Composting and vermicomposting experiences in the treatment and bioconversion of asphaltens from the Prestige oil spill

This work illustrates the effectiveness of composting and vermicomposting in degrading fuel-in-water emulsions from oil spills (chapapote), and the isolation of potentially useful microorganisms for its biodegradation. Firstly, an alternative to the biodegradation of asphaltens from the Prestige oil spill (still present in some chapapote rafts in the Cantabrian coast) by means of the application of composting techniques to a microbial partnership acclimated to fuel-oil is offered. Our aim is that, after a relatively short period of time, the microorganisms can obtain its source of carbon and energy from asphaltens. The addition of metabolic co-substrates, like cow bed and potato peelings, allows the fragmentation of complex compounds into smaller structures, susceptible to further degradation. Afterwards, a maturation of the compost by means of a treatment with earthworms (Eisenia foetida) is necessary. Thus, through the vermicomposting it will be possible to obtain a valued product, useful in the processes of ground amendment, with little presence of asphaltens and occluded polycyclic aromatic hydrocarbons, rich in humus, and with an important bacterial flora of Bacillus genera, so that it can be typical of co-activators and accelerating products in composting processes. Along with this article, we show some parameters that control the evolution of the compost products (evolved gases, acidity, temperature and humidity); the chemical and microbiological analytical results; and the germination assays of vermicomposting. Results reveal that by using microorganisms living in either earthworm intestines (Stenotrophomonas maltophilia) or vermiculture substrates (Scedosporium apiospermium), it is possible to degrade and to eliminate the polycyclic asphaltens into CO(2) and H(2)O, helped by evaporation, dissolution and/or photo-oxidation processes. The obtained end product has contents of interesting vegetal nutrients and, mainly, it displays very high germination indices.     

Profiling bacterial survival through a water treatment process and subsequent distribution system

AIMS: To profile fractions of active bacteria and of bacteria culturable with routine heterotrophic plate count (HPC) methods through a typical water treatment process and subsequent distribution system. In doing so, investigate how water treatment affects both bacterial abundance and diversity, and reveal the identities of active bacteria not detected by traditional HPC culture. METHODS AND RESULTS: Profiling active fractions was performed by flow cytometric cell sorting of either membrane-intact (BacLight kit) or enzymatically active (carboxyfluorescein diacetate, CFDA) bacteria, followed by eubacterial 16S rDNA-directed PCR and denaturing gradient gel electrophoresis (DGGE). Water treatment significantly reduced active bacterial numbers detected by the BacLight kit and CFDA assay by 2.89 and 2.81 log respectively. Bacterial diversity was also reduced from > 20 DGGE bands in the active fractions of reservoir water to only two bands in the active fractions of finished water. These two bands represented Stenotrophomonas maltophila, initially culturable by HPC, and a Burkholderia-related species. Both species maintained measurable traits of physiological activity in distribution system bulk water but were undetected by HPC. CONCLUSIONS: Flow cytometric cell sorting with PCR-DGGE, to assess water treatment efficacy, identified active bacteria from a variety of major phylogenetic groups undetected by routine HPC. Following treatment S. maltophila and a Burkholderia-related species retained activity and entered distribution undetected by HPC. SIGNIFICANCE AND IMPACT OF THE STUDY: Methods used here demonstrate how water treatment operators can better monitor water treatment plant efficacy and assess distribution system instability by the detection and identification of active bacteria recalcitrant to routine HPC culture.     

Municipal solid waste management through vermicomposting employing exotic and local species of earthworms

A comparative study was conducted between exotic and local (epigeic--Eisenia fetida and anaecic--Lempito mauritii, respectively) species of earthworms for the evaluation of their efficacy in vermicomposting of municipal solid waste (MSW). Vermicomposting of MSW for 42 days resulted in significant difference between the two species in their performance measured as loss in total organic carbon, carbon-nitrogen ratio (C:N) and increase in total Kjeldahl nitrogen, electrical conductivity and total potassium and weight loss of MSW. The change in pH and increase in number of earthworms and cocoons and weight of earthworms were non-significant.     

