Optimisation of Nitrogen Supply from Sugarcane Residues in the Wet Tropics

Retention of sugarcane leaves and tops on the soil surface after harvesting has almost completely replaced burning of crop residues in the Australian sugar industry. Long term retention of residue is believed to improve soil fertility to the extent that nitrogen (N) fertilizer applications might be reduced by up to 40 kg N/ha/y. However, the fate of N in the extreme environment of the wet tropics is not known with certainty. Indices of potential N mineralisation and nitrification were developed and indicate that potential N fertility is greater in the wet tropics compared to more southern cane growing areas, and is enhanced under residue retention. Field results from the wet tropics support this prediction, but indicate high soil ammonium-N concentrations relative to nitrate-N.

Multiple roles of charged amino acids in cytoplasmic loop 7 for expression and function of the multidrug and organic anion transporter MRP1 (ABCC1)

Multidrug resistance protein MRP1 mediates the ATP-dependent efflux of many chemotherapeutic agents and organic anions. MRP1 has two nucleotide binding sites (NBSs) and three membrane spanning domains (MSDs) containing 17 transmembrane helices linked by extracellular and cytoplasmic loops (CL). Homology models suggest that CL7 (amino acids 1141-1195) is in a position where it could participate in signaling between the MSDs and NBSs during the transport process. We have individually replaced eight charged residues in CL7 with Ala, and in some cases, an amino acid with the same charge, and then investigated the effects on MRP1 expression, transport activity, and nucleotide and substrate interactions. A triple mutant in which Glu(1169), Glu(1170), and Glu(1172) were all replaced with Ala was also examined. The properties of R1173A and E1184A were comparable with those of wild-type MRP1, whereas the remaining mutants were either poorly expressed (R1166A, D1183A) or exhibited reduced transport of one or more organic anions (E1144A, D1179A, K1181A, (1169)AAQA). Same charge mutant D1183E was also not expressed, whereas expression and activity of R1166K were similar to wild-type MRP1. The moderate substrate-selective changes in transport activity displayed by mutants E1144A, D1179A, K1181A, and (1169)AAQA were accompanied by changes in orthovanadate-induced trapping of [alpha-(32)P]azidoADP by NBS2 indicating changes in ATP hydrolysis or release of ADP. In the case of E1144A, estradiol glucuronide no longer inhibited trapping of azidoADP. Together, our results demonstrate the extreme sensitivity of CL7 to mutation, consistent with its critical and complex dual role in both the proper folding and transport activity of MRP1.

Fast pyrolysis and nitrogenolysis of biomass and biogenic residues:production of a sustainable slow release fertiliser

