Morphological development and gibberellin production by different strains of Gibberella fujikuroi in shake flasks and bioreactor

Strain H-984 of G. fujikuroi grown for 38h in a shake flask with medium containing 20g glucose l^–1, 3g yeast extract l^–1, 2.5g NH4NO3 l^–1, 0.5g KH2PO4 l^–1, 0.1g MgSO4 l^–1, 1g CaCO₃ l^–1, and inoculated into a bioreactor with medium containing 60g glucose l^–1; 1g NH_4Cl l^–1; 3g KH2PO4 l^–1 and 1.5g MgSO₄ l^–1 produced 1100mg gibberellic acid l–1.     

Vermicompost as a soil supplement to improve growth, yield and fruit quality of tomato (Lycopersicum esculentum)

The effects of earthworm-processed sheep-manure (vermicompost) on the growth, productivity and chemical characteristics of tomatoes (Lycopersicum esculentum) (c.v. Rio Grande) were investigated in a greenhouse experiment. Five treatments were applied combining vermicompost and soil in proportions of 0:1, 1:1, 1:2, 1:3, 1:4 and 1:5 (v/v). Growth and yield parameters were measured 85 days and 100 days after transplanting. Addition of vermicompost increased plant heights significantly, but had no significant effect on the numbers of leaves or yields 85 days after transplanting. Yields of tomatoes were significantly greater when the relationship vermicompost:soil was 1:1, 1:2 or 1:3, 100 days after transplanting. Addition of sheep-manure vermicompost decreased soil pH, titratable acidity and increased soluble and insoluble solids, in tomato fruits compared to those harvested from plants cultivated in unamended soil. Sheep-manure vermicompost as a soil supplement increased tomato yields and soluble, insoluble solids and carbohydrate concentrations.     

Limitations of a calculated N mineralization potential in studies of the N mineralization process

Soil is often incubated under controlled conditions to assess its capacity to mineralize nitrogen and to define the N mineralization potential (N_o) by fitting a negative exponential curve to N mineralization data. The specificity of N_o for a given soil and its relevance in N mineralization studies was examined as part of an overall study of the N mineralization process. Soil mixed with an equal amount of sand was aerobically incubated at 35 °C and leached at specific time intervals. Upon leaching, ammonium and nitrate were measured in the extract.It was found that N mineralization data did not always follow first-order kinetics making it difficult to calculate N_o. The computed N_o value was influenced by the shape of the curve, the duration of the incubation experiment and was reciprocally related to the N mineralization constant (k_exp). N_o did not always give an adequate indication of the amount of N mineralized and was not soil specific as the time of sampling largely affected its size. The usefulness of N_o in the simulation of the N mineralization process with a k_exp value valid for all soils was limited and a k_exp value specific for each soil was required. A value combining the soil specific N_o and k_exp values and reflecting the amount of N mineralized over one year was proposed as a suitable alternative to the use of N_o in comparative studies of the N mineralization process. It was concluded that a calculated N_o could not be used in studies comparing the N mineralization of different soils. In addition, the simulation of the N mineralization required the use of the soil specific k_exp and could not be carried out with a k_exp valid for all soils.     

CD4-Positive T Cells and M2 Macrophages Dominate the Peritoneal Infiltrate of Patients with Encapsulating Peritoneal Sclerosis

Background

Encapsulating peritoneal sclerosis (EPS) is a severe complication of peritoneal dialysis (PD). Previously, it has been shown that infiltrating CD4-positive T cells and M2 macrophages are associated with several fibrotic conditions. Therefore, the characteristics of the peritoneal cell infiltrate in EPS may be of interest to understand EPS pathogenesis. In this study, we aim to elucidate the composition of the peritoneal cell infiltrate in EPS patients and relate the findings to clinical outcome.

Study Design, Setting, and Participants

We studied peritoneal membrane biopsies of 23 EPS patients and compared them to biopsies of 15 PD patients without EPS. The cellular infiltrate was characterized by immunohistochemistry to detect T cells(CD3-positive), CD4-positive (CD4+) and CD8-positive T cell subsets, B cells(CD20-positive), granulocytes(CD15-positive), macrophages(CD68-positive), M1(CD80-positive), and M2(CD163-positive) macrophages. Tissues were analysed using digital image analysis. Kaplan-Meier survival analysis was performed to investigate the survival in the different staining groups.