Quality assessment of compost prepared from fly ash and crop residue

Fly ash was co-composted with wheat straw and 2% rock phosphate (w/w) for 90 days and different chemical and microbiological parameters monitored to evaluate its effect on the composting process. Fly ash addition at 20% level resulted in the lowest C/N of 16.4:1 and highest available and total phosphorus. Increasing the addition of fly ash from 40 to 60% (w/w) did not exert any detrimental effect on either C:N or the microbial population.     

Biomass yield and nitrogen fixation of legumes monocropped and intercropped with rye and rotation effects on a subsequent maize crop

The research is focused on an ecologically sound and highly productive cultivation system for fodder and/or biomass for thermal power generation on the basis of winter legumes and maize as subsequent summer crop, managed without additional nitrogen fertiliser. Therefore the yield of biomass and N-fixing capacity of a winter pea (Pisum sativum L.) and crimson clover (Trifolium incarnatum L.) monocropped and intercropped with rye (Secale cereale L.) were examined for five years in a field trial. In mid-June above-ground biomass of winter crops was removed and maize transplanted. The winter crops achieved maximum dry matter yield about three to five weeks before maturity. Mixed stands yielded more biomass than pure stands and exhibited greater yield stability. The relative advantage of intercropping, expressed as land equivalent ratio (LER), determined for intercropped winter pea/rye were 1.1 to 1.2 and for crimson clover/rye 1.3. At maturity, the amount of fixed nitrogen ranged between 178 kg N for crimson clover and 242 kg N ha^-1 for winter pea, respectively. At the end of anthesis (middle of June, harvesting stage for silage fodder) 75% and 88% of the total fixed nitrogen was achieved, for clover and pea, respectively. In intercropping the amount of fixed nitrogen was lower than in pure stands due to a lower seed density of the legume; however, the N-fixing efficiency was greater than in pure stands. N-release of the winter pea in a pure stand produced a maximum yield in maize (Zea mays L.) without additional N-fertiliser. An additional N mineral fertilisation of 75 to 150 kg N and 75 to 225 kg N was necessary to achieve maximum yields in maize following intercropped winter pea and crimson clover, respectively. Legumes in mixed stands with rye resulted in lower amounts of residual nitrogen after maize harvest. The beneficial effect of legumes on maize can be divided into N-effects and rotation effects. Both effects were positive regarding winter pea. The rotation effect of crimson clover in pure stands on maize was negative. Allelopathic effects and the high sensitivity of crimson clover to mineral nitrogen in the soil, released by residues of the preceding crop, winter rape (Brassica rapa L.), were discussed as the reason for this observation. The combination of the winter pea in pure stand and maize achieved the highest total biomass yield from winter and summer crops, unfertilised (156 dt ha^-1 dry). The combinations of intercropped legumes and maize produced biomass yields of 142 to 145 dt ha^-1. Because winter pea is highly susceptible to lodging, intercropping with low seed density of rye is recommended (3/4 winter pea, 1/4 rye). The rye crop prevents lodging by providing support and high rates of N-fixation are achieved with high seed density of pea. Intercropping with crimson clover and rye should be based on high seed densities of legumes, too because rye is highly competitive within those mixtures. This revised version was published online in August 2006 with corrections to the Cover Date.     

Nitrogen mineralization and denitrification as influenced by crop residue particle size

