Loop-mediated isothermal amplification for the rapid, sensitive, and specific detection of the O9 group of Salmonella in chickens

In the present study, the loop-mediated isothermal amplification (LAMP) assay was developed to amplify the fragments of the O9 Salmonella-specific insertion element and evaluated in the laboratory for its potential use in a field situation, such as poultry farms. Among the bacteria tested, a positive reaction was observed only for 128 strains of 6 serovars of the O9 group Salmonella, such as Enteritidis (SE) and Pullorum. The detection limit of the LAMP assay was 10(3)CFU/ml, which was more sensitive than that of the polymerase chain reaction (PCR) assay with the same target gene (10(6)CFU/ml). The final results were obtained within 30 min for the LAMP assay, while the PCR assay needed a total of 120 min. When the LAMP assay was applied to the enrichment broth mixed with cecal dropping samples either spiked with SE in vitro or excreted by SE-inoculated hens, the results were comparable to those of the conventional plating method including 2 separate enrichments. In conclusion, the LAMP assay developed in the present study is an effective method for the specific detection of the O9 group Salmonella serovars, including SE.     

Concentration of macrophage colony-stimulating factor (M-CSF) in bovine peripheral blood during pregnancy

Macrophage colony-stimulating factor (M-CSF) is a hemopoietic cytokine with a primary role in placental physiology. Gene expression of M-CSF in the bovine endometrium shows a temporal upward trend during early and mid pregnancy. This study determined the plasma M-CSF levels during pregnancy using ELISA. In experiment 1, to investigate the relationship between the concentration of M-CSF in peripheral blood and pregnancy, the plasma M-CSF levels were determined in 125 pregnant and 21 non-pregnant Japanese Black cows. The pregnant animals were divided into nine groups based on the month of pregnancy. An ELISA for bovine M-CSF established previously was used according to the authors' instructions. In experiment 2, the plasma M-CSF level was determined to investigate the temporal changes in its concentration in the peripheral blood during pregnancy. In experiment 1, the plasma M-CSF level varied from month to month during pregnancy; the mean level in the first-month of pregnancy was significantly higher than those in the third and last months of pregnancy and non-pregnancy (P<0.05). In experiment 2, the plasma M-CSF level varied with the day of pregnancy (P<0.05). The mean level of plasma M-CSF decreased gradually until 6 weeks of pregnancy; it appeared to increase during weeks 7-9, then varied with several small peaks until 27 weeks of pregnancy and finally decreased gradually until parturition. These results suggest that the plasma M-CSF level may be related to changes in the uterus and placenta as pregnancy progresses.     

Physiological and immunological responses to weaning and transport in the young pig: modulation by administration of porcine somatotropin

To examine the effects of exogenous porcine (p) ST on measures of stress and immune function in weaned pigs with or without transport, pigs (20 +/- 1 d of age) received daily injections of pST (0.5 mg/kg; n = 16) or saline (n = 16) for 5 d. On d 5, a blood sample was collected immediately before injection. At 4 h postinjection, pigs were weighed, sampled for blood, injected with di-nitrophenyl-conjugated keyhole limpet hemocyanin, and weaned. One half of the pigs in each group were transported for 3 h before placement in the nursery. Pigs were weighed, and blood was collected on 1, 7, and 14 d postweaning. Statistical significance was set at P < 0.05. Serum IGF-I concentrations were increased by pST and decreased by weaning, but not affected by transport. The free cortisol index was elevated in all pigs 1 d postweaning, although less in transported versus nontransported pigs. By 7 d postweaning, the free cortisol index returned to prewean values. Serum concentrations of immunoglobulin (Ig) G increased in all pigs by 14 d postweaning, but were not affected by pST or transport. Serum IgM concentrations were elevated at 7 and 14 d postweaning. Before weaning and again 1 d postweaning, pigs treated with pST had greater concentrations of IgM than did control animals. Circulating neutrophils increased in pST-treated pigs 4 h after the final pST injection. Improved immune function in weaned pigs by pST may lead to greater health and growth in a commercial setting.     

Intraradical hyphae phosphatase of the arbuscular mycorrhizal fungus, Gigaspora margarita

An alkaline phosphatase in the intraradical hyphae of arbuscular mycorrhizal fungi was found to be closely related to an improvement of plant growth. To detect the phosphatase activity in a crude extract of mycorrhizal roots, phosphatase isozymes in mycorrhizal and non-mycorrhizal onion roots were compared with those in Gigaspora margarita by electrophoresis. A mycorrhiza-specific band was found when the phosphatase was stained under alkaline conditions. To clarify the origin of this phosphatase, the phosphatase extracted from intraradical hyphae was also compared with the phosphatase from mycorrhizal roots by electrophoresis. The intraradical hyphae was isolated from mycorrhizal roots by enzyme digestion followed by Percoll gradient centrifugation. The soluble protein was extracted from the hyphae by ultra-sonication after treatment with chitinase. A phosphatase in the hyphal soluble protein showed a similar, but slightly higher, relative mobility on the gel, compared with the mycorrhiza-specific phosphatase from roots. By adding the hyphal extract to the root extract, the relative mobility of the mycorrhiza-specific phosphatase was slightly changed and became identical to that of the phosphatase in the hyphae. This indicated that the specific band of phosphatase found in the crude extract from mycorrhizal roots was of intraradical hyphal origin. Received: 16 April 1997     

Correction to: A new and simple method for measuring in situ field-saturated hydraulic conductivity using a falling-head single cylinder

Paddy and Water Environment - Hydraulic properties of soil play important roles in water and temperature regimes. Measuring hydraulic properties has been studied for decades in the laboratory and...     

