Chemical characterization of fatty acids, alkanes, n-diols and alkyl esters produced by a mixed culture of Trichoderma koningii and Penicillium janthinellum grown aerobically on undecanoic acid, potatoe dextrose and their mixture.

Little is known about the mixed fungal synthesis of high-value aliphatics derived from the metabolism of simple and complex carbon substrates. Trichoderma koningii and Penicillium janthinellum were fed with undecanoic acid (UDA), potatoe dextrose broth (PDB), and their mixture. Pyrolysis Field Ionization Mass Spectrometry (Py-FIMS) together with (1)H and (13)C Nuclear Magnetic Resonance (NMR) characterized CHCl3 soluble aliphatics in the fungal cell culture. Data from NMR and Py-FIMS analysis were complementary to each other. On average, the mixed fungal species produced mostly fatty acids (28% of total ion intensity, TII) > alkanes (2% of TII) > n-diols (2% of TII) > and alkyl esters (0.8% of TII) when fed with UDA, PDB or UDA+PDB. The cell culture accumulated aliphatics extracellularly, although most of the identified compounds accumulated intracellularly. The mixed fungal culture produced high-value chemicals from the metabolic conversion of simple and complex carbon substrates.

Metabolism of nC11 fatty acid fed to Trichoderma koningii and Penicillium janthinellum II: Production of intracellular and extracellular lipids.

Little is known about the fungal metabolism of nC10 and nC11 fatty acids and their conversion into lipids. A mixed batch culture of soil fungi, T. koningii and P. janthinellum, was grown on undecanoic acid (UDA), a mixture of UDA and potato dextrose broth (UDA+PDB), and PDB alone to examine their metabolic conversion during growth. We quantified seven intracellular and extracellular lipid classes using Iatroscan thin-layer chromatography with flame ionization detection (TLC-FID). Gas chromatography with flame ionization detection (GC-FID) was used to quantify 42 individual fatty acids. Per 150 mL culture, the mixed fungal culture grown on UDA+PDB produced the highest amount of intracellular (531 mg) and extracellular (14.7 mg) lipids during the exponential phase. The content of total intracellular lipids represented 25% of the total biomass-carbon, or 10% of the total biomass dry weight produced. Fatty acids made up the largest class of intracellular lipids (457 mg/150 mL culture) and they were synthesized at a rate of 2.4 mg/h during the exponential phase, and decomposed at a rate of 1.8 mg/h during the stationary phase, when UDA+PDB was the carbon source. Palmitic acid (C16:0), stearic acid (C18:0), oleic acid (C18:1), linoleic acid (C18:2) and vaccenic acid (C18:1) accounted for >80% of the total intracellular fatty acids. During exponential growth on UDA+PDB, hydrocarbons were the largest pool of all extracellular lipids (6.5 mg), and intracellularly they were synthesized at a rate of 64 µg/h. The mixed fungal species culture of T. koningii and P. janthinellum produced many lipids for potential use as industrial feedstocks or bioproducts in biorefineries.

Wheat straw biomass: a resource for high-value chemicals.

