Carbon limitations to nitrous oxide emissions in a humid tropical forest of the Brazilian Amazon

The availability of labile organic C for microbial metabolic processes could be an important factor regulating N₂O emissions from tropical soils. We explored the effects of labile C on the emissions of N₂O from a forest soil in the State of Rondônia in the southwestern quadrant of the Brazilian Amazon. We measured emissions of N₂O from a forest soil after amendments with solutions containing glucose, water only or NO₃^–. Addition of glucose to the forest soil resulted in very large increases in N₂O emissions whereas the water only and NO₃^– additions did not. These results suggest a strong C limitation on N₂O production in this forest soil in the southwestern Amazon.     

Quantitative aspects of solid-state C-NMR spectra of humic substances from soils of volcanic systems

Cross-Polarisation Magic Angle Spinning Carbon-13 Nuclear Magnetic Resonance spectroscopy (CPMAS ¹³C-NMR) represents one of the most powerful tools to investigate soil organic matter (SOM) mainly because of its inherent capacity to provide a semi-quantitative evaluation of carbon distribution. A critical parameter during acquisition of CPMAS ¹³C-NMR spectra is the contact time required to obtain the cross-polarisation between proton and carbon nuclei. The procedure to evaluate the best contact time for the acquisition of a quantitative CPMAS ¹³C-NMR spectrum is to perform Variable Contact Time (VCT) experiments. In this work the structural features of a number of purified humic substances from Italian and Costarican volcanic soils were investigated by CPMAS ¹³C-NMR spectroscopy after having performed preliminary VCT experiments. The VCT experiments showed that the average contact times vary according to the origin and chemical structure of the humic material. The optimal contact times (OCT) for nine humic samples were between 250 and 800 μs These values were different from the time of 1000 μs that is commonly applied as the best average contact time for humic materials. Moreover, by comparing the NMR data to those obtained by elemental analysis (C/H ratio), it appeared that the efficiency of the cross-polarisation between protons and carbons, and hence the contact time, is affected not only by the number of protons, but also by their distribution over the molecules. The evaluation of errors in quantitative estimation of the different carbons revealed that the use of OCT generally reduced by half the loss of signals occurring when the average contact time of 1000 μs is used in CPMAS ¹³C-NMR spectra of humic substances.     

Integration of TGF-beta and Ras/MAPK signaling through p53 phosphorylation

During development and tissue homeostasis, cells must integrate different signals. We investigated how cell behavior is controlled by the combined activity of transforming growth factor-beta (TGF-beta) and receptor tyrosine kinase (RTK) signaling, whose integration mechanism is unknown. We find that RTK/Ras/MAPK (mitogen-activated protein kinase) activity induces p53 N-terminal phosphorylation, enabling the interaction of p53 with the TGF-beta-activated Smads. This mechanism confines mesoderm specification in Xenopus embryos and promotes TGF-beta cytostasis in human cells. These data indicate a mechanism to allow extracellular cues to specify the TGF-beta gene-expression program.     

Effects of fractions of coal-derived humic substances on seed germination and growth of seedlings (Lactuga sativa and Lycopersicum esculentum)

Summary Lettuce and tomato seeds were treated in Petri dishes with a humic acid derived from an oxidized coal and with fractions of the same humic acid obtained by ultrafiltration through membranes of known molecular cutoff and by extracting with buffers set at pH 4 and pH 5. The unfractionated humic acid was applied at 40, 100, 1000, and 5000 mg l^-1 whereas the humic fractions were applied at 40, 100, and 200 mg l^-1. Germination parameters such as the number of total germinated seeds, the velocity of seed germination, the fresh weight and dry weight of total seedlings were measured and related to the chemical and physicochemical properties of the humic material. No increase in the germination percentage or the germination rate was observed for either lettuce or tomato seeds. The fresh weight of total seedlings and per seedling increased in treatments with unfractionated humic acid with increasing concentrations for both lettuce and tomato plants without showing signs of growth inhibition up to 5000 mg l^-1. This was attributed to cell elongation and more efficient water uptake. For the lettuce, the fresh weight both of total seedlings and per seedling was enhanced by treating the seeds with fractions of low molecular weight and high content of acidic functions, whereas the dry weight both of total seedlings and per seedling did not change with the humic fraction used. For the tomato seeds in contrast, the dry weight both of total seedlings and per seedling was increased by the use of unfractionated humic acid and by some of the humic fractions. An uptake of humic material by growing tomato seedlings was inferred.     