Results

The cellular infiltrate in EPS biopsies was dominated by mononuclear cells. For both CD3 and CD68, the median percentage of area stained was higher in biopsies of EPS as opposed to non-EPS patients (p<0.001). EPS biopsies showed a higher percentage of area stained for CD4 (1.29%(0.61-3.20)) compared to CD8 (0.71%(0.46-1.01), p = 0.04), while in the non-EPS group these cells were almost equally represented (respectively 0.28%(0.05-0.83) versus 0.22%(0.17-0.43), p = 0.97). The percentage of area stained for both CD80 and CD163 was higher in EPS than in non-EPS biopsies (p<0.001), with CD163+ cells being the most abundant phenotype. Virtually no CD20-positive and CD15-positive cells were present in biopsies of a subgroup of EPS patients. No relation was found between the composition of the mononuclear cell infiltrate and clinical outcome.

Conclusions

A characteristic mononuclear cell infiltrate consisting of CD4+ and CD163+ cells dominates the peritoneum of EPS patients. These findings suggest a role for both CD4+ T cells and M2 macrophages in the pathogenesis of EPS.     

Hemizygous deletion of CTGF/CCN2 does not suffice to prevent fibrosis of the severely injured kidney

BackgroundConnective Tissue Growth Factor (CTGF/CCN2) is an important mediator of kidney fibrosis. Previous observations indicated that attenuation of CCN2 expression sufficed to alleviate early kidney damage. However, little is known about the role of CCN2 in fibrosis of severely damaged and more chronically injured kidneys. Therefore, we examined the effects of CCN2 haploinsufficiency on the progression of renal scarring in long-term STZ-induced diabetic nephropathy, in a more advanced stage of obstructive nephropathy following unilateral ureteric obstruction (UUO), and in severe aristolochic acid (AA)-induced tubulotoxic nephritis.MethodsWild-type (WT, CCN2^+/+) and hemizygous CCN2^+/? C57Bl/6 mice were studied. In the diabetes experiment, streptozotocin-injected and control mice were followed for 6 months, with regular blood pressure, glycaemia and albuminuria recordings. In the UUO experiment, the left ureter was obstructed for 14 days with the contralateral kidney serving as control. For the AA experiment, mice were followed for 25 days after 5 intraperitoneal injections with AA and compared to control mice injected with buffer alone. Organs were harvested for histology, mRNA and protein measurements. Collagen content was determined by HPLC and expressed as hydroxyproline/proline ratio.ResultsCCN2 expression was significantly increased in the damaged as compared to control kidneys. In all three models, CCN2 levels in the damaged kidneys of CCN2^+/? mice averaged about 50% of those in damaged WT kidneys. After 6 months of diabetes, albuminuria was increased 2.5-fold in WT mice, compared to 1.5-fold in CCN2^+/? mice, mesangial matrix was expanded 5-fold in WT and 4.4-fold in CCN2^+/? mice and the glomerular basement membrane was thickened 1.3-fold in WT and 1.5-fold in CCN2^+/? mice (all differences between WT and CCN2^+/? mice are NS). Tubular damage and interstitial fibrosis scores were also not different between Wt and CCN2^+/? mice in the diabetes (1.8 vs. 1.7), UUO (2.8 vs. 2.6), and AA (1.4 vs. 1.2) models, as was the case for macrophage influx and collagen content in these three models.ConclusionUnlike in mild and relatively early STZ-induced diabetic nephropathy, scarring of severely and chronically damaged kidneys is not attenuated by a 50% reduction of CCN2 to (near) normal levels. This suggests that CCN2 is either redundant in severe and chronic kidney disease, or that it is a limiting factor only at subnormal concentrations requiring further reduction by available or emerging therapies to prevent fibrosis of the severely injured kidney.     

Phylogenetic and Multivariate Analyses To Determine the Effects of Different Tillage and Residue Management Practices on Soil Bacterial Communities