Two methods are proposed for increasing the commercial value of wheat straw based on its chemical constituents. The first method involves the determination and extraction of the major organic components of wheat straw, and the second involves those found and extracted in the aqueous and viscous biooils derived from the straw by fast pyrolysis. We used pyrolysis-field ionization mass spectrometry to identify the fine chemicals, which have high commercial values. The most abundant organic compounds in the wheat straw and biooil used as precursors for green chemicals are N-heterocycles (16 to 29% of the Total Ion Intensities, TII) and fatty acids (19 to 26% of TIIs), followed by phenols and lignins (12 to 23% of TIIs). Other important precursors were carbohydrates and amino acids (1 to 8% TIIs), n-alkyl benzenes (3 to 5% of TIIs), and diols (4 to 9% TIIs). Steroids and flavonoids represented 1 to 5% of TIIs in the three materials. Examples of valuable chemical compounds that can be extracted from the wheat straw and biooils are m/z 256, 270, 278, 280, 282 and 284, which are the n-C16 and n-C17 fatty acids respectively, and the C18:3, C18:2 and C18:1 unsaturated fatty acids. In particular, the C18:2 (linoleic acid) is present at a concentration of 1.7% of TIIs. Pyrazole, pyrazine, pyridine, indoles, quinolines, carbazoles, and their identified derivatives are found in relatively high concentrations (1 to 8% of TIIs). Other useful compounds are sterols such as m/z 412 (stigmasterol), m/z 414 (ß-sitosterol), and steroids such m/z 394 (stigmastatriene), m/z 398 (stigmastene) and m/z 410 (stigmastadienone). Relative to the wheat straw, the relative concentration of all flavonoids such as m/z 222 (flavone) and m/z 224 (flavonone) doubled in the biooils. The conversion of wheat straw by fast pyrolysis, followed by chemical characterization with mass spectrometry, and extraction of fine chemicals, opens up new possibilities for increasing the monetary value of crop residues.

Production of a refined biooil derived by fast pyrolysis of chicken manure with chemical and physical characteristics close to those of fossil fuels.

The chemical and physical properties of raw biooils prevent their direct use in combustion engines. We processed raw pyrolytic biooil derived from chicken manure to yield a colorless refined biooil with diesel qualities. Chemical characterization of the refined biooil involved elemental and several spectroscopic analyses. The physical measurements employed were viscosity, density and heat of combustion. The elemental composition (% wt/wt) of the refined biooil was 82.7 % C, 15.3 % H, 0.2 % N and 1.8 % O, no S. Its viscosity was 0.006 Pa.s and a heat of combustion of 43 MJ kg(-1). The refined biooil fraction contains n-alkanes, ranging from n-C(14) to n-C(27), alkenes varying from C(10:1) to C(22:1), and long-chain alcohols. The refined biooil makes a good diesel fuel due to its chemical and physical properties.

Separation and identification of heterocyclic nitrogen compounds in biooil derived by fast pyrolysis of chicken manure.

N-heterocyclics were separated from a biooil, generated by the pyrolysis of chicken manures by column chromatography over neutral alumina and silica, and identified by Pyrolysis Field Ionization Mass Spectrometry (Py-FIMS) and Electrospray Ionization Mass Spectrometry (ESI-MS). Identities of chemical structures, whose presence was indicated by ESI-MS, were confirmed by comparing the Collision-Induced Dissociations (CID's) mass spectra of unknown and standards. The following seven base structures were identified: pyrazine, benzoquinoline, carbazole, phenylpyridine, indole, pyrazole and pyridine. Available hydrogens bonded to ring carbons and nitrogens on the seven N-heterocyclics were increasingly substituted by alkyl groups, mainly methylene groups (m/z 14) to yield mono-, di-, tri- methyl N-heterocyclics. In some instances, longer alkyl chains, such as ethyl, propyl, up to heptyl groups were the substituents.

Chemical composition of acid-base fractions separated from biooil derived by fast pyrolysis of chicken manure.

Our earlier investigations on the chemical composition of biooils derived by the fast pyrolysis of chicken manure revealed the presence of more than 500 compounds. In order to simplify this heterogeneous and complex chemical system, we produced four biooil fractions namely strongly acidic fraction A, weakly acidic fraction B, basic fraction C and neutral fraction D on the basis of their solubilities in aqueous solutions at different pHs. The yield (wt/wt.%) for fraction A was 3%, for fraction B 21.3%, for fraction C 2.4% and for fraction D 32.4%, respectively. The four fractions were analyzed by elemental analyses, Fourier Transform infrared spectrophotometry (FTIR), (1)H and (13)C nuclear magnetic spectroscopy (NMR), and electrospray ionization mass spectrometry (ESI-MS). The major components of the four fractions were saturated and unsaturated fatty acids, N-heterocyclics, phenols, sterols, diols and alkylbenzenes. The pH separation system produced fractions of enhanced chemical homogeneity.

