Effect of Zn-tolerant bacterial strains on growth and Zn accumulation in Orychophragmus violaceus

Several Zn-tolerant bacterial strains were isolated from heavy-metal contaminated sludge, and their effects on root elongation, mobility, and accumulation of Zn in Orychophragmus violaceus were studied. The isolated strains included Bacillus subtilis, B. cereus, Flavobacterium sp. and Pseudomonas aeruginosa which were capable of stimulating root elongation in O. violaceus seedlings either in the presence or absence of Zn. The four bacterial strains significantly increased the concentration of water-extractable Zn compared with axenic soil. In addition, the four Zn-tolerant bacteria significantly increased the shoot biomass and Zn accumulation in O. violaceus compared to non-inoculated plants. The bacterial strains displayed different capacities to enhance plant Zn accumulation. Flavobacterium sp. was identified as the best candidate for enhancing Zn accumulation in plants, increasing Zn accumulation up to 1.21- and 1.19-fold in shoots and roots, respectively, compared to non-inoculated plants. It was indicated that Zn-tolerant bacteria played an important role in influencing the availability of water-soluble Zn in soil and Zn accumulation by plants. This study provides insight into the development of plant–microbe systems for phytoremediation.     

Effect of Microbial Activity and Nitrogen Mineralization on Free-living Nitrogen Fixation in Permanent Grassland Soils

Free‐living nitrogen (N) fixation can be important for sustainable soil fertility, particularly in extensively managed soils with low abundance of leguminous plant species. However, the factors affecting free N₂‐fixation in situ are still poorly documented. We investigated the role of microbial active biomass activity, particularly substrate‐induced respiration (SIR) and net N mineralization, on the free‐living N₂ fixation in soils under a semi‐natural grassland ecosystem in France. Analysis of replicated bulk soil and rhizospheric soil samples obtained from sites experiencing contrasting grazing regimes revealed highly significant negative relationships (P < 0.01) between free‐living N₂‐fixation and SIR or N‐mineralization with a significant rhizosphere effect. The study has demonstrated that the activity of free‐living N₂‐fixers is more important in soils having low active microbial biomass and low N‐mineralization rates in these permanent grasslands.     

Hepatitis E virus: Animal reservoirs and zoonotic risk

Hepatitis E virus (HEV) is a small, non-enveloped, single-strand, positive-sense RNA virus of approximately 7.2 kb in size. HEV is classified in the family Hepeviridae consisting of four recognized major genotypes that infect humans and other animals. Genotypes 1 and 2 HEV are restricted to humans and often associated with large outbreaks and epidemics in developing countries with poor sanitation conditions, whereas genotypes 3 and 4 HEV infect humans, pigs and other animal species and are responsible for sporadic cases of hepatitis E in both developing and industrialized countries. The avian HEV associated with Hepatitis-Splenomegaly syndrome in chickens is genetically and antigenically related to mammalian HEV, and likely represents a new genus in the family. There exist three open reading frames in HEV genome: ORF1 encodes non-structural proteins, ORF2 encodes the capsid protein, and the ORF3 encodes a small phosphoprotein. ORF2 and ORF3 are translated from a single bicistronic mRNA, and overlap each other but neither overlaps ORF1. Due to the lack of an efficient cell culture system and a practical animal model for HEV, the mechanisms of HEV replication and pathogenesis are poorly understood. The recent identification and characterization of animal strains of HEV from pigs and chickens and the demonstrated ability of cross-species infection by these animal strains raise potential public health concerns for zoonotic HEV transmission. It has been shown that the genotypes 3 and 4 HEV strains from pigs can infect humans, and vice versa. Accumulating evidence indicated that hepatitis E is a zoonotic disease, and swine and perhaps other animal species are reservoirs for HEV. A vaccine against HEV is not yet available.     

Clonal growth and fragment regeneration of Rumex obtusifolius L.

Clonal growth and fragment regeneration of Rumex obtusifolius L. were analysed in two dif ferent studies. Clonal growth system was des cribed by a morphological study of underground structure of different-aged individuals, using maximal branching order in the stem system as an age estimator. Glasshouse experiments were conducted, testing the regenerative capacity of different below-ground parts and the estab lishment of above root-collar fragments planted at different depths under contrasting water regimes. Results showed the presence of a ‘phalanx’ type clonal growth system in grassland populations of Rumex. The main structure in volved in clonal growth proved to be the stem system; the region above the root collar was also the only part able to regrow after damage. Stem fragment regeneration occurred to depths of 15 cm, but was prevented in soils maintained at waterlogging and field capacity. The significance of these results in relation to nonchemical con trol of Rumex populations in grasslands is discussed.     