The production of agricultural and horticultural products requires the use of nitrogenous fertiliser that can cause pollution of surface and ground water and has a large carbon footprint as it is mainly produced from fossil fuels. The overall objective of this research project was to investigate fast pyrolysis and in-situ nitrogenolysis of biomass and biogenic residues as an alternative route to produce a sustainable solid slow release fertiliser mitigating the above stated problems. A variety of biomasses and biogenic residues were characterized by proximate analysis, ultimate analysis, thermogravimetric analysis (TGA) and Pyrolysis – Gas chromatography – Mass Spectroscopy (Py–GC–MS) for their potential use as feedstocks using beech wood as a reference material. Beech wood was virtually nitrogen free and therefore suitable as a reference material as added nitrogen can be identified as such while Dried Distillers Grains with Solubles (DDGS) and rape meal had a nitrogen content between 5.5wt.% and 6.1wt.% qualifying them as high nitrogen feedstocks. Fast pyrolysis and in-situ nitrogenolysis experiments were carried out in a continuously fed 1kg/h bubbling fluidized bed reactor at around 500°C quenching the pyrolysis vapours with isoparaffin. In-situ nitrogenolysis experiments were performed by adding ammonia gas to the fast pyrolysis reactor at nominal nitrogen addition rates between 5wt.%C and 20wt.%C based on the dry feedstock’s carbon content basis. Mass balances were established for the processing experiments. The fast pyrolysis and in-situ nitrogenolysis products were characterized by proximate analysis, ultimate analysis and GC– MS. High liquid yields and good mass balance closures of over 92% were obtained. The most suitable nitrogen addition rate for the in-situ nitrogenolysis experiments was determined to be 12wt.%C on dry feedstock carbon content basis. However, only a few nitrogen compounds that were formed during in-situ nitrogenolysis could be identified by GC–MS. A batch reactor process was developed to thermally solidify the fast pyrolysis and in-situ nitrogenolysis liquids of beech wood and Barley DDGS producing a brittle solid product. This was obtained at 150°C with an addition of 2.5wt% char (as catalyst) after a processing time of 1h. The batch reactor was also used for modifying and solidifying fast pyrolysis liquids derived from beech wood by adding urea or ammonium phosphate as post processing nitrogenolysis. The results showed that this type of combined approach was not suitable to produce a slow release fertiliser, because the solid product contained up to 65wt.% of highly water soluble nitrogen compounds that would be released instantly by rain. To complement the processing experiments a comparative study via Py–GC–MS with inert and reactive gas was performed with cellulose, hemicellulose, lignin and beech wood. This revealed that the presence of ammonia gas during analytical pyrolysis did not appear to have any direct impact on the decomposition products of the tested materials. The chromatograms obtained showed almost no differences between inert and ammonia gas experiments indicating that the reaction between ammonia and pyrolysis vapours does not occur instantly. A comparative study via Fourier Transformed Infrared Spectroscopy of solidified fast pyrolysis and in-situ nitrogenolysis products showed that there were some alterations in the spectra obtained. A shift in frequencies indicating C=O stretches typically related to the presence of carboxylic acids to C=O stretches related to amides was observed and no double or triple bonded nitrogen was detected. This indicates that organic acids reacted with ammonia and that no potentially harmful or non-biodegradable triple bonded nitrogen compounds were formed. The impact of solid slow release fertiliser (SRF) derived from pyrolysis and in-situ nitrogenolysis products from beech wood and Barley DDGS on microbial life in soils and plant growth was tested in cooperation with Rothamsted Research. The microbial incubation tests indicated that microbes can thrive on the SRFs produced, although some microbial species seem to have a reduced activity at very high concentrations of beech wood and Barley DDGS derived SRF. The plant tests (pot trials) showed that the application of SRF derived from beech wood and barley DDGS had no negative impact on germination or plant growth of rye grass. The fertilizing effect was proven by the dry matter yields in three harvests after 47 days, 89 days and 131 days. The findings of this research indicate that in general a slow release fertiliser can be produced from biomass and biogenic residues by in-situ nitrogenolysis. Nevertheless the findings also show that additional research is necessary to identify which compounds are formed during this process.

Pyrolysis and gasification of biomass and acid hydrolysis residues

This research was carried for an EC supported project that aimed to produce ethyl levulinate as a diesel miscible biofuel from biomass by acid hydrolysis. The objective of this research was to explore thermal conversion technologies to recover further diesel miscible biofuels and/or other valuable products from the remaining solid acid hydrolysis residues (AHR). AHR consists of mainly lignin and humins and contains up to 80% of the original energy in the biomass. Fast pyrolysis and pyrolytic gasification of this low volatile content AHR was unsuccessful. However, successful air gasification of AHR gave a low heating value gas for use in engines for power or heat with the aim of producing all the utility requirements in any commercial implementation of the ethyl levulinate production process. In addition, successful fast pyrolysis of the original biomass gave organic liquid yields of up to 63.9 wt.% (dry feed basis) comparable to results achieved using a standard hardwood. The fast pyrolysis liquid can be used as a fuel or upgraded to biofuels. A novel molybdenum carbide catalyst was tested in fast pyrolysis to explore the potential for upgrading. Although there was no deoxygenation, some bio-oil properties were improved including viscosity, pH and homogeneity through decreasing sugars and increasing furanics and phenolics. AHR gasification was explored in a batch gasifier with a comparison with the original biomass. Refractory and low volatile content AHR gave relatively low gas yields (74.21 wt.%), low tar yields (5.27 wt.%) and high solid yields (20.52 wt.%). Air gasification gave gas heating values of around 5MJ/NM3, which is a typical value, but limitations of the equipment available restricted the extent of process and product analysis. In order to improve robustness of AHR powder for screw feeding into gasifiers, a new densification technique was developed based on mixing powder with bio-oil and curing the mixture at 150°C to polymerise the bio-oil.

