Enhanced phagocytic activity of neutrophils caused by administration of egg white derivatives (EWD) in cats injected with cyclophosphamide (CPA)

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.     

Effects of 6-hydroxydopamine on the development of the immune system 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.     

Protective effects of sugar cane extracts (SCE) on Eimeria tenella infection in chickens

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.     

Detection of mitogen-induced lymphocyte proliferation by bromodeoxyuridine (BrdU) incorporation in the chicken

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.     

Immunostimulating and growth-promoting effects of sugar cane extract (SCE) 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.     

A split-root apparatus for examining the effects of hydraulic lift by trees on the water status of neighbouring crops

We describe a split-root system for examining the effects of hydraulic lift by trees on crop growth. In this system, upper lateral tree roots were grown in a container set on the ground through which the taproot of the tree could penetrate into the moist soil below. The container, with a radius of 0.5 m and a height of 0.20 m, consisted of two compartments divided by a waterproof barrier. A markhamia tree (Markhamia lutea (Benth.) Schumann) and upland rice (Oryza sativa (L.)) plants were planted in one compartment, with only rice plants planted in the other compartment. Irrigation of the container was ceased at the start of the experiment. The stomatal conductance of the rice plants in the associated side, in which both trees and rice plants were grown, declined more rapidly during the first drying period than in the rice-only compartment, suggesting that there was competition for water between the tree and the crop plants. However, during the later drying period, the rice plants in the associated side were green and viable, while those in the rice-only side became desiccated. Rice roots were seen intermingling with tree roots, and the soil water content in the associated site tended to be higher than in the rice-only side. It is likely that hydraulic lift occurred in the associated side and that water that had been transferred to the surface roots was released into the soil, enabling the rice plants in this compartment to remain viable. This novel system is useful for examination of the effects of hydraulic lift by trees on the growth of neighbouring crops. This revised version was published online in June 2006 with corrections to the Cover Date.     

Methane emissions from different vegetation zones in a Qinghai-Tibetan Plateau wetland

We measured methane (CH₄) emissions in the Luanhaizi wetland, a typical alpine wetland on the Qinghai-Tibetan Plateau, China, during the plant growth season (early July to mid-September) in 2002. Our aim was to quantify the spatial and temporal variation of CH₄ flux and to elucidate key factors in this variation. Static chamber measurements of CH₄ flux were made in four vegetation zones along a gradient of water depth. There were three emergent-plant zones (Hippuris-dominated; Scirpus-dominated; and Carex-dominated) and one submerged-plant zone (Potamogeton-dominated). The smallest CH₄ flux (seasonal mean=33.1 mg CH₄ m⁻² d⁻¹) was observed in the Potamogeton-dominated zone, which occupied about 74% of the total area of the wetland. The greatest CH₄ flux (seasonal mean=214 mg CH₄ m⁻² d⁻¹) was observed in the Hippuris-dominated zone, in the second-deepest water area. CH₄ flux from three zones (excluding the Carex-dominated zone) showed a marked diurnal change and decreased dramatically under dark conditions. Light intensity had a major influence on the temporal variation in CH₄ flux, at least in three of the zones. Methane fluxes from all zones increased during the growing season with increasing aboveground biomass. CH₄ flux from the Scirpus-dominated zone was significantly lower than in the other emergent-plant zones despite the large biomass, because the root and rhizome intake ports for CH₄ transport in the dominant species were distributed in shallower and more oxidative soil than occupied in the other zones. Spatial and temporal variation in CH₄ flux from the alpine wetland was determined by the vegetation zone. Among the dominant species in each zone, there were variations in the density and biomass of shoots, gas-transport system, and root-rhizome architecture. The CH₄ flux from a typical alpine wetland on the Qinghai-Tibetan Plateau was as high as those of other boreal and alpine wetlands.     

Development of enzyme-linked immunosorbent assays for conglutinin, mannan-binding protein, and serum amyloid-P component in bovine sera.

Sandwich ELISA were developed to quantitatively determine conglutinin (CG), mannan-binding protein (MBP), and serum amyloid-P component (SAP) in the sera of cattle. The ELISA system was found to have high repeatability for quantitation of these serum proteins at concentration as low as 5 ng/ml. From results obtained for 10 healthy cows aged 2 to 7 years, mean +/- SD serum concentrations were 56.5 +/- 14.4 micrograms of CG/ml, 2.37 +/- 0.87 micrograms of MBP/ml, and 11.14 +/- 3.92 micrograms of SAP/ml, respectively. Values in 6 healthy heifer calves aged 6 months were 3.45 +/- 1.22 micrograms/ml for CG, 1.71 +/- 0.96 micrograms/ml for MBP, and 5.45 +/- 2.75 micrograms/ml for SAP, respectively. Concentrations in 9 healthy bullocks aged 6 months were 1.83 +/- 0.66 micrograms/ml for CG, 1.04 +/- 0.63 micrograms/ml for MBP, and 4.9 +/- 1.13 micrograms/ml for SAP, respectively.