An Original Halo-Alkaline Protease from <Emphasis Type="Italic">Bacillus halodurans</Emphasis> Strain US193: Biochemical Characterization and Potential Use as Bio-Additive in Detergents

Over a hundred of halophilic/halotolerant microorganisms were screened for alkaline protease production. The bacterium showing the highest enzyme production was characterized and identified as Bacillus halodurans US193 on the basis of 16S rRNA gene analysis. It was alkalophilic, thermophilic and halotolerant since it grew optimally at pH 9.7 and 50?°C with tolerance of up to 125 g NaCl l^?1. The alkaline protease was purified 4.9 times with about 40186.1 U/mg as specific activity. It exhibited optimal activity at pH 10, 70?°C and 0.25 M NaCl with perfect stability at wide ranges of pH (6–12), temperatures (30–60?°C) and NaCl concentrations (0–2 M). The serine alkaline protease maintained high stability in the presence of Cu²⁺, Mg²⁺, Ba²⁺ and Ca²⁺ ions, various organic solvents [50% (v/v)] and ionic and non ionic detergent additives. In addition, it was more compatible with various commercialized detergents than other reported detergent proteases, and was very efficient in blood stain removal. These findings let B. halodurans US193 alkaline protease be an ideal candidate for many industrial processes at harsh conditions, especially as a bio-additive in detergent industry.     

Production of Curdlan Grown on Cassava Starch Waste Hydrolysates

Cassava starch waste hydrolysates (CSWHs) with different degrees of polymerisation, i.e., CSWHs-1, CSWHs-2 and CSWHs-3, were prepared through the hydrolysis of cassava starch waste with thermostable a-amylase from Thermococcus sp. HJ21. The prepared CSWHs were then used as a carbon source for curdlan production with Alcaligenes faecalis ATCC 31749. The amount of curdlan produced and the glucosyltransferase activity during curdlan synthesis increased more obviously when CSWHs-2 was used as the carbon source than when glucose was used. Using both carbon sources, the maximum curdlan production was observed at day 5, and the maximum glucosyltransferase activity was observed at day 4. Glucosyltransferase activity decreased thereafter, and biomass continued to increase until the end of the experiment (day 6). Results indicated that the enhanced curdlan production with CSWHs as the carbon source was highly correlated with glucosyltransferase activity.     

Effect of pretreatment techniques on food waste solubilization and biogas production during thermophilic batch anaerobic digestion

The purpose of this study was to optimize the alkaline, ultrasonication, and thermal pretreatment in order to enhance the solubilization of food waste (FW) for the production of volatile fatty acids, hydrogen, and methane in thermophilic batch anaerobic digestion. Initially, the effect of pretreatment techniques in the acidogenic phase was studied, and the optimal combinations of different conditions were determined. It was found that each pretreatment technique affected food waste solubilization differently. Alkaline pretreatment increased hydrogen yield in the acidogenic sludge by four times over control. COD solubilization was increased by 47 % when FW pre-heated at 130 °C for 60 min. Ultrasonication at 20 kHz and 45 min reduced processing time to 38 h from the 60–80 h needed in normal operation. Response surface methodology (RSM) was used to optimize a combination of alkaline, ultrasonication, and thermal pretreatment. Optimized conditions were applied to methanogenic single-stage thermophilic AD process, and their impact on biogas production was monitored. Results showed that FW heated at 130 °C for 50 min geminates biogas production compared to control experiment. In conclusion, a short thermal pretreatment regime could significant affect biogas production in single-stage thermophilic AD.     

