Oxidative responses in ferret macrophages

Although the basic function of T and B lymphocytes in ferrets has been known for some time, the function of mononuclear phagocytes has not been described in this species. The present study has characterised basic oxidative responses in ferret macrophages, and has investigated the effects of endogenous and exogenous modulators of macrophage function on oxidative capacity in vitro. Macrophages derived from the blood or lungs of ferrets were shown capable of generating the reactive oxygen intermediate (ROI) molecules superoxide and hydrogen peroxide, and secreting a lysosomal enzyme (acid phosphatase), in response to appropriate stimuli. A T cell supernatant (derived from mitogen-stimulated peripheral blood lymphocytes) was able to activate both blood- and lung-derived macrophages for enhanced ROI production, while specific ROI inhibitors (superoxide dismutase and catalase) were able to partially ablate ROI activity. The accumulation of nitrite in culture supernatants, as an indicator for the production of reactive nitrogen intermediates, could not be demonstrated by ferret macrophages derived from either tissue source. In contrast to the enhancing effects of TCS on the oxidative function of blood-derived macrophages, exposure to bacterial LPS caused marked suppression of ROI and lysosomal enzyme production by these cells. Finally, the generation of superoxide anion, following phagocytosis of live or heat-killed Mycobacterium bovis or zymosan, indicated that ROI production in response to phagocytic stimulation was relatively weak in ferret blood-derived macrophages. These results are discussed in relation to the study of immune function in a novel species, and with particular reference to research into tuberculosis (Tb), since ferrets are important wildlife vectors of bovine Tb in New Zealand.     

Effects of organic forms of zinc on growth performance,tissue zinc distribution,and immune response of weanling pigs

This study was conducted to determine the effect of zinc level and source on growth performance, tissue Zn concentrations, intracellular distribution of Zn, and immune response in weanling pigs. Ninety-six 3-wk-old crossbred weanling pigs (BW = 6.45 +/- 0.17 kg) were assigned to one of six dietary treatments (four pigs per pen, four replicates per treatment) based on weight and litter origin. Treatments consisted of the following: 1) a corn-soybean meal-whey diet (1.2% lysine) with a basal level of 80 ppm of supplemental Zn from ZnSO4 (control; contained 104 ppm total Zn); 2) control + 80 ppm added Zn from ZnSO4; 3) control + 80 ppm added Zn from Zn methionine (ZnMet); 4) control + 80 ppm added Zn from Zn lysine (ZnLys); 5) control + 40 ppm added Zn from ZnMet and 40 ppm added Zn from ZnLys (ZnML); and 6) control + 160 ppm added Zn from ZnSO4. Zinc supplementation of the control diet had no effect on ADG or ADFI. Gain efficiency was less (P < 0.05) for pigs fed 80 ppm of Zn from ZnSO4 than for control pigs and pigs fed 160 ppm of Zn from ZnSO4. Organ weights, Zn concentration, and intracellular distribution of Zn in the liver, pancreas, and spleen were not affected (P = 0.12) by Zn level or source. Skin thickness response to phytohemagglutinin (PHA) was not affected (P = 0.53) by dietary treatment. Lymphocyte proliferation in response to PHA was greater (P < 0.05) in pigs fed ZnLys than in pigs fed the control diet or the ZnML diet; however, when pokeweed mitogen was used, lymphocyte proliferation was greatest (P < 0.05) in pigs fed the ZnMet diet than pigs fed the control, ZnLys, ZnML, or 160 ppm ZnSO4 diets. Antibody response to sheep red blood cells was not affected by dietary treatments. Supplementation of 80 ppm of Zn from ZnSO4 or ZnMet and 160 ppm of Zn from ZnSO4 decreased (P < 0.05) the antibody response to ovalbumin on d 7 compared with control pigs, but not on d 14. Phagocytic capability of peritoneal exudate cells was increased (P < 0.05) when 160 ppm of Zn from ZnSO4 was supplemented to the diet. The number of red blood cells ingested per phagocytic cell was increased (P < 0.05) in pigs fed the diet supplemented with a combination of ZnMet and ZnLys and the diet with 160 ppm of Zn from ZnSO4. Results suggest that the level of Zn recommended by NRC for weanling pigs was sufficient for optimal growth performance and immune responses, although macrophage function may be enhanced at greater levels of Zn. Source of Zn did not alter these measurements.     

