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Isolation and comparison of somatic and excretory-secretory antigens for serological diagnosis in Trichuris suis
1999
Jee, C.H. | Lee, C.S. | Park, S.J. (Chungbuk National University, Cheongju (Korea Republic). College of Veterinary Medicine)
Swine shipworm(Trichuris suis) is cosmopolitan nematode which can cause serious pathology in immature stage(larva2~larva5) of infected pigs, such as anorexia, diarrhea, anemia, and death in heavy infections. In this larval stages, it is very difficult to diagnose the infection of whipworm and to differentiate from other common swinegstrointestinal disorders such as 21 day scours which are associated with TGE virus, rota virus, coccidium, and the stress of weaning. In this experiment, the isolated antigens of Trichuris spp. were carried out to examine the structure and specificity of antigens and to select the resonable antigens which would be used in serological diagnosis by electrophoresis, Western blotting, ELISA. The results of this experiment were as follews: 1. The common fractions of each Trichuris suis antigen were identified 28,32,45, 80kDa by SDS-PAGE with silver stain and four major fractions could be detected in positive swine sera by Western blot analysis. 2. The OD(optical density) values of somatic and excretory-secretory antigens which were reacted against positive(negative) sera from pigs infected with Trichuris suis by ELISA reader were; 1) OD values(mean+_SD) of adult somatic antigen against positive(negative) sera were O.30+_0.12(0.09+_0.006) and third-stage larva of somatic antigen were 0.28+_0.038(0.10+_0.005). And OD values of excretory-secretory antigens of adult and third-stage larva were 0.24+_0.031(0.11+_0.005) and 0.08+_0.013(0.10+_0.003), respectively. 2) OD values of adult somatic, larval somatic antigen and adult excretory-secretory antigen response to positive sera were significantly (p0.01) associated with negative swine sera. And the Cut-off OD values(minimum positive value) were determined to be mean negative value plus 3 SD that would minimized the risk of false positives. 3. The OD values of somatic antigens of T suis and T vulpis against swine positive(negative) sera were 0.30+_0.120(0.09+_0.006) and 0.25+_0.141(0.09+_0.003). These data mean that the somatic antigens of T suis and T vulpis were able to diagnose T vulpis infection in dogs as well as T suis infection in pigs. These results suggest that somatic antigen of third-stage larva and excretory-secretory antigen of adult T suis could be used the diagnostic antigen by serological test(ELISA) in immature Trichuris spp. infection.
显示更多 [+] 显示较少 [-]Specific detection of Salmonella serogroup D1 by polymerase chain reaction(PCR) for sefA gene
1999
Jun, M.H. | Kim, T.J. | Chang, K.S. | Kang, K.I. | Kim, K.H. | Kim, H.S. | Shin, K.S. | Kim, C.J. (Chungnam National University, Taejon (Korea Republic). College of Veterinary Medicine) | Kim, K.S. (National Veterinary Research and Quarantine Service, Anyang (Korea Republic).) | Yoo, S.S. (Taejon City Institute of Health and Environment, Taejon (Korea Republic).)
Sal enteritidis thin fimbriae, SEF14, were found to be restricted to the predominantly poultry-associated members of the Salmonella serogroup D1 that are considered as the important pathogens in poultry industry. SefA together with sefB and sdfC encode the proteins involved in SEF14 biosynthesis. In order to develop the rapid and specific detection methods for Salmonella serogroup D1, a PCR technique for the am;lification of sefA gene was established, and its specificity and sensitivity were investigated with various microorganisms. The bacterial genomic DNA was extracted by colony-picking and rapid boiled-lysate technique. In comparison of SefI and SefII primers used in the PCR. SefI primer for sefA gene of 513bp showed higher specificity than that of SefII. The established PCR was s sensitive as to detect 1pg of Sal enteritidis DNA. When 73 strains in 28 genera including the reference strains and the field isolates of various Salmonella serotypes, Bacillus subtilis, Bordetella bronchiseptica, E coli, Listeria spp., Micrococcus luteus, Rhodococcus equi, Staphylococcus spp., Streptococcus spp., Vibrio parahemolyticus, Yersinia spp. were studied, the established PCR yielded specifically positive results with only Salmonella serogroup D1. The results suggested that the PCR for sefA gene could be a potential candidate among the specific detection methods for Salmonella serogroup D1.
