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Considering the usefulness of the immunopotentiator from Pantoea agglomerans 1 (IP-PA1), which is a purified lipopolysaccharide (LPS) derived from symbiotic gram-negative bacteria of food crops, in controlling immunosuppression in poultry husbandry, in this study, we examined its immune-recovery effects in dexamethasone-treated stressed chicken models. Three-week-old chickens daily administered 10 microg/kg of dexamethasone for 35 days to induce stress showed more whole body weight loss; relative thymic, bursal, and splenic weight losses; and decrease in the number of peripheral blood lymphocytes, as compared with the control chickens on day 35; the IP-PA1-pretreated, dexamethasone-treated chickens showed reduced weight losses. Five- to eight-week-old chickens administered 5 mg/kg of dexamethasone showed excessive apoptosis of thymic and bursal lymphocytes 24 hr after a single dexamethasone treatment; apoptosis was inhibited in the IP-PA1-pretreated, dexamethasone-treated chickens. Chickens daily administered 10 microg/kg of dexamethasone for 35 days and injected with commercial Salmonella Enteritidis (SE) vaccine or sheep red blood cells (SRBC) on days 7 and 21 showed about 8- or 2-fold lower antibody production in response to SE or SRBC, respectively, as compared with the control chickens on day 35; the antibody production in response to SE or SRBC was increased in the IP-PA1-pretreated, dexamethasone-treated chickens. These results indicate that IP-PA1 exerts inhibitory effects on dexamethasone-induced immunosuppression and that it may be useful in controlling immunosuppression in poultry husbandry.

A total of 230 Salmonella isolates representing 33 serotypes originated from food (pork, beef, chicken meat, duck meat, and shrimp), domestic animals (pig, chicken, and duck), and human (children with diarrhea) in the Mekong Delta, Vietnam were examined for the antimicrobial resistance to 10 antibiotics. Of the 230 Salmonella isolates examined, 49 (21.3%) showed antimicrobial resistance. Thirty-eight isolates (16.5%) were resistant to oxytetracycline, 26 (11.3%) to chloramphenicol, 17 (7.4%) to nalidixic acid, 16 (7.0%) to streptomycin, 5 (2.2%) to kanamycin, and 4 (1.7%) to ampicillin. No isolate showed resistance to gentamicin, cefazolin, ceftriaxone, and ciprofloxacin. Among the resistant isolates, nineteen isolates were resistant to one antimicrobial agent, 10 to two, 15 to three, 3 to four, and 2 to five antimicrobial agents. The resistance rate of Salmonella isolates from the Mekong Delta, Vietnam to these antimicrobial agents seems to be relatively lower than the results of developed countries and even those of the neighboring countries.

Anti-bacterial activity of two synthesized oligopeptides, RLYLRIGRR-NH2 (peptide A) and RLRLRIGRR-NH2 (peptide B), both which based on a putative active site of defensin, an anti-bacterial peptide from the beetle Allomyrina dichotoma, was examined by macroscopic and histopathologic assessment during the course of infection in mice inoculated with methicillin-resistant Staphylococcus aureus (MRSA) in vivo. Both peptides A and B decreased the mortality of mice inoculated with MRSA. Peptides A and B decreased the macroscopical and histopathological lesions by MRSA infection in mice even seven days after the challenge. The anti-bacterial activity of peptides A and B has a therapeutic effect on MRSA infection in mice even seven days after being challenged.

Neutrophil elastase (NE) released from neutrophils during inflammation is related to tissue disturbance and organ failure. We investigated the effects of an orally active NE inhibitor, ONO-6818, on acetic acid induced colitis in Syrian hamsters. The ulcer area, hemoglobin level in the colonic lumen, NE activity, and tissue myeloperoxidase (MPO) activity in the colitis control animals were significantly increased compared to the normal control ones. Either oral or subcutaneous treatment with ONO-6818 had significant inhibitory effects on the ulcer area, hemoglobin level and NE activity in the colonic lumen, but ONO-6818 did not have a significant inhibitory effect on tissue MPO activity. We conclude that NE is closely related to the development of inflammation in acetic acid-induced colitis in Syrian hamsters and that the condition is improved by the inhibition of NE.

