Comparative evaluation of virulence and pathology of Streptococcus suis serotypes 2 and 9 in experimentally infected growers

Streptococcus (S.) suis is an invasive porcine pathogen causing meningitis, septicemia, arthritis and other diseases. Studies on pathogenesis as well as vaccine trials have focused on serotype 2 strains, which are worldwide the most prevalent among invasive isolates. However, in Europe serotype 9 strains also contribute substantially to S. suis-associated invasive diseases of piglets. The objective of this study was to determine the virulence of an MRP* SLY+ serotype 9 S. suis strain in comparison to an MRP+ EF+ SLY+ serotype 2 strain. Experimental intranasal and intravenous infections of 7-8 weeks old SPF piglets were investigated with regard to clinic and pathology. In contrast to the virulent serotype 2 strain, the serotype 9 strain did not cause disease with clinical manifestations after intranasal administration. However, histological screenings of these animals revealed pathological lesions, such as mild focal suppurative meningitis. Clinical manifestations related to meningitis, arthritis and serositis could be induced by intravenous application of this serotype 9 strain. Bacteriological culture and immunohistochemistry of the brain confirmed association with the S. suis challenge strains in all cases with clinical manifestations. Interestingly, expression of MRP within meningitis lesions was demonstrated for both pathotypes via immunohistochemistry. In conclusion, this study demonstrates that MRP* SLY+ serotype 9 strains are less virulent for growers than MRP+ EF+ SLY+ serotype 2 strains. Thus, intravenous application of this serotype 9 strain is required to evaluate heterologous protection in the course of vaccine development based on serotype 2 strains in the future.     

Rapid testing leads to the underestimation of the scrapie prevalence in an affected sheep and goat flock

To obtain a more detailed understanding of the prevalence of classical scrapie infections in a heavily affected German sheep flock (composed of 603 sheep and 6 goats), we analysed 169 sheep and 6 goats that carried the genotypes susceptible to the disease and that were therefore culled following discovery of the index case. The initial tests were performed using the Biorad TeSeE ELISA and reactive results were verified by official confirmatory methods (OIE-immunoblot and/or immunohistochemistry (IHC)) to demonstrate the deposition of scrapie-associated PrP(Sc) in the brain stem (obex). This approach led to the discovery of 40 additional subclinically scrapie-infected sheep. Furthermore, peripheral lymphatic and nervous tissue samples of the 129 sheep and 6 goats with a negative CNS result were examined by IHC in order to identify any preclinical infections which had not already spread to the central nervous system (CNS). Using this approach we found 13 additional sheep with PrP(Sc) depositions in the gut-associated lymph nodes (GALT) as well as in the enteric nervous system. Moreover, in most of these cases PrP(Sc) was also deposited in the spleen and in the retropharyngeal and superficial cervical lymph nodes. Taken together, these results show a 30.3% infection prevalence in this scrapie-affected flock. Almost 7.4% of the infected animals harboured PrP(Sc) exclusively in the peripheral lymphatic and nervous tissue and were therefore missed by the currently used testing strategy.     

Effective stiffness prediction of GLT beams based on stiffness distributions of individual lamellas

Wood Science and Technology - The mechanical properties of structural timber—particularly in terms of stiffness and strength—are subject to high variability, which also affects the...     

Microbial reaction in activity, biomass, and community structure after long-term continuous mixing of a grassland soil

Long-term continuous mixing at 40% water holding capacity (WHC) or as slurry at 400% WHC should result in increased soil organic matter decomposition rates in comparison to a control treatment at 40% WHC, but may have strong impacts on soil microbial indices for activity, biomass, and community structure. The amount of extractable inorganic N (NO₃-N+NH₄-N) accumulated in the soil solution after 40 weeks of incubation at 25 °C was 3% of total N in the control treatment and 4% in the two continuous mixing treatments. However, in the treatment mixing at 40% WHC, this 33% increase compared to the control treatment might be explained solely by the decrease in microbial biomass N. In the control treatment, microbial indices decreased in the order microbial biomass C (−10%), microbial biomass N (−40%), ergosterol (−45%) and ATP (−60%). In the treatment mixing at 40% WHC, all four microbial biomass indices were significantly lower than the respective index in the control treatment. This was especially true for microbial biomass N. In the treatment mixing as slurry, only the contents of microbial biomass C and ATP were significantly lower in comparison to the control treatment. The correspondence analysis ordination biplot of the phospholipid fatty acid (PLFA) profiles showed distinct clusters for the three treatments at the end of the incubation. The strongest relative decline of 64% was observed for the fungi-specific PLFA 18:3ω6 in the treatment mixing as slurry in comparison to the control treatment. The content of total bacterial PLFA decreased only by 23%. The differences between the control treatment and the treatment mixing at 40% WHC were less apparent. Fungi represent on average 21% of total microbial biomass C at the end of the incubation if the ergosterol content is recalculated into fungal biomass C. In accordance with this percentage, 22% of the group-specific PLFA could be attributed to fungi.     

