To investigate the regional distribution and relative frequency of the neurotensin-, pancreatic polypeptide(PP)- and gastrin/cholecystokinin(Gas/CCK)-immunoreactive cells in the gastrointestinal tract of the bullfrog(Rana catesbeiana) with developmental stages, group of bullfrogs subdivided into the tadpole withhindlegs, metamorphosed bullfrog with tail, 2 weeks after metamorphosed bullfrog and adult bullfrog, were stained by immunohistochemical methods(PAP methods). Neurotensin-immunoreactive cells were observed from the pylorus of the metamorphosed bullfrog with tail, but these cells were not detected after that periods. PP-immunoreactive cells were dectected from the adult bullfrog in the pylorus, duodenum and ileum. These cells were most predominant in the pylorus. Gas/CCK-immunoreactive cells were observed from the adult bullfrog in the pylorus. According to these results, most of immunoreactive cells in the gastrointestinal tract of the bullfrog were appeared after the complete metamorphosed periods, in which the complete differentiation of structure of gastrointestinal tract were occurred, and variable changes of the regional distribution and relative frequency with developmental stages were observed.

A 474-base pair segment covering the hypervaible region of VP2 gene from six Korean infectious bursal disease virus(K-IBDV) isolates(K1, K2, SH/92, 225, 269, 310) and one attenuated IBDV(DAE) were amplified using RT-PCR, sequenced, and compared with published sequences for IBDV. K-IBDV isolates(K1, K2, SH/92, 225, 269) and foreigh very virulent(vv) IBDV strains had 94.93-100% amino cid sequence similarity. K-IBDV isolate 310 and other K-IBDV isolates had 84.31%-86.07% amino acid sequence similarity. Attenuated strain(DAE), like other attenuated strain, has substitution at positions 279(D to N) and 284(A to T) as well as in the serine-rich heptapeptide region. Five K-IBDV isolates except 310 isolate share unique amino acid residues at positions 222(A), 256(I), 294(I) which are not present in other standard and attenuated strains. At the two hydrophilic region, K-IBDV isolates except 310 isolate had identical amino cids comparing with Belgium vv IVDV 894VB but had four amino acid substitutions comparing with Chinese vv IBDV F9502. The SWSASGS heptapeptide is conserved in all KIBDV isolates. The sequence of K-IBDV isolate 310 was markedly different from other IBDV strains, evolving from a separate lineage than the others. By phylogenetic analysis, Five K-IBDV isolates except 310 isolate were categorized in one group with foreign vv IBDV isolates but K-IBDV isolate 310 was categorized ina separate group which was differentiated form other compared IBDV strains

The purpose of this study was to evaluate effect of chitosan-oligosaccharides(CHIOL) on hydrophobicity of pathogenic E coli including a field isolate from suckling piglet with diarrhea, E coli-0157:H7, and E coli-O149:K88ac. E coli field isolate appeared adhesion of 100% to n-hexadecne between 0.00125% and 0.05% CHIOL. E coli-O157:H7 occurred adhesion of 69% and 64% under the level of 0.00125% and 0.025% CHIOL, respectively. E coli-O149:K88ac showed adhesion of 100% in higher than 0.025% CHIOL. For cationic action, the adhesion of E coli isolate and E coli-O149:K88ac to n-hexadecane were inhibited at level of higher than 10mM Ca2+ but did not induce any difference among the concentrations used(p0.01). However,the adhesion of E coli-O157:H7 to n-hexadecane was inhibited at level of higher than 50mM Ca2+. In a field trial, control piglets showed average mortality of up to 58% during 3 days after the onset of diarrhea. In contrast, the prevalence of E coli-induced diarrhea in CHIOL-treated groups without mortality was dropped down to average 34% on the 1st day after the treatment of CHIOL, and average 2% on the 4th day. After then, piglets with diarrhea was not present. In conclusion, the low concentrations of CHIOL were most likely to associate with the enhancement of hydrophobicit to pathogenic E coli. Calcium inhibited the hydrophobicity of E coli by CHIOL. These results suggested that CHIOL could be played an efficient and reliable role in treating enteric colibacillosis of piglets.

