Development and characterization of mouse monoclonal antibodies reactive with chicken CD80

This study was carried out to develop and characterize mouse monoclonal antibodies (mAbs) against chicken CD80 (chCD80). A recombinant plasmid containing a chCD80/horse IgG4 fusion gene was constructed and expressed in CHO cells to produce recombinant chCD80/IgG4 protein. Chicken CD80 was purified from the chCD80/IgG4 fusion protein following enterokinase digestion, and used to immunize BALB/c mice, resulting in 158 hybridomas that produced mAbs against chCD80. Three mAbs with high binding specificity for recombinant chCD80/IgG4-transfected CHO cells were identified by flow cytometry, and one of these (#112) was selected for further characterization. Immunoprecipitation of CD80/IgG4-CHO cell extract, or lipopolysaccharide (LPS)-treated monocytes identified 35.0 kDa proteins. Immunohistochemical analysis revealed chCD80-expressing cells exclusively in the bursal follicles at the outer portion of the cortex, and throughout the red pulp and the outer boundary of the white pulp in the spleen. By immunofluorescence microscopy, chCD80 was observed on intestinal dendritic cells. LPS treatment of bursa or spleen monocytes for 24 or 48 h increased chCD80 expression. Finally, addition of chCD80 mAb to Con A-stimulated spleen cells inhibited the expression of major histocompatibility complex class II antigens and IL-2-driven proliferation of lymphoblast cells. In summary, these chCD80 mAbs will serve as valuable immunological reagents for basic and applied poultry immunology research.     

Development and characterization of mouse monoclonal antibodies reactive with chicken interleukin-2 receptor αlpha chain (CD25)

This study was carried out to develop and characterize mouse monoclonal antibodies (mAbs) against chicken CD25 (chCD25), the alpha chain of the interleukin-2 (IL-2) receptor. A recombinant chimeric chCD25/IgG4 fusion protein was expressed in Chinese hamster ovary (CHO) cells and isolated from spent cell culture medium by protein G affinity chromatography. Purified chCD25 protein was used to immunize mice, from which 54 stable hybridomas secreting chCD25 mAbs were produced. Two mAbs, chCD25-32 and chCD25-54, with high binding affinity for chCD25-expressing CHO cells were selected for further characterization. By flow cytometry, both mAbs detected cells in the spleen, bursa of Fabricius, intestinal duodenum, and immunostained established chicken T cell, B cell, and macrophage cell lines. Both mAbs reacted with a 55 kDa protein on Western blots of lysates from concanavalin A (Con A)-stimulated spleen mononuclear cells. Intraperitoneal injection of chickens with bacterial lipopolysaccharide increased the percentage of chCD25(+) spleen cells by approximately 4-fold compared with untreated animals. In vitro stimulation of spleen cells with Con A increased the percentage of chCD25(+) cells by up to 50-fold compared with cells treated with medium alone. Finally, the chCD25-32 mAb suppressed IL-2-driven spleen cell proliferation and reduced IL-2-induced nitric oxide production. These mAbs may be useful for future investigation of chicken regulatory T cells.     

Genetic characterization of H7N2 influenza virus isolated from pigs

Because pigs have respiratory epitheliums which express both α2-3 and α2-6 linked sialic acid as receptors to influenza A viruses, they are regarded as mixing vessel for the generation of pandemic influenza viruses through genetic reassortment. A H7N2 influenza virus (A/swine/KU/16/2001) was isolated from pig lungs collected from the slaughterhouse. All eight genes of the influenza virus were sequenced and phylogenetic analysis indicated that A/swine/KU/16/2001 originated in Hong Kong and genetic reassortment had occurred between the avian H7N2 and H5N3 influenza viruses. The first isolation of H7 influenza virus in pigs provides the opportunity for genetic reassortment of influenza viruses with pandemic potential and emphasizes the importance of surveillance for atypical swine influenza viruses.     

Creation of metal-independent hyperthermophilic L-arabinose isomerase by homologous recombination

Hyperthermophilic L-arabinose isomerases (AIs) are useful in the commercial production of D-tagatose as a low-calorie bulk sweetener. Their catalysis and thermostability are highly dependent on metals, which is a major drawback in food applications. To study the role of metal ions in the thermostability and catalysis of hyperthermophilic AI, four enzyme chimeras were generated by PCR-based hybridization to replace the variable N- and C-terminal regions of hyperthermophilic Thermotoga maritima AI (TMAI) and thermophilic Geobacillus stearothermophilus AI (GSAI) with those of the homologous mesophilic Bacillus halodurans AI (BHAI). Unlike Mn(2+)-dependent TMAI, the GSAI- and TMAI-based hybrids with the 72 C-terminal residues of BHAI were not metal-dependent for catalytic activity. By contrast, the catalytic activities of the TMAI- and GSAI-based hybrids containing the N-terminus (residues 1-89) of BHAI were significantly enhanced by metals, but their thermostabilities were poor even in the presence of Mn(2+), indicating that the effects of metals on catalysis and thermostability involve different structural regions. Moreover, in contrast to the C-terminal truncate (Δ20 residues) of GSAI, the N-terminal truncate (Δ7 residues) exhibited no activity due to loss of its native structure. The data thus strongly suggest that the metal dependence of the catalysis and thermostability of hyperthermophilic AIs evolved separately to optimize their activity and thermostability at elevated temperatures. This may provide effective target regions for engineering, thereby meeting industrial demands for the production of d-tagatose.     