显示更多 [+] 显示较少 [-]A study on the drug residues in the raw milk collected over the withdrawal period after mastitis treatment using TTC-II test and delvotest SP
1999
Kang, J.H. | Kim, J.S. | Lee, W.C. (KonKuk University, Seoul (Korea Republic). School of Veterinary Medicine)
From July of 1997 to June of 1998, total 279 raw milk samples over wkthdarwal period after mastitis treatment from dairy farms located in the provinces of Kyonggi and Choongchung were collected to test drug residues. Each sample was tested by TTC-II test and Delvotest SP. Among the total 152 raw milk samples of cow treated by beta-lactams, 32 samples(21.2%) were positive on the Delvotest and 15 samples(9.9%) showed positive on the TTC-II test. Also, from the total 37 samples treated by sulfonamides, 5 samples(13.5%) were positive on the Delvotest and 3 samples(8.1%) showed positive on the TTC-II test. For the total 55 raw milk samples of cow treated by tetracyclines, 9 samples(16.4%) were positive on the Delvotest and 5 samples(9.1%) showed positive on the TTC-II test. In addition, from the total 35 samples treated by aminoglycosides, 7 samples(20.0%) were positive on the Delvotest and 5 samples(14.3%) showed positive on the TTC-II test. Our study shows that it is possible that drugs are to be detected by the drug residues test of and individual raw milk even over the withdrawal period after mastitis treatment and the raw milk of bulk tank.
显示更多 [+] 显示较少 [-]Polymerase chain reaction for the detection of Toxoplasma gondii in the blood of cats
1999
Suh, M.D. | Joo, B.H. (Gyeongsang National University, Chinju (Korea Republic). College of Veterinary Medicine)
This study was conducted to detect the toxoplasma-specific DNA in peripheral blood collected from cats experimentally infected with Toxoplasma gondii (RH strain) and from domiciled cats by B1 gene-base polymerase chain reaction (PCR). The sensitivity of oligonucleotide primer, T-1 & T-2, designed from toxoplasma B1 gene amplification method was compared with parasite detection by mouse inoculation(MI). And also, latex agglutination test(LAT) and indirect fluorescent antibody test(IFAT) were conducted to detect the fluctuation of serum antibodies compared with the detection of toxoplasma by PCR and MI. Toxoplasma B1 gene PCR was shown consistently high sensitivity and the results obtained by PCR agreed completely with those from MI. All blood samples collected before infection with T gondii gave negative results by PCR and MI. Also, toxoplasma B1 gene PCR was not cross reaction with Neospora caninum DNA and normal cat leucocyte as cnotrols. The toxoplasma-specific DNA was detected by PCR in blood of 5 cats experimentally infected with T gondii 6 days after infection and the detection of this specific-DNA was long lastedin blood for 64 days after infeciton. The detection of toxoplasma-specific DNA by PCR could be identified as few as 10 tachyzoites and the isolation of T gondii by MI could be isolated as few as 1 tachyzoite from tenfold serial dilution of T gondii with normal cat blood, respectively. In healthy domiciled cats, the toxoplasma-specific DNA and the parasite were detected and isolated in blood from 3 of 56(5.3%) cats by both PCR and MI, respectively. In the results of antibody test from the total 56 heads of healthy domiciled cats, the positive rates are 15(26.7%) by LAT and 19(33.9%) by IFAT. These results suggest that PCR detection of toxoplasma can be applied as a sensitive and specific diagnostic and research tool.
