Characterization of the in vitro core surface proteome of Mycoplasma mycoides subsp. mycoides, the causative agent of contagious bovine pleuropneumonia

Contagious bovine pleuropneumonia (CBPP), caused by Mycoplasma mycoides subsp. mycoides (Mmm) is a severe cattle disease, present in many countries in sub-Saharan Africa. The development of improved diagnostic tests and vaccines for CBPP control remains a research priority. Polyacrylamide gel electrophoresis and mass spectrometry were used to characterize the Triton X-114 soluble proteome of nine Mmm strains isolated from Europe or Africa. Of a total of 250 proteins detected, 67 were present in all strains investigated. Of these, 44 were predicted to be lipoproteins or cytoplasmic membrane-associated proteins and are thus likely to be members of the core in vitro surface membrane-associated proteome of Mmm. Moreover, the presence of all identified proteins in other ruminant Mycoplasma pathogens were investigated. Two proteins of the core proteome were identified only in other cattle pathogens of the genus Mycoplasma pointing towards a role in host–pathogen interactions. The data generated will facilitate the identification and prioritization of candidate Mycoplasma antigens for improved control measures, as it is likely that surface-exposed membrane proteins will include those that are involved in host–pathogen interactions.     

Late watergrass (Echinochloa phyllopogon): mechanisms involved in the resistance to fenoxaprop-p-ethyl

Fenoxaprop-p-ethyl (FE), 2-[4-[(6-chloro-2-benzoxazolyl)oxy]phenoxy] propanoate, ethyl ester (R), is an aryloxyphenoxypropionate herbicide for postemergence control of annual and perennial grasses in paddy fields; its site of action is acetyl-coenzyme A carboxylase (ACCase), an enzyme in fatty acids biosynthesis. The possible mechanism(s) of resistance to FE in a resistant biotype of Echinochloa phyllopogon was examined, namely, absorption, translocation, and metabolism of FE and ACCase susceptibility to fenoxaprop acid (FA). Studies of the in vitro inhibition of ACCase discounted any differential active site sensitivity as the basis of resistance to FE. There were differences in absorption rates between biotypes from 3 to 48 h after application (HAA). Biotypes did not differ in either the amounts or the rates of FE translocated; 98% of applied [14C]FE remaining in the treated leaf. However, there was a good correlation between the rate of herbicide metabolism and the plant resistance. The R biotype produced 5-fold less FA and approximately 2-fold more nontoxic (polar) metabolites 48 HAA than the S biotype. Moreover, the higher rate of GSH conjugation in the resistant biotype as compared to the susceptible one indicates that GSH and cysteine conjugation is the major mechanism of resistance of the R biotype against FE toxicity.     

Novel ferrihydrite sand coating process as a first step for designed technosols

Purpose

The production of technosols to remediate polluted or sealed urban soils to sustain new green areas is mainly empirical. For this, our research aims to contribute with the scientific knowledge base for purpose designing of technosols. Since iron minerals play an important role for many different functions of soils, we simplified a technique to incorporate and stabilize iron minerals in a substrate: a sand coated with an amorphous iron (hydr)oxide, a 2-line ferrihydrite (2L-FH).

Materials and methods

The 2L-FH was precipitated by neutralization of a concentrated FeCl₃ solution. The suspension was homogeneously mixed with the sand and the mixture was dried at 35 °C. The mechanical stability of the 21 2L-FH-coated sand was determined by shaking the aggregates in water for 0, 1, 10, 100, and 1 000 min. The degree of coating detachment and the properties of the coating after shaking were characterized through (a) Fe content, (b) zeta-potential and particle size of the detached particles, (c) the specific surface area (SSA) of the coated sand, and (d) its surface structure using scanning electron microscopy (SEM). A phosphate adsorption isotherm was performed to measure the P-sorption capacity of the shaken samples and to test the 2L-FH-quartz attachment stability against the surface charge reduction of the 2L-FH associated with P adsorption.

