1.Preparation and characterization of follwing the national standard anti-Brucella abortus serum, bovine.
Cui LI ; Fushi GUAN ; Zhihong DAI ; Hui JIANG ; Fang WEN ; Lianshou LU ; Zaishi WANG
Chinese Journal of Biotechnology 2011;27(5):812-816
To prepare anti-Brucella abortus serum used for calibrate the agglutination test follwing the national standard, 4 anti-Brucella abortus sera were obtained from 4 cows infected with Brucella abortus naturally. By potency testing, the third serum was selected. Sterility, vaccum degree, residual moisture, uniformity and stability of this standard material were tested and proved to meet the national standard. Referring to the international standard, RBT (Rose-Bengal plate agglutination test), SAT (standard tube agglutination) and CFT (complement fixation test) titers of this standard material were measured to be 1:160 "+" 1:2 400 "++" and 1:800 "++", which are identical with the collaborative assay results. International unit of the standard material is 4 000 IU/mL.
Agglutination Tests
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veterinary
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Animals
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Antibodies, Bacterial
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blood
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Antigens, Bacterial
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immunology
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Brucella abortus
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immunology
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Brucellosis, Bovine
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diagnosis
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Cattle
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China
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Complement Fixation Tests
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veterinary
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Immune Sera
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Reference Standards
2.Isolation and characterization of Streptococcus sp. from diseased flounder (Paralichthys olivaceus) in Jeju Island.
Gun Wook BAECK ; Ji Hyung KIM ; Dennis Kaw GOMEZ ; Se Chang PARK
Journal of Veterinary Science 2006;7(1):53-58
Streptococcus sp. is gram-positive coccus that causes streptococcal infections in fish due to intensification of aquaculture and caused significant economic losses in fish farm industry. A streptococcal infection occurred from cultured diseased olive flounder (Paralichthys olivaceus) in May, 2005 at a fish farm in Jeju Island, Korea. The diseased flounder exhibited bilateral exophthalmic eyes and rotten gills; water temperature was 16~18oC when samples were collected. Of the 22 fish samples collected, 3 samples were identified as Lactococcus garvieae and 18 samples were identified as Streptococcus parauberis by culture-based, biochemical test. Serological methods such as slide agglutination, hemolysis and antimicrobial susceptibility test were also used as well as multiplex PCRbased method to simultaneously detect and confirm the pathogens involved in the infection. S. parauberis and L. garvieae have a target region of 700 and 1100 bp., respectively. One fish sample was not identified because of the difference in the different biochemical and serological tests and was negative in PCR assay. In the present study, it showed that S. parauberis was the dominant species that caused streptococcosis in the cultured diseased flounder.
Agglutination Tests/veterinary
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Animals
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Aquaculture
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DNA, Bacterial/chemistry/genetics
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Fish Diseases/*microbiology
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*Flounder
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Hemolysis
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Korea
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Microbial Sensitivity Tests/veterinary
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Polymerase Chain Reaction/veterinary
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Streptococcal Infections/microbiology/*veterinary
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Streptococcus/genetics/*isolation&purification
3.Development of a monoclonal antibody-based co-agglutination test to detect enterotoxigenic Escherichia coli isolated from diarrheic neonatal calves.
Brajesh C VARSHNEY ; N M PONNANNA ; Pranati A SARKAR ; Pragna REHMAN ; Jigar H SHAH
Journal of Veterinary Science 2007;8(1):57-64
Escherichia coli (E. coli) strains were collected from young diarrheic calves in farms and field. Strains that expressed the K99 (F5) antigen were identified by agglutination tests using reference antibodies to K99 antigen and electron microscopy. The K99 antigen from a selected field strain (SAR-14) was heat-extracted and fractionated on a Sepharose CL-4B column. Further purification was carried out by sodium deoxycholate treatment and/or ion-exchange chromatography. Monoclonal antibodies to purified K99 antigen were produced by the hybridoma technique, and a specific clone, NEK99-5.6.12, was selected for propagation in tissue culture. The antibodies, thus obtained, were affinity-purified, characterized and coated onto Giemsastained Cowan-I strain of Staphylococcus aureus (S. aureus). The antibody-coated S. aureus were used in a coagglutination test to detect K99+ E. coli isolated from feces of diarrheic calves. The specificity of the test was validated against reference monoclonal antibodies used in co-agglutination tests, as well as in ELISA. Specificity of the monoclonal antibodies was also tested against various Gram negative bacteria. The developed antibodies specifically detected purified K99 antigen in immunoblots, as well as K99+ E. coli in ELISA and co-agglutination tests. The co-agglutination test was specific and convenient for large-scale screening of K99+ E. coli isolates.