Managing the crop residue particle size has the potential to affect N conservation in agricultural systems. We investigated the influence of barley (Hordeum vulgare) and pea (Pisum sativum) crop residue particle size on N mineralization and denitrification in two laboratory experiments. Experiment 1: ¹⁵N-labelled ground (3 mm) and cut (25 mm) barley residue, and microcrystalline cellulose+glucose were mixed into a sandy loam soil with additional inorganic N. Experiment 2: inorganic¹⁵ N and C₂H₂ were added to soils with barley and pea material after 3, 26, and 109 days for measuring gross N mineralization and denitrification.Net N immobilization over 60 days in Experiment 1 cumulated to 63 mg N kg^-1 soil (ground barley), 42 (cut barley), and 122 (cellulose+glucose). More N was seemingly net mineralized from ground barley (3.3 mg N kg^-1 soil) than from cut barley (2.7 mg N kg^-1 soil). Microbial biomass peaked at day 4 with the barley treatments and at day 14 with the cellulose+glucose whereafter the biomass leveled out at values 79 mg C kg^-1 (ground), 104 (cut), and 242 (cellulose+glucose) higher than for the control soil. Microbial growth yields were similar for the two barley treatments, ca. 60 mg C g^-1 substrate C added, which was lower than the 142 mg C g^-1 C added with cellulose+glucose. This suggests that the 75% (w/w) holocelluloses and sugars contained with the barley material remained physically protected despite grinding. In Experiment 2 gross mineralization on day 3 was 4.8 mg N kg^-1 d^-1 with ground pea, twice as much as for all other treatments. On day 26 the treatment with ground barley had the greatest gross N mineralization. In static cores ground barley denitrified 11-fold more than did cut barley, whereas denitrification was similar for the two pea treatments. In suspensions denitrification was similar for the two treatments both with barley and pea residue.We conclude that the higher microbial activity associated with the initial decomposition of ground plant material is due to a more intimate plant residue-soil contact. On the long term, grinding the plant residues has no significant effect on N dynamics.     

Utility of some nitrogen-fixing microorganisms in the phyllosphere of crop plants

Summary The utility of spraying some known N₂-fixing microorganisms on rice leaves grown both in N-less sand culture and under field conditions was examined. The effect was compared with that of spraying a phyllosphere N₂-fixing isolate of Klebsiella, KUPBR₂, and application of nitrogenous fertilizers. All the growth parameters studied including dry weight and N-content were enhanced. Under field conditions number of tillers was increased by 26% withKlebsiella pneumoniae M5al and by 65% with Aphanothece. The dry weight of the plants was enhanced by 61–119%. The yield per 10 m² was almost doubled with Aphanothece, Beijerinckia 8007,Mycobacterium flavum, K. pneumoniae M5al and KUPBR₂. The increases observed withStreptomyces sp. G12 though less spectacular was significant at 1% level with respect to several growth parameters.K. pneumoniae M5al,M. flavum andStreptomyces sp. G12 exhibited nitrogenase activity both in laboratory culture and in association with rice plants.     

Association of earthworm-denitrifier interactions with increased emission of nitrous oxide from soil mesocosms amended with crop residue

Earthworm activity is known to increase emissions of nitrous oxide (N(2)O) from arable soils. Earthworm gut, casts, and burrows have exhibited higher denitrification activities than the bulk soil, implicating priming of denitrifying organisms as a possible mechanism for this effect. Furthermore, the earthworm feeding strategy may drive N(2)O emissions, as it determines access to fresh organic matter for denitrification. Here, we determined whether interactions between earthworm feeding strategy and the soil denitrifier community can predict N(2)O emissions from the soil. We set up a 90-day mesocosm experiment in which (15)N-labeled maize (Zea mays L.) was either mixed in or applied on top of the soil in the presence or absence of the epigeic earthworm Lumbricus rubellus and/or the endogeic earthworm Aporrectodea caliginosa. We measured N(2)O fluxes and tested the bulk soil for denitrification enzyme activity and the abundance of 16S rRNA and denitrifier genes nirS and nosZ through real-time quantitative PCR. Compared to the control, L. rubellus increased denitrification enzyme activity and N(2)O emissions on days 21 and 90 (day 21, P = 0.034 and P = 0.002, respectively; day 90, P = 0.001 and P = 0.007, respectively), as well as cumulative N(2)O emissions (76%; P = 0.014). A. caliginosa activity led to a transient increase of N(2)O emissions on days 8 to 18 of the experiment. Abundance of nosZ was significantly increased (100%) on day 90 in the treatment mixture containing L. rubellus alone. We conclude that L. rubellus increased cumulative N(2)O emissions by affecting denitrifier community activity via incorporation of fresh residue into the soil and supplying a steady, labile carbon source.     

