Modelization of anaerobic processes during co-digestion of slowly biodegradable substrates.

The influence of the soluble substrates over the anaerobic processes has been extensively investigated, but little is known about the effects of particulate substrate. The biodegradation of these substrates starts with the hydrolytic step, this process is slower than the other ones involved in the biodegradation of particulate substrates and usually becomes the rate-limiting step. This study investigate the effect of the initial total solids (TS) concentration on the anaerobic co-digestion of two slowly biodegradable organic substrates. The wastes mixtures were prepared at different dilutions in the range from 10% to 28% TS. From these experiments it was observed that as TS concentration increased, the methane production decreased. These results were modelled and it was observed that neither hydrolysis nor fermentation stages controlled the methane production rate. Being a substrate inhibition event experienced at the methanogenic stage the responsible of the lower methane production when operating at high TS concentrations.

Dark fermentation from real solid waste. Evolution of microbial community.

The purpose of this paper was to study the evolution of microbial community and its relation to the hydrogen production (HP) steps in thermophilic-dry dark fermentation from real organic fraction of municipal solid waste (OFMSW). Nine organic loading rates (OLRs) (from 9 to 220 g TVS/l/d) were investigated. Population dynamics study showed that increasing OLR (between 9 and 110 g TVS/l/d) resulted in an increase in the relations between Eubacteria:Archaea and hydrolytic-acidogenic bacteria (HABs):acetogens. This was strongly influenced by the microbial content of the OFMSW. The presence of acetogens and Archaea was due to contribution of these microorganisms in the substrate (the biogas produced was methane-free). The maximum value of hydrolysis (63±7%) was observed at 110 g TVS/l/d OLR according to maximum HP and HAB activity. The highest average values of acidification yields (57-60%) were achieved for OLR between 28 and 43 g TVS/l/d.

Optimisation of the two-phase dry-thermophilic anaerobic digestion process of sulphate-containing municipal solid waste: population dynamics.

Microbial population dynamics and anaerobic digestion (AD) process to eight different hydraulic retention times (HRTs) (from 25d to 3.5d) in two-phase dry-thermophilic AD from sulphate-containing solid waste were investigated. Maximum values of gas production (1.9 ± 0.2 l H2/l/d; 5.4 ± 0.3 l CH4/l/d and 82 ± 9 ml H2S/l/d) and microbial activities were obtained at 4.5d HRT; where basically comprised hydrolysis step in the first phase (HRT=1.5d) and acidogenic step finished in the second phase as well as acetogenic-methanogenic steps (HRT=3d). In the first phase, hydrolytic-acidogenic bacteria (HABs) was the main group (44-77%) and Archaea, acetogens and sulphate-reducing bacteria (SRBs) contents were not significant; in the second phase (except to 2d HRT), microbial population was able to adapt to change in substrate and HRTs to ensure the proper functioning of the system and both acetogens and Archaea were dominated over SRBs. Decreasing HRT resulted in an increase in microbial activities.

Optimisation of single-phase dry-thermophilic anaerobic digestion under high organic loading rates of industrial municipal solid waste: population dynamics.

Different high feed organic loading rates (OLRs) (from 5.7 g to 46.0 g TVS/l/d) or hydraulic retention times (HRTs) (from 15 d to 2 d) in single-phase dry-thermophilic anaerobic digestion (AD) of organic fraction municipal solid waste (OFMSW) were investigated. The specific gas production (SGP) values (0.25-0.53 m(3)/kg TVS) and the percentages of Eubacteria, Archaea, H2-utilising methanogens (HUMs) and acetate-utilising methanogens (AUMs) were stable within the ranges 80.2-91.1%, 12.4-18.5%, 4.4-9.8% and 5.5-10.9%, respectively. A HUM/AUM ratio greater than 0.7 seems to be necessary to maintain very low partial pressures of H2 required for dry AD process. Increasing OLR resulted in an increase in all the populations, except for propionate-utilising acetogens (PUAs). Optimal conditions were obtained at 3d HRT (OLR=30.7 g TVS/l/d), which is lower than the doubling time of acetogens and methanogens. The methane production (MP) was clearly higher than those reported in AD of OFMSW.

Hydrogen production from the organic fraction of municipal solid waste in anaerobic thermophilic acidogenesis: influence of organic loading rate and microbial content of the solid waste.