Agglutination Tests/methods/*veterinary
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Animals
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*Animals, Newborn
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Antibodies, Monoclonal/*immunology
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Antigens, Surface/immunology/isolation & purification
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Bacterial Toxins/immunology/isolation & purification
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Cattle
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Cattle Diseases/*immunology/*microbiology
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Chromatography, Gel/veterinary
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Chromatography, Ion Exchange/veterinary
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Chromatography, Liquid/veterinary
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Diarrhea/immunology/*veterinary
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Electrophoresis, Polyacrylamide Gel/veterinary
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Enzyme-Linked Immunosorbent Assay/veterinary
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Escherichia coli/*immunology
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Escherichia coli Infections/immunology/*veterinary
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Immunoblotting/veterinary
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Staphylococcus aureus
4.Experimental evaluation of pathogenicity of Lactococcus garvieae in black rockfish (Sebastes schlegeli).
Sung Hyun KANG ; Gee Wook SHIN ; Yong Seung SHIN ; K J PALAKSHA ; Young Rim KIM ; Hyang Hee YANG ; Eun Young LEE ; Eung Goo LEE ; Nam Eung HUH ; Oh Myung JU ; Tae Sung JUNG
Journal of Veterinary Science 2004;5(4):387-390
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
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Animals
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Bacterial Capsules
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DNA, Bacterial/genetics/isolation&purification
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Fish Diseases/*microbiology/mortality
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Fishes
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Fluorescent Antibody Technique, Indirect/veterinary
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Gram-Positive Bacterial Infections/microbiology/mortality/*veterinary
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Lactococcus/*pathogenicity
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Polymerase Chain Reaction/veterinary
5.Seroprevalence of Toxoplasma gondii Infection in Police Dogs in Shenyang, Northeastern China.
Cheng Wu LIU ; Na YANG ; Jian Bin HE ; Ming Yang MU ; Min YANG ; Ning SUN ; Hong Kui LI
The Korean Journal of Parasitology 2013;51(5):579-581
In recent years, worldwide surveys of Toxoplasma gondii infection in dogs have been reported. However, only limited surveys of T. gondii infection in police dogs have been available, including China. In the present study, we report the seroprevalence of T. gondii in police dogs in Shenyang, northeastern China. Sera from 291 police dogs were examined for T. gondii antibodies with the modified agglutination test (MAT), and 30.9% animals were tested seropositive. The results of the present study indicated a relatively high prevalence of T. gondii infection in police dogs in Shenyang, China.
Agglutination Tests/veterinary
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Animals
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Antibodies, Protozoan/*blood
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China/epidemiology
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Dog Diseases/*epidemiology/parasitology
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Dogs
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Female
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Male
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Seroepidemiologic Studies
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Toxoplasma/*immunology/isolation & purification
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Toxoplasmosis, Animal/*epidemiology/parasitology
6.Capsular polysaccharide typing of domestic mastitis-causing Staphylococcus aureus strains and its potential exploration of bovine mastitis vaccine developmen. I. capsular polysaccharide typing, isolation and purification of the strains.
Hong Ryul HAN ; Son Il PAK ; Seung Won KANG ; Woo Seog JONG ; Cheol Jong YOUN
Journal of Veterinary Science 2000;1(1):53-60
One hundred seven isolates of Staphylococcus aureus from bovine mastitis were investigated for colony morphology in serum-soft agar (SSA), autoagglutination in salt, and capsular serotype. Capsular polysaccharide (CP) was purified and quantified from the extracts of clinical isolates. Overall, 89 isolates (83.2%) were diffuse in the SSA, without any difference in the proportion of diffuse colony between type 5 and type 8 strains. Some strains exhibited compact colonies in the SSA and expressed CP as determined by an enzyme-linked immunosorbent assay, indicating that compact morphology does not exclude encapsulation. The majority of the strains (11/12) showed autoagglutination in the salt aggregation test. The serotype 336 accounted for 46.7% of the isolates followed by serotype 5 (12.1%) and serotype 8 (12.1%). Particularly, twenty-six (24.3%) isolates reacted with two serotypes; 7 for type 8/336 and 19 for type 5/336. Five isolates (4.7%) were nontypeable with monoclonal antibodies specific for CP serotype 5, 8, or 336. The CP concentration in culture supernatants varied with the serotypes, and the total amount of CP produced by cells grown in a liquid medium was much less than that produced by cells grown on a solid medium. The Western blotting indicated that the CP bands of S. aureus serotype 5 and 8 were ranged in the molecular mass of 58-84 kilodalton (kDa), with additional bands in the region of approximately >or= 48 or