A novel colitis model using Syrian hamsters was developed. Colitis was induced by intracolonic administration of 1% acetic acid, and the ulcer area, tissue myeloperoxidase (MPO) activity, and luminal neutrophil elastase (NE) activity of the colon were determined at 1, 3, 8, 24 and 48 hr after colitis induction. The histopathological changes of the colon were also examined in this model. An increase of tissue MPO activity and NE activity was evident at 3 hr after induction of colitis, peaked at 24 hr, and decreased subsequently. The increase of luminal NE activity was well correlated with the colonic ulcer area. In histopathological examination, ulceration, erosion, crypt abscesses, neutrophil infiltration, hemorrhage, and edema were seen. The effects of prednisolone were examined to evaluate the adequacy of our colitis model. Syrian hamsters were treated orally with prednisolone at 18 and 1 hr before and at 6 hr after induction of colitis, and the ulcer area, tissue MPO activity, and luminal NE activity were evaluated at 24 hr after colitis induction. Prednisolone therapy had little effect on the tissue MPO activity. However, the NE activity of the prednisolone-treated group was significantly decreased. In addition, although prednisolone did not significantly decrease the ulcer area, a tendency toward decrease was noted. We conclude that this new model of experimental colitis in Syrian hamsters is useful for investigating the pathophysiology of colitis, especially useful for studying the relationship between colitis and NE activity.

We previously reported that synthetic peptides, RLYLRIGRR-NH2 (peptide A) and RLRLRIGRR-NH2 (peptide B), derived from the beetle Allomyrina dichotoma defensin, showed antimicrobial activity against both Gram-positive and negative bacteria and suppressed lipopolysaccharide (LPS)-induced tumor necrosis factor-alpha (TNF-alpha) mRNA expression in a murine macrophage cell line. In this study, inhibitory effects of these peptides in LPS-induced mouse peritoneal macrophage activation were investigated. The supplement of peptide A to macrophages cultured with LPS resulted in a significant decrease in nitric oxide and TNF-alpha production. Furthermore, NF-kappaB activation was also blocked by addition of peptide A. These results indicated that peptide A blocked macrophage activation induced by LPS.

The novel peptides based on a putative active site of defensin, an anti-bacterial peptide from the beetle Allomyrina dichotoma, were synthesized. These synthetic oligopeptides exhibited strong anti-bacterial activity in vitro, even against antibiotic-resistant pathogenic bacteria. Then, anti-bacterial activity of two newly synthesized peptides, RLYLRIGRR-NH(2) (peptide A) and RLRLRIGRR-NH(2) (peptide B) was also examined by macroscopic and histopathologic assessment during the course of infection in mice inoculated with antibiotic-resistant pathogenic Escherichia coli (E. coli) in vivo. Peptide B decreased the mortality of mice inoculated with antibiotic-resistant pathogenic E. coli. The results of macroscopic and histopathologic examinations revealed that peptide B could protect the mice from infection. In contrast, peptide A failed to protect mice from infection with antibiotic-resistant pathogenic E. coli. Also, modified peptides A and B produced no toxicity or side effects in mice. These results suggest that peptide B is useful for developing novel antibiotics against antibiotic-resistant pathogenic bacteria.

The effects of oral administration of sugar cane extracts (SCE) on Eimeria tenella oocysts infection in chickens were studied with 2 different experiments. In Experiment 1, 3-week-old inbred chickens (MHC; H.B15) were inoculated into the crop with SCE (500 mg/kg/day) for 1 day or 3 consecutive days, and then challenged with E. tenella sporulated oocysts (2 x 10(4) cells/chicken). In Experiment 2, 1-week-old chickens were orally administered SCE at the same dose for 3 consecutive days, and then initially infected with E. tenella sporulated oocysts (2 x 10(3) cells/chicken). At 2 and 3 weeks of age, these chickens were immunized intravenously with the mixed antigens of sheep red blood cells (SRBC) and Brucella abortus (BA). At 4 weeks of age, chickens were challenged with E. tenella sporulated oocysts (1 x 10(5)/chicken). Challenged chickens with E. tenella oocysts showed markedly decreased body weight gain/day, severe hemorrhage and great number of shedding oocysts in feces and high lesion scores. Oral administration of SCE and initial infection with oocysts (2 x 10 (3)/chicken) resulted in a remarkable improvement in body weight gain/day, hemorrhage, the number of shedding oocysts and lesion score, compare to other infected groups. In addition, SCE-inoculated chickens with the initial infection showed a significant increase in antibody responses against SRBC and BA and also improvement in decreased relative proportions of Bu-1a(+) and CD4( )cells in cecal tonsil lymphocytes of E. tenella-challenged chickens. Cecal tissues of chickens administered SCE and initially infected with E. tenella oocysts showed lower numbers of schizonts, gametocytes and oocysts than those of infected control chickens. These results suggest that SCE have immunostimulating and protective effects against E. tenella infection in chickens.