Since Robert Koch found tubercle bacilli in 1882, the studies on tubercle bacilli of human and animal had been carried out. Being old tuberculin(OT) introduced in 1890, the specificity of the diagnosis of tuberculosis has been improved by continual uses of heat concentrated synthetic medium(HCSM) and purified protein derivatives(PPD) tuberculin. Now, two types of tuberculin test are used worldwidly; the single intradermal test(SIT) using bovine tuberculin and the single intradermal comparative tuberculin test(SICTT) using avian and bovine tuberculins. In the SICTT, each countries have used with different combination of both avain and bovine tuberculins' titers. However, this kinds of studies have not reported in Korea. Therefore, the studies on the combination of their tuberculins' titers were performed through intradermal test of guinea pigs sensitized with either Mycobacterium bovis or M avium and were examined in 10 cattles of SIT positive reactors. Also, IFN-y assay, the latest diagnostic method of bovine tuberculosis, was experimentally applied to SIT positive reactors. For determinint the optimal titers, sensitized guinea pigs with M bovis and M avium were intradermally injected avain and bovine tuverculin. In guinea pigs sensitized with M bovis, bovine tuberculin 50 T.U. showed significant difference from all tested concentrations of avian tuberculin(p0.05). In guinea pigs sensitized with M avium, there is significantly different between bovine tuberculin and avian tuberculin by 25 T.U.(p0.01). Therefore, optimal titers of bovine and avian PPD tuberculins' titers for the SICTT in Korea were 5,000 and 2,500 tuberculin units, respectively, and the swelling differences between bobine and avian site in SIT positive reactors were above 3mm. Also, in IFN-y assay, the 9 SIT positive reactors were showed all the positive reactions.

Atrial natriuretic peptide(ANP) is a hormone with potent natriuretic, diuretic and relaxing properties on vascular smooth muscle. Specific chemical modulator in response for the ANP secretion has not been found yet. Therefore, we have investigated the role of Ca2+ responsible for the regulation of ANP induced by protein kinase C(PKC) on mechanically stretch-induced ANP secretion in the rat atria. The results obtained were as follows; 1. ANP secretion and ANP concentration were increased to more in Ca2+-free buffer than in the Kreb-Henseleit buffer on mechanically stretch-induced ANP secretion(p0.05), but extracellular fluid translocation(ECF) was not significant. Phorbol 12-myristate 13-acetate(PMA, 10-7M) induced ANP secretion and ANP concentration in Ca2+-free buffer shown to more accentuate on mechanically stretch-induced ANP secretion than in the Ca2+-free buffer(p0.05), but ECF translocation was not significant. 2. In the presence of ryanodine(3*10-6M), PMA(10-7M) induced ANP secretion and ANP concentration in the Kreb-Henseleit buffer were shown to more increase on mechanically stretch-induced ANP secretion than in the ryanodine(3*10-6M) with the Kreb-Henseleit buffer(p0.05), but ECF translocation was not significant. 3. In the presence of ryanodine(3*10-6M), PMA(10-7M) induced ANP secretion and ANP concentration in the Ca2+-free buffer was shown to more increase on mechanically stretch-induced ANP secretion than in the ryanodine(3*10-6M) with the Ca2+-free buffer on mechanically induced ANP secretion(p0.05), but ECF translocation was not significant. The results suggest that PKC-induced ANP secretion may not be related to the change of Ca2+ on mechanically induced ANP secretion in the rat atria.