Results and discussion

A reduced Fe loss (30 %) and smaller sizes of the coating detached particles in the sample shaken for 1 000 min indicate that a fractioning and reattachment of these aggregates occurred during the agitation process, resulting in a smoother surface (SEM), and a larger SSA and P-sorption capacity. The coated shaken samples showed P-adsorption capacities (5.3–6.34 μmol P g^?1) comparable to high loadings of phosphate in soils, and low detachment of Fe (7–14 %) in spite of negativity surface charge increase.

Conclusions

The practical novel coating process along with the 1 000-min shaking produced a mechanical resistant and P-adsorptive effective coated sand that could sustain the needs of plants in further experiments.

     

Plant‐compositional effects on surface runoff and sediment yield in subalpine grassland

Soil erosion such as sheet erosion is frequently encountered in subalpine grassland in the Urseren Valley (Swiss Central Alps). Erosion damages have increased enormously in this region during the last 50 y, most likely due to changes in land‐use practices and due to the impact of climatic changes. In order to estimate the effect of vegetation characteristics on surface runoff and sediment loss, we irrigated 22 pasture plots of 1 m² during 1 h at an intense rain rate of 50 mm h^–1 in two field campaigns using a portable rain simulator. The rain‐simulation plots differed in plant composition (herb versus grass dominance) and land‐use intensity but not in plant cover (>90%) nor in soil conditions. Prior to the second rain‐simulation campaign, aboveground vegetation was clipped in order to simulate intense grazing. The generated surface runoffs, sediment loss, relative water retention in the aboveground vegetation, and changes in soil moisture were quantified. Runoff coefficient varied between 0.1% and 25%, and sediment loss ranged between 0 and 0.053 g m^–2. Thus, high infiltration rates and full vegetation cover resulted in very low erosion rates even under such extreme rain events. Surface runoff did not differ significantly between herb‐ and grass‐dominated plots. However, clipping had a notable effect on surface runoff in the test plots under different land‐use intensity. In plots without or with intensive use, surface runoff decreased after clipping whereas in extensively used plots, surface runoff increased after the clipping. This opposite effect was mainly explained by higher necromass and litter presence at the extensively used plots after the clipping treatment. The results obtained here contribute to a better understanding of the importance of vegetation characteristics on surface‐runoff formation, thus, on soil‐erosion control. Overall, we delineate vegetation parameters to be crucial in soil‐erosion control which are directly modified by the land‐use management.     

Effects of olive oil mill wastewater on chemical,microbiological, and physical properties of soil incubated under four different climatic conditions

The objective of this study was to understand the degradation of the organic matter of olive mill wastewater (OMW) and its phytotoxic and water repellent effects in dependence on four different climatic conditions. We hypothesized that warm conditions with sufficient soil moisture ensure optimal biological activity and thus minimize negative effects of the OMW treatment. Therefore, OMW-treated soil was incubated for 60 days under four climatic conditions. During incubation, we monitored pH, contents of nitrate, manganese and phenolic compounds, soil respiration, soil water repellency, and δ¹³C. Additionally, calorific value and thermal stability of the soil organic matter at the beginning and end of incubation were determined. Soil samples of the wet-cold and moist-warm incubation were tested for phytotoxicity using a seed germination bioassay with Lepidium sativum. As a function of climatic conditions, positive and negative effects, e.g., addition of nutrients, phytotoxicity, and soil water repellency, were observed. Under dry-hot conditions, the soil was still water repellent after 60 days of incubation whereas the wet-hot, moist-warm, and wet-cold incubation show that soil would stay wettable if soil moisture before OMW treatment would be sufficient. Thus, the impact of OMW treatment on soil quality strongly depends on the environmental conditions which should favor an enhancement of microbial activity to minimize negative effects.     