It has been suggested that the sympathetic nervous system communicates with lymphocytes expressing cell surface receptors for neurotransmitters such as norepinephrine (NE), on the basis of the finding that neurotransmitters modify immune responses in mammalian species. We confirmed that chicken lymphocytes in the brusa of Fabricius, thymus and spleen expressed beta-adrenergic receptor (beta-AR) mRNA from embryonic day (E) 10 and that intracellular cAMP level was elevated by NE, suggesting that lymphocytes express functional beta-AR on their surface at an early embryonal stage. To clarify whether the nervous system is involved in the development of the immune system, the effects of 6-hydroxydopamine (6-OHDA), one of sympathectomizing agents, on chicken lymphocytes was investigated. A single injection of 6-OHDA at a dose of 400 microg into a chicken embryo was carried out at E7 or 14 (as referred to E7 group and E14 group, respectively). NE level and the relative proportion of Bu-1a(+), CD4(+) and CD8(+) cells in the spleen of 3-week-old chickens were not altered by 6-OHDA treatment. However, the proliferative responses and expression of IL-2 mRNA in spleen cells cultured with pokeweed mitogen were reduced in E7 group compared with those of control. Furthermore, in CD8(+) spleen cells of E14 group of 3-week-old chickens, the expression of beta-AR mRNA and the relative increase of intracellular cAMP stimulated with NE were significantly decreased. These results suggest that the sympathetic nervous system affects the development of the immune system.

The effects of sugar cane extracts (SCE) on immune responses in chickens were studied. Two- or 10-month-old chickens orally administered SCE (500 mg/kg/day), for 3 consecutive days before immunized with sheep red blood cells, Brucella abortus and Salmonella Enteritidis organisms, showed significantly increased and prolonged antibody responses to these antigens, compared to control chickens without SCE. Furthermore, chickens orally administered SCE also revealed enhanced delayed type hypersensitivity responses to human gamma globulin. These results indicated that SCE has immunostimulating and adjuvant effects in chickens.

Polymorphonuclear cells of the peripheral blood in the chicken significantly increased their phagocytosis when cultured with sugar cane extract (SCE; 250-1,000 microg/ml) for 24 hr. Chickens orally administered SCE (500 mg/kg/day) for 3 or 6 consecutive days at 1 week of age showed significantly higher body weight and gain in body weight/day and a lower food conversion ratio within the growing period of 6 weeks than physiological saline-administered control chickens. Furthermore, oral administration of SCE also resulted in significantly higher immune responses against sheep red blood cells and Brucella abortus. These results suggest that SCE has immunostimulating and growth promoting effects in chickens.

3H-thymidine (3H-TdR) incorporation assay has been generally used to measure lymphocyte proliferation in the chicken. Disadvantages of this assay are that radioisotope is biological hazard to the person and environment and that it can not measure which subset of lymphocytes proliferates. In this study, bromodeoxyuridine (BrdU) incorporation assay by flow cytometry was compared with 3H-TdR incorporation assay. As a result, BrdU incorporation assay showed a strong correlation with 3H-TdR incorporation assay, and it could be applied simultaneously to detect BrdU incorporation and expression of cell surface marker antigens. These results suggest that the BrdU incorporation assay by flow cytometry is useful to analyze lymphocyte proliferation in detail.