The conversion of chicken manure to bio-oil by fast pyrolysis. III. Analyses of chicken manure, bio-oils and char by Py-FIMS and Py-FDMS.

Fast pyrolysis of chicken manure produced the following three fractions: bio-oil Fraction I, bio-oil Fraction II, and a char. In a previous investigation we analyzed each of the four materials by curie-point pyrolysis-gas chromatography/mass spectrometry (CpPy-FDMS). The objective of this article is to report on the analyses of the same chicken manure and the three fractions derived from it by fast pyrolysis. We now used pyrolysis-field ionization mass spectrometry (Py-FIMS) to characterize the three fractions. In addition, the two bio-oil materials were analyzed by pyrolysis-field desorption mass spectrometry (Py-FDMS). The use of both Py-FIMS and Py-FDMS produced signals over significantly wider mass ranges than did CpPy-GC/MS, and so allowed us to identify considerably larger numbers of constituents in each material. Individual compounds identified in the mass spectra were classified into the following twelve compound classes: (a) low molecular weight compounds (< m/z 62); (b) carbohydrates; (c) phenols + lignin monomers; (d) lignin dimers; (e) n-alkylbenzenes; (f) N-heterocyclics; (g) n-fatty acids; (h) n-alkanes; (i) alkenes; (j) sterols; (k) n-diols and (l) high molecular weight compounds (> m/z 562). Of special interest were the high abundances of low-molecular weight compounds in the two bio-oils which constituted close to one half of the two bio-oils. Prominent among these compounds were water, ammonia, acetic acid, acetamide, propyl radical, formamide and hydrogen cyanide. The main quantitative differences between the two bio-oils was that bio-oil Fraction I, as analyzed by the two mass spectrometric methods, contained lower concentrations of low-molecular weight compounds, carbohydrates, and N-heterocyclics than bio-oil Fraction II but was richer in lignin dimers, n-alkylbenzenes and aliphatics (n-fatty acids, n-alkanes, alkenes, and n-diols). Of special interest were the N-heterocyclics in the two bio-oils such as pyrazole, pyrazoline, substituted pyrroles, pyridine and substituted pyridines, substituted methoxazole, substituted pyrazines, indole and substituted indoles. Fatty acids in all four materials ranged from n-C(9) to n-C(33), alkanes from n-C(9) to n-C(40), alkenes from C(10:1) to C(40:1) and diols from n-C(7) to n-C(29). The chicken manure, bio-oil Fraction I, and char each contained about 4% sterols with cholesterol, ethylcholestriene, ergosterol, ethylcholestene, ethylcholesterol and beta -sitosterol as major components. Semi-quantitative estimates of the total materials identified by Py-FIMS were: chicken manure: 61.1%; bio-oil Fraction I: 81.3%; bio-oil Fraction II: 78.6%; char: 61.3%; and by Py-FDMS were: bio-oil Fraction I: 65.4%; bio-oil Fraction II: 70.0%.

The conversion of chicken manure to biooil by fast pyrolysis I. Analyses of chicken manure, biooils and char by 13C and 1H NMR and FTIR spectrophotometry.