Aerobic thermophilic bacteria enhance biogas production

The enhancing effect of aerobic thermophilic (AT) bacteria on the production of biogas from anaerobically digested sewage sludge (methanogenic sludge) was investigated. Sewage sludge (5%, w/w) was incubated at 65°C with shaking for a few months to prepare the AT seed sludge. AT sludge was prepared by incubation of the AT seed sludge (5%, v/v) and sewage sludge (5%, w/w) at 65°C with shaking. The addition of this AT sludge (1.2% ± 0.5% of total volatile solids) to methanogenic sludge enhanced the production of biogas. The optimum volume of the addition and the pretreatment temperature of the AT sludge for optimum biogas production were 5% (v/v) and 65°C. Batch-fed anaerobic digestion was covered with the addition of various AT sludges. The AT sludge prepared with the AT seed sludge improved the biogas production by 2.2 times relative to that from the sewage sludge addition. The addition of sludge without AT seed sludge weakly enhanced biogas production. An aerobic thermophilic bacterium (strain AT1) was isolated from the AT seed sludge. Strain AT1 grew well in a synthetic medium. The production of biogas from the anaerobic digestion of sewage sludge was improved by the addition of 5% (v/v) AT1 bacterial culture compared with that from the sewage sludge addition. The addition of AT1 culture reduced the volatile solids by 21%, which was higher than the 12.6% achieved with the sewage sludge addition. The AT1 bacterial culture enhanced the biogas production more than the AT seed sludge. The phylogenetic analysis of the 16S rRNA gene revealed that strain AT1 is closely related to Geobacillus thermodenitrificans (100% sequence similarity). The improvement in the production of biogas with the AT sludge could be caused by thermophilic bacterial activity in the AT sludge.     

Fungal survival during anaerobic digestion of organic household waste

Anaerobic digestion of organic waste yields energy rich biogas and retains nutrients (N, P, K, S, etc.) in a stabilised residue. For the residue to be used as a soil fertiliser, it must be free from pollutants and harmful microorganisms. Fungal survival during sanitation and anaerobic treatment of source-separated organic household waste and during aerobic storage of the residue obtained was investigated. Decimal reduction times were determined for inoculated fungi (Aspergillus flavus and Aspergillus fumigatus, Penicillium roqueforti, Rhizomucor pusillus, Thermoascus crustaceus and Thermomyces lanuginosus). Several different fungal species were found after waste sanitation treatment (70 degrees C, 1 h), with Aspergillus species dominating in non-inoculated waste. Anaerobic waste degradation decreased the diversity of fungal species for processes run at both 37 and 55 degrees C, but not total fungal colony forming units. Fungi surviving the mesophilic anaerobic digestion were mainly thermotolerant Talaromyces and Paecilomyces species. T. crustaceus and T. lanuginosus were the only inoculated fungi to survive the thermophilic anaerobic degradation process. Aerobic storage of both types of anaerobic residues for one month significantly decreased fungal counts.     

Low‐concentration hydrogen peroxide decontamination for Bacillus spore contamination in buildings

Remediation and recovery efforts after a release of Bacillus anthracis (anthrax) spores may be difficult and costly. In addition, response and recovery technologies may be focused on critical resources, leaving the small business or homeowner without remediation options. This study evaluates the efficacy of relatively low levels of hydrogen peroxide vapor (HPV) delivered from off‐the‐shelf equipment for the inactivation of Bacillus spores within an indoor environment. Decontamination evaluations were conducted in a house using both Bacillus atrophaeus var. globigii (Bg; as surrogates for B. anthracis) inoculated on the carpet and galvanized metal as coupons and Geobacillus stearothermophilus (Gs) as biological indicators on steel. The total decontamination time ranged from 4 to 7 days. Using the longer exposure times, low concentrations of HPV (average levels below 20 parts per million) effectively inactivated Bg and Gs spores on the materials tested. The HPV was generated with commercial humidifiers and household‐strength hydrogen peroxide solutions. The presence of home furnishings did not have a significant impact on HPV efficacy. This simple, inexpensive, and effective decontamination method could have significant utility for remediation following a B. anthracis spore release, such as following a terrorist attack.     

Optimization of Polyhydroxyalkanoates Production from Thermus thermophilus HB8 Using Response Surface Methodology

The thermophilic bacterium Thermus thermophilus HB8 was used for the overproduction of polyhydroxyalkanoates (PHAs) using a mathematical approach for the first time for optimization of process variables. In addition, the combined effect of nitrogen and phosphate concentrations on PHAs production was also investigated. A five-level-three-factor central composite rotary design was employed in combination with response surface methodology (RSM) to optimize the process variables for the production of PHAs in Thermus thrermophilus HB8. The three independent variables studied in the work were cultivation time, C/N ratio and phosphate concentration. Two second-order polynomial equations were obtained for biomass and PHA production by multiple regression analysis using RSM. The statistical analyses of the results showed that all the three variables had significant impact both on the cell growth and polymer accumulation. The model predicted a maximum PHA production of 0.47?g/L which represents the 42?% of dry cell weight (DCW) after 55?h of cultivation and with on setting the C/N ratio at 9:1?g/g and phosphate concentration at 20?mM. Verification of the predicted value resulted into a PHA production of 0.44?g/L (40.36?% of DCW).     