MytiLec,a Mussel R-Type Lectin,Interacts with Surface Glycan Gb3 on Burkitt’s Lymphoma Cells to Trigger Apoptosis through Multiple Pathways

MytiLec; a novel lectin isolated from the Mediterranean mussel (Mytilus galloprovincialis); shows strong binding affinity to globotriose (Gb3: Galα1-4Galβ1-4Glc). MytiLec revealed β-trefoil folding as also found in the ricin B-subunit type (R-type) lectin family, although the amino acid sequences were quite different. Classification of R-type lectin family members therefore needs to be based on conformation as well as on primary structure. MytiLec specifically killed Burkitt''s lymphoma Ramos cells, which express Gb3. Fluorescein-labeling assay revealed that MytiLec was incorporated inside the cells. MytiLec treatment of Ramos cells resulted in activation of both classical MAPK/ extracellular signal-regulated kinase and extracellular signal-regulated kinase (MEK-ERK) and stress-activated (p38 kinase and JNK) Mitogen-activated protein kinases (MAPK) pathways. In the cells, MytiLec treatment triggered expression of tumor necrosis factor (TNF)-α (a ligand of death receptor-dependent apoptosis) and activation of mitochondria-controlling caspase-9 (initiator caspase) and caspase-3 (activator caspase). Experiments using the specific MEK inhibitor U0126 showed that MytiLec-induced phosphorylation of the MEK-ERK pathway up-regulated expression of the cyclin-dependent kinase inhibitor p21, leading to cell cycle arrest and TNF-α production. Activation of caspase-3 by MytiLec appeared to be regulated by multiple different pathways. Our findings, taken together, indicate that the novel R-type lectin MytiLec initiates programmed cell death of Burkitt’s lymphoma cells through multiple pathways (MAPK cascade, death receptor signaling; caspase activation) based on interaction of the lectin with Gb3-containing glycosphingolipid-enriched microdomains on the cell surface.     

Elevated atmospheric CO2 concentration alters the effect of phosphate supply on growth of Japanese red pine (Pinus densiflora) seedlings

We demonstrated that the inorganic phosphate (P(i)) requirement for growth of Japanese red pine (Pinus densiflora Sieb. & Zucc.) seedlings is increased by elevated CO(2) concentration ([CO(2)]) and that responses of the ectomycorrhizal fungus Pisolithus tinctorius (Pers.) Coker & Couch to P(i) supply are also altered. To investigate the growth response of non-mycorrhizal seedlings to P(i) supply in elevated [CO(2)], non-mycorrhizal seedlings were grown for 73 days in ambient or elevated [CO(2)] (350 or 700 micromol mol(-1)) with nutrient solutions containing one of seven phosphate concentrations (0, 0.02, 0.04, 0.06, 0.08, 0.10 and 0.20 mM). In ambient [CO(2)], the growth response to P(i) was saturated at about 0.1 mM P(i), whereas in elevated [CO(2)], the growth response to P(i) supply did not saturate, even at the highest P(i) supply (0.2 mM), indicating that the P(i) requirement is higher in elevated [CO(2)] than in ambient [CO(2)]. The increased requirement was due mainly to an altered shoot growth response to P(i) supply. The enhanced P(i) requirement in elevated [CO(2)] was not associated with a change in photosynthetic response to P(i) or a change in leaf phosphorus (P) status. We investigated the effect of P(i) supply (0.04, 0.08 and 0.20 mM) on the ectomycorrhizal fungus P. tinctorius in mycorrhizal seedlings grown in ambient or elevated [CO(2)]. Root ergosterol concentration (an indicator of fungal biomass) decreased with increasing P(i) supply in ambient [CO(2)], but the decrease was far less in elevated [CO(2)]. In ambient [CO(2)] the ratio of extramatrical mycelium to root biomass decreased with increasing P(i) supply but did not change in elevated [CO(2)]. We conclude that, because elevated [CO(2)] increased the P(i) requirement for shoot growth, the significance of the ectomycorrhizal association was also increased in elevated [CO(2)].     

两种功能高分子材料对沙土物理性质影响的比较

采用SEM,透水系数法、加压板法等现代测试手段,观察与分析高分子吸水树脂、固沙植被多功能高分子复合材料表面微观结构形态,比较高分子吸水树脂与固沙植被多功能高分子复合材料改良沙土后沙土的透水、保水等特性及沙土水分的有效利用情况.实验结果证明固沙植被多功能高分子复合材料比高分子吸水树脂改良沙土的效果好,使沙土的气、液、固三相比有利植物的生长,能保持高的水有效利用率.本实验结果为固沙植被多功能高分子复合材料的应用提供了新的依据.