Hydrogen production (HP) from the organic fraction of municipal solid waste (OFMSW) under thermophilic acidogenic conditions was studied. The effect of nine different organic loading rates (OLRs) (from 9 to 220 g TVS/l/d) and hydraulic retention times (HRTs) (from 10d to 0.25 d) was investigated. Normally, butyrate was the main acid product. The biogas produced was methane- and sulfide-free at all tested OLR. Increasing the OLR resulted in an increase in both the quantity and quality of hydrogen production, except at the maximum OLR tested (220 g TVS/l/d). The maximum hydrogen content was 57% (v/v) at an OLR of 110 g TVS/l/d (HRT=0.5 d). HP was in the range of 0.1-5.7 l H2/l/d. The results have clearly shown that the increase in OLR was directly correlated with HP and microbial activity. The bacterial concentration inside the reactor is strongly influenced by the content of microorganisms in the OFMSW.

Kinetics of mesophilic anaerobic digestion of the organic fraction of municipal solid waste: Influence of initial total solid concentration.

The anaerobic mesophilic degradation of municipal waste has been studied in discontinuous lab-reactors with two different initial concentrations of OFMSW: R20, with 931.1mg DOC/L (20% TS), and R30, with 1423.4 mg DOC/L (30% TS). The anaerobic digestion process was favoured when it was carried on material with a total solids content of 20% in comparison to a similar process with 30% TS. A higher level of organic matter, in terms of DOC and VFA, 18.18% and 8.09% respect, was removed in the system with the lower amount of solids. The kinetics parameters showed higher active biomass and a higher coefficient for the production of methane at the lower loading. The highest maximum specific growth rate for the microorganisms (mu(MAX)) in the reactors with 20% TS, a value of 0.192 d(-1), was achieved in comparison to 0.131 d(-1) in the reactors with 30% TS. The inverse of the F:M ratio was higher in the R20 system with a value of 0.0104 (cf. 0.0006 in R30) and, as a result, the R30 system required longer hydrolytic and acidogenic stages. X(V0)/Y(X/S) and the yield coefficient for product generation (alpha(P/S)) were higher, by around 53%, for the R20 digester.

Anaerobic digestion of municipal solid wastes: dry thermophilic performance.

The purpose of this study was to analyze the performance of two laboratory-scale reactors (5.0L) treating organic fraction of municipal solid waste (OFMSW): source sorted OFMSW (SS_OFMSW) obtained from a university restaurant and mechanically selected municipal fraction (MS_OFMSW) obtained from a Municipal Treatment Plant placed in Cadiz-Spain. Discontinuous reactors operated at thermophilic (55 degrees C) and dry (20% total solid) conditions. Different decomposition patterns were observed: (1) the SS_OFMSW exhibited the classical waste decomposition pattern with a fast start up phase beginning within 0-5 days and 20-30 and a subsequent stabilization phase. The VS removal was 45% with a cumulative biogas of 120L in approx. 60 days; (2) the MS_OFMSW showed a methanogenic pattern throughout the whole experimental period (60 days) and this gave higher levels of organic biodegradation (56%VSr) and biogas production (82L). Both processes were completed and a high level of cumulative methane production was achieved in less than 60 days, proximally 25-30L.

Thermophilic anaerobic digestion of source-sorted organic fraction of municipal solid waste.

The influence of different organic fraction of municipal solid wastes during anaerobic thermophilic (55 degrees C) treatment of organic matter was studied in this work: food waste (FW), organic fraction of municipal solid waste (OFMSW) and shredded OFMSW (SH_OFMSW). All digester operated at dry conditions (20% total solids content) and were inoculated with 30% (in volume) of mesophilic digested sludge. Experimental results showed important different behaviours patterns in these wastes related with the organic matter biodegradation and biogas and methane production. The FW reactor showed the smallest waste biodegradation (32.4% VS removal) with high methane production (0.18 LCH4/gVS); in contrast the SH_OFMSW showed higher waste biodegradation (73.7% VS removal) with small methane production (0.05 LCH4/g VS). Finally, OFMSW showed the highest VS removal (79.5%) and the methane yield reached 0.08 LCH4/g VS. Therefore, the nature of organic substrate has an important influence on the biodegradation process and methane yield. Pre-treatment of waste is not necessary for OFMSW.

Effect of substrate concentration on dry mesophilic anaerobic digestion of organic fraction of municipal solid waste (OFMSW).