显示更多 [+] 显示较少 [-]Sex determination of bovine embryos with hamster H-Y antibody and by polymerase chain reaction
1999
Yu, I.J. | Kim, Y.J. (Chonbuk National University, Chonju (Korea Republic). Department of Obstetrics, College of Veterinary Medicine) | Lee, K.K. (Korea Institute of Science and Technology, Taejon (Korea Republic). Korea Research Institute of Bioscience and Biotechnology)
To determine sex of bovine embryos using hamster histocompatibility Y(H-Y) antibodies, bovine compact morulae were incubated for 6 hours in TCM199 supplemented with 10% hamster H-Y antiserum and the embryos with developmental arrest were diagnosed as male embryos, while the embryos showing development during the incubation as femalle embryos. This presumptive embryo sexing was confirmed by polymerase chain reaction(PCR)method. 1. In the result of hamster sperm cytotoxicity test to measure H-Y antibody titer, the rate of dead sperm was considerably lower in H-Y antiserum absorbed with hamster male splenocytes than in H-Y antiserum absorbed with hamster female splenocytes or H-Y antiserum umabsorbed with splenocytes(p0.01). 2. The rate of oocytes fertilized in vitro and the rate of blastocysts of the fertilized oocytes were 58.5% and 32.4%, respectively. The rate of blastocysts on day 8 was 15.9%, denoting the highest rate during whole culture period posterior to in vitro fertilization(IVF). 3. The bovine 16 cell and compact morulae embryos incubated in the medium supplemented with hamster H-Y antibodies showed 37.1% and 48.9% of developmental arrest which were diagnosed as male, respectively, and rates of redeveloped embryos from the arrested were 24.1% in 16 cell and 44.3% in compact morulae embryos, respectively, denoting higher rater of sex determination and rate of redevelopment in compact morulae than 16 cell embryos. 4. Bovine compact morulae of Korean cattle and Holstein were treated with hamster H-Y antibodies for sex determination and the rates fo developmental arrest(deagnosed as male) were 48.4% for Korean cattle and 47.9% for Holstein, respectively. The rates of redeveloped embryos to blastocyst after treatment were 42.6% for Korean cattle and 41.8% for Holstein, respectively, ahowing no sighificant differences of sex determination and redevelopment between both breed. 5. The sex determination of bovine cmbryos(Korean cattle Holstein) using hamster H-Y antibodies was diagnosed by PCR for confirmation, denoting the rates of 86.1% for Korean cattle and 85.9% for Holstein male embryos, respectively, and the rates of 91.9% for Korean cattle and 90.1% for Holstein female embryos, respectively, with no significant differences of sex determination between both breed. These results indicated that hamster H-Y antibodies can be usable for sex determination of bovine embryos of Korean cattle and Holstein, the viability of bovine embryos was sustained while being cultured in the medium supplemented with hamster H-Y antibodies of appropriate titer and sex determination of bovine embryos by PCR can be feasible for confirmation.
显示更多 [+] 显示较少 [-]Seroprevalence and epidemiological analysis of porcine reproductive and respiratory syndrome virus in Korea
1999
Park, C.K. | Chang, C.H. | Kang, Y.B. (National Veterinary Reseach and Quarantine Service, Anyang (Korea Republic).) | Lee, C.H. (Cheju National University, Cheju (Korea Republic). Department of Veterinary Medicine) | Lyoo, Y.S. (Kon-Kuk University, Seoul (Korea Republic). School of Veterinary Medicine) | Kim, H.S. (Chungnam National University, Taejon (Korea Republic). College of Veterinary Medicine)
A nation wide sero-epidemiological survery of porcine reproductive and respiratory syndrome(PRRS) was carried out to analyze the current status of the PRRS virus infections in the field using the indirectr immunofluorescent antibody assay(IFA) with the field isolate PL96-1. Since the first report of the antibody detection to PRRSV in 1993, the prevalence of seropositive pigs has increased dramatically and the data indicate that over 21% of the pigs and around 60% of the farms showed seropositives to the PRRS virus. A slightly higher positive rate was recognized in breeders than fattenings and it might be due to the higher age at the time of testings. No significant regionl defferences were detected in the sero-epidemiological survey. Higher sero-positive rate in growers indicatesthat PRRSV infection in the field was common after weaning(around 40 days). However, the number of seropositive pigs were declined in fattening pigs. Sows showed around 26% of sero-positive rate that there is a higher chance of continuous virus circulation in the infected farms. Low rate of sero-positivity in boars(9.8%) implies that there is high demand in proper controlmeasures to prevent virus spreading through breeding procedures such as natural or artificial insemination Therefore it was concluded that PRRSV infection in domestic swine herds is endemic and the positive rate and economic loses will be increased by spontaeous infections in naive farms.