Mass Production of Juveniles of the Fat Snook Centropomus parallelus in Brazil

A pilot‐scale trial to rear fat snook Centropomus parallelus through larval, weaning and nursery phases was conducted in Florianópolis, Brazil. Eggs (96% fertilization) from captive broodstock, induced to spawn using 50 μg/kg LHRHa were stocked in two 4,000‐L cylindrical fiberglass tanks at a mean density of 19.2 eggs/L. Nannochloropsis oculata was stocked with the eggs and maintained at a mean density of 0.5–1.0 ± 10⁶ cells/mL up to 31 dph (31 dph). Hatching averaged 90%. Larvae were fed rotifers Brachionus rotundiformis enriched with an oil emulsion from 3 dph to 36 dph (30–40 rotifers/mL) and Artermia meta‐nauplii enriched with Selco from 22 dph to 60 dph (mean 2.9 meta‐nauplii/mL). Weaning began at 45 dph with an artificial dry diet NRG (50% protein), supplied together with concentrated and enriched Artemia meta‐nauplii. No critical period of mortality was observed during larval rearing. During the 43 days of weaning and nursery, less than 1% mortality was recorded. Food conversion rate during nursery was 1.17, with a change in the coefficient of variation of mean total length of 1.3%. Specific total growth rate in weight was 13.0 %/d and mean growth in total length and total weight were 0.65 mm/d and 24.0 mg/d over the whole rearing trial respectively. Mean total length and total weight of juveniles were 57.6 ± 0.1 mm and 2.11 ± 0.12 g, respectively, and the length‐weight relationship was W = 8.29931 ± 10–⁵ TL^3.049607 (r= 0.9986). on 88 dph when the trial was terminated. The condition factor on 88 dph was 1.104. On 88 dph a total of 35,000 juveniles were harvested, overall survival was 25.5% with mean final density of 4.4 fishn and biomass of 6.9 kg/m³. The present trial demonstrated the feasibility of mass production of fat snook. Possible improvements necessary for commercial cultivation of fat snook C. parallelus are discussed.     

Rapid isolation of Brachyspira hyodysenteriae and Brachyspira pilosicoli from pigs

The aim of this study was to compare and evaluate the time required to isolate Brachyspira hyodysenteriae and Brachyspira pilosicoli from porcine faeces. This was done using previously described selective media (spectinomycin) S400, (colistin, vancomycin and spectinomycin) CVS and (spectinomycin, vancomycin, colistin, spiramycin and rifampin with swine faecal extract) BJ, compared with the method based on blood agar modified medium, with spectinomycin and rifampin (BAM-SR), including a pre-treatment step. Fourteen spirochaetal strains were obtained in pure cultures after 5 days (48 h in BAM-SR primary plate and three passages every 24 h in brain heart infusion (BHI) without antibiotics) pre-treating simulated samples in brain heart infusion broth with spectinomycin (400 microg/ml) and rifampin (15 microg/ml), before streaking on the selective BAM-SR medium. Spirochaetes from samples in S400, CVS and BJ, with and without pre-treatment, were obtained in pure cultures only after repeatedly transferring on plates of the same selective medium requiring 15-18 days according to the strain. BAM-SR used after the pre-treatment step showed a detection limit ranging from 3.5 x 10(2) to 6.7 x 10(7) cells/g faeces and was the only method able to support the growth of spirochaetes after 48 h.     

Linoleic acid increases adhesion,chemotaxis, granule release,intracellular calcium mobilisation,MAPK phosphorylation and gene expression in bovine neutrophils

Neutrophils are critical to the innate immune response; therefore, the proper function of neutrophils is critical to avoid the development of certain diseases. Linoleic acid, a polyunsaturated long-chain fatty acid, is one of the most abundant long-chain fatty acids found in the plasma of cows after giving birth. In this study, we evaluated the effects of linoleic acid treatment on bovine neutrophil adhesion, chemotaxis, metalloproteinase (MMP)-9 release, CD11b expression, intracellular calcium mobilisation, mitogen-activating protein kinase (MAPK) phosphorylation and COX-2 and IL-8 expression. Bovine neutrophils isolated from healthy heifers were incubated with different concentrations of linoleic acid, and then neutrophil responses were evaluated. Our results show that the treatment of neutrophils with 100 μM linoleic acid increased their adhesion to the bovine endothelial cell line CPA47. The results of a transwell migration assay revealed that linoleic acid could also promote the chemotaxis of bovine neutrophils. Furthermore, linoleic acid treatment increased MMP-9 activity and CD11b cell surface expression in neutrophils. Fifty and 100 μM linoleic acid also increased intracellular calcium mobilisation in neutrophils loaded with Fluo-4 AM dye. Linoleic acid also rapidly (2–5 min) stimulated the phosphorylation of ERK1/2 and p38 MAPK as evaluated by immunoblot. Finally, COX-2 and IL-8 mRNA expression increased after 2 h of linoleic acid treatment. In conclusion, linoleic acid stimulates adhesion, chemotaxis, granule release and intracellular responses in bovine neutrophils.     