Microbial decomposition of fuel oil after compost addition to soil

The effects of adding straw and bark compost at different nitrogen levels to fuel oil contaminated soil was investigated using the CHCl₃ fumigation-extraction method to measure the soil microbial biomass, The addition of these substrates decreased the percentage of extractable fuel oil, increased the percentage of fuel oil mineralized to CO₂, but also increased the fraction of unrecovered fuel oil residues. The different N levels had no significant effects on fuel oil decomposition in the presence of the compost substrates. The addition of the compost substrates increased markedly the content of microbial biomass in contrast to the fuel oil addition which had no effect on the biomass level indicating toxic properties.     

FTIR‐spectroscopic characterization of humic acids and humin fractions obtained by advanced NaOH,Na4P2O7, and Na2CO3 extraction procedures

Aim of our study was the development of the methodological basis for the characterization of humic fractions of a long‐term field experiment. Humic acids (HAs) were extracted from three layers of a nontilled soil using three different extractants (1 M NaOH, 0.1 M Na₄P₂O₇, 1 M Na₂CO₃), and the humin fraction was enriched. NaOH as extractant for FTIR analysis of humic substances yields higher resolved IR spectra, especially in the important regions of stretching vibrations including aromatic and aliphatic groups and in the fingerprint area including amides, aliphats, and aromats than the other extractants. The NaOH extraction has lower extraction yields as compared to Na₄P₂O₇ and Na₂CO₃ and represents a different part of the soil organic matter (SOM). This is reflected by lower C : N ratios and higher E₄ : E₆ and fulvic acid–to–humic acid ratios as compared to the other extractants. The FTIR band areas of HA fraction obtained by NaOH showed an increase of the aromatic and carbonyl groups and a decrease of amide groups with increasing soil depth. Aliphatic groups showed contradicting results: The bands of the stretching vibrations increased, and the band of the bending vibrations decreased. We assume that band interactions in the bending vibrations were responsible for that phenomenon under the assumption of an increase of aliphatic groups with increasing soil depth. The IR bands of the enriched humin fraction showed a decreasing trend in case of both aliphatic bands deriving from stretching vibrations and an increase of aromatic characteristics with depth. Our study led to the conclusion that HA fractions obtained by 1 M NaOH represent a small and dynamic fraction indicated by the measured yields in combination with values of N_t, C : N, E₄ : E₆ ratios, and ratios of fulvic acids (FA) to HA. The humin fraction has a high contribution to the total organic C and represents a more stabilized fraction of SOM which still shows changes in its aromatic and aliphatic characteristics with soil depth.     

Adenylates as an estimate of microbial biomass C in different soil groups

Adenylate (i.e. adenosine tri- (ATP), di- (ADP) and monophosphates (AMP)) and microbial biomass C data were collected over a wide range of sites including forest floor layers and forest, grassland and arable soils. Microbial biomass C was measured by fumigation extraction and adenylates after alkaline Na₃PO₄/DMSO/EDTA extraction and HPLC detection. Our aims were (1) to test whether the sum of adenylates is a better estimate for microbial biomass than the determination of ATP, (2) to compare our conversion values with those proposed by others, and (3) to analyse whether soil properties or land use form affect the relationships between ATP, adenylates and microbial biomass C. A close relationship was found between microbial biomass C and ATP (r=0.96), but also with the sum of adenylates (r=0.96) within all appropriately conditioned soil samples (n=112). In the mineral soil (n=98), the geometric means of the ATP-to-microbial biomass C ratio and the adenylates-to-microbial biomass C ratio were 7.4 and 11.4 μmol g⁻¹, respectively. The mean ratios did not differ significantly between the different texture classes and land use forms. In the forest floor, the ATP-to-microbial biomass C ratio and the adenylates-to-microbial biomass C ratio were both roughly two-thirds of those of the mineral soil. The average adenylate energy charge (AEC) of all soil samples was 0.79 and showed a strong negative relationship with the soil pH (r=−0.69). However, the AEC is presumably only indirectly affected by the soil pH.     