Survival of selected pathogens in diluted sludge of a thermophilic waste treatment plant and in NaCl-solution under aerobic and anaerobic conditions

Decimal reduction times (DRT or D-value) of Campylobacter jejuni, Salmonella enterica (formerly Salmonella choleraesuis) serovar Senftenberg, Escherichia coli, and Listeria monocytogenes were determined in two different matrices, diluted fermenter sludge (DFS) and 0.95% NaCl-solution (NaCl) at 50 degrees C, both under aerobic and anaerobic conditions. Depending on aeration, matrix composition, and the respective organism, the D-values varied between 10min and more than 15h. Generally the viability of bacteria decreased faster in DFS compared to NaCl-solution and under aerobic compared to anaerobic conditions. After 24h no viable cells could be detected in DFS, both under aerobic as well as under anaerobic conditions, whereas viable cells were still found in NaCl solutions. In both matrices the detection limits determined by means of PCR-based and classical microbiological methods were compared and pointed to lower detection limits of the latter methods. Results of the present investigation show that test organisms were far from surviving several days in DFS whereas hydraulic retention times normally used for thermophilic anaerobic digestion are in the range of 2 weeks. However, an underestimation of survival rates of the test organisms seems probable when applying aerobic standard methods.     

Effect of temperature on VFA’s and biogas production in anaerobic solubilization of food waste

The effectiveness of methane fermentation treatment used in food waste processing is currently limited by solubilization and acidogenesis. In efforts to improve the treatment process, this study examined the effects of temperature on solubilization and acidogenesis. The solubilization rate of food waste, which was based on suspended solid removal, was 47.5%, 62.2%, 70.0%, 72.7%, 56.1% and 45.9% at 15 °C, 25 °C, 35 °C, 45 °C, 55 °C and 65 °C, respectively. Solubilization rate was accelerated from the middle to late experimental periods under mesophilic (35 °C and 45 °C) conditions. In contrast, overall solubilization rate was significantly lower under thermophilic (55 °C and 65 °C) conditions than under mesophilic conditions, although solubilization occurred rapidly in the early experimental period. The production of biogas was high under mesophilic conditions of 35 °C and 45 °C, at 64.7 and 62.7 mL/g-VS, respectively, while it was scarce under thermophilic conditions. Solubilization of food waste was accelerated under both mesophilic and thermophilic conditions; however, solubilization rate was observed to be particularly high under mesophilic conditions, and a shortening of the hydraulic retention time is expected under thermophilic conditions.     

Prediction of temperature and thermal inertia effect in the maturation stage and stockpiling of a large composting mass

A macroscopic non-steady state energy balance was developed and solved for a composting pile of source-selected organic fraction of municipal solid waste during the maturation stage (13,500 kg of compost). Simulated temperature profiles correlated well with temperature experimental data (ranging from 50 to 70 degrees C) obtained during the maturation process for more than 50 days at full scale. Thermal inertia effect usually found in composting plants and associated to the stockpiling of large composting masses could be predicted by means of this simplified energy balance, which takes into account terms of convective, conductive and radiation heat dissipation. Heat losses in a large composting mass are not significant due to the similar temperatures found at the surroundings and at the surface of the pile (ranging from 15 to 40 degrees C). In contrast, thermophilic temperature in the core of the pile was maintained during the whole maturation process. Heat generation was estimated with the static respiration index, a parameter that is typically used to monitor the biological activity and stability of composting processes. In this study, the static respiration index is presented as a parameter to estimate the metabolic heat that can be generated according to the biodegradable organic matter content of a compost sample, which can be useful in predicting the temperature of the composting process.     