Fast pyrolysis of chicken manure produced two biooils (Fractions I and II) and a residual char. All four materials were analyzed by chemical methods, 13C and 1H Nuclear Magnetic Resonance Spectrometry (13C and 1H NMR), and Fourier Transform Infrared Spectrosphotometry (FTIR). The char showed the highest C content and the highest aromaticity. Of the two biooils Fraction II was higher in C, yield and calorific value but lower in N than Fraction I. The S and ash content of the two biooil fractions were low. The Cross Polarization Magic Angle Spinning (CP-MAS) 13C NMR spectrum of the initial chicken manure showed it to be rich in cellulose, which was a major component of sawdust used as bedding material. Nuclear Magnetic Resonance (NMR) spectra of the two biooils indicated that Fraction I was less aromatic than Fraction II. Among the aromatics in the two biooils, we were able to tentatively identify N-heterocyclics like indoles, pyridines, and pyrazines. FTIR spectra were generally in agreement with the NMR data. FTIR spectra of both biooils showed the presence of both primary and secondary amides and primary amines as well as N-heterocyclics such as pyridines, quinolines, and pyrimidines. The FTIR spectrum of the char resembled that of the initial chicken manure except that the concentration of carbohydrates was lower.

The conversion of chicken manure to biooil by fast pyrolysis II. Analysis of chicken manure, biooils, and char by curie-point pyrolysis-gas chromatography/mass spectrometry (Cp Py-GC/MS).

The initial chicken manure and the three fractions derived from it by fast pyrolysis, that is, the two biooils Fractions I and II as well as the residual char were analyzed by Curie-point pyrolysis-gas chromatography/mass spectrometry (Cp Py-GC/MS). The individual compounds identified were grouped into the following six compound classes: (a) N-heterocyclics; (b) substituted furans; (c) phenol and substituted phenols; (d) benzene and substituted benzenes; (e) carbocyclics; and (f) aliphatics. Of special interest were the relatively high concentrations of N-heterocyclics in biooil Fraction II which was obtained in the highest yield and had the highest calorific value. Prominent N-heterocyclics in biooil Fraction II were methyl-and ethyl-substituted pyrroles, pyridines, pyrimidine, pyrazines, and pteridine. Also noteworthy was the high abundance of aliphatics in biooil Fraction I and the char. The alkanes and alkenes in biooil Fraction I ranged from n-C7 to n-C18 and C7:1 to C18:1, respectively, and those in the char from n-C7 to n-C19 and C7:1 to C19:1, respectively. The N-heterocyclics in the two biooil Fractions came from the chicken manure, from proteinaceous materials during fast pyrolysis or were formed during the fast pyrolysis manure conversion by the Maillard reaction which involved the formation of N-heterocyclics by amino acids interacting with sugars.

Moist olive husks addition to a silty clay soil: influence on microbial and biochemical parameters.

To investigate the effects of moist olive husks (MOH-residues) on soil respiration, microbial biomass, and enzymatic (o-diphenoloxidase, beta-glucosidase, dehydrogenase and alkaline phosphatase) activities, a silty clay soil was incubated with 0 (control), 8 x 10(3) (D), 16 x 10(3) (2D) and 80 x 10(3) (10D) kg ha-1 of MOH-residues on a dry weight basis. Soil respiration and microbial biomass data indicated that the addition of MOH-residues strongly increased microbial activity proportionally to the amounts added. Data of qCO2 suggested that the respiration to biomass ratio of the microbial population was strongly modified by MOH-residues additions during the first 90 days of incubation. The qCO2 data suggested a low efficiency in energy yields from C oxidation during the first 2 months of soil incubation. qFDA seemed to be relatively unaffected for treatments D and 2D as compared to the control, but was significantly lowered by the application of 10D, showing the lowest hydrolytic activity of microbial biomass in this treatment up to 360 days of incubation. o-Diphenoloxidase activity was delayed, and this delay was extended with the addition of larger quantities of MOH-residues. Alkaline phosphatase, beta-glucosidase and dehydrogenase activities were in line with the findings on microbial biomass changes and activities. The biological and biochemical data suggest that the addition of a large quantity of MOH-residues (80 x 10(3) kg ha-1) strongly modifies the soil characteristics affecting the r- and K-strategist populations, and that these changes last for at least the 360 days of incubation. The data also suggest that application rates exceeding 16 x 10(3) kg ha-1 are not recommended until the agro-chemical and -physical functions of the soil are further studied.