To examine in vivo effects of egg white derivatives (EWD), the numbers of peripheral blood cells and neutrophil phagocytosis were evaluated in cats injected intramuscularly with cyclophosphamide (CPA). There were no changes in the number of red blood cells (RBC) or packed cell volume (PCV) values regardless of oral administration of EWD or injection of CPA, but the numbers of platelets, white blood cells (WBC) and neutrophils in cats administered EWD significantly increased (p<0.05 to 0.01) when compared with those in control cats which received saline solution. In addition, the administration of EWD resulted in a significant enhancement in the phagocytic activity of neutrophils (p<0.01) when compared to control cats, suggesting that EWD has a stimulating effect on leukocyte progenitors. The numbers of platelets, WBC and neutrophils, and the phagocytic activity of neutrophils in cats injected with CPA alone were significantly lower (p<0.05 to 0.01) than those in control cats. However, co-administration of EWD to cats injected with CPA resulted in a significant increase in the numbers of platelets, WBC and neutrophils (p<0.05 to 0.01), and in the phagocytic response of neutrophils (p<0.01) when compared to cats injected with CPA alone. Therefore, these results suggest that co-administration of EWD may be effective in reducing some possible side effects in animals treated with immunosuppressive or antitumor agents.

The effects of the lipopolysaccharide-protein complex (LPS) and crude capsular antigen (CCA) prepared from Pasteurella multocida serotype A isolated from a duck in the Philippines, on antibody responses to sheep red blood cells (SRBC) and Brucella abortus (BA) and delayed type hypersensitivity (DTH) responses to bovine serum albumin (BSA) in the chickens were studied. Chickens injected subcutaneously with LPS and CCA at 1 and 2 weeks of age and immunized intravenously with the mixed antigens of SRBC and BA, at 3 and 4 weeks of age showed significantly increased antibody responses against both SRBC and BA, when evaluated at 7 days after each immunization. In addition, these chickens sensitized intramuscularly with the emulsion of BSA in complete Freund's adjuvant at 5 weeks of age, and then injected into the wattle with BSA at 7 weeks of age also showed significantly increased DTH responses against BSA, when evaluated at 24 and 48 hr after challenge. These results indicate that LPS and CCA of P. multocida serotype A have a property enhancing humoral and cell-mediated immune responses.

The effects of four antigenic fractions of Pasteurella multocida serotype A isolated from a duck in the Philippines on the phagocytic activity of chicken peripheral blood leukocytes were studied by a flow cytometer. These fractions were the lipopolysaccharide-protein complex (LPS), crude capsular antigen (CCA), ribosomal fraction (RS) and outer cell layer (OCL). Among these four antigens, only CCA but not LPS RS and OCL, significantly increased the phagocytic activities of mononuclear cells (MNC) and polymorphonuclear cells (PMN). This result indicates that CCA has an immunological property enhancing the phagocytic activities of MNC and PMN.

To estimate canine interleukin-8 (cIL-8) levels in blood plasma samples, a sandwich enzyme linked immunosorbent assay (ELISA) was established. For the development of the sandwich ELISA, polyclonal anti-cIL-8 (capturing), biotinylated anti-cIL-8 (developing) antibodies and glutathione-S-transferase/cIL-8 (GST/cIL-8) fusion protein as an antigen were used. cIL-8 in the fusion protein of GST/cIL-8 was detected in a dose dependent manner. The lowest limit of GST/cIL-8 detectable by this method was 2 ng/ml of GST/cIL-8 (containing; 0.470 ng/ml of cIL-8). IL-8 levels in the plasma samples from apparently healthy dogs were less than 0.470 ng/ ml. Higher levels of IL-8 were detected in the plasma samples of dogs with cystitis, dermatitis, and gastric cancer. These results suggest that the determination of cIL-8 by the sandwich ELISA is useful in diagnosis of inflammatory diseases in dogs.

