Biodegradation of humic substances by microscopic filamentous fungi: chromatographic and spectroscopic proxies

Purpose

The study of interactions between humic substances (HSs) and soil filamentous fungi is the key to understanding the sustainable soil functioning. The present work aims to examine the decomposition of HSs by filamentous dark-pigmented fungus Alternaria alternatа under the laboratory conditions and to determine the effect of easily assimilable organic carbon on this process. Analyzing such polydisperse substances like HSs by a complex integrated methodology makes it possible to explore the data on their decomposition by microorganisms.

Materials and methods

To achieve the aforementioned goals, we used chromatographic and spectroscopic approaches: low-pressure size-exclusion and hydrophobic interaction chromatography accompanied by absorption and fluorescence spectroscopy. To determine the effect cometabolism conditions produced on HS decomposition, two types of carbon substrates were added to the nutrient media: easily assimilable organic carbon (standard 0.3% or reduced 0.03% sucrose content) and hardly assimilable organic carbon (HSs), as well as their combinations. Five HS samples of different organic matter origin have been inspected: potassium humates (HPs) and humic acids (HAs) from coal, peat, and lignosulfonate. Correlation matrix and principal component analysis (PCA) were calculated for comprehensive data analysis.

Results and discussion

Transformations of the investigated HSs under fungal cultivation lead to the increase in the low molecular weight fraction, rise of hydrophilic fraction, enlargement of absorbance ratio A₂₅₀/A₃₆₅, shortening of the emission wavelength of the humic-type fluorescence, and growth in the fluorescence quantum yield measured with excitation at 355 nm. A positive correlation was observed between the accumulation of fungal biomass and the degree of HS decomposition. PCA analysis confirms that the difference in the results of HS decomposition largely depends on the sucrose content and the nature of HSs. We divided all the HS samples into four groups according to the degree of HS decomposition: original HS solutions, HPs altered using fungal cultivation at 0.03% sucrose, HAs after fungal cultivation at 0.03% sucrose, and finally, HSs (both HPs and HAs) after fungal cultivation at 0.3% sucrose.

Conclusions

In the laboratory experiments, we showed that (1) the isolated HAs were more effectively degraded than the parent HPs, and this process was more pronounced at a reduced sucrose content, and (2) the decomposition of stable organic compounds (HSs) was activated by the easily assimilable carbon sources (especially 0.3% sucrose) being present. We assume that it is the easily assimilable organic carbon that most likely triggers the HS degradation working as the priming effect in natural environments.

     

Arsenic-contaminated soils phytotoxicity studies with sunflower and sorghum

Background, Aim and Scope  Environmental pollution caused by arsenic (As) is a major ecological problem. There has been intense worldwide effort to find As-hyperaccumulating plants that can be used in phytoremediation—the green-plant-assisted removal of chemical pollutants from soils. For phytoremediation, it is natural to prefer cultivated rather than wild plants, because their agriculture is well known. This study was conducted to evaluate the tolerance of common sunflower(Helianthus annuus L.) and sugar sorghum(Sorghum saccharatum Pers.) for soil-As contents of 10–100 mg As kg^-1 soil, with sodium arsenite as a model contaminant. Methods  Plants were grown in a growth chamber for 30 days. Microfield experiments were conducted on experimental plots. To study the phytoremediation effect of the auxins indole-3-acetic acid (IAA) and 2,4-dichlorophenoxyacetic acid (2,4-D), we treated 1- and 3-day-old plant seedlings with water solutions of the auxins (concentrations 10^-5, 10^-7, and 10^-9 g l^-1). The soil and plant-biomass samples were analyzed for total As by using the color reaction of ammonium molybdate with As. Results and Discussion  Phytotoxicity studies showed that 100 mg As kg^-1 soil poisoned sunflower and sorghum growth by 50%. There was a linear correlation between soil-As content and As accumulation in the plants. Laboratory experiments showed that the soil-As content was reduced two- to threefold after sunflower had been grown with 10–100 mg As kg^-1 soil for 30 days. Treatment of sunflower and sorghum seedlings with IAA and 2,4-D at a concentration of 10^-5 g l^-1 in microfield experiments enhanced the phytoremediation two- to fivefold as compared with untreated control plants. The best results were obtained with 3-day-old seedlings. Conclusion, Recommendation and Outlook  (a) Sunflower and sorghum are good candidates to remediate As-polluted soils. (b) Phytoremediation can be improved with IAA or 2,4-D. (c) Mixed cropping of sorghum and sunflower may be another way of improving phytoremediation.     