Co-composting of paper mill sludge and hardwood sawdust under two types of in-vessel processes.

Recycling of organic residues by composting is becoming an acceptable practice in our society. Co-composting dewatered paper mill sludge (PMS) and hardwood sawdust, two readily available materials in Canada, was investigated using uncontrolled and controlled in-vessel processes. The composted materials were characterized for total C and N, water-soluble, acid-hydrolyzable, and non-hydrolyzable N, extractable lipids, and by Fourier Transform Infrared (FT-IR) spectrophotometry. In the controlled scale process, the loss of organic matter was approximately 65% higher than in the uncontrolled process. After undergoing initial fluctuations in N fractions during the first two days of composting, by the end of the process, concentrations of water-soluble N decreased while those of acid-hydrolyzable and nonhydrolyzable N increased in the controlled process, whereas in the uncontrolled process, water-soluble N increased, but N in the other two fractions decreased continuously, indicating that the biochemical transformations of organic matter were not completed. Data on extractable lipids and FT-IR spectra suggest that the compost produced from the controlled process was bio-stable after 14 days, while the uncontrolled process was not stabilized after 18 days. In addition, FT-IR data suggest the biological activity during composting centered mainly on the degradation of aliphatic structures while aromatic structures were preserved. The co-composting of the PMS and hardwood sawdust can be successfully achieved if aeration, moisture, and bio available C/N ratios are optimized to reduce losses of N.

Extractable soil lipids and microbial activity as affected by Bt and non Bt maize grown on a silty clay loam soil.

Pyrolysis-gas (Py-GC) chromatography was used to characterize extractable lipids from Bt and non-Bt maize shoots and soils collected at time of harvesting. Py-GC-MS (mass spectrometry) showed that the concentrations of total alkenes identified in non-Bt shoots and soils were 47.9 and 21.3% higher than in Bt maize shoots and soils, respectively. N-alkanes identified were of similar orders of magnitude in Bt and non-Bt maize shoots, but were 28.6% higher in Bt than in non-Bt soils. Bt maize shoots contained 29.7% more n-fatty acids than non-Bt maize shoots, whereas the concentrations of n-fatty acids in Bt soils were twice as high as those in non-Bt soils. Concentrations of unsaturated fatty acids in Bt maize shoots were 22.1% higher than those in non-Bt maize shoots, while concentrations of unsaturated fatty acids were 22.5% higher in non-Bt than in Bt soils. The cumulative CO2-C evolved from soils under Bt and non-Bt crops was 30.5% lower under Bt as compared to non-Bt crops, whereas when maize shoots were added to Bt and non-Bt soils, the decrease in CO2-C evolved were 16.5 and 23.6%, respectively. Our data showed that the cultivation of Bt maize significantly increased the saturated to unsaturated lipid ratios in soils which appeared to negatively affect microbial activity.

Composting duck excreta enriched wood shavings: C and N transformations and bacterial pathogen reductions.

Composting of agricultural and domestic wastes is used increasingly to reduce weight, volume, and odor; destroy animal and plant pathogens; and improve the quality of end-products to be used as soil amendments and growth substrates. The objective of this study was to investigate the transformation of C and N and the survival of bacterial populations and pathogenic bacteria during in-vessel composting of duck excreta enriched wood shavings. Two feedstocks, collected on different dates, were composted (C1 and C2) in an enclosed hall system equipped with an electromechanical turner. Temperature was continuously recorded, whereas moisture content and bacterial counts were determined twice a week. Data showed that, although the N content of C2 was only half of that of C1, both materials were fully biostabilized at the end of the composting period as indicated by extractable lipid ratios. In the compost with the low C/N ratio (C1), all bacterial populations were eliminated, whereas fecal streptococci, total coliforms, and gram-negative bacteria were still present in C2 at the end of the composting period. Our results emphasize that the composting of manures and other organic wastes needs to be properly managed to stabilize C and N and to eliminate or reduce bacterial populations.