Polyclonal anti-canine interleukin-8 (cIL-8) antibodies were raised in rabbits immunized with cIL-8 expressed by E. coli. Polyclonal antibodies were purified by affinity chromatography. In the enzyme linked immunosorbent assay (ELISA), the resulting anti- cIL-8 antibodies showed relatively high reactivities with cIL-8 in the fusion proteins of glutathione-S-transferase/cIL-8 (GST/cIL-8) and maltose binding protein/cIL-8 (MBP/cIL-8), but negligible ones with MBP. Furthermore, Western blotting analysis using these polyclonal antibodies showed distinct bands for cIL-8, GST/cIL-8, and MBP/cIL-8. These antibodies also bound to recombinant human IL-8 (rhIL-8) in ELISA but not in Western immunoblotting. The rHIL-8 (50-800 ng/ml) was chemoattractant for canine neutrophils in a dose dependent manner, but the anti-cIL-8 antibodies did not show the inhibitory effect on the chemotactic activity of rhIL-8 of canine neutrophils, when tested by the chemotaxis assay using Boyden chambers. In addition, GST/cIL-8 and rhIL-8 induced strong and rapid shape change responses of canine neutrophils. However, the anti-cIL-8 antibodies inhibited shape change responses induced by GST/cIL-8 but not by rhIL-8.

Antibody responses to the hapten dinitrophenyl (DNP) and the concentration of catecholamine in chickens received a single injection of 6-Hydroxydopamine (6-OHDA) into the chorioallantoic cavity at the embryonal stage were evaluated. A significantly enhanced anti-DNP IgA response and a markedly decreased level of noradrenaline in the peripheral blood were observed in chickens treated with 400 micrograms of 6-OHDA at 14 days of incubation. These results suggested the immunomodulatory influence of the sympathetic nervous system.

A cell line, FBM-17, was established from a fetal bovine thymus and passaged for more than a year. Electron microscopic examination revealed that the cytoplasm contained many phagosomes and a significant amount of rough endoplasmic reticula. In addition, FBM-17 cells showed non-specific esterase, phagocytosis and surface expression of Fc receptors for IgG and classes I and II MHC antigens. These observations confirmed that this cell line had the morphologic and functional features as a macrophage. MHC expression markedly increased by treatment with recombinant bovine gamma-interferon, showing the cell line to be sensitive to bovine gamma-interferon and functional. These present results suggest that the FBM-17 is a functional bovine macrophage cell line. This line gamma-interferon, showing the cell line to be useful for clarification of roles of macrophages in immunoregulation in cattle.

Immunostimulative effects of chicken egg white derivatives (EWD) on phagocytic responses of peripheral blood mononuclear cells (MNC) and polymorphonuclear cells (PMN) in dogs were evaluated by flow cytometric analysis. Peripheral blood leukocytes (PBL) cultured with EWD showed the enhanced phagocytic response. The response was maximal when PBL were cultured with 100 - 400 micrograms/ml of EWD for 3 - 12 hr. Furthermore, significantly increased phagocytic responses were also induced even when PBL were cultured with protein components (200 micrograms/ml) of EWD such as conalbumin, flavoprotein and ficin-papain inhibitor for 3 hr. In addition, the enhancing effect of EWD on the phagocytic responses was also observed in MNC cultured with EWD (200 micrograms/ml) for 4 hr but not in PMN cultured with EWD in the same procedures. The supplement of the supernatant (20%) of MNC cultured with EWD (200 micrograms/ml) for 24 hr at 37 degrees C to PBL and MNC resulted in the enhancement of their phagocytic responses. In contrast, the supernatant of PMN cultured with EWD for 24 hr at 37 degrees C did not show any significant enhancing effect on the phagocytic responses of PBL, MNC and PMN. These results suggest that EWD has an enhancing effect on phagocytosis of MNC and PMN, which may be mediated through active humoral substances produced by EWD-stimulated MNC.

Chemotactic activities in the culture supernatants of Cos7 cells transfected with a cloned canine IL-8 cDNA (pcIL-8SR alpha 14) were evaluated by using mononuclear cells (MNC) and polymorphonuclear cells (PMN) from the peripheral blood of dogs. The culture supernatants of Cos7 cells were collected 66 hr after the transfection of pcIL-8SR alpha 14 (Cos7/cIL-8). Chemotactic activities in the culture supernatants for MNC and PMN were determined as migration distances in Millipore membrane filters in a modified Boyden's chamber method. Peroxidase staining for MNC was effective not only for cells in cytospun smears but also for cells migrated in the filters. PMN in cytospun smears were well stained by peroxidase staining, whereas migrated PMN in the filters were stained weakly. Chemotactic activities in the culture supernatant of Cos7/cIL-8 cells for both MNC and PMN were significantly higher than those of control Cos7 cells. In addition, the culture supernatant of Cos7/cIL-8 cells was chemotactic for peroxidase negative nonadherent MNC (lymphocytes), but not for peroxidase positive adherent MNC (monocytes). This Cos7/cIL-8 supernatant also showed chemotactic activities for neutrophils in a dose dependent manner. These results suggest that the culture supernatant of Cos7/cIL-8 is chemotactic for lymphocytes as well as for neutrophils.