Degradability of soils under oak and pine in Central Spain

The purpose of this research was to study the influence of the vegetation on the soil C pool of forests of pines (Pinus sylvestris) and oaks (Quercus pyrenaica), located in Central-Western Spain. Horizons from selected soils located in these forests were sampled, and the soil organic C (SOC) was determined. In addition, in vitro incubation experiments were carried out, under controlled conditions, to monitor the stability of SOC against the microbial activity. Soil humus fractions were isolated following a classical procedure of chemical fractionation using alkaline solutions, before and after the incubation experiment. A deeper O horizon was found under the pine forest than under oak one; however, higher SOC content was found in the oak site than that under pine one. During the in vitro mineralization process, a lower CO₂ production by the soil sample from pine forest was observed, in relation to that emitted by the oak soil. In addition, a lower humification degree was estimated for the soil humus under pines than for that under oaks. In conclusion, replacement of oaks by pines produced a decrease in SOC accumulation and a lower quality of humus in the forest soils.     

Removal of Copper and Cadmium Ions from Diluted Aqueous Solutions by Low Cost and Waste Material Adsorbents

The sorption of copper and cadmium ions using activated carbon,kaolin, bentonite, diatomite and waste materials such as compost,cellulose pulp waste and anaerobic sludge as sorbents is reported. Equilibrium isotherms were obtained for the adsorption of these metals in single and binary solutions. Bentonite presented the highest adsorption capacities for both copper and cadmium. A competitive uptake was observed when both metals are present; copper being preferentially adsorbed by all materials with theexception of anaerobic sludge. Equilibrium data were fitted toLangmuir and Freundlich models, with satisfactory results for most of the adsorbent-metal systems studied.Of all the adsorbents studied, bentonite and compost presentedthe highest removal efficiencies, reaching 99% for copper whencadmium is also present, for initial solution concentrations ofup to 100 mg L^-1. Anaerobic sludge has a greater preferencefor cadmium, even in the presence of copper, with removal efficiencies of 98% for similar concentrations to those mentioned above.     

Water mobility in multicomponent model media as studied by 2H and 17O NMR

Molecular mobility of water was studied in a microbiological media containing complex and heterogeneous mixtures of cellulose, l-sorbose, and orange serum broth (OSB) using (2)H and (17)O high-resolution NMR. All mixtures showed Lorentian (17)O NMR spectra but complex (2)H NMR line shapes. Sorbose, when solubilized, caused line-narrowing where as cellulose-OSB mixtures showed wide peaks with flat plateaus. Presence of liquid or solvent water had a profound effect on a marked increase in T(2) relaxation time observed in sorbose-containing samples. (17)O NMR data were not composition dependent, while (2)H NMR data were highly sorbose dependent.     

