Decreased colonization of chicks by Salmonella enterica serovar Gallinarum expressing mannose-sensitive FimH adhesin from Salmonella enterica serovar Enteritidis

To investigate the role of non-hemagglutinating type 1 fimbriae in the pathogenesis of Salmonella Gallinarum, the isogenic mutant elaborating type 1 fimbriae with mannose-sensitive (MS) variant of the FimH adhesin from Salmonella Enteritidis and the mutant strain with no FimH expression were constructed. Their binding to chicken leukocytes in vitro and invasiveness in 1-day-old chicks were studied. Our results demonstrated that S. Gallinarum type 1 fimbriae with an endogenous variant of the FimH adhesin mediated mannose-resistant (MR) binding to avian leukocytes and did not bind to human epithelial cells. However, after allelic replacement of the FimH, mutated fimbriae with S. Enteritidis variant of the FimH adhesin bound to both cell types in a mannose-dependent manner. In chick model, S. Gallinarum expressing wild-type FimH variant colonized cecal tonsils and bursa of Fabricius more effectively and invaded the spleen and liver in greater numbers than S. Gallinarum fimH knockout strain or mutant expressing MS FimH variant from S. Enteritidis. The invasive potential of the latter was greatly reduced in chicks since no viable bacteria expressing MS variant of the adhesin could be recovered from intestinal lymphoid tissues or liver over a 6 days course of infection. Together, these results demonstrate that the S. Gallinarum type 1 fimbriae with the endogenous MR variant of the FimH protein increase systemic dissemination of S. Gallinarum and colonization of internal organs in chicks indicating the importance of these adhesive structures in the virulence of S. Gallinarum.     

Competitive interactions for food resources between the invasive Amur sleeper (Perccottus glenii) and threatened European mudminnow (Umbra krameri)

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Effect of tillage and crop management on runoff,soil erosion and organic carbon loss

Proper management of soil organic matter is an important issue in the context of sustainable agriculture. The intensification of production and the loss of organic carbon associated with agriculture reduce the efficiency of production and the quality of the environment, especially in relation to areas exposed to erosion. The aim of this study was to determine the impact of specific tillage systems and plant cover on the organic carbon losses, as well as on runoff and soil losses, over a 6-year study period following the introduction of no-till. The first factor in the experiment was the tillage system: conventional tillage (CT) and no-till (NT). The second factor was plant cover: horse bean, spring wheat and winter oilseed rape. The results showed that runoff was 4.3 ± 0.6% higher under NT than under CT, while soil loss was 66.8 ± 2.7% lower under NT than under CT. Compared to CT, NT limited the total organic carbon losses by an average of 46.0 ± 2.9% and organic carbon bound with sediment losses by 53.2 ± 0.7%, whereas for dissolved organic carbon, there were no significant differences for the tillage systems. The anti-erosion effectiveness of NT was lower in the first year, but it increased in subsequent years after the introduction of this tillage system. Plant cover also had a significant impact on organic carbon losses and soil protection. The crops were ranked according to runoff, soil losses and organic carbon losses in the following order from lower to higher losses: winter oilseed rape > spring wheat > horse bean.     

Differentiation of Salmonella Gallinarum biovar Gallinarum from Salmonella Gallinarum biovar Pullorum by PCR‐RFLP of the fimH gene

In our studies on FimH adhesins expressed by different Salmonella serovars, we cloned and sequenced the fimH genes from Salmonella enterica ssp. Enterica ser. Gallinarum biovar Gallinarum and S. enterica ssp. Enterica ser. Gallinarum biovar Pullorum. Comparison of the nucleotide sequences revealed the presence of a single‐nucleotide polymorphism (SNP) at position 544 bp from the A of the start codon of the fimH open reading frame (ORF). Further analysis of the restriction enzyme sites in fimH gene showed that the SNP at this position is responsible for a sequence specifically recognized by SacI in S. Gallinarum biovar Gallinarum only, making it possible to differentiate both biovars with the use of polymerase chain reaction‐restriction fragment length polymorphism (PCR‐RFLP). Digestion of PCR amplicons of the fimH gene from S. Gallinarum biovar Gallinarum strains with SacI gave two DNA fragments of 554 and 472 bp and only one fragment of 1026 bp for S. Gallinarum biovar Pullorum. This allows a clear differentiation between these two biovars.     

Bovine herpesvirus type 1 infection of bovine bronchial epithelial cells increases neutrophil adhesion and activation

Respiratory infection of cattle with bovine herpesvirus type 1 (BHV-1) predisposes cattle to secondary pneumonia with Mannheimia haemolytica as part of the bovine respiratory disease complex (BRD). One cell type that has received limited investigation for its role in the inflammation that accompanies BRD is the respiratory epithelial cell. In the present study we investigated mechanisms by which BHV-1 infection of respiratory epithelial cells contributes to the recruitment and activation of bovine polymorphonuclear neutrophils (PMNs) in vitro. Primary cultures of bovine bronchial epithelial (BBE) cells were infected with BHV-1 and assessed for cytokine expression by real-time PCR. We found that BHV-1 infection elicits a rapid IL-1, IL-8 and TNF-α mRNA response by BBE cells. Bovine PMNs exhibited greater adherence to BHV-1 infected BBE cells than uninfected cells. The increased adherence was significantly reduced by the addition of an anti-IL-1β antibody or human soluble TNF-α receptor (sTNF-αR). Pre-incubation of bovine PMNs with conditioned media from BHV-1 infected BBE cells increased PMN migration, which was inhibited by addition of an anti-IL-1β antibody, sTNF-αR, or an IL-8 peptide inhibitor. Conditioned media from BHV-1 infected BBE cells activated bovine PMNs in vitro as demonstrated by PMN shape change, production of reactive oxygen species and degranulation. PMNs also exhibited increased LFA-1 expression and susceptibility to M. haemolytica LKT following incubation with BHV-1 infected BBE cell conditioned media. Our results suggest that BHV-1 infection of BBE cells triggers cytokine expression that contributes to the recruitment and activation of neutrophils, and amplifies the detrimental effects of M. haemolytica LKT.     