Delayed-type hypersensitivity (DTH) responses to rat hepatocytes (HCs) in mice were investigated by flow cytometric analysis and immunohistochemistry with monoclonal antibodies directed against murine class II, CD4, and CD8 antigens. Mice were immunized subcutaneously (s.c.) with 10(6) rat HCs (referred to as s.c.-immunized mice), and control mice were injected s.c. with sterile Hanks' solution (non-immunized mice). Four days later, 10(5) rat HCs were injected into the footpad of s.c.-immunized mice and non-immunized mice. The DTH response in s.c.-immunized mice significantly increased after challenge when compared to that in non-immunized mice. The numbers of class II+, CD4+, and CD8+ cells in the footpad, and CD4+ and CD8+ cells in the inguinal lymph node of s.c.-immunized mice significantly increased during the DTH response. An increase in the number of CD4+ cells in the footpad of s.c.-immunized mice after challenge was more significant than that of non-immunized mice. The number of CD4+ cells increased more markedly in the footpad of s.c.-immunized mice as compared to that of CD8+ cells. Furthermore, immunohistochemical studies of the footpad of s.c.-immunized mice showed more severe infiltration of CD4+ cells rather than CD8+ cells at the injection site of rat HCs. These results suggest that the DTH response in the footpad of mice immunized with rat HCs is associated with severe infiltration of CD4+ cells.

The delayed type hypersensitivity (DTH) response to xenogeneic hepatocytes (HCs) was investigated in mice received subcutaneous (s.c.), intrasplenic (i.s.), or intravenous (i.v.) transplantation of rat HCs. The DTH response in mice preimmunized i.s. or i.v. with rat HCs (1 x 10(6) cells) was significantly lower than that in mice preimmunized s.c. with the same doses of rat HCs. Co-transfer of spleen cells from i.s. immunized mice with spleen cells from s.c. immunized mice to naive recipient mice did not suppress the DTH response induced by transfer of spleen cells from s.c. immunized mice. On the other hand, co-transfer of spleen cells from i.v. immunized mice with spleen cells from s.c. immunized mice suppressed the DTH response to rat HCs in recipients. Furthermore, the levels of DTH responses in recipients transferred with spleen cells from mice sensitized i.s. or i.v. with rat HCs, immunized s.c. with rat HCs 6 hr after transfer, and challenged with rat HCs 7 days later was almost similar to those in recipients transferred with spleen cells from s.c. immunized mice. These results suggest that antigen-specific suppression of DTH responses to rat HCs in mice is associated with the presence of suppressive spleen cells induced by i.s. or i.v. immunization with rat HCs.

The serum concentration of a bovine mannan-binding protein reactive with a Ra chemotype strain of Salmonella Typhimurium in sera from cows with or without mastitis were determined by sandwich enzyme-linked immunosorbent assay. From the results obtained for 10 healthy cows aged 2 to 7 years, mean +/- SD serum concentrations of MBP in bovine sera were 77.5 +/- 33.1 micrograms/ml. Concentrations in 7 healthy heifer calves aged 6 months were 52.2 +/- 10.2 micrograms/ml, whereas those in 7 healthy bullocks 65.8 +/- 21.8 micrograms/ml. Concentrations in 4 cows aged 4 to 7 years with mastitis were slightly lower (34 to 72 micrograms/ml). After recovery, the serum concentrations rose to the normal concentrations in healthy cows. These findings indicate that the serum concentrations of bovine MBP decrease during mastitis, suggesting that bovine MBP may not be an acute phase reactant.