Expression of recombinant Toxoplasma gondii P24

The gene encoding Toxoplasma gondii P24 has been reported previously. To determine the function of P24 against immune systems in the near future, we prepared recombinant P24 antigens using Escherichia coli, insect cells infected with recombinant baculovirus and mammalian cells infected with recombinant vaccinia virus. The P24 antigens derived from E. coli, insect cells and mammalian cells were detected with mouse immune sera against P24 or T. gondii homogenates by Western blot analysis; these corresponded to the authentic P24 and secreted into the supernatants of the insect and mammalian cell cultures. These proteins were not effected by tunicamycin treatment in cultured cells, indicating that recombinant P24 did not contain N-linked sugars. Recombinant P24 was separated by two-dimensional electrophoresis and analyzed by Western blotting. From these results, P24 was acidic protein and had identical isoelectric point with the authentic P24.     

Use of a von Frey device for evaluation of pharmacokinetics and pharmacodynamics of morphine after intravenous administration as an infusion or multiple doses in dogs

OBJECTIVE: To evaluate the pharmacokinetics and pharmacodynamics of morphine after IV administration as an infusion or multiple doses in dogs by use of a von Frey (vF) device. ANIMALS: 6 dogs. PROCEDURE: In the first 2 crossover experiments of a 3-way crossover study, morphine or saline (0.9%) solution was administered via IV infusion. Loading doses and infusion rates were administered to attain targeted plasma concentrations of 10, 20, 30, and 40 ng/mL. In the third experiment, morphine (0.5 mg/kg) was administered IV every 2 hours for 3 doses. The vF thresholds were measured hourly for 8 hours. Plasma concentrations of morphine were measured by high-pressure liquid chromatography. RESULTS: No significant changes in vF thresholds were observed during infusion of saline solution. The vF thresholds were significantly increased from 5 to 8 hours during the infusion phase, corresponding to targeted morphine plasma concentrations > 30 ng/mL and infusion rates > or = 0.15 +/- 0.02 mg/kg/h.The maximal effect (EMAX) was 78 +/- 11% (percentage change from baseline), and the effective concentration to attain a 50% maximal response (EC50) was 29.5 +/- 5.4 ng/mL. The vF thresholds were significantly increased from 1 to 7 hours during the multiple-dose phase; the EC50 and EMAX were 23.9 +/- 4.7 ng/mL and 173 +/- 58%, respectively. No significant differences in half-life, volume of distribution, or clearance between the first and last dose of morphine were detected. CONCLUSIONS AND CLINICAL RELEVANCE: Morphine administered via IV infusion (0.15 +/- 0.02 mg/kg/h) and multiple doses (0.5 mg/kg, IV, every 2 hours for 3 doses) maintained significant antinociception in dogs.     

Coccidiosis immunization: effects of mushroom and herb polysaccharides on immune responses of chickens infected with Eimeria tenella

An experiment was conducted to investigate the effects of polysaccharide extracts (E) of two mushrooms, Lentinus edodes (LenE) and Tremella fuciformis (TreE), and an herb, Astragalus membranaceus (AstE), on the immune responses of chickens infected with Eimeria tenella. A total of 180 broiler chickens were assigned to nine groups: three groups were fed with each of the extracts (LenE, TreE, and AstE), three groups were fed with the extracts and immunized with live oocyst vaccine (LenE+V, TreE+V, and AstE+V), a group was immunized with the vaccine only, and there were two controls (E. tenella-infected and noninfected groups). The oocyst vaccine was given at 4 days of age, and the extracts (1 g/kg of the diet) were supplemented from 8 to 14 days of age. At 18 days of age, all birds except those in the noninfected group were infected with 9 x 10(4) sporulated oocysts. The results showed that at 7 days postinfection (p.i.), birds fed the extracts without vaccination had lower body weight (BW) gain than those given the vaccine only. However, the extracts in conjunction with the vaccine significantly enhanced BW gain of the infected chickens compared with the vaccine group. Of the three extracts, LenE and TreE showed a better growth-promoting effect. The extracts largely increased oocyst excretion of droppings during the primary response postvaccination. The cecal peak oocyst output and lesion scores measured at 7 days p.i. were higher in the groups fed the extracts than in the group immunized with the vaccine only, whereas those of the groups fed with the extracts and immunized with the vaccine were not significantly different from the vaccine group. Of the three extracts, both LenE- and AstE-fed groups showed lower cecal oocyst output. Thus, as compared with the extracts, the live, attenuated vaccine showed better results with significantly increased immune response in coccidial infected birds. The polysaccharide extracts may prove useful against avian coccidiosis, and, particularly when they are used in conjunction with vaccine, they have shown preliminary promise against the experimental coccidial infection.     