Bacterial communities are important not only in the cycling of organic compounds but also in maintaining ecosystems. Specific bacterial groups can be affected as a result of changes in environmental conditions caused by human activities, such as agricultural practices. The aim of this study was to analyze the effects of different forms of tillage and residue management on soil bacterial communities by using phylogenetic and multivariate analyses. Treatments involving zero tillage (ZT) and conventional tillage (CT) with their respective combinations of residue management, i.e., removed residue (−R) and kept residue (+R), and maize/wheat rotation, were selected from a long-term field trial started in 1991. Analysis of bacterial diversity showed that soils under zero tillage and crop residue retention (ZT/+R) had the highest levels of diversity and richness. Multivariate analysis showed that beneficial bacterial groups such as fluorescent Pseudomonas spp. and Burkholderiales were favored by residue retention (ZT/+R and CT/+R) and negatively affected by residue removal (ZT/−R). Zero-tillage treatments (ZT/+R and ZT/−R) had a positive effect on the Rhizobiales group, with its main representatives related to Methylosinus spp. known as methane-oxidizing bacteria. It can be concluded that practices that include reduced tillage and crop residue retention can be adopted as safer agricultural practices to preserve and improve the diversity of soil bacterial communities.Agricultural sustainability is linked to soil management and efficient use of natural and economic resources (25, 53). Sustainable handling of resources can be obtained by applying conservation agricultural practices, i.e., reduced tillage, crop residue retention, and crop rotation (26). Reduced tillage and crop residue retention have been proposed, as they facilitate water infiltration, reduce erosion, improve soil structure, increase soil organic matter and carbon content, and moderate soil temperatures (13, 16, 30, 33, 56). Compared with conventional tillage and crop residue removal, these practices can also decrease production costs by reducing the use of heavy machinery, fuels, water, and fertilizers (19, 23). The positive effect of these practices seems to be correlated with the improvement of soil structure and a higher availability of organic substrates for microorganisms (3, 30). Improved soil structure allows better soil aeration and diffusion of water and nutrients through the soil profile, while the retention of crop residues enhances microbial activity and the soil microbial biomass content (12, 28). These improvements in soil quality can also increase soil microbial diversity, thus protecting crops against pests and diseases through competition for soil nutrients (8).Until now, most research has focused on microbial communities affected by agricultural practices, i.e., tillage and residue management, by using indicators such as plate counting and microbial biomass or by analyzing denaturing gradient gel bacterial banding patterns (21, 22, 37). Salles et al. (46) reported the use of canonical correspondence analysis on denaturing gradient gel electrophoresis banding pattern data to understand the effect of crop and land history on Burkholderia communities. However, few studies have applied phylogenetic and multivariate analyses to understand the effect of soil management practices, i.e., tillage and residue management, on microbial communities.It is necessary to interpret the changes in microbial communities as a function of contextual environmental parameters to analyze the effect of anthropogenic activities on microbial communities (42). Once modifications in microbial communities are interpreted as a function of contextual environments, it becomes possible to determine the kind of organisms that dominate such environments and to establish whether specific practices could lead to changes in beneficial or nonbeneficial microorganisms for agro-ecosystems. Changes in microbial communities can then be related to food production, soil quality, and greenhouse gas emissions (19, 20, 36).Govaerts et al. (19, 20, 21, 22) had previously characterized the soils used in this study. They showed that soils under zero tillage (ZT) and crop residue retention (+R) have better soil quality, crop yields, and catabolic diversity and a higher diversity of microflora groups than do soils under conventional tillage (CT) with or without crop residue retention (−R). The aim of this study was to complement the results of Govaerts et al. (19, 20, 21, 22) by using phylogenetic approaches and the additive main effect and multiplicative interactions (AMMI) model (18, 60) to analyze the effect of the above treatments on soil bacterial communities.     

Residue fractionation and decomposition: The significance of the active fraction

This paper describes an incubation experiment with homogeneously ¹⁴C labeled maize-straw and its insoluble fraction. The role of the soluble fraction in the decomposition process was assessed, using three independently measured characteristics: (1) fractionation of the maize-straw, resulting in kinetically different fractions; (2) microbial biomass C and its ¹⁴C activity determined by a fumigation extraction method, and (3) the ¹⁴C activity of the released CO₂-C. The fumigation extraction method was proved to be useful from 9 days after the application of the maize-straw onwards. The fractionation method yielded a soluble (48%), a (hemi) cellulosic (47%), and a lignin fraction (1%). Nine days after addition of either the complete residue or its insoluble fraction, the microbial biomass C increased from 53 to 337 and 217 mg C kg^-1 dry soil, respectively. Similar values were maintained up to day 40. The large increase in microbial activity was accompanied by a N-immobilization of 65 and 29 mg N Kg^-1 dry soil for the maize-straw treatment and its insoluble fraction, respectively, resulting in biomass C/N values of 5.5 and 5.6 A genuine priming effect (10 and 7% of the total CO₂-C production) on the mineralization of native soil organic C was caused by an increase in decomposition of the native C rather than by an increase in turnover of the microbial biomass in the soil amended with maize straw. The soluble fraction caused a priming effect on the decomposition of the less decomposable cell-wall fraction. Calculations by nonlinear regression confirmed this observation.     