Method development for the analysis of smokeless powders and organic gunshot residue by ultra performance liquid chromatography with tandem mass spectrometry

The goal of this project was to develop a rapid separation and detection method for analyzing organic compounds in smokeless powders and then test its applicability on gunshot residue (GSR) samples. In this project, a total of 20 common smokeless powder additives and their decomposition products were separated by ultra performance liquid chromatography (UPLC) and confirmed by tandem mass spectrometry (MS/MS) using multiple reaction monitoring mode (MRM). Some of the targeted compounds included diphenylamines, centralites, nitrotoluenes, nitroglycerin, and various phthalates. The compounds were ionized in the MS source using simultaneous positive and negative electrospray ionization (ESI) with negative atmospheric pressure chemical ionization (APCI) in order to detect all compounds in a single analysis. The developed UPLC/MS/MS method was applied to commercially available smokeless powders and gunshot residue samples recovered from the hands of shooters, spent cartridges, and smokeless powder retrieved from unfired cartridges. Distinct compositions were identified for smokeless powders from different manufacturers and from separate manufacturing lots. The procedure also produced specific chemical profiles when tested on gunshot residues from different manufacturers. Overall, this thesis represents the development of a rapid and reproducible procedure capable of simultaneously detecting the widest possible range of components present in organic gunshot residue.^

Factors and mechanisms controlling soil organic carbon turnover in subsoils of agricultural sites

Two-third of the terrestrial C is stored in soils, and more than 50% of soil organic C (SOC) is stored in subsoils from 30 – 100 cm. Hence, subsoil is important as a source or sink for CO2 in the global carbon cycle. Especially the stable organic carbon (OC) is stored in subsoil, as several studies have shown that subsoil OC is of a higher average age than topsoil OC. However, there is still a lack of knowledge regarding the mechanisms of C sequestration and C turnover in subsoil. Three main factors are discussed, which possibly reduce carbon turnover rates in subsoil: Resource limitation, changes in the microbial community, and changes in gas conditions. The experiments conducted in this study, which aimed to elucidate the importance of the mentioned factors, focused on two neighbouring arable sites, with depth profiles differing in SOC stocks: One Colluvic Cambisol (Cam) with high SOC contents (8-12 g kg-1) throughout the profile and one Haplic Luvisol (Luv) with low SOC contents (3-4 g kg-1) below 30 cm depth. The first experiment was designed to gain more knowledge regarding the microbial community and its influence on carbon sequestration in subsoil. Soil samples were taken at four different depths on the two sites. Microbial biomass C (MBC) was determined to identify depth gradients in relation to the natural C availability. Bacterial and fungal residues as well as ergosterol were determined to quantify changes in the in the microbial community composition. Multi-substrate-induced-respiration (MSIR) was used to identify shifts in functional diversity of the microbial community. The MSIR revealed that substrate use in subsoil differed significantly from that in topsoil and also differed highly between the two subsoils, indicating a strong influence of resource limitations on microbial substrate use. Amino sugar analysis and the ratio of ergosterol to microbial biomass C showed that fungal dominance decreased with depth. The results clearly demonstrated that microbial parameters changed with depth according to substrate availability. The second experiment was an incubation experiment using subsoil gas conditions with and without the addition of C4 plant residues. Soil samples were taken from topsoil and subsoil of the two sites. SOC losses during the incubation, were not influenced by the subsoil gas conditions. Plant-derived C losses were generally stronger in the Cam (7.5 mg g-1), especially at subsoil gas conditions, than in the Luv (7.0 mg g-1). Subsoil gas conditions had no general effects on microbial measures with and without plant residue addition. However, the contribution of plant-derived MBC to total MBC was significantly reduced at subsoil gas conditions. This lead to the conclusion that subsoil gas conditions alter the metabolism of microorganisms but not the degradation of added plant residues is general. The third experiment was a field experiment carried out for two years. Mesh bags containing original soil material and maize root residues (C4 plant) were buried at three different depths at the two sites. The recovery of the soilbags took place 12, 18, and 24 months after burial. We determined the effects of these treatments on SOC, density fractions, and MBC. The mean residence time for maize-derived C was similar at all depths and both sites (403 d). MBC increased to a similar extent (2.5 fold) from the initial value to maximum value. This increase relied largely on the added maize root residues. However, there were clear differences visible in terms of the substrate use efficiency, which decreased with depth and was lower in the Luv than in the Cam. Hence freshly added plant material is highly accessible to microorganisms in subsoil and therefore equally degraded at both sites and depths, but its metabolic use was determined by the legacy of soil properties. These findings provide strong evidence that resource availability from autochthonous SOM as well as from added plant residues have a strong influence on the microbial community and its use of different substrates. However, under all of the applied conditions there was no evidence that complex substrates, i.e. plant residues, were less degraded in subsoil than in topsoil.