Transmission of Mycobacterium bovis from experimentally infected ferrets to non-infected ferrets (Mustela furo)

AIMS: To demonstrate the transmission of Mycobacterium bovis infection from experimentally infected ferrets (Mustela furo) to non-infected ferrets in a laboratory setting, using three different isotypes of M. bovis, and to observe ferret behaviour that might be implicated in disease transmission. METHODS: Three female ferrets, each experimentally infected with a unique strain of M. bovis, were housed together with six female and two male non-infected ferrets in an isolation facility. Transmission of infection was monitored clinically, serologically (using an ELISA test), bacteriologically, histologically, and by isotype analysis of M. bovis isolates using spoligotyping to determine whether or not transmission of each strain occurred. Ferret behaviour was observed using a time-lapse video recorder. RESULTS: Transmission of M. bovis infection was confirmed in two male and four female ferrets. Isotype analysis showed that of the experimentally infected females, one did not infect any other ferret, another transmitted M. bovis to one ferret before it died prematurely 49 days post-infection, and the third, which was cannibalised, appears to have transmitted M. bovis to both males and three females. However, two of these latter three females had died before the event of cannibalism took place. One female was infected with two strains. Several behavioural interactions were observed that could have resulted in M. bovis transmission, including den sharing, sniffing of orifices and faeces, cannibalism and aggressive breeding behaviour. CONCLUSIONS: Horizontal transmission of M. bovis infection was demonstrated in ferrets under experimental housing conditions. Routes of transmission may involve cannibalism and factors such as den sharing, playing, fighting, mating, and sniffing of faeces.     

Amelioration of calcareous saline sodic soils through phytoremediation and chemical strategies

Abstract. The worldwide occurrence of saline sodic and sodic soils on more than half a billion hectares warrants attention for their efficient, inexpensive and environmentally acceptable management. These soils can be ameliorated by providing a source of calcium (Ca²⁺) to replace excess sodium (Na⁺) from the cation exchange sites. Although chemical amendments have long been used to ameliorate such soils, the chemical process has become costly during the last two decades in several developing countries. As a low‐cost and environmentally acceptable strategy, the cultivation of certain salt tolerant forage species on calcareous sodic and saline sodic soils, i.e. phytoremediation, has gained interest among scientists and farmers in recent years. In a field study conducted at three calcareous saline sodic sites (pH_s=8.1–8.8, EC_e=7.8–12.5 dS m^–1, SAR=30.6–76.1) in the Indus Plains of Pakistan, we compared chemical and phytoremediation methods. There were four treatments; two involved plants: Kallar grass (Leptochloa fusca (L.) Kunth), and sesbania (Sesbania bispinosa (Jacq.) W. Wight). The other two treatments were uncropped: soil application of gypsum and an untreated control. All treatments were irrigated with canal water (EC=0.22–0.28 dS m^–1). The plant species were grown for one season (5–6 months). Sesbania produced more forage yield (34 t ha^–1) than Kallar grass (23 t ha^–1). Phytoremediation and chemical treatments resulted in similar decreases in soil salinity and sodicity, indicating that phytoremediation may replace or supplement the more costly chemical approach. The soil amelioration potential of sesbania was similar to that of the Kallar grass, which suggests that moderately saline sodic calcareous soils can be improved by growing a forage legume with market value.     

Trans-endocytosis of CD80 and CD86: a molecular basis for the cell-extrinsic function of CTLA-4

Cytotoxic T lymphocyte antigen 4 (CTLA-4) is an essential negative regulator of T cell immune responses whose mechanism of action is the subject of debate. CTLA-4 shares two ligands (CD80 and CD86) with a stimulatory receptor, CD28. Here, we show that CTLA-4 can capture its ligands from opposing cells by a process of trans-endocytosis. After removal, these costimulatory ligands are degraded inside CTLA-4-expressing cells, resulting in impaired costimulation via CD28. Acquisition of CD86 from antigen-presenting cells is stimulated by T cell receptor engagement and observed in vitro and in vivo. These data reveal a mechanism of immune regulation in which CTLA-4 acts as an effector molecule to inhibit CD28 costimulation by the cell-extrinsic depletion of ligands, accounting for many of the known features of the CD28-CTLA-4 system.     