CO2 fertilization does not affect biomass production and nutritive value of a C4 tropical grass in short timeframe

Increased atmospheric carbon dioxide (CO₂) is a consequence of recent anthropogenic environmental changes, and few studies have evaluated its effects on tropical grasses used in Brazilian pastures, the main feed source for major part of ruminant livestock. This study evaluated forage production, chemical composition, in vitro total gas production and organic matter degradability of Brachiaria brizantha under contrasting CO₂ atmospheric conditions in a free air carbon dioxide enrichment (FACE) facility. The forage plants were sown in each of the 12 octagonal rings of the FACE facility: six under ambient atmospheric CO₂ concentration of approximately 390 μmol/mol, hereafter referred to as control (CON) plots, and other six rings enriched with pure CO₂ flux to achieve a target CO₂ concentration of 550 μmol/mol, hereafter called elevated CO₂ (eCO₂) plots. Soil samples were collected to determine carbon and nitrogen concentrations. After seventy days of sowing, a standardization cutting was performed and then at regular intervals of 21 days the forage was harvested (ten harvest dates) and forwarded to laboratorial analyses. Forage above‐ground biomass production (dry matter (DM): 6,143 vs. 6,554 kg/ha), as well as morphological characteristics (leaves: 71% vs. 68%; stem: 28% vs. 31%), chemical composition (crude protein: 162.9 vs. 161.8; neutral detergent fibre: 663.8 vs. 664.3; acid detergent fibre: 369.5 vs. 381; lignin: 60.1 vs. 64.1 g/kg DM; total C: 45.9 vs. 45.9; total N: 2.8 vs. 2.8; total S: 0.2% vs. 0.2%), organic matter in vitro degradability (573.5 vs. 585.3 g/kg), methane (5.7 vs. 4.3 ml/g DM) and total gas (128.3 vs. 94.5 ml/g DM) production did not differ significantly between CON and eCO₂ treatments (p > .05). The results indicated that at least under short‐term enrichment, B. brizantha was not affected by eCO₂.     

Interactions of three s-triazines with humic acids of different structure

The adsorption of three s-triazines (atrazine, terbutylazine, and simazine) on three different humic acids (HAs), before and after acidic hydrolysis, was studied at four diverse pH values from 3 to 4.5. The Freundlich sorption affinity (K(f)) and intensity (1/n) of s-triazines were related to the chemical and structural composition of HA and used in a multivariate statistical analysis. At low pH, the sorption values for s-triazines were not directly justified by the content of carboxyl groups in original HA, while only an increase of phenolic carbons in hydrolyzed HA supported the increased s-triazine adsorption. The structural composition of both s-triazines and humic samples explained 86% of the significance in multivariate analyses, whereas the role of pH remained hidden in only 14% of the statistical significance. Adsorption of s-triazines was mainly related to carbon content, hydrophobicity, and aromaticity of HA, thereby implying a predominant binding role of weak dispersive forces. Steric properties explained the larger adsorption affinity of atrazine and terbutylazine with respect to simazine. The occurrence of charge-transfer interactions between atrazine and mainly hydrolyzed HA was further suggested by the increased number of free radicals detected in atrazine-HA complexes at different pH values. Charge-transfer interactions were postulated to occur when HA conformations became progressively controlled by aromatic components. This work indicates that humic matter rich in hydrophobic and aromatic constituents is more likely to adsorb s-triazines and reduce their environmental mobility.     

Quantification of EGFR family in canine mammary ductal carcinomas in situ: implications on the histological graduation

Veterinary Research Communications - The epithelial growth factor receptors are transmembrane proteins with an important role in the neoplastic progression of tumors, and in this context, DCIS is...