Dual layer solid phase extraction based on molecular imprinting technology: Seeking a route to enhance selectivity for trace analysis of pesticide residues in olive oil.

Aiming to introduce a multiresidue analysis for the trace detection of pesticide residues belonging to organophosphorus and triazine classes from olive oil samples, a new sample preparation methodology comprising the use of a dual layer of “tailor-made” molecularly imprinted polymers (MIPs) SPE for the simultaneous extraction of both pesticides in a single procedure has been attempted. This work has focused on the implementation of a dual MIP-layer SPE procedure (DL-MISPE) encompassing the use of two MIP layers as specific sorbents. In order to achieve higher recovery rates, the amount of MIP layers has been optimized as well as the influence of MIP packaging order. The optimized DL-MISPE approach has been used in the preconcentration of spiked organic olive oil samples with concentrations of dimethoate and terbuthylazine similar to the maximum residue limits and further quantification by HPLC. High recovery rates for dimethoate (95%) and terbuthylazine (94%) have been achieved with good accuracy and precision. Overall, this work constitutes the first attempt on the development of a dual pesticide residue methodology for the trace analysis of pesticide residues based on molecular imprinting technology. Thus, DL-MISPE constitutes a reliable, robust, and sensitive sample preparation methodology that enables preconcentration of the target pesticides in complex olive oil samples, even at levels similar to the maximum residue limits enforced by the legislation.

A survey on the composition of wines made with grapes produced by an organic system.

Summary There is concern about the health problems caused by pesticides in humans, which has led some grape producers to adopt organic procedures in their vineyards, and a certain amount of these grapes are directed to winemaking. Despite the approval awarded to this organic grape production by the certified organizations, there has been a demand to carry out a survey to determine the physicochemical composition of the wine derived from these products. Some of these wines were made from a single grape variety and others from more than one. For this survey, the samples consisted of five bottles of each type of wine, acquired from wineries and supermarkets in the Serra Gaúcha region, RS, Brazil. The analyses were carried out by physicochemical methods: volatile compounds by gas chromatography; minerals and trace elements by inductively coupled plasma optical emission spectrometry; and pesticide residues by liquid chromatography-mass spectrometry. The results showed that in general the physicochemical composition of these wines was within the limits established by Brazilian legislation. The mineral and trace element concentrations were very low and pesticide residues were not detected (MRL = 10 μg.kg?1) in any of the wines. Resumo: Há preocupação em relação a problemas causados por pesticidas no ser humano, o que levou uma parcela de viticultores a adotar procedimentos de agricultura orgânica em seus vinhedos. Assim, devido a essa preocupação, eles estão produzindo uva pelo sistema orgânico, sendo que uma parte dessa produção é direcionada à elaboração de vinho. Apesar de essas uvas terem sido aprovadas por entidades certificadoras, houve demanda para a realização de um levantamento para determinar a composição dos vinhos delas derivados. Portanto, cinco garrafas de cada tipo de vinho foram coletadas em vinícolas e supermercados da Serra Gaúcha, Rio Grande do Sul, sendo uma parte de vinhos varietais e outra, de cortes de diferentes variedades. As análises foram feitas por métodos físico-químicos: compostos voláteis, por cromatografia gasosa; minerais e elementos-traço, por espectrometria de emissão ótica com plasma acoplado indutivamente, e resíduos de pesticidas, por cromatografia líquida-espectrometria de massa. Os resultados mostram que a composição físico-química desses vinhos e dos minerais situou-se, em geral, dentro dos limites da legislação brasileira. As concentrações de minerais e de elementos-traço foram muito baixas, e não foram detectados resíduos de pesticidas (LMR = 10 μg.kg?1) em nenhum vinho.