Use of random amplified polymorphic DNA (RAPD) to identify races 1, 2, 4 and 8 of Fusarium oxysporum f. sp. dianthi in Italy

The RAPD fingerprinting procedure was used in combination with pathogenicity assays on differential cultivars to characterize a representative collection of 72 Fusarium spp. isolates of different geographic origin collected from diseased carnation. In F. oxysporum f. sp. dianthi, isolates were grouped according to the physiologic race: group 1 included isolates of race 4; group 2 was formed by isolates of race 2 and single representatives of races 5 and 6; group 3 included isolates of races 1 and 8. No correlation was found between RAPD data and geographic origin of the isolates tested: representatives of race 2 isolated in Italy, Israel and Japan had the same amplification profile. Three isolates which showed a low level of pathogenicity on all carnation cultivars tested shared an identical amplification pattern and are probably saprophytic F. oxysporum. Finally, two F. redolens isolates from Japan and seven non-pathogenic isolates of F. proliferatum collected from diseased carnation in Italy, Israel and The Netherlands were clearly distinguishable according to their RAPD fingerprint. The results are discussed in relation to previous studies on the genetic diversity of F. oxysporum f. sp. dianthi and to the development of forma specialis- and pathotype-specific diagnostic tools.     

Rumen-protected choline administration to transition cows: effects on milk production and vitamin E status

We investigated the influence of rumen-protected choline (RPC) supplementation on milk production, lipid metabolism and vitamin E status in dairy cows receiving a silage-based diet. Twenty-six Italian Holstein multiparous cows were assigned by weight and average production in the previous lactation, to one of two groups: control (no RPC supplementation) and RPC (supplemented with 20 g/day rumen-protected choline chloride). Treatment began 14 days before expected calving and continued for 30 days after parturition. Choline administration significantly increased milk production during the first month of lactation and also the concentration (and total secretion) of choline in milk, but did not affect fat or protein concentrations in milk, or plasma levels of glucose, beta-hydroxybutyrate, cholesterol and non-esterified fatty acids (NEFA). However, around parturition, NEFA concentrations in plasma were lower in treated animals than in controls, suggesting improved lipid metabolism as a result of choline supplementation. Choline supplementation also increased alpha-tocopherol plasma concentrations, suggesting a novel aspect in dairy cows.     

Characteristic fragment ions from lignin and polysaccharides in ToF–SIMS

Different model compounds for lignin, hemicelluloses and pectins were studied by time-of-flight secondary-ion mass spectrometry (ToF–SIMS). Mass spectra of Klason lignin from normal and compression spruce wood, aspen wood and wheat straw were compared. Spectra of brominated spruce and aspen wood sections showed fragment ions attributed to brominated guaiacyl and syringyl units in lignin at m/z 215, 217, 229 and 231, and m/z 245, 247, 249 and 261, respectively. Spectra of mono-, oligo- and polysaccharides showed fragment ions at m/z 127 and 145 characteristic for hexose units, and ions at m/z 115 and 133 characteristic for pentose units. The same ions were detected in spectra of delignified spruce and aspen wood sections. Labelling of anionic groups by Sr²⁺ ions followed by ToF–SIMS analysis showed that pectins were present at specific locations on the surfaces of spruce and aspen wood sections still after delignification with hydrogen peroxide in acetic acid.     

Effect of emerged shipment on the physiological condition of the adductor muscle in adult giant lion's paw scallop Nodipecten subnodosus (Sowerby 1835)

Currently, there is an increasing interest in shipping live adult scallops to markets and broodstock to hatcheries; nevertheless, information about the shipping effect on live scallops physiology is very scarce. In a previous study, a method in emersion was developed to ship scallop seed out of water, but it was not known whether this method is useful to transport adult organisms. As a consequence, the purpose of this study was to evaluate the effects of transportation by the emersion method on the physiological status of the adductor muscle of adult giant lion's paw scallop. Live specimens were packaged in a container and transported in emersion for 11 h. Six scallops were frozen in a farm and a similar number were frozen as soon as they arrived to the laboratory. Physiological indices were determined in each lot and the survival was estimated 24 h after re‐immersion. As a result of the transportation, a significant loss (P<0.05) of total carbohydrates, glycogen, adenosine 5′‐triphosphate and adenilated energetic charge, and a significant increase (P<0.05) in the free amino acids concentration were observed. Eighty‐eight per cent survival was achieved; therefore, we conclude that this method is appropriate for shipping live adult scallops of Nodipecten subnodosus.