Effects of food deprivation and particle size of ground wheat on digestibility of food components in broilers fed on a pelleted diet

The first aim of the experiment was to study the effect of wheat (Triticum aestivum) particle size on the digestibility of starch in a pelleted diet given to broilers. The second aim was to study the consequences of food deprivation before the excreta collection period (from 21 to 24 d). Wheat from a strong hardness cultivar was incorporated at 546.1 g/kg in diets. The other main ingredients were soybean meal (353.5 g/kg) and rapeseed oil (55.0 g/kg). Diets were given as pellets. The experimental design was a 2 x 2 factorial design testing two particle sizes of wheat flour and two procedures of a balance experiment (with or without food deprivation). Birds given diet C (wheat coarse grinding before pelleting) had significantly greater gizzard weight than birds fed on diet F (wheat fine grinding before pelleting). Starch digestibility value was significantly increased when birds were fed on diet F. This effect was halved by food deprivation. No significant effect of grain particle size was observed for protein and lipid digestibility values. However, food deprivation decreased apparent protein digestibility, with an effect which was more pronounced for fine than for coarse grinding. AMEN of the diet was significantly improved by fine grinding of wheat and decreased by food deprivation. However, no significant differences in growth performance were induced by differences in wheat grinding. No significant effect of grinding was observed on the water excretion:feed intake ratio. No significant difference was observed for vent score between treatments. There was over-excretion of starch in the first hours of refeeding following food deprivation.     

Effects of beta-glucan obtained from the Chinese herb Astragalus membranaceus and lipopolysaccharide challenge on performance, immunological, adrenal, and somatotropic responses of weanling pigs

A total of 108 crossbred piglets (7.75 +/- 0.24 kg of BW) weaned at 28 d was used to study the interactive effects of beta-glucan obtained from the Chinese herb Astragalus membranaceus (AM) and Escherichia coli lipopolysaccharide (LPS) challenge on performance, immunological, adrenal, and somatotropic responses of weaned pigs. The treatments were in a 2 x 3 factorial arrangement; main effects were level of Astragalus membranaceus glucan (AMG; 0, 500, or 1,000 mg/kg; as-fed basis) and presence of immunological challenge (with or without LPS). The experiment included six replicate pens per treatment and three pigs per pen. Lipopolysaccharide challenges were conducted on d 7 and 21 of the trial. Blood samples were obtained from the vena cava from one pig per pen at 3 h after LPS challenge to determine plasma responses. Weight gain and feed:gain ratio were unaffected by glucan. However, there was a quadratic effect on feed intake (P < 0.05): pigs fed 500 mg of glucan/kg had the highest feed intake. Immunological challenge with LPS decreased weight gain (P = 0.02). An interaction (P = 0.01 to 0.09) between AMG and LPS was observed for glucose, IL-1beta, PGE2, and cortisol. Astragalus membranaceus glucan had a quadratic effect on the plasma concentrations of glucose, IL-1beta, PGE2, and cortisol (P < 0.05) after both LPS challenges. Plasma concentrations of glucose, IL-1beta, PGE2, and cortisol (P < 0.05) were all increased in LPS-challenged pigs compared with the control pigs after both LPS challenges. The IGF-I concentrations were less for LPS-challenged pigs than for unchallenged pigs. The lymphocyte proliferation response of peripheral blood induced by 5 microg of concanavalin A/mL (P < 0.01) and IL-2 bioactivity (P < 0.05) increased linearly with increasing addition of glucan. Pigs challenged with LPS had greater T-lymphocyte proliferation (P = 0.06) and IL-2 bioactivity (P = 0.07) than unchallenged pigs after the first immunological challenge but not after the second. In conclusion, although glucan did not improve pig performance under the conditions of the present experiment, when included at 500 mg/kg, it decreased the release of inflammatory cytokine and corticosteroid and improved the lymphocyte proliferation response of weanling piglets via enhanced IL-2 bioactivity.