Fast Detection and Chemical Characterization of Gunshot Residues by CMV-GC-MS and LIBS

Gunshot residue (GSR) is the term used to describe the particles originating from different parts of the firearm and ammunition during the discharge. A fast and practical field tool to detect the presence of GSR can assist law enforcement in the accurate identification of subjects. A novel field sampling device is presented for the first time for the fast detection and quantitation of volatile organic compounds (VOCs). The capillary microextraction of volatiles (CMV) is a headspace sampling technique that provides fast results (< 2 min. sampling time) and is reported as a versatile and high-efficiency sampling tool. The CMV device can be coupled to a Gas Chromatography-Mass Spectrometry (GC-MS) instrument by installation of a thermal separation probe in the injection port of the GC. An analytical method using the CMV device was developed for the detection of 17 compounds commonly found in polluted environments. The acceptability of the CMV as a field sampling method for the detection of VOCs is demonstrated by following the criteria established by the Environmental Protection Agency (EPA) compendium method TO-17. The CMV device was used, for the first time, for the detection of VOCs on swabs from the hands of shooters, and non-shooters and spent cartridges from different types of ammunition (i.e., pistol, rifle, and shotgun). The proposed method consists in the headspace extraction of VOCs in smokeless powders present in the propellant of ammunition. The sensitivity of this method was demonstrated with method detection limits (MDLs) 4-26 ng for diphenylamine (DPA), nitroglycerine (NG), 2,4-dinitrotoluene (2,4-DNT), and ethyl centralite (EC). In addition, a fast method was developed for the detection of the inorganic components (i.e., Ba, Pb, and Sb) characteristic of GSR presence by Laser Induced Breakdown Spectroscopy (LIBS). Advantages of LIBS include fast analysis (~ 12 seconds per sample) and good sensitivity, with expected MDLs in the range of 0.1-20 ng for target elements. Statistical analysis of the results using both techniques was performed to determine any correlation between the variables analyzed. This work demonstrates that the information collected from the analysis of organic components has the potential to improve the detection of GSR.

Carbon in humic fractions of organic matter in soil treated with organic composts under mango cultivation.

Soil organic matter (SOM) plays a key role in maintaining the productivity of tropical soils, providing energy and substrate for the biological activity and modifying the physical and chemical characteristics that ensure the maintenance of soil quality and the sustainability of ecosystems. This study assessed the medium-term effect (six years) of the application of five organic composts, produced by combining different agro-industrial residues, on accumulation and chemical characteristics of soil organic matter. Treatments were applied in a long-term experiment of organic management of mango (OMM) initiated in 2005 with a randomized block design with four replications. Two external areas, one with conventional mango cultivation (CMM) and the other a fragment of regenerating Caatinga vegetation (RCF), were used as reference areas. Soil samples were collected in the three management systems from the 0.00-0.05, 0.05-0.10, and 0.10-0.20 m layers, and the total organic carbon content and chemical fractions of organic matter were evaluated by determining the C contents of humin and humic and fulvic acids. Organic compost application significantly increased the contents of total C and C in humic substances in the experimental plots, mainly in the surface layer. However, compost 3 (50 % coconut bagasse, 40 % goat manure, 10 % castor bean residues) significantly increased the level of the non-humic fraction, probably due to the higher contents of recalcitrant material in the initial composition. The highest increases from application of the composts were in the humin, followed by the fulvic fraction. Compost application increased the proportion of higher molecular weight components, indicating higher stability of the organic matter.

Characterisation of indoor air organic pollutants in Brisbane

The occurrence and levels of airborne polycyclic aromatic hydrocarbons and volatile organic compounds in selected non-industrial environments in Brisbane have been investigated as part of an integrated indoor air quality assessment program. The most abundant and most frequently encountered compounds include, nonanal, decanal, texanol, phenol, 2-ethyl-1-hexanol, ethanal, naphthalene, 2,6-tert-butyl-4-methyl-phenol (BHT), salicylaldehyde, toluene, hexanal, benzaldehyde, styrene, ethyl benzene, o-, m- and pxylenes, benzene, n-butanol, 1,2-propandiol, and n-butylacetate. Many of the 64 compounds usually included in the European Collaborative Action method of TVOC analysis were below detection limits in the samples analysed. In order to extract maximum amount of information from the data collected, multivariate data projection methods have been employed. The implications of the information extracted on source identification and exposure control are discussed.