To compare a chicken serum mannan-binding protein (MBP) with a chicken Ra-reactive factor (RaRF), both serum proteins were isolated by combination of different chromatography. In SDS-PAGE, both purified proteins were resolved into a major protein band with a molecular weight of 34,000 and a few minor protein bands. In Western blotting with both purified proteins, only a single band with a molecular weight of 34,000 was detected with anti-chicken serum MBP antibody, indicating that the major protein band of chicken serum MBP and RaRF are antigenically identical. The combining sites of these two proteins were most reactive with N-acetylmannosamine followed by mannose, N-acetylglucosamine, and L-fucose. These findings suggest that chicken serum MBP may be identical to its corresponding bactericidal factor RaFR in terms of antigenicity and monosaccharide specificity.

The effect of vitamin A deficiency and vitamin A replacement on spermatogenesis in golden hamsters was studied using a light microscope. Male golden hamsters were fed a vitamin A deficient (VAD) diet from 3 weeks of age. Hamsters with a VAD diet reached maximum body weight at about 13 weeks. After 17 weeks, the body weight of the hamsters began to decrease. When their body weight decreased to 70 g, only Sertoli cells, spermatogonia, and a few spermatocytes were present within the seminiferous tubules. Administering retinol acetate (vitamin A) combined with a conventional diet to the VAD hamsters induced a reinitiation of spermatogenesis with stage-synchronization. At 9, 10, and 11 weeks after vitamin A replacement, the testes with active spermatogenesis possessed only a few successive stages of the seminiferous epithelial cycle.

A Ca(2+)-dependent agarose-binding protein was purified from chicken serum by affinity chromatography on SeaKem agarose gels followed by gel filtration on Sephacryl S-300. Purified protein was eluted in a symmetrical peak at the position of molecular weight of 146,000. In sodium dodecyl sulfate-polyacrylamide gel electrophoresis, it was resolved into a single protein band with a molecular weight of 39,000 under nonreducing and of 41,000 under reducing conditions. Thus, the agarose-binding protein is suggested to be composed of noncovalently-associated identical subunits which contain intrachain disulfide bonds.

The distribution of lymphocytes in lymphoid organs of germfree (GF) and conventional (CV) chickens was compared immunohistochemically with several monoclonal antibodies. The lymphoid follicles in the cecal tonsils of GF chickens were more poorly developed than in those of CV chickens, and IgG and IgA positive cells were also not found in GF chickens. The number of both B and T cells in the villous region of the cecal tonsils in GF chickens was less than that in CV chickens. In the spleen of GF chickens, the distribution of B cells was similar, but T cells were distributed more widely in CV chickens than in GF chickens. There was no clear difference in the distribution of lymphoid cells in the bursa of Fabricius and the thymus. These results suggest that the development of gut-associated lymphoid tissues is affected by intestinal flora.

A Ca(2+)-dependent phosphorylcholine (PC)-binding protein in normal chicken serum was purified by affinity chromatography on p-aminophenyl PC-Sepharose 4B followed by gel filtration on Sephacryl S-300. In gel filtration, the isolated PC-binding protein was eluted in a symmetrical protein peak at the position of approximately 100,000. On sodium dodecyl sulfate-polyacrylamide gel electrophoresis, the protein was resolved into two protein bands of 31,000 and 38,000 under nonreducing conditions and of 40,000 and 46,000 under reducing conditions. These results suggest that chicken serum PC-binding protein may be composed of two different subunits which contain intrachain disulfide bonds.

The localization of lymphocytes in the lymphoid organs of cyclophosphamide (Cy) treated chickens and untreated control chickens was compared immunohistochemically using a variety of monoclonal antibodies (CT3, 2-6, 11-39, TCR1, TCR2, TCR3, L22, 11G, 3E8, B-4D-4, A-13). In the Harderian glands of Cy treated chickens, an an increase of T cells was observed, though T cells were a few in untreated controls. These increased T cells consisted of CD4 positive or CD8 positive cells. Further, these T cells were stained with TCR2 or TCR3 antibody, and a small number of cells were stained with TCR1 antibody. In other lymphoid organs such as the bursa of Fabricius, thymus, spleen and cecal tonsils, B lymphocyctes severely decreased or disappeared in Cy treated chickens, though no significant alteration in T cell distribution was observed.

