Class P dye-decolorizing peroxidase gene: Degenerated primers design and phylogenetic analysis.

Dye-decolorizing peroxidases (DyPs) were classified as a new family of heme peroxidase in 2007. Produced by various bacteria, they are the first bacterial enzymes known able to degrade lignin and dyes, for which their application in waste treatment and pretreatment of lignocellulosic biomass could be envisaged. In this work, a PCR primer pair was created and tested that enabled the detection and quantification of a wide range of bacterial genes of P class DyP in complex matrices. In addition, a phylogenetic tree was built with all available sequences of DyP genes available, offering a first overview of their presence in the bacteria kingdom.

Isolation of bacterial strains able to metabolize lignin and lignin-related compounds.

UNLABELLED: In this study, we identified five strains isolated from soil and sediments able to degrade kraft lignin, aromatic dyes and lignin derivatives. Using 16S rRNA gene sequencing, the isolates were identified as Serratia sp. JHT01, Serratia liquefacien PT01, Pseudomonas chlororaphis PT02, Stenotrophomonas maltophilia PT03 and Mesorhizobium sp. PT04. All the isolates showed significant growth on lignin with no water-extractable compounds. Synthetic aromatic dyes were used to assess the presence of oxidative enzymes. All the isolates were able to use the thiazine dye Methylene blue and the anthraquinone dye Remazol Brilliant Blue R as the sole carbon source. Guaiacol, veratryl alcohol and biphenyl were also mineralized by all the strains isolated. These results suggest they could be used for the treatment of aromatic pollutants and for the degradation of the lignocellulosic biomass. SIGNIFICANCE AND IMPACT OF THE STUDY: The valorization of waste lignin and lignocellulosic biomass by biocatalysis opens up new possibilities for the production of value-added substituted aromatics, biofuel and for the treatment of aromatic pollutants. Bacteria with ligninolytic potential could be a source of novel enzymes for controlled lignin depolymerization. In this work, five soil bacteria were isolated and studied. Every isolate showed significant growth on lignin and was able to degrade several lignin monomers and ligninolytic indicator dyes. They could thus be a source of novel ligninolytic enzymes as well as candidates for a bacterial consortium for the delignification of lignocellulosic biomass.

Anaerobic co-digestion of waste activated sludge and greasy sludge from flotation process: batch versus CSTR experiments to investigate optimal design.

In this study, the maximum ratio of greasy sludge to incorporate with waste activated sludge was investigated in batch and CSTR experiments. In batch experiments, inhibition occurred with a greasy sludge ratio of more than 20-30% of the feed COD. In CSTR experiments, the optimal greasy sludge ratio was 60% of the feed COD and inhibition occurred above a ratio of 80%. Hence, batch experiments can predict the CSTR yield when the degradation phenomenon are additive but cannot be used to determine the maximum ratio to be used in a CSTR configuration. Additionally, when the ratio of greasy sludge increased from 0% to 60% of the feed COD, CSTR methane production increased by more than 60%. When the greasy sludge ratio increased from 60% to 90% of the feed COD, the reactor yield decreased by 75%.

Sulphur fate and anaerobic biodegradation potential during co-digestion of seaweed biomass (Ulva sp.) with pig slurry.

Seaweed (Ulva sp.) stranded on beaches were utilized as co-substrate for anaerobic digestion of pig slurry in three-month co-digestion tests in pilot scale anaerobic digesters in the laboratory. The methanogenic potential of Ulva sp. was low compared to that of other potential co-substrates available for use by farmers: 148 N m3CH4/t of volatile solids or 19 N m3CH4/t of crude product. When used as a co-substrate with pig manure (48%/52% w/w), Ulva sp. seaweed did not notably disrupt the process of digestion; however, after pilot stabilisation, biogas produced contained 3.5% H2S, making it unsuitable for energy recovery without treatment. Sequentially addition of the sulphate reduction inhibitor, potassium molybdate, to a final concentration of 3mM, temporarily reduced H2S emissions, but was unable to sustain this reduction over the three-month period. According to these pilot tests, the use of seaweed stranded on beaches as co-substrate in farm-based biogas plants shows some limitations.