Mesophilic and thermophilic anaerobic biodegradability of water hyacinth pre-treated at 80 °C

Water hyacinth (Eichornia crassipes) is a fast growing aquatic plant which causes environmental problems in continental water bodies. Harvesting and handling this plant becomes an issue, and focus has been put on the research of treatment alternatives. Amongst others, energy production through biomethanation has been proposed. The aim of this study was to assess the anaerobic biodegradability of water hyacinth under mesophilic and thermophilic conditions. The effect of a thermal sludge pre-treatment at 80 °C was also evaluated. To this end, anaerobic biodegradability tests were carried out at 35 °C and 55 °C, with raw and pre-treated water hyacinth. According to the results, the thermal pre-treatment enhanced the solubilisation of water hyacinth (i.e. increase in the soluble to total chemical oxygen demand (COD)) from 4% to 12% after 30 min. However, no significant effect was observed on the methane yields (150–190 L CH₄/kg volatile solids). Initial methane production rates for thermophilic treatments were two fold those of mesophilic ones (6–6.5 L vs. 3–3.5 L CH₄/kg COD·day). Thus, higher methane production rates might be expected from thermophilic reactors working at short retention times. The study of longer low temperature pre-treatments or pre-treatments at elevated temperatures coupled to thermophilic reactors should be considered in the future.     

High rate mesophilic, thermophilic, and temperature phased anaerobic digestion of waste activated sludge: a pilot scale study

The paper reports the findings of a two-year pilot scale experimental trial for the mesophilic (35°C), thermophilic (55°C) and temperature phased (65+55°C) anaerobic digestion of waste activated sludge. During the mesophilic and thermophilic runs, the reactor operated at an organic loading rate of 2.2 kgVS/m(3)d and a hydraulic retention time of 20 days. In the temperature phased run, the first reactor operated at an organic loading rate of 15 kgVS/m(3)d and a hydraulic retention time of 2 days while the second reactor operated at an organic loading rate of 2.2 kgVS/m(3)d and a hydraulic retention time of 18 days (20 days for the whole temperature phased system). The performance of the reactor improved with increases in temperature. The COD removal increased from 35% in mesophilic conditions, to 45% in thermophilic conditions, and 55% in the two stage temperature phased system. As a consequence, the specific biogas production increased from 0.33 to 0.45 and to 0.49 m(3)/kgVS(fed) at 35, 55, and 65+55°C, respectively. The extreme thermophilic reactor working at 65°C showed a high hydrolytic capability and a specific yield of 0.33 g COD (soluble) per gVS(fed). The effluent of the extreme thermophilic reactor showed an average concentration of soluble COD and volatile fatty acids of 20 and 9 g/l, respectively. Acetic and propionic acids were the main compounds found in the acids mixture. Because of the improved digestion efficiency, organic nitrogen and phosphorus were solubilised in the bulk. Their concentration, however, did not increase as expected because of the formation of salts of hydroxyapatite and struvite inside the reactor.     

Solubilization and methanogenesis of a particulate industrial waste: Impact of solids loading and temperature

Effective anaerobic treatment of particulate wastes requires solubilization and acid formation prior to methanogenesis. In this case study of a particulate waste from a corn-processing industry, the influence of solids loading in solubilization, acid formation and methanogenesis was studied under mesophilic (35°C) and thermophilic (60°C) conditions. The waste was concentrated by centrifugation to initial suspended solids concentrations (TSS_i) of 150 to 350 g/L (15% to 35%). Anaerobic batch tests were conducted for 20 days, and significant solubilization of the particulate organic matter occurred in all cases. The thermophilic systems were more effective than the mesophilic systems with respect to solubilization of particulates, volatile solids destruction, acetic acid uptake, and methane generation. Methanogenesis appreared to be a rate-limiting step at higher TSS_i values, indicated by accumulation of volatile organic acids in the batch systems. Slower rates of methane production led to identification of the limiting solids loading for both temperature regimes. The results of this study can be used to evaluate the limitations of a single stage system for anaerobic treatment of organic particulate industrial wastes.     

Method for determination of methane potentials of solid organic waste

A laboratory procedure is described for measuring methane potentials of organic solid waste. Triplicate reactors with 10 grams of volatile solids were incubated at 55 degrees C with 400 ml of inoculum from a thermophilic biogas plant and the methane production was followed over a 50-day period by regular measurements of methane on a gas chromatograph. The procedure involves blanks as well as cellulose controls. Methane potentials have been measured for source-separated organic household waste and for individual waste materials. The procedure has been evaluated regarding practicality, workload, detection limit, repeatability and reproducibility as well as quality control procedures. For the source-separated organic household waste a methane potential of 495 ml CH4/g VS was found. For fat and oil a lag-phase of several days was seen. The protein sample was clearly inhibited and the maximal methane potential was therefore not achieved. For paper bags, starch and glucose 63, 84 and 94% of the theoretical methane potential was achieved respectively. A detection limit of 72.5 ml CH4/g VS was calculated from the results. This is acceptable, since the methane potential of the tested waste materials was in the range of 200-500 ml CH4/g VS. The determination of methane potentials is a biological method subject to relatively large variation due to the use of non-standardized inoculum and waste heterogeneity. Therefore, procedures for addressing repeatability and reproducibility are suggested.     