The effects of recombinant bovine interferon alpha 1 (rBoIFN alpha 1) on the incidence of bovine respiratory disease (BRD) were examined in 60 young male cattle. Each group of clinically healthy 30 male cattle was injected intramuscularly with either rBoIFN alpha 1 (5 mg/head) or physiological saline 5 times in 6 months. They were examined daily for the clinical symptoms of respiratory distress. The clinical signs of respiratory distress in rBoIFN alpha 1-treated cattle were significantly less severe than those of the placebo group. The incidence rate, relapse rate and mean sick days of respiratory disease in the rBoIFN alpha 1-treated group were 23%, 14% and 4.43 days, respectively, whereas those in the placebo group were 80%, 46% and 6.42 days, respectively. Furthermore, the mean increase of body weights in rBoIFN alpha 1-treated group and placebo group were 1.365 kg/day and 1.340 kg/day, respectively. These results suggest that rBoIFN alpha 1 has a potentially preventive effect on the incidence of BRD.

To isolate the major carbohydrate-binding proteins in pooled normal chicken sera, affinity chromatographies on different affinity gels were carried out. From the results obtained, normal chicken sera were found to contain serum proteins which were Ca(2+)-dependently and -independently reactive with mannose, N-acetylglucosamine, N-acetylgalactosamine, L-fucose, melibiose, lactose, unsubstituted Sepharose 4B, and SeaKem HE agarose. They were electrophoretically and antigenically found to be mainly IgM and IgG.

Changes in the levels of plasma cortisol, progesterone and total testosterone were examined in developing hairless and haired dogs. Cortisol levels in the hairless dogs seemed to be higher than those in haired dogs within the age of 4-5 weeks. No apparent changes were seen in the level of plasma progesterone between the groups of hairless females and haired females. Total testosterone levels in hairless males showed to be significantly lower than those in haired males at the age of 13-21 weeks.

To study the roles of conglutinin (Kg), mannan-binding protein (MBP), and serum amyloid P component (SAP) in the protection of cattle against infections, the concentrations of these proteins in the sera from cows with mastitis were determined by sandwich enzyme-linked immunosorbent assays. The Kg and MBP concentrations in the sera from cows with mastitis were much lower than those from uninfected (or clinically healthy) ones, and increased after recovery. No significant difference in the SAP concentrations was found between healthy and infected cows. With the sera from cows experimentally infected with bovine leukemia virus, the Kg concentrations were also lower than those from uninfected ones, whereas the MBP concentrations were not. From these findings, the Kg concentration is suggested to be serologically a possible indicator for clinical diagnosis of treatment for mastitis although both Kg and MBP concentrations were found to decrease in cows with mastitis and to increase after recovery.

Binding specificities of calcium-dependent and -independent bovine IgM and IgG reactive with unsubstituted Sepharose 4B were determined by competitive binding assays. The binding of 125I-labeled calcium-dependent bovine IgM to unsubstituted Sepharose 4B was most effectively inhibited by lactose which was about 100 times more reactive than galactose and melibiose. Other inhibitors were much less potent. With 125I-labeled calcium-independent bovine IgM and IgG, on the other hand, lactose was as potent as galactose. Melibiose was about 10 times less potent than lactose and galactose, whereas other mono- and disaccharides were much less potent. From these findings, the combining site of calcium-dependent bovine IgM reactive with unsubstituted Sepharose 4B may be specific for lactose, whereas those of calcium-independent bovine IgM and IgG specific for galactose.

Calcium-dependent and -independent bovine IgG reactive with invertase were isolated by affinity chromatography on invertase-coupled Sepharose 4B and their specificities were determined by competitive binding assays. The binding of 125I-labeled calcium-dependent and -independent bovine IgG reactive with invertase to invertase-coupled Sepharose 4B were most effectively inhibited by invertase and mannan. The binding was more weakly inhibited by mannose, glucose, and methyl alpha-D-mannopyranoside, whereas that was much more weakly inhibited by other monosaccharides and glycoproteins. These findings suggest that the combining sites of calcium-dependent and -independent bovine IgG reactive with invertase may be specific for invertase and/or mannan.