Improved enantioselective conversion of styrene epoxides and meso-epoxides through epoxide hydrolases with a mutated nucleophile-flanking residue

In epoxide hydrolase from Agrobacterium radiobacter (EchA), phenylalanine 108 flanks the nucleophilic aspartate and forms part of the substrate-binding pocket. The influence of mutations at this position on the activity and enantioselectivity of the enzyme was investigated. Screening for improved enantioselectivity towards para-nitrophenyl glycidyl ether (pNPGE) using spectrophotometric progress curve analysis yielded five different mutants with 3- to 7-fold improved enantioselectivity. The increase in enantioselectivity was in most cases the result of an enhanced catalytic efficiency toward the preferred enantiomer. Several mutations at position F108 resulted in a higher activity toward cis-disubstituted meso-epoxides, which were converted to a single product enantiomer. Mutant F108C converted cis-2,3-epoxybutane to (2R,3R)-2,3-butanediol of >99% ee with a 7-fold improved activity, and mutant F108A hydrolyzed cyclohexene oxide to (1R,2R)-1,2-cyclohexanediol of >99% ee with a more than 150-fold higher activity than wild-type enzyme. It is concluded that single amino acid substitutions in the active site of epoxide hydrolase can result in enzyme variants with catalytic properties that are suitable for preparative scale production of (S)-epoxides and chiral vicinal diols in high yield and with excellent ee.     

Field evaluation of heat units for predicting crop residue carbon and nitrogen mineralization

More efficient fertilization practices could conceivably be developed if nutrient availability from manures and crop residues could be accurately predicted under field conditions. Heat units were recently found useful for describing the combined thermal and temporal relationships with papermill sludge C and N mineralization. This study was conducted to describe C and N mineralization of corn (Zea mays L. cv. ‘King 1113’), lupin (Lupinus albus L. cv. ‘ultra’), and potato (Solanum tuberosum L. cv. ‘russet burbank’) residues and to field test the utility of heat units in predicting these processes. Residues of each crop were applied to soil microplots in May, June, and July of 1987. Carbon mineralization was monitored by periodic capture of evolved CO₂ in alkali traps. Nitrogen mineralization-immobilization processes were followed by frequent sampling for KCl extractable inorganic N. Net N mineralization from the potato residue occurred almost immediately after residue application, apparently owing to its relatively low C/N of 15/1. Net N mineralization from the corn and lupin residues began 119, 99, and 317 d after application in the May, June, and July application months, respectively. These respective times represented 2346, 1990, and 2360 degree days after application. Heat units appear to provide a mathematically simple, pragmatic approach for predicting crop residue N mineralization under field climatic conditions.     

Analysis of the occurrence and distribution of tobacco rattle virus in field soil and disease in a subsequent tulip crop

The occurrence and distribution of tobacco rattle virus (TRV) in field plots was determined by soil bait-testing and disease incidence in tulips subsequently grown on these plots was studied. The virus occurred in patches, calculated as 1.5 m × 3.6 m. The presence of virus was not correlated with numbers of potential vector trichodorid nematodes. Of three trichodorid nematode species present, only Paratrichodorus teres transmitted TRV which, as with virus isolates obtained in bait-tests and from infected tulips, reacted in serological tests with an antiserum prepared against a Dutch isolate of pea-early browning virus (PEBV). Virus prevalence in a subsequent tulip crop was 0.8% and in a sample of tulip plants, virus was recovered only from plants showing virus symptoms. Plots from which TRV was recovered in bait-tests yielded significantly more virus diseased tulips than plots which tested negative for virus. Growing bait-plants in field-plots, as compared with greenhouse tests using soil collected as a series of sub-samples, resulted in an underestimate of the occurrence of TRV.     

Penetration of cover crop roots through compacted soils

Tap-rooted species may penetrate compacted soils better than fibrous-rooted species and therefore be better adapted for use in “biological tillage”. We evaluated penetration of compacted soils by roots of three cover crops: FR (forage radish: Raphanus sativus var. longipinnatus, cv. ‘Daikon’), rapeseed (Brassica napus, cv. ‘Essex’), two tap-rooted species in the Brassica family, and rye (cereal rye: Secale cereale L., cv. ‘Wheeler’), a fibrous-rooted species. Three compaction levels (high, medium and no compaction) were created by wheel trafficking. Cover crop roots were counted by the core-break method. At 15–50 cm depth under high compaction, FR had more than twice and rapeseed had about twice as many roots as rye in experiment 1; FR had 1.5 times as many roots as rye in experiment 2. Under no compaction, little difference in root vertical penetration among three cover crops existed. Rapeseed and rye root counts were negatively related to soil strength by linear and power functions respectively, while FR roots showed either no (Exp.1) or positive (Exp. 2) relationship with soil strength. We conclude that soil penetration capabilities of three cover crops were in the order of FR > rapeseed > rye.     