Impact of nitrate-enhanced leachate recirculation on gaseous releases from a landfill bioreactor cell.

This study evaluates the impact of nitrate injection on a full scale landfill bioreactor through the monitoring of gaseous releases and particularly N(2)O emissions. During several weeks, we monitored gas concentrations in the landfill gas collection system as well as surface gas releases with a series of seven static chambers. These devices were directly connected to a gas chromatograph coupled to a flame ionisation detector and an electron capture detector (GC-FID/ECD) placed directly on the field. Measurements were performed before, during and after recirculation of raw leachate and nitrate-enhanced leachate. Raw leachate recirculation did not have a significant effect on the biogas concentrations (CO(2), CH(4) and N(2)O) in the gas extraction network. However, nitrate-enhanced leachate recirculation induced a marked increase of the N(2)O concentrations in the gas collected from the recirculation trench (100-fold increase from 0.2 ppm to 23 ppm). In the common gas collection system however, this N(2)O increase was no more detectable because of dilution by gas coming from other cells or ambient air intrusion. Surface releases through the temporary cover were characterized by a large spatial and temporal variability. One automated chamber gave limited standard errors over each experimental period for N(2)O releases: 8.1 +/- 0.16 mg m(-2) d(-1) (n = 384), 4.2 +/- 0.14 mg m(-2) d(-1) (n = 132) and 1.9 +/- 0.10 mg m(-2) d(-1) (n = 49), during, after raw leachate and nitrate-enhanced leachate recirculation, respectively. No clear correlation between N(2)O gaseous surface releases and recirculation events were evidenced. Estimated N(2)O fluxes remained in the lower range of what is reported in the literature for landfill covers, even after nitrate injection.

The efficiency of biological aerobic treatment of piggery wastewater to control nitrogen, phosphorus, pathogen and gas emissions.

Due to the water pollution and in order to reduce the nitrogen load applied on soils, biological nitrogen removal treatment of piggery wastewaters was developed in Brittany (France), with 250-300 units running. Four types of treatment processes were built including a biological reactor allowing to remove about 60-70% of the nitrogen content as gas by nitrification/denitrification. The addition of different mechanical separators (screw-press, centrifuge decanter ...) led to concentration of phosphorus in an exportable solid phase, allowing a reduction up to 80% of the phosphorus applied locally on soils. Moreover, a reduction of the gaseous emissions was observed using this management process as compared to conventional management (storage + land spreading) including ammonia (up to 68%) and greenhouse gases (55%). Finally, the level of enteric and pathogenic bacteria was also decreased with the treatment process as compared to conventional management systems. However, in spite of these results, the significant cost of the treatment must be underlined and alternative systems including anaerobic digestion will have to be studied.

Long term fate of slurry derived nitrogen in soil: a case study with a macro-lysimeter experiment having received high loads of pig slurry (Solepur).

In intensive livestock production areas, land application remains the traditional management of manure and slurries for nutrient recycling. For sustainable agriculture there is fear, however, that this practice may have detrimental effects, particularly on the depletion of Soil Organic Matter associated with pig slurry applications. We investigated the long-term fate of nitrogen in a reconstituted soil having received high doses of pig slurry during 5 years (1991-1995). After 5 years of intensive application rates (nearly 1000 m(3)yr(-1)), the N and C content of the soil profile (0-20 cm) had increased by about 60% and 50%, respectively. These results confirm previous findings although it seems that the particularly high rates of application may explain, in part, the relatively important N incorporation in soil. Pig slurry applications ceased in 1995 and nitrogen content in soil and drainage water have been monitored. Apparent mineralization rates were calculated from the decrease in N content of the soil. This analysis indicated that more than 50% of the added N stored in the soil at the end of the applications would eventually be mineralized, leaving nearly 50% of the stored N to be immobilized in the soil. These results are the first published of their kinds, as most reports never examine the fate of applied pig slurry N after halting applications. In addition the few reports on long-term experiments suggest that Soil Organic Matter following pig slurry applications may be unstable. Our analysis tends to show the contrary. However, this conclusion must be tempered because data on nitrate leachate patterns suggest that soil management such as ploughing and sowing may actually trigger mineralization that could eventually deplete nitrogen stored following applications.