Emissions of carbon dioxide, methane and nitrous oxide from soil receiving urban wastewater for maize (Zea mays L.) cultivation

In the Darmstadt long-term fertilization trial, the application of composted cattle farmyard manure without (CM) and with (CMBD) biodynamic preparations was compared to mineral fertilization with straw return (MIN). The present study was conducted to investigate the effects of spatial variability, especially of soil pH in these three treatments, on soil organic matter and soil microbial biomass (C, N, P, S), activity (basal CO₂ production and O₂ consumption), and fungal colonization (ergosterol). Soil pH was significantly lower in the MIN treatments than in the organic fertilizer treatments. In the MIN treatments, the contents of soil organic C and total N were also significantly lower (13% and 16%, respectively) than those of the organic fertilizer treatments. In addition, the total S content increased significantly in the order MIN < CM < CMBD. The microbial biomass C content was significantly lower (9%) in the MIN treatments than in the organic fertilizer treatments. Microbial biomass N and biomass P followed microbial biomass C, with a mean C/N ratio of 7.9 and a mean C/P ratio of 23. Neither the microbial biomass C to soil organic C ratio, the metabolic quotient qCO₂, nor the respiratory quotient (mol CO₂/mol O₂) revealed any clear differences between the MIN and organic fertilizer treatments. The mean microbial biomass S content was 50% and the mean ergosterol content was 40% higher in the MIN treatments compared to the organic fertilizer treatments. The increased presence of saprotrophic fungi in the MIN treatments was indicated by significantly increased ratios of ergosterol-to-microbial biomass C and the microbial biomass C/S ratio. Our results showed that complex interactions between the effects of fertilizer treatments and natural heterogeneity of soil pH existed for the majority of microbial biomass and activity indices.     

Poultry slaughter wastewater treatment with an up-flow anaerobic sludge blanket (UASB) reactor

Removal of organic material from poultry slaughter wastewater as determined by changes in biological oxygen demand (BOD5) was investigated by adding three different types of inoculum combining cow manure, yeast extract or hydraulic residence time as variables with response vector of reduction of BOD5. In a 3-l reactors, a 95% removal of BOD5 from poultry slaughter wastewater was obtained with organic loading rates up to 31 kg BOD5 m(-3) d(-1) without loss of stability. This 95% removal was obtained between 25 and 39 degrees C with a hydraulic residence time between 3.5 and 4.5 h. The growth of the consortium of micro-organisms in the reactor followed a first-order kinetic with a constant specific growth rate of 0.054 h(-1). It was concluded that an inoculum from cow manure added with nutrients and yeast extract allowed a 95% removal of BOD5 from poultry slaughter wastewater at ambient temperatures within a hydraulic residence time of 4 h, sharply reducing possible environmental hazards.     

Mineralization of 14C-labelled maize in alkaline saline soils

The turnover of organic material determines the availability of plant nutrients in unfertilized soils, and this applies particularly to the alkaline saline soil of the former Lake Texcoco in Mexico. Uniformly labelled [¹⁴C] maize and its neutral detergent fibre (NDF) fraction, mainly containing cellulose and hemi-cellulose, were added to these soils to investigate dynamics of C and N and the importance of the NDF fraction. Soil with electrolytic conductivity (EC) of 1.2, 3.2, 24.6 and 32.7 dS m^–1 was incubated aerobically, while CO₂ and ¹⁴CO₂ production, and inorganic N dynamics (NH₄ ⁺, NO₂ ^–, NO₃ ^–) were monitored. The amount of ¹⁴C-labelled maize mineralized after 97 days was >500 mg C kg^–1 dry soil (D.S.) of the 1000 mg C kg^–1 D.S. added in soils with EC 24.6 dS m^–1, but only 257 mg C kg^–1 D.S. in soil with EC 32.7 dS m^–1. The decomposition of the NDF fraction showed a lag, greatest in the soil with the largest EC and the amount of ¹⁴C-labelled NDF fraction mineralized after 97 days was > 300 mg C kg^–1 D.S. in soils with EC 3.2 dS m^–1, but in the soil with EC 32.7 dS m^–1 it was only 118 mg C kg^–1D.S. Application of ¹⁴C-labelled maize and the NDF fraction induced a priming effect, most accentuated at the onset of the incubation. The ratio between the amount of CO₂ produced due to the priming effect and the ¹⁴CO₂ produced was 16-times larger when 250 mg maize-C kg^–1 D.S. was added and only 3-times when 2000 mg maize-C kg^–1 D.S. was added. Oxidation of NO₂ ^– occurred in soil with EC 32.7 dS m^–1 as witnessed by decreases in concentration of NO₂ ^– and increases in concentration of NO₃ ^–. It was found that EC affected the decomposition of maize, the NDF fraction and the priming effect. Decomposition of cellulose and oxidation of NO₂ ^– occurred in soil with EC 32.7 dS m^–1 although cellulolytic micro-organisms and autotrophic NO₂ ^– oxidizers could previously not be isolated from this soil.