Compost and P amendments for stimulating microorganisms and maize growth in a saline soil from Pakistan in comparison with a nonsaline soil from Germany

A 92 d greenhouse pot experiment with maize (Zea mays L.) was carried out with a strongly saline soil from Pakistan (P‐s) in comparison with a nonsaline soil from Germany (G‐s) similar in pH and texture. The aim was to evaluate salinity effects on the decomposition of compost and effects of compost and P amendments on (1) plant growth and (2) microbial‐biomass formation. The yield of maize shoot‐C and root‐C increased in both soils in the order nonamended control < +triple superphosphate (TSP) (A1) < +compost (A2) < +(compost + TSP) (A3) < +TSP‐enriched compost (A4). In comparison with the control, the highest yield in treatment A4 was nearly doubled on the G‐s, but was increased more than 8‐fold on the saline P‐s. Averaging the three compost treatments, 32% of the compost added was decomposed in the German soil and 36% in the Pakistani soil on the basis of the compost recovered as particulate organic matter. These data were roughly in agreement with the CO₂‐evolution data. This indicates that the decomposition of compost was not affected by salinity. Compost‐derived CO₂ was mainly evolved until day 32, the root‐derived CO₂ from day 74 until the end of the experiment. The addition of compost resulted in higher contents of microbial biomass C and biomass P, but also in that of NaHCO₃‐extractable P. These three properties were significantly interrelated (r = 0.64–0.85), but on a lower level of significance than the relationships between shoot‐C, root‐C, and NaHCO₃‐extractable P (r = 0.90–0.93). Applying compost enriched with TSP (incubation of compost and TSP for 24 h) provided considerably more P to plants and microorganisms than the separate addition of these two components. The results suggest that the role of the microbial biomass as a sink and source for available P deserves further attention.     

Application of multielement stable isotope ratio analysis to the characterization of French, italian, and spanish cheeses

The stable isotope ratios (delta13C, delta15N, and delta34S of casein and delta13C and delta18O of glycerol) measured by IRMS of French, Italian, and Spanish cheeses are presented and discussed. Variability factors such as animal-feeding regimen, geographical origin, and climatic and seasonal conditions were studied to check the possibilities of cheese characterization offered by each isotopic parameter. Delta13C values of both casein and glycerol appeared to be strongly correlated to the amount of maize in the animal diet. Delta15N and delta34S of casein proved to be mostly influenced by the geoclimatic conditions of the area (aridity, closeness to the sea, altitude). Delta18O of glycerol was more dependent on the geographical origin of the cheeses and on climatic/seasonal parameters. By applying a multivariate stepwise canonical discriminant analysis, good discrimination possibilities for the different European cheeses were obtained, confirmed by the classification analysis, when >90% of the samples were correctly reclassified.     

Soil protozoa and forest tree growth: non-nutritional effects and interaction with mycorrhizae

Mycorrhizal (Lactarius rufus Fr.) and non-mycorrhizal Norway spruce seedlings (Picea abies Karst.) were grown in a sand culture and inoculated with protozoa (naked amoebae and flagellates) extracted from native forest soil or with protozoa grown on agar cultures. A soil suspension from which the protozoa were eliminated by filtration or chloroform fumigation was used as a control. After 19 weeks of growth in a climate chamber at 20–22°C, the seedlings were harvested. Protozoa reduced the number of bacterial colony-forming units extracted from the rhizoplane of both non-mycorrhizal and mycorrhizal seedlings and significantly increased seedling growth. However, concentrations of mineral nutrients in needles were not increased in seedlings with protozoan treatment. It is concluded that the increased growth of seedling was not caused by nutrients released during amoebal grazing on rhizosphere micro-organisms. The protozoa presumably affected plant physiological processes, either directly, via production of phytohormones, or indirectly, via modification of the structure and performance of the rhizosphere microflora and their impact on plant growth. Mycorrhizal colonization significantly increased the abundance of naked amoebae at the rhizoplane. Our observations indicate that protozoa in the rhizosphere interact significantly with mycorrhizae.