Removal of Heavy Metals from Soil Components and Soils by Natural Chelating Agents. Part II. Soil Extraction by Sugar Acids

Aqueous solutions of the natural chelatingagents D-gluconic acid and D-glucaric acid (D[+]-saccharic acid) were tested for their ability to remove heavy metals (Cd, Cr, Cu,Ni, Pb, Zn) from a soil polluted by long-term application of sewage sludge. Batch equilibrium experiments were performed undervariation of fundamental process parameters, i.e. pH value, sugaracid concentration, batch solution volume, solid:liquid ratioand number of treatment cycles.The extractability of heavy metals was low under near-neutral andslightly basic pH conditions. It increased drastically between pH12.0 and 13.0. Pb and Cu were preferentially extracted metals.Compared with the extraction efficiency of pH adequate puresodium hydroxide solutions, the sugar acids enhanced thesolubilisation of Pb and Cr especially. The metal depletion fromsoil was the highest when applying 20 or 50 g L^-1 solutionsof the chelating agents. Under strongly basic conditions solid:liquid ratios of 1:10 or 1:20 were proofed to be advantageous.Except Ni, multi-step extraction improved the metal removalstrongly. This effect was the greatest for Cr extraction. Underoptimised conditions the following metal extraction degrees wereachieved with strongly alkaline D-gluconic acid solutions: Ni 43%%, Cr 60%%, Cd 63%%,Zn 70%%, Pb 80%%, and Cu 84%%.     

Homogeneous inoculation vs. microbial hot spots of isolated strain and microbial community: What is the most promising approach in remediating 1,2,4-TCB contaminated soils?

We investigated the effectiveness of different inoculation approaches in enhancing the mineralization of [U-¹⁴C] labeled 1,2,4-trichlorobenzene (1,2,4-TCB) in soil. Inoculation was conducted with a soil-borne 1,2,4-TCB mineralizing microbial community (MC) as well as the Bordetella sp. strain F2 originally isolated from this community as the key degrader organism (IS). Both were applied either via liquid medium (LM) or attached on clay particles (CP). Fluorescence in-situ hybridization in combination with ¹⁴C-1,2,4-TCB mineralization measurements as well as measurements of ¹⁴C-residues in soil were used to investigate the bioaugmentation efficiency of the different approaches. Bordetella sp. cell numbers increased about 2-5 times during the incubation process, indicating that the bacteria could survive and develop in the new soil habitat. While the native soil showed negligible 1,2,4-TCB mineralization rates, soil inoculated with the MC attached on CP showed the highest 1,2,4-TCB mineralization rate per Bordetella cell, whereas the other inoculum approaches showed an increased but lower contaminant mineralization. Additionally, the MC-CP approach showed the highest cumulative 1,2,4-TCB mineralization as well as the highest formation of bound ¹⁴C-residues which is most likely equivalent to ¹⁴C incorporated into the microbial biomass. Thus, our results allow the conclusion that the application of a specific microbe-clay-particle-complex is the most promising approach for an accelerated in-situ mineralization of chemicals in agricultural soils.     

FEASIBILITY FOR ULTRASOUND‐GUIDED INJECTION OF THE COLLATERAL LIGAMENTS OF THE DISTAL INTERPHALANGEAL JOINT IN HORSES

Desmitis of the collateral ligament of the distal interphalangeal joint is a cause of lameness in performance horses. The objective of this prospective, experimental, ex vivo feasibility study was to evaluate the success of ultrasound‐guided injection of the collateral ligaments of the distal interphalangeal joint in the equine forelimb. Seventy‐six ultrasound‐guided dye injections of the collateral ligament of the distal interphalangeal joint were performed on horses’ cadaver limbs. The hooves were sectioned transversely to verify the location of the dye relative to the collateral ligaments and surrounding structures. Evaluations of transverse sections were performed independently by two experienced observers. A scoring system was used to assess injection of the collateral ligament of the distal interphalangeal joint at the proximal, middle, and distal aspect over the length of the ligament. The collateral ligament was injected at any point in 97.4% of cases. The ligament was injected over the entire scored length in 43.2% of cases (32/74), over two scored length areas in 45.9% of cases (34/74), and in one area in 10.8% of cases (8/74). The distal interphalangeal joint and the common digital extensor tendon were also injected in 81.6% (62/76) and 43.4% (33/76) of the cases, respectively. Use of the ultrasound had a positive and negative predictive value of 98% and 9%, respectively. In this study, ultrasound guidance was useful for confirming injection of the collateral ligament of the distal interphalangeal joint but did not prevent injecting the distal interphalangeal joint and the common digital extensor tendon.