Improved gas chromatographic method for quantitation of deoxynivalenol in wheat, corn, and feed

A gas chromatographic (GC) method is described to determine deoxynivalenol in wheat and corn at levels as low as 20 ppb. Ground samples are extracted with water, adsorbed onto a Clin Elut column, extracted with ethyl acetate, and passed through a silica gel Sep-Pak cartridge. The final extract is then derivatized with N-heptafluorobutyrylimidazole and quantitated by GC using an electron capture detector. Recoveries are greater than 85% for spiked samples at levels of 50-1000 ppb. Results for wheat, corn, and mixed feed samples are given as well as the results of an interlaboratory study on a naturally contaminated wheat sample.     

Comparative effects of applying leguminous and non-leguminous green manures and inorganic N on biomass yield and nitrogen uptake in flooded rice (<Emphasis Type="Italic">Oryza sativa</Emphasis> L.)

The beneficial role of green manures in rice production is generally ascribed to their potential of supplying plant nutrients, particularly nitrogen (N). However, the mechanisms through which green manures enhance the crop productivity are poorly understood. Pot experiments were conducted using a ¹⁵N-tracer technique: (1) to compare the biomass production potential of sesbania (Sesbania aculeata Pers.) and maize (Zea mays L.) as green manuring crops for lowland rice and (2) to compare the effect of the two types of green manure and inorganic N on the dry matter accumulation and N uptake by two rice (Oryza sativa L.) cultivars, viz. IR-6 and Bas-370. Although maize produced three times higher shoot biomass compared with sesbania, the latter showed higher N concentration; and thus the total N yield was similar in the two types of plants. Applying the shoot material of the two plants to flooded rice significantly enhanced the dry matter yield and N uptake by the two rice cultivars, the positive effects generally being more pronounced with sesbania than with maize amendment. The difference in the growth-promoting potential of the two plant residues was related more to an increased uptake of the native soil N rather than to their direct role as a source of plant-available N. A positive added nitrogen interaction (ANI) was observed due to both plant residues, the effect was much more pronounced with the application of sesbania than with maize residues. In both rice cultivars, inorganic N also caused a substantial ANI, particularly at higher application rate. Losses from the applied N were 2–3 times lower from sesbania, compared with maize treatment. Green manuring with sesbania also caused much lower N losses than the inorganic N applied at equivalent or higher rates. The overall benefit of green manuring to rice plants was higher than inorganic N applied at comparable rates. The two rice cultivars differed in their response to green manuring, IR-6 generally being more responsive than Bas-370.     

Regeneration in sugarcane via somatic embryogenesis and genomic instability in regenerated plants

In the present study, embryogenic calli of sugarcane variety BL4 were induced using 2,4-dichlorophenoxyacetic acid (2,4-D) and kinetin in different concentrations and combinations. In contrast to earlier studies, embryogenic callus sectors were identified and isolated microscopically within 1–2 weeks. Subsequently, 51 media formulations were used for regeneration of proliferated embryogenic callus, using MS medium supplemented with three different cytokinins [kinetin, 6-Benzylamino purine (BAP), and thidiazuron (TDZ)] and auxins (IAA/NAA and IBA) in different combination and concentrations. After acclimatization, the genomic DNA of regenerated plants was studied to explore the insertion polymorphism of transposable elements in order to ascertain the variation among somaclones. Though low concentration of kinetin with 2,4-D was found supportive to embryogenic callus development, the highest number of regenerated plantlets was observed using BAP (1 μM), however the plantlets had very low fresh weight (2.2 g). Conversely, TDZ alone supported a significant increase in the number of plantlets regenerated (38–40) with higher fresh weight. The somaclones generated during this study showed considerable positional polymorphism of activator-like transposable elements possibly due to the stress associated with in vitro culture. This study provides a procedure to produce regenerated sugarcane plants from embryogenic callus in a relatively short time.     

Comparative evaluation of different therapeutic protocols for contagious caprine pleuropneumonia in Himalayan Pashmina goats

Tropical Animal Health and Production - Therapeutic management of contagious caprine pleuroneumonia (CCPP) involves mostly the use of oxytetracycline followed by enrofloxacin and rarely tylosin. In...