Avian pathogenic Escherichia coli (APEC) is a causative agent for a number of extra intestinal diseases and account for significant losses to the poultry industry. Since protective immunity against APEC is largely directed to virulence antigens, we have individually expressed four different viulence antigens, papA, papG, IutA, and CS31A, using an attenuated Salmonella Typhimurium and a plasmid pBB244. Following oral immunization of mice with combination of two or four of these strains, serum IgG and mucosal IgA responses were elicited against each antigen represented in the mixture. The antigen-specific mucosal IgA responses were significantly higher in the group of mice immunized with the heat-labile Escherichia coli enterotoxin B subunit (LTB) strain than those in the group of mice immunized without the LTB strain. While, there was no significant difference between these two groups in antigen-specific serum IgG responses. The results showed that LTB could act as mucosal immune adjuvant. To assess the nature of immunity, the distribution of antigen-specific IgG isotypes was analyzed. All groups promoted Th1-type immunity as determined by the IgG2a/IgG1 ratio. Thus, our findings provided evidence that immunization with a combination of several vaccine strains is one of the strategies of developing effective vaccines against APEC.

Lactobacillus salivarius JWS 58 (JWS 58) and Lactobacillus plantarum JWS 1354 (JWS 1354) are isolated from duck intestine and have ability to produce bacteriocin. The objective of this study was to evaluate the immunomodulatory effects of JWS 58 and JWS 1354. The nitric oxide (NO) and cytokines (IL-1β and TNF-α) were measured in C57BL/6 mouse peritoneal macrophages to determine immune enhancing effects of JWS 58 and JWS 1354. A Listeria (L.) monocytogenes challenge mice model was used to evaluate immune enhancement ability of JWS 58 and JWS 1354 in vivo. The results showed that JWS 58 and JWS 1354 increased the production of NO or cytokines by peritoneal macrophages and that oral administration of viable probiotic strains in mice elicited the immuno-modulatory effect upon L. monocytogenes challenge. JWS 1354 showed stronger immune enhancing effects than JWS 58. Collectively, this study demonstrated that Lactobacillus strain JWS 58 and JWS 1354 possess immune enhancing effect. Furthermore, two stains are expected to use feed supplement to prevent diseases by pathogenic bacteria through releasing bacteriocin and enhancing host immune responses in animal.

Ontogeny, distributions and relative frequencies of somatostatin-immunoreactive cells were investigated in the proventriculus of the chicken embryos with incubation periods. Samples were taken from 10 groups(10 days of incubation to hatching) and studied by immunogistochemical methods. The findings were as follows. Somatostatin-immunoreactive cells were observed from 12 days of incubation in the proventricular glands and after that increased with incubation periods. The first observation time of these cells in the epithelium were at 15 days of incubation in the basal portion but in 16 and 17 days of incubation, no immunoreactive cells were observed in the epithelium but after that a few immunoreactive cells were observed in the basal portion and gastric gland regions. The shpaes of these cells were spherical to spindle in the proventricular glands and spherical to round in the epithlium and gastric gland.

The localization of substance P(SP), calcitonin gene-related peptide(CGRP) and three calcium binding proteins, calbindin D-28k(CB), calretinin(CR) and parvalbumin(PA) was immunohistochemically examined in the myenteric and submucous plexuses of Korean native goat duodenum. In the neurons of myenteric and submucous plexuses of duodenum, immunoreactivities of SP, CGRP and CB were confirmed in both nerve cell bodies and fibers. In contrast, CR immunoreactivity was found only in nerve fibers of myenteric plexuses, while PA immunoreactivity was found only in nerve cell bodies of submucoun plexuses. In the inner circular muscle layer, dense SP-like immunoreactive fibers were prominent but only a few CGRP-like immunoreactivities were observed. Most of SP-and CGRP-like immunoreactive neurons of both plexuses colocalized with CB. This result showed that SP and CGRP may have a important role for the movement of small intestine. The colocalizations of CB with SP or CGRP in myenteric and submucous plexuses suggest that CB may serve neuromodulatory role for SP-and CGRP-immunoreactecd neurons on the movement of intestinal wall.

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.

This study was carried out to nvestigate the NPY-immunohistochemical characteristics of the olfactory bulb in the striped field mouse(Apodemus agrarius). The animals were anesthesized with thiopental sodium and perfused with 4% paraformaldehyde through left ventricle and aorta. Brains were removed and tranfered 10%, 20% and 30% sucrose. Sections were then cut on a cryostat into 40 micro meter-thick. The tissue immunostained with avidin-biotinylated complex method. The main olfactory bulb consisted of seven circumferential laminae: and olfactory nerve fiber layer, a glomerular layer with glomeruli surrounding by periglomerular cells, an external plexiform layer having granule and tufted cells, a mitral cell layer, a narrow internal plexiform layer, a granule cell layer forming several cell rows and alayer of white matter. The accessory olfactory bulb had four layers: an olfactory or vomeronasal nerve fiber layer, a glomerular layer consisting of small glomeruli, a mixed layer not distinguishing the external plexiform/mitral cell/granule cell layers and a granule cell layer. Most of NPY-immunoreactive(NPY-IR) neurons in main olfactory bulb were localized in the deeper portion of granule cell layer, white matter and anterior olfactory nucleus. In addition, some NPY-IR neurons were identified in the external plexiform layer. The shape of NPY-IR neurons of all olfactory bulb were predominant round or oval, sometime multipolar in shape. And most NPY-IR processes were parallel to long axis of white matter. In accessory olfactory bulb, NPY-IR neurons were not found in all region.

The regional distribution and relative frequency of the endocrine cells in the pancreas of the bean goose were investigated by immunohistochemical methods using 6 types of the specific antisera. Spindle shpaed serotonin-immunoreactive cells were detected in the exocrine portions. Spherical or spindle shpaed glucagon-immunoreactive cells were observed in the exocrine and dark and mammalian type islets. In the dark type islets, numerous cells were dispersed throughout whole islets but they were located in the peripheral regions of the mammalian type islets. No glucagon-immunoreactive cells were detected in light type islets. Round or spherical shpaed insulin-immunoreactive cells were observed in the exocrine and dark, light and mammalian type islets. They were observed in the exocrine regions with a few numbers. Extremely rare cells were detected in central portion of the dark type islets but moderate to numerous cells were found in the central regions of the mammalian and light type islets, respectively. Spherical or spindle shaped somatostatin-immunoreactive cells were observed in the exocrine and dark, light and mammalian type islets. A few single cells were detected in the exocrine portions. In the dark type islets, numerous cells were dispersed throughout whole islets but a few to moderate numbers of cells were located in the peripheral regions of the light and mammalian type islet. Moderate numbers of the bovine pancreatic polypeptide-immunoreactiv ecells were found in the exocrime portions with round, spherical or spindle shape. But no bovine Sp-1/chromogranin-immunoreactive cells were observed in this study.

This study was carried out to produce IgY against B. bronchiseptica, parvovirus and distemper virus that are major pathogens in alimentary and/or respiratory diseases of dogs. In the comparison of adjuvants, ISA70 was the best in the rapid induction and maintence of antibody titers. Agglutination antibody titers against B. bronchiseptica were 1:1,280~1:10,240 in sera and 1:160~1:1,280 in egg yolk. Hemagglutination inhibition(HI) titers against parvovirus in sera and egg yolk were 1:80~1:320 and 1:64~1:256, respectively. Virus neutralization titers against canine distemper was 1:8~1:64 in sera and egg yolk.