Differentiation of Salmonella Gallinarum biovar Gallinarum from Salmonella Gallinarum biovar Pullorum by PCR-RFLP of the fimH gene

In our studies on FimH adhesins expressed by different Salmonella serovars, we cloned and sequenced the fimH genes from Salmonella enterica ssp. Enterica ser. Gallinarum biovar Gallinarum and S. enterica ssp. Enterica ser. Gallinarum biovar Pullorum. Comparison of the nucleotide sequences revealed the presence of a single-nucleotide polymorphism (SNP) at position 544 bp from the A of the start codon of the fimH open reading frame (ORF). Further analysis of the restriction enzyme sites in fimH gene showed that the SNP at this position is responsible for a sequence specifically recognized by SacI in S. Gallinarum biovar Gallinarum only, making it possible to differentiate both biovars with the use of polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP). Digestion of PCR amplicons of the fimH gene from S. Gallinarum biovar Gallinarum strains with SacI gave two DNA fragments of 554 and 472 bp and only one fragment of 1026 bp for S. Gallinarum biovar Pullorum. This allows a clear differentiation between these two biovars.     

Wetland macrophyte decomposition under different nutrient conditions: Relationships between decomposition rate, enzyme activities and microbial biomass

We used oligotrophic, P-limited herbaceous wetlands of northern Belize as a model system, on which to document and explain how changes in nutrient content along a salinity gradient affect activities of extracellular enzymes involved in macrophyte decomposition. To determine what is more important for decomposition, the initial litter quality, or site differences, we used reciprocal litter placement in a combined “site quality” and “litter quality” experiment running from August 2003 to April 2004. The experiment was set up in long-term control and nutrient addition plots (P, N, and NP) established in 2001 in 15 limestone-based inland marshes with a wide range of water conductivities (200-6000 μS) and a uniform pH (7.0-7.7) dominated by emergent macrophytes, Eleocharis spp. There were no differences among the plots in total sediment N and water NH₄-N, but total and KCl-extractable sediment P and water PO₄-P were significantly higher in P and NP plots throughout the duration of the experiment. The initial litter N content was slightly but significantly different between control and N plots versus P and NP plots (5.7 and 7.1 mg g⁻¹, respectively). The difference was much bigger for litter P content, 0.1 and 0.7 mg g⁻¹, respectively. Enzyme activities of alkaline phosphatase, leucine-aminopeptidase, arylsulfatase, and β-glucosidase were measured fluorometrically in Eleocharis litter in both the litterbag experiment and the naturally decomposing material. Total phospholipid fatty acid (PLFA) content in litter samples was used as a measure of microbial biomass present. Phosphatase always exhibited the highest activity of the enzymes studied, followed by leucine-aminopeptidase, arylsulfatase and β-glucosidase. There were no significant differences between enzyme activities from litterbags and the unconfined litter. Phosphatase activity was significantly suppressed in P-addition plots under all salinity levels while the activities of the remaining enzymes were significantly higher in P-enriched plots. There was a strong correlation between decomposition coefficient k-values and most of the enzymes as well as between the amount of PLFA and enzyme activities. PLFA, arylsulfatase, and litter C/P were the best predictors of k-values.     

Activity Disequilibrium between 234U and 238U Isotopes in Southern Baltic

The aim of the work was to determine the concentration of ²³⁴U and ²³⁸U and calculate the values of the ²³⁴U/²³⁸U activity ratio in samples of living organisms, bottom water, surface and interstitial water and also sediments from the various regions of southern Baltic Sea. The knowledge of ²³⁴U/²³⁸U activity ratio in ecosystem allows getting know about the mechanisms and processes of uranium transport and origin.The activities of the analyzed uranium radionuclides in samples were measured using alpha spectrometry. The results of researches revealed diversified concentrations of uranium in the sediments of the southern Baltic Sea (sea and coastal waters) and increase of uranium with sediment depth, suggesting the diffusion of uranium from sediments to water through interstitial water and diagenesis processes in sediment material. The nuclides of uranium ²³⁴U and ²³⁸U were radioactive state equilibrium in most of the sediments. The values of the ²³⁴U/²³⁸U activity ratio oscillate around one. In bottom, surface, interstitial water and living organisms mean values of the ²³⁴U/²³⁸U activity ratio are between 1.12 and 1.15. Higher uranium concentration was observed in samples of sediments and seawater after flood in 1997 and torrential rainfalls in 2000–2001. River waters are characterized by higher ²³⁴U/²³⁸U activity ratio. The values of the ²³⁴U/²³⁸U activity ratio equal 1.27 in sediments and 1.34–1.38 in seawater indicate the influence of fresh waters.