Slope stability considering multi-fissure seepage under rainfall conditions.

Fissures form the channel for rainwater infiltration, which accelerate the infiltration of rainwater into slope bodies, hence its important impact on the seepage field and stability of the slope. In this paper, taking one landslide of Liang-Wan freeway as the research object, firstly, the equivalent permeability coefficient method is used to homogenize the fissured soil. Then considering the boundary conditions of rainfall infiltration and groundwater level, a fluid-structure coupling model is established based on saturated-unsaturated seepage theory, and evolution characteristics of seepage, displacement and stress of the slope are studied. Based on these, the slope stability coefficient is determined. The results show that the rising rate of pore water pressure and volume water content of topsoil increases when multi-fissure seepage is considered, and the pore water velocity is larger in the local seepage range of fissures. With the increase of buried depth, the closer to groundwater level, the influence of multi-fissure seepage gradually weakens. The theoretical calculation results of slope displacement are more consistent with the field monitoring results. With the increase of rainfall time, the stability coefficient of slope decreases gradually, and the rate and range of decrease are greater.

Metalloproteases secreted by Actinobacillus suis.

Actinobacillus suis secretes metalloproteases into its medium. These secreted proteins, when concentrated by precipitation with 70% (NH4)2SO4 or methanol, displayed proteolytic activity at >200 kDa molecular mass bands in 10% polyacrylamide gels copolymerized with bovine casein (1%). They showed activity in a broad pH range (from pH 5 to pH 10) and were inhibited by 20 mM EDTA or EGTA, but could be reactivated by calcium. They were found heat stable at 40 degrees C, 50 degrees C, 60 degrees C, and 70 degrees C, but their activity diminished at 80 degrees C or higher. They degraded pig and bovine IgG and cross-reacted with a polyclonal serum against a high molecular mass secreted protease from A. pleuropneumoniae. Extracellular proteases could play a role in diseases caused by A. suis.

Adherence of Actinobacillus pleuropneumoniae to swine-lung collagen.

Actinobacillus pleuropneumoniae serotype 1 adhered to immobilized swine-lung collagen. Bacteria bound to collagen type I, III, IV and V. At 5 min incubation, 30 % of bacteria adhered to collagen, reaching saturation in around 90 min. Treatment of bacteria with divalent-metal chelators diminished their attachment to collagen, and Ca(2+) but not Mg(2+) increased it, suggesting Ca(2+) dependence for adherence. Proteolytic enzymes drastically reduced bacterial adherence to collagen, showing that binding involved bacterial surface proteins. Porcine fibrinogen, haemoglobin and gelatin partially reduced collagen adhesion. A 60 kDa outer-membrane protein of A. pleuropneumoniae recognized the swine collagens by overlay. This membrane protein was apparently involved in adhesion to collagen and fibrinogen, but not to fibronectin and laminin. Antibodies against the 60 kDa protein inhibited the adhesion to collagen by 70 %, whereas pig convalescent-phase antibodies inhibited it by only 40 %. Serotypes 1 and 7 were the most adherent to pig collagen (taken as 100 %); serotypes 6 and 11 were the lowest (approximately 50 %), and neither showed the 60 kDa adhesin to biotinylated collagens. By negative staining, cells were observed initially to associate with collagen fibres in a polar manner, and the adhesin was detected on the bacterial surface. The results suggest that swine-lung collagen is an important target for A. pleuropneumoniae colonization and spreading, and that the attachment to this protein could play a relevant role in pathogenesis.

Actinobacillus pleuropneumoniae metalloprotease: cloning and in vivo expression.

The complete amino acid and nucleotide sequence of a secreted metalloprotease produced by Actinobacillus pleuropneumoniae serotype 1 is reported. A clone showing proteolytic activity in cell-free culture media was selected from a genomic library of A. pleuropneumoniae serotype 1 in pUC 19. The sequence obtained contained an open reading frame encoding a protein with 869 amino acids. This protein was identified as a zinc neutral-metalloprotease belonging to the aminopeptidase family, with a predicted molecular weight of approximately 101 kDa. This sequence showed high homology with other predicted or sequenced aminopeptidases reported for different Gram-negative bacteria. Expression of the protease was observed in lung tissue from pigs that died of porcine pleuropneumonia suggesting a role in pathogenesis.

Actinobacillus pleuropneumoniae: virulence and gene cloning

Actinobacillus pleuropneumoniae is the causal agent of porcine contagious pleuropneumonia(PCP). The infection produces important economic losses in porciculture due to its high morbidity and mortality. Survivors are asymptomatic carriers infectious to other pig and have low alimentary conversion. The causative agent possesses several virulence factor: adhesion fimbriae, lipopolysaccharide of the outer membrane, capsule, and cytolysins. In addition, our group has reported secretion proteases of a wide pH range of activity. These proteases degrade different substrates such as porcine gelatin, hemoglobin and IgA, and bovine or human hemoglobin. To control PCP dissemination, farmers require serodiagnostic tests which detect carriers and discriminate between vaccinated and infected animal. Bacterines used as immunogens are serotype specific and do not prevent the infection. Genes have been cloned that codify a cohemolysin, cytolysins, and an iron-binding protein. We have cloned A. pleuropneumoniae genes using the expression plasmids pUC19 and Bluescript, in Escherichia coli Q358 and DH5alpha; the screening for antigen production was made in four gropus of pigs (vaccinated, experimentally infected, naturally infected, and from alaughterhouses); two E. coli clones expressed polypeptides recognized by sera from all the groups