Effects of long-term treatment with acetylene on nitrogen-fixing microorganisms.

Long periods of experimental incubation with acetylene led to a multifold enhancement of acetylene-reducing activity in Anabaena cylindrica, Anabaenopsis circularis, Rhodospirillum rubrum, and Azotobacter vinelandii. Rates of acetylene reduction showed a gradual increase and reached a peak after 2 to 6 h of continuous incubation under acetylene. Thereafter, enzyme activity rapidly declined. A similar enhancement of ethylene production was observed when pretreatment with acetylene was interrupted periodically by a brief exposure to ambient (or oxygen-free) atmosphere without acetylene although the decline of acetylene-reducing activity was less rapid. Pretreatment with acetylene depressed photosynthetic 14CO2 fixation and 15N2 incorporation in Anabaena cylindrica. It is concluded that assessments based on long-term experimental incubation with acetylene may grossly overestimate the actual quantities of fixed nitrogen in the field.     

Effects of long-term treatment with acetylene on nitrogen-fixing microorganisms.

Long periods of experimental incubation with acetylene led to a multifold enhancement of acetylene-reducing activity in Anabaena cylindrica, Anabaenopsis circularis, Rhodospirillum rubrum, and Azotobacter vinelandii. Rates of acetylene reduction showed a gradual increase and reached a peak after 2 to 6 h of continuous incubation under acetylene. Thereafter, enzyme activity rapidly declined. A similar enhancement of ethylene production was observed when pretreatment with acetylene was interrupted periodically by a brief exposure to ambient (or oxygen-free) atmosphere without acetylene although the decline of acetylene-reducing activity was less rapid. Pretreatment with acetylene depressed photosynthetic 14CO2 fixation and 15N2 incorporation in Anabaena cylindrica. It is concluded that assessments based on long-term experimental incubation with acetylene may grossly overestimate the actual quantities of fixed nitrogen in the field.     

Nitrogen use in mixed tree crop plantations with a legume cover crop

Plant and Soil - In a multi-strata agroforestry system in central Amazonia, we studied the nitrogen (N) use of two indigenous fruit tree species, Theobroma grandiflorum Willd. (ex Spreng.) K....     

Cowpea N rhizodeposition and its below-ground transfer to a co-existing and to a subsequent millet crop on a sandy soil of the Sudano-Sahelian eco-zone

Nitrogen (N) rhizodeposition by cowpea (Vigna unguiculata (L.) Walp) is potentially a large N source in cropping systems of Sub-Saharan Africa. A field experiment was conducted to measure cowpea N rhizodeposition under the conditions of the Sudano-Sahelian zone using direct ¹⁵N labelling techniques to trace the amount of deposition and its transfer to associated and subsequent crops. Half of the total cowpea crop N was located below-ground at plant maturity, which exceeded 20 kg?N ha^?1 when intercropped with millet. Only 15% of the below-ground cowpea N was recovered in roots, while 85% was found in the rhizodeposited pools. The experiment demonstrated that direct below-ground N transfer occurred from cowpea to millet in intercrop at a rate of 2 kg?N ha^?1 over the growing season. Forty percent of the 25 kg below-ground N that the cowpea crop left at harvest were identifiable in the top 0.30 m soil in the beginning of the next planting season 7 months later; a pool still present at the end of that second season. Thus, the subsequent crop of millet (Pennisetum glaucum (L.) R. Br.) only recovered 2.5 kg?N ha^?1 from the below-ground cowpea pre-crop N during this growth season. The role and potential of cowpea as N provider has been underestimated in the past by ignoring the large proportion of N contained in its rhizodeposits. However, information is needed to determine how losses of the rhizodeposited N can be minimized to fully harness the potential of cowpea as N provider in agro-ecosystems of the region.