We have studied, by a nonisotopic in situ end-laveling(ISEL) technique, frequency of apoptosis in the external granular layer(EGL) of the cerebellum of immature mice by Y-rays irradiation from 60Co or diagnostic ultrasound exposure. The total number of normal cells and cells showing morphological features of apoptosis were counted. The frequency of apoptotic cells was expressed as a percentage of the total number of cells in EGL. The extent of changes following 200 cGy(1090 cGy/min) was studied at 2, 4, 6, 8, 12, or 24 hours after exposure. The maximal frequency was found 6~8 hours after exposure. The immature mice that received 18, 36, 54, 108, 198, 396, cGY of y-rays or diagnostic ultrasound(7.5MHz, 4.2mW, Ispta=7.9mW/cm2, Ispta=114.3W/cm2) for 10 or 30 minutes were examined 6 hours after irradiation. Measurements performed after y-ray irradiation showed a dose-related increase in apoptotic cells in each of the mice studied. The dose-response curves were analyzed by a linear-quadratic model;frequency of apoptotic cell in the EGL was y=(0.1349+_0.01175)D+(-0.0001522+_0.0000334)D2+0.048(r2=0.981, D-dose in cGy). In the experiment of ultrasound exposure, the frequency of apoptotic cell was 0.106+_0.130(10 minutes exposure) and 0.167+_0.220(30 minutes exposure). We estimated the relative dose of the yield from the experiment with ultrasound by substituting the yield from ultrasound exposure into the curve from the y-irradiation. The relative dose of ultrasound exposure compared with y-irradiation were 0.432 cGY(10 minutes exposure) and 0.885 cGY(30 minutes exposure). We have found that there is no evidence to indicate that diagnostic ultrasound involves a significant risk.

Swine plasminogen is not activated by staphylokinase of Staphylococcus aureus. In this study, the activation of swine plasminogen by staphylokinase of Staph hyicus subsp. hyicus was investigated and the effect of EDTA(disodium) on plasminogen activation was also studied. When the activation of swine plasminogen by staphylokinase of Staph hyicus subsp. hyicus was examined in fresh swine plasma, swine plasminogen could be weakly activated. However, when EDTA was added to the swine plasma, plasminogen activation was markedly enhanced, but this enhancement was not observed on bovine fibrin-dog plasminogen agar plate containing EDTA. Chicken and bovine plasminogens were not activated by staphylokinase of Staph hyicus subsp. hyicus. Using fresh swine plasma agar containing 0.07% EDTA, staphylokinase activity was detected in 96.3% of Staph hyicus subsp. hyicus strains isolated from pigs and in none of the chicken and bovine strains.

In order to study the effects of Jengjengamiyjintang on the duodenal ulcer induced by HCI-aspirin in rats, the changes of histological profiles, goblet cells(PAS-positive cells), and the distribution and frequency of cholecystokinin(CCK)-8 and serotonin-producing gastro-entero-endocrine cells were observed after oral administration of Jengjengamiyjintang. Histologically, very severe injury to duodenal epithelium were observed in control groups and theses injuries were increased with time intervals. But in the Jengjengamiyjintang administrated groups, no gross lesion of ulcer were demonstrated and histologically minor injury to the mucosal epithelium were observed. PAS-positive cells were increased in the Jengjengamiyjintang administrated groups compared to that of control groups. Severe degranulation of CCK-8- and serotonin-immunoreactive cells were observed in control groups but these phenomenon was seldom in the Jengjengamiyjintang administrated groups. Serotonin-immunoreactive cells were significantly decreased in control groups but increased in Jengjengamiyjintang administrated groups compared with control groups. According to these result, it is suggested that Jengjengamiyjintang would accelarat the healing of the duodenal ulcer but the functional mechanisms were unknown.

DNA hybridization assay using probes prepared from liver was carried out to identify species characterization of the domestic animals. Gel electrophoresis showed that the target DNA extracted from raw muscle were 1kb and uniform pattern while fragments size of heated muscle were irrgular. Hybridizatino was performed by adding 200ng/ml probe in hybridization solution and incubating for 12 hours at 68 degrees centigrade. To obtain good discrimination, applied washing buffer and washing step differently depending on the species. The probes of pig, horse and dog formed the specific hybrids with each target DNA respectively. Although cross reaction ws detected in cattle, goat and sheep but signal intensity among these species made the discrimination possible each other. Such pattern was the same in the cases of chicken, turkey and duck. The hybridization pattern of heated muscle was similar to that of raw muscle in general, but the signal intensity was inferior to that of raw muscle. Species identification between closely related animal species, hybridized using the target DNA of such closely related animal species as a blocking agent, remarkable increase of discrimination from the evident decrease of non specific reaction compared with the control group. In addition, in the admixture where certain meat was included in the beef, pork, chicken meat, we colud find whether any unjust meat was admixed or not. In this case, detection limit of certain meat in admixture was 1%.

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.