Anaerobic co-digestion of dairy cow manure and high concentrated food processing waste

Anaerobic co-digestion of dairy manure (DM) and concentrated food processing wastes (FPW) under thermophilic (55 °C) and mesophilic (35 °C) temperatures, and fertilizer value of the effluent were investigated in this study. Two types of influent feedstock were utilized: 100 % DM and a 7:3 mixture (wet weight basis) of DM and FPW. The contents of the FPW, as feedstock were 3:3:3:1 mixture of cheese whey, animal blood, used cooking oil and residue of fried potato. Four continuous digestion experiments were carried out in 10 L digesters. Co-digestion under thermophilic temperature increased methane production per digester volume. However, co-digestion at 35 °C was inhibited. Total Kjeldahl nitrogen (N) recovered after digestion ranged from 73.1 to 91.9 %, while recoveries of ammonium nitrogen (NH₄-N) exceeded 100 %. The high recovery of NH₄-N was attributed to mineralization of influent organic N. The mixture of DM and FPW showed greater recoveries of NH₄-N after digestion compared to DM only, reflecting its greater organic N degradability. The ratios of extractable to total calcium, phosphorus and magnesium were slightly reduced after digestion. These results indicate that co-digestion of DM and FPW under thermophilic temperature enhances methane production and offers additional benefit of organic fertilizer creation.     

Carbon mineralization dynamics in soils amended with meat meals under laboratory conditions

Meat and bone meal (MBM) is obtained from the wastes produced during slaughtering operations. Its high concentration of N and P makes it interesting as an organic fertiliser but its use in soil has been barely studied previously. In this work four laboratory experiments were performed to study the influence of different variables (MBM composition, rate of application, temperature of incubation and the type of soil) on C mineralization dynamics of MBM in agricultural soils. The total CO2-C evolved (as % of added C) after 2 weeks ranged between 10% and 20%. The kinetics of mineralization were rapid, with C evolved as CO2 within the first 4 days representing more than 50% of total C mineralized. A linear correlation was found between the rate of application (added-C) and CO2-C evolved (r2: 0.997; P<0.001). A temperature coefficient (Q10) was used to assess the difference in biological activity at 5 degrees C intervals. Q10, which ranged from 1.0 to 2.7 (250h), was higher for the lower temperature range (Q10 (15-20 degrees C)>Q10 (20-25 degrees C)) and it was found to be related to the soil properties. Finally, the mineralization process was found to be highly dependent upon the different soil factors, although no simple linear correlation was found between mineralization and soil properties.     

Kinetics of microbial landfill methane oxidation in biofilters

A methane oxidizing biofilter system fitted to the passive venting system of a harbor sludge landfill in Germany was characterized with respect to the the methanotrophic population, methane oxidizing capacity, and reaction kinetics. Methanotrophic cell counts stabilized on a high level with 1.3 x 10(8) to 7.1 x 10(9) cells g dw(-1) about one year after first biofilter operation, and a maximum of 1.2 x 10(11) cells g dw(-1). Potential methane oxidizing activity varied between 5.3 and 10.7 microg h(-1) g dw(-1). Cell numbers correlated well with methane oxidation activities. Extrapolation of potential activities gave methane removal rates between 35 and 109 g CH4 h(-1) m(-3), calculated for 30 degrees C. Optimum temperature was 38 degrees C for freshly sampled biofilter material and 22 degrees C for a methanotrophic enrichment culture grown at 10 degrees C incubation temperature. Substrate kinetics revealed the presence of a low-affinity methane oxidizing community with a high Vmax of 1.78 micromol CH4 h(-1) g ww(-1) and a high K(M) of 15.1 microM. K(MO2) for methane oxidation was 58 microM. No substantial methane oxidizing activity was detected below 1.7-2.6 vol.-% O2 in the gaseous phase. Methane deprivation led to a decrease in methane oxidation activity within 5-9 weeks but could still be detected after 25 weeks of substrate deprivation and was fully restored within 3 weeks of continuous methane supply. Very high salt loads are leached from the novel biofilter material, expanded clay, yielding electric conductivity values of up to 15 mS cm(-1) in the leachate. Values > 6 mS cm(-1) were shown to depress methane consumption. Water retention characteristics of the material proved to be favourable for methane oxidizing systems with a gas permeable volume of 78% of bulk volume at field capacity water content. Correspondingly, no influence of water content on methane oxidation activity could be detected at water contents between 2.5 and 20 vol.-%.     