显示更多 [+] 显示较少 [-]Effects of forskolin on secrtion of insulin like growth factor-1 in the perfused rat liver model
1999
Kang, C.W. | Lee, H.I. (Chonbuk National University, Chonju (Korea Republic). Bio-Safety Research Institute) | Lee, D.Y. (Chonbuk National University, Chonju (Korea Republic). Department of Pediatrics, Medical School)
The insulin-like growth factor-I(IGF-I) is an important metabolic factor involved in cell growth and metabolism. Although secretion of IGF-I in rat liver is regulated by growth hormone, the effects of forskolin, adenylate cyclase activator, on secretion of IGF-I have not been reported. Therefore, a modified perfused rat liver model was used to investigate the regulatory effects of forskolin on IGF-I secretion in this experiment. The results were summerized as follows: 1. Modified perfused rat liver model was not changed to aspartate aminotransferase(AST), alanine aminotransferase(ALT) and lactic dehydrogenase(LDH) secretion in time. 2. The IGF-I secretion in hepatic cell was increased by forskolin(10-5, 10-6 and 10-7M) in a dose-dependent manner as compared with those of the controls, and significantly increased by 10-5 and 10-6M forskolin(p0.05). 3. Secretion of glucose in hepatic cell significantly was decreased by 10-5M forskolin as compared with those of controls(p0.05). These results suggest that forskolin may be involved in the regulation of IGF-I secretion in the perfused rat liver.
显示更多 [+] 显示较少 [-]Numerical and morphologic changes of ovarian follicles in each stage of estrus cycle in rats
1999
Lee, Y.J. (Gyeongsang National University Hospital Institute of Animal Medicine, Chinju (Korea Republic). Department of Histopathology) | Kwak, S.D. (Gyeongsang National University, Chinju (Korea Republic). College of Veterinary Medicine)
The effects of electroacupuncture and laser at Da-zhui and Shen-shu on hematologic changes and blood concentration of endocrine substances in dogs
1999
Cho, Y.S. | Bae, C.S. (Konkuk University, Seoul (Korea republic). Faculty of Veterinary Medicine)
This study was carried out (a) to investigate the variations of blood chemistry and (b) to examine the secretion trend of endocrine substances in a dog model after electroacupuncture and laser stimulation at different time period(9 to 11a.m. and 6 to 8p.m.). Two acupuncture points; Da Zhui(GV-14) and Shen Shu(BL-23) were electroacupunctured for 20 minutes with 2Vol, 20Hz and irradiated for 5 minutes with 8,000Hz. Before stimulation and after a lapse of time(10-minutes, 30-minutes and 60-minutes) all dogs were checked the following parameters; cortisol, ACTH, RBC, hemoglobin, hematocrit, WBC, Ca, P, SGPT, SGOT and creatinine. The results were as follow:The levels of cortisol and ACTH have been increased 10 minutes after the stimulation of the electroacupuncture and laser. The higher levels of cortisol and ACTH have been decreased to keep the normal levels from 30 minutes after the stimulation of the electroacupuncture and laser. The RBC, hemoglobin, hematocrit and WBC showed the physiological phenomena in the electroacupuncture and laser stimulation. In sero chemical analysis, calcium, phosphate, SGOT, SGPT and creatinine levels were within normal physiological ranges.
显示更多 [+] 显示较少 [-]Genetic studies of blood markers in Cheju horses
1999
Cho, G.J. | Kim, T.S. | Um, Y.H. (Korea racing Association, Gwacheon (Korea Republic).) | Kim, B.H. (Kyungpook National University, Taegu (Korea Republic). College of Veterinary Medicine) | You, J.S. (Cheju Province Institute for Livestock Promotion, Cheju (Korea Republic).)
The present study was carried out to investigate the blood markers of Cheju horses. The red cell types (blood groups) were tested from 73 Cheju native horses and 118 Cheju racehorses by serological procedures with 23 reagents. The blood group phenotypes observed with high frequency were Pb(34.3%), Qc(56.2%), Qb(15.1%) and genotypes Dbcm/dghm(12.3%), Dde/dghm(9.6%), Dad/bcm(6.8%), Dcgm/de(6.8%) in Cheju native horses, while Aa(63.6%), Pa(44.9%), P-(28.8%), Qabc(36.4%), Dbcm/cgm(14.4%), Dbcm/bcm(10.2%), Dbcm/de(7.6%), Dbcm/dghm(5.1%), Dde/dk(5.1%) in Cheju racehorses. Alleles observed with high frequency were Ab(0.128), Ac(0.169), Dad(0.103), Dadn(0.075), Ddghm(0.226), Pb(0.316), Qc(0.494) in Cheju native horses and Aa(0.529), Dbcm(0.306), P-(0.531), Qabc(0.197), Q-(0.504) in Cheju racehorses. No specific variation of blood groups and allele frequencies of C, K, U system were observed in Cheju native horses and Cheju racehorses. The mean heterozygosity in Cheju native horses and Cheju racehorses was observed 0.5344 and 0.5102, respectively.
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