w) in a final concentration (T-1, n=10), a diet mixed with SuperFeedTM as 3% (w/w) in a final concentration (T-2, n=10), or a diluted PowerFeelTM solution (1:500, v/v) as drinking water (T-3, n=10), respectively. A group (n=10) served as a non-treated control. Proportions of expressing CD2+ and CD8+ cells increased significantly (p<, 0.05) at 8-week post-application. Mean antibody titers of each group against HCV gradually increased to higher levels after vaccination and with challenge of the virulent virus. In conclusion, the IAMC-treated diets can be helpful for the improvement of growth in pigs with proper vaccination program, while the IAMC-treated diets have no effects on the clinical protection against hog cholera.
Alkalies/immunology/*pharmacology ; Animals ; Antibodies, Viral/blood ; Classical Swine Fever/*immunology/prevention & control ; Classical swine fever virus/*immunology ; Flow Cytometry/veterinary ; Fluorescent Antibody Technique, Indirect/veterinary ; HLA Antigens/immunology ; Minerals/immunology/*pharmacology ; Swine ; Vaccination/*veterinary ; Vaccines, Attenuated/immunology ; Viral Vaccines/*immunology

Porcine proliferative enteropathy(PPE) is an enteric disease been caused by Lawsonia intracellularis. It has become one of the critical problems in the pig industry. To investigate the prevalence of PPE in Korea, serum samples of 828 pigs from 65 herds were tested using indirect immunofluorescence antibody technique(IFA). The infection rate in individual pigs varied from 44 to 69%, whereas 100% in pig farms. The infection frequency was 57, 44.9, and 59.4% according to age respectively. Administration of tylosin in feed at a concentration of 110 ppm for 14 days reduced the infection rate of the farms. These data indicated that the high prevalence of PPE may be controlled by tylosin.
Administration, Oral ; Animal Feed ; Animals ; Anti-Bacterial Agents/*therapeutic use ; Antibodies, Bacterial/blood ; Enteritis/epidemiology/prevention&control/*veterinary ; Fluorescent Antibody Technique, Indirect/veterinary ; Gram-Negative Bacterial Infections/prevention&control/*veterinary ; Korea/epidemiology ; *Lawsonia Bacteria/drug effects/growth&development/immunology ; Seroepidemiologic Studies ; Swine ; Swine Diseases/*epidemiology/microbiology/prevention&control ; Tylosin/*therapeutic use

Porcine circovirus (PCV) type2 was isolated using primary porcine kidney cells from lymph node of piglets with typical PMWS. The presence of the virus was identified by PCR using primers specific to PCV type2. The ORFs 1 and 2 were amplified by PCR using primers corresponding to the target genes of the PCV type 2. Cloned genes were inserted into the baculovirus expression vector and PCV recombinant proteins were expressed using baculovirus expression system. Recombinant protein expression was determined by indirect immunofluorescent assay (IFA) and immunoblotting using polyclonal antiserum to PCV. ORF1 gene expressed two proteins with approximately 17 kDa and 31 kDa proteins in the baculovirus system. Recombinant protein of the ORF2 was similar to that of the native virus except minor bands with different molecular weight were detected. Recombinant protein expressed in the baculovirus system showed at least two glycosylation sites based on the tunicamycin treatment. Recombinant protein of the ORF2 assembled virus-like particle in recombinant virus infected insect cells.
Animals ; Baculoviridae/*genetics ; Blotting, Western ; Circoviridae Infections/*veterinary/*virology ; Circovirus/*classification/genetics/isolation & purification/ultrastructure ; Cloning, Molecular ; Fluorescent Antibody Technique, Indirect ; Lymph Nodes/virology ; Microscopy, Electron ; Open Reading Frames ; Palatine Tonsil/virology ; Polymerase Chain Reaction/methods/veterinary ; Recombinant Proteins/analysis ; Swine ; Swine Diseases/*virology ; Transfection ; Tunicamycin/pharmacology ; Viral Proteins/*analysis