Coal fly ash as raw material for the manufacture of geopolymer-based products

In this work coal fly ash has been employed for the synthesis of geopolymers. Two different systems with silica/alumina ratios stoichiometric for the formation of polysialatesiloxo (PSS, SiO2/Al2O3=4) and polysialatedisiloxo (PSDS, SiO2/Al2O3=6) have been prepared. The alkali metal hydroxide (NaOH or KOH) necessary to start polycondensation has been added in the right amount as concentrated aqueous solution to each of the two systems. The concentration of each alkali metal solution has been adjusted in order to have the right liquid volume to ensure constant workability. The systems have been cured at four different temperatures (25, 40, 60, and 85 degrees C) for several different times depending on the temperature (16-672 h at 25 degrees C; 72-336 h at 40 degrees C; 16-120 h at 60 degrees C and 1-6h at 85 degrees C). The products obtained in the different experimental conditions have been submitted to the quantitative determination of the extent of polycondensation through mass increase and loss on ignition, as well as to qualitative characterization by means of FT-IR spectroscopy. Furthermore, physico-structural and mechanical characterization has been carried out through microscopic observations and the determination of unconfined compressive strength, elasticity modulus, apparent density, porosity and specific surface area. The results have indicated that the systems under investigation are suited for the manufacture of pre-formed building blocks at room temperature.     

Microbial biomass in a soil amended with different types of organic wastes.

Application of different types of organic wastes may have a marked effect on soil microbial biomass and its activity. The objective of this study was to quantify the amount of microbial biomass in a loamy-clayey soil, amended with different types of organic waste residues (composts of municipal solid waste of different ages, sewage sludge and farmyard manure) and incubated for 8 weeks at 25 degrees C and two-thirds of field capacity, using the fumigation-extraction method. Both microbial biomass-C and -N (BC and BN, respectively) appeared to be dependent on the type of organic waste residues, on their degree of stability, and on their chemical characteristics. In general, organic wastes increased the microbial biomass-C content in the soil and the microbial BC was positively correlated with the organic C content, the C/N, neutral detergent fibre/N (NDF/N) and acid detergent fibre/N (ADF/ N) ratios. The microbial biomass content decreased according to the period of incubation, especially when the compost used was immature. The microbial biomass-N was positively correlated with the total N and percentage of hemicellulose. The microbial biomass-C was linearly related with the microbial biomass-N and the ratio BC/BN was exponentially related with the BC.     

Influence of dairy manure addition on the biological and thermal kinetics of composting of greenhouse tomato plant residues

A laboratory-scale bioreactor was used to investigate the influence of dairy manure addition (as an inoculum and a carbon source) on the biological and thermal kinetics of the composting process of tomato plant residues-wood shavings mixture. Urea was added (as a nitrogen source) to correct the initial C:N ratio to 30:1 and the initial moisture content was also adjusted to 60%. The result of this study indicated that manure addition to the tomato residues-wood shavings mixture is a good source of macro and micronutrients required for supporting the composting microorganisms. Manure addition increased the rate of temperature increase and the duration of maximum temperature and reduced the lag and the peak time, all of which resulted in a significant reduction in the retention time. However, thermophilic temperature (> or = 40 degrees Celsius) was only achieved with 30%, 40% and 50% manure addition for 3, 7 and 9h. Total carbon reductions were in the range of 9.4-10.8% and TKN reductions were in the range of 3.4-6.0%. Neither the nitrogen nor the moisture content were limiting factors as the C:N ratio remained in the range of 26:1 to 28:1 and the moisture content remained within the optimum range of 58-61%. The maximum temperature of each mixture correlated with the reduction of total carbon, but carbon availability was a limiting factor in these experiments. In order to attain and sustain a thermophilic phase during the composting process, the addition of a readily available carbon source to the tomato should be investigated and carbon type (carbohydrates, proteins and fats) should be taken into account.