对妊娠母猪应用的理想蛋白质模式及其对氨基酸的需要量

由于蛋白质摄入量受限而用于胎儿组织（McPherson等,2004）和乳腺实质组织（Kim等,1999;Ji等,2006）生长以及猪乳合成（Revell等,1998;Jones和Stahly,1999）所需的营养增加,导致妊娠后期和哺乳阶段的母猪处于营养分解状态。母体处于分解状态对胎儿和新生仔猪的生长不利,也增加了它们的患病率和死亡率（Wu等,2006）。在蛋白质摄入量受限的情况下,尤其是在妊娠期和哺乳期,向母猪提供获得最大利用效率的理想平衡氨基酸是重要的。     

Effects of corn dried distiller's grains with solubles and enzyme premix supplements on growth performance,carcass characteristics and meat quality parameters in finishing pigs

The objective of the present study was to investigate the effects of corn dried distiller's grains with solubles (DDGS) and enzyme premix (mannanase + phytase) supplementation on the growth performance, carcass and meat quality parameters in finishing pigs. Sixty hybrid pigs (L × LW × D) with initial weight of 63.92 ± 1.50 kg were used in a 3 × 2 factorial design with main effects of DDGS levels (0, 10 and 20%) and enzyme premix levels (0% vs. 0.14%). Average daily gain (ADG, P < 0.01) and average daily feed intake (ADFI, P < 0.05) were decreased due to an increased level of DDGS additive while the feed conversion ratio was improved (P < 0.05) by adding enzyme premix. The diet cost/gain (won/kg) was saved (P < 0.01) due to an increased level of DDGS additive. There were no significant differences in carcass characteristics and meat quality parameters of Longissimus dorsi muscle by DDGS level and enzyme premix. Palmitoleic acid, oleic acid and monounsaturated fatty acid (MUFA) decreased (P < 0.05) according to DDGS level. The results indicate that DDGS may be used in feeds for finishing pig as a replacement of corn and soybean meal without affecting their carcass characteristics and meat quality.     

Biocontrol of mildew with Bacillus subtilis in bitter gourd (Momordica charantia L.) seeds during germination

The bitter gourd seed has a thick, hard seed coat. Mildew often occurs during germination and causes uneven and low rates of seed germination. However, the problems caused by mildew can be overcome by treating seeds with hot water, by soaking in water, or by using microorganisms. Seeds of the ‘Ching Pi’ bitter gourd were treated in a water bath at 60 °C for 10 min and then soaked in tap water at 25 °C for 24 h. The resulting germination percentage was 86.7%, and the resulting percentage of mildewed seeds was 10%. The biocontrol potential of three commercially available Bacillus subtilis solutions was examined. For seeds primed with Huodijun B. subtilis solution, the germination percentage was 73.3% and the mildewed percentage 6%. In dual cultures, the antibiotic content in the Huodijun B. subtilis solution was significantly greater than in Yunghsing and Huolibao, the other B. subtilis solutions examined. B. subtilis effectively inhibited the growth of Rhizoctonia solani mycelium and caused abnormal mycelial growth.     

The effect of leaf water status on stomatal activity,transpiration and nitrate reductase of sweet potato

The diffusive resistance, transpiration, and nitrate reductase activity were examined in two field grown sweet potato (Convolvulus batatas L.) genotypes. A relationship between leaf water potential and activity of each measured physiological parameter was established.Slight increases in leaf diffusive resistance were observed as leaf water potential decreased, although complete closure of stomata due to water stress was not observed. No genotypic difference was ascertained in the relationship between leaf water potential and diffusive resistance. The stomatal response to humidity deficit is genotype-dependent, and gives a variation in water transport efficiency.The activity of leaf nitrate reductase was adversely affected by leaf water potential. Distinct genotypic differences were observed in the nitrate reductase activity. The data indicated that the stomata of field grown sweet potato are relatively insensitive to water stress, although the leaf nitrate reductase activity is influenced by it. Therefore, measurement of leaf diffusive resistance may not be a reliable estimate of plant water status and related physiological activity.