The influence of total solid contents during anaerobic mesophilic treatment of the organic fraction of municipal solid waste (MSW) has been studied in this work. The work was performed in batch reactors of 1.7L capacity, during a period of 85-95 days. Two different organic substrate concentrations were studied: 931.1 mgDOC/L (20% TS) and 1423.4 mgDOC/L (30% TS). Experimental results showed that the reactor with 20% total solids content had significantly higher performance. Thus, the startup phase ended at 14 days and the total DOC removal was 67.53%. The startup in reactor R30 ended at 28 days obtaining 49.18% DOC removal. Also, the initial substrate concentration contributed substantially to the amount of methane in the biogas. Hence, the total methane production in the methanogenic phase was 7.01 L and 5.53 L at the end of the experiments for R20 and R30, respectively.

Influence of total solid and inoculum contents on performance of anaerobic reactors treating food waste.

The aim of this paper was to analyze the biomethanization process of food waste (FW) from a university campus restaurant in six reactors with three different total solid percentages (20%, 25% and 30% TS) and two different inoculum percentages (20-30% of mesophilic sludge). The experimental procedure was programmed to select the initial performance parameters (total solid and inoculum contents) in a lab-reactor with V: 1100mL and, later, to validate the optimal parameters in a lab-scale batch reactor with V: 5000mL. The best performance for food waste biodegradation and methane generation was the reactor with 20% of total solid and 30% of inoculum: give rise to an acclimation stage with acidogenic/acetogenic activity between 20 and 60 days and methane yield of 0.49L CH4/g VS. Also, lab-scale batch reactor (V: 5000mL) exhibit the classical waste decomposition pattern and the process was completed with high values of methane yield (0.22L CH4/g VS). Finally, a protocol was proposed to enhance the start-up phase for dry thermophilic anaerobic digestion of food waste.

Composting potential of different inoculum sources in the modified SEBAC system treatment of municipal solid wastes.

The aim of the work described here was to analyse the biomethanization process for three types of organic fraction of municipal solid waste (OFMSW) using two different inoculum sources in a sequential leach-bed anaerobic reactor under thermophilic (55 degrees C) and dry conditions (20%TS). The OFMSWs studied were: separately collected food waste (SC_OFMSW) from restaurants, synthetic waste (ST_OFMSW) and mechanically selected municipal waste (MS_OFMSW). The inoculum studied were digested mesophilic sludge (SLUDGE) and digested SC_OFMSW. The results show that SLUDGE gave the best performance and this was therefore selected for the rest of the experiments. Three assays were performed to analyse the biomethanization processes of three types of organic waste. The results suggest that all three wastes give rise to an acclimation stage with acidogenic/acetogenic activity between days 5 and 20-30 and a stabilization phase associated with methanogenic activity. In conclusion, a mixture of OFMSW (regardless of its origin) and SWINE arranged in layers in the reactor, as well as SLUDGE used with an inoculum source, enhanced the fast start up phase of a modified sequential leach-bed system under dry thermophilic conditions.

Performance of anaerobic thermophilic fluidized bed in the treatment of cutting-oil wastewater.

This paper examines the effect of organic loading rate on the removal efficiency of COD and TOC anaerobic thermophilic fluidized bed reactor (AFBR) in the treatment of cutting-oil wastewater at different hydraulic retention time (HRT) conditions. The essays are development at laboratory scale using a porous support medium. The AFBR reactor was subjected to a programme of steady-state operation over a range of hydraulic retention times, HRTs, in the range 12-2h and organic loading rates, OLRs, between 11.9 and 51.3kgCOD/m(3)d. The highest efficiency was 95.9% for an OLR of 13kgCOD/m(3)d and HRT of 11h. Over an operating period of 92 days, an OLR of 51.3kgCOD/m(3)d was achieved with 67.1% COD removal efficiency (71.3% TOC) in the experimental AFBR reactor. Although the level of biogas generation was not high, the anaerobic fluidized bed technology provided significant advantages over the conventional physico-chemical treatment applied in the factory. The effluent had a better quality (lower organic loading) and it was possible to reuse it in different applications in the factory (e.g., irrigation of gardens). The biological treatment did not lead to the generation of oily sludge, which is considered as hazardous waste by legislation. Furthermore, a continuous stream is produced and this reduced the impact of large flows discharged 4-5 times per week to the urban collector and MWWTP (municipal wastewater treatment plant).

Dry-thermophilic anaerobic digestion of organic fraction of the municipal solid waste: focusing on the inoculum sources.

The effect of inoculum source on anaerobic thermophilic digestion of separately collected organic fraction of municipal solid wastes (SC_OFMSW) has been studied. Performance of laboratory scale reactors (V: 1.1 L) were evaluated using six different inoculums sources: (1) corn silage (CS); (2) restaurant waste digested mixed with rice hulls (RH_OFMSW); (3) cattle excrement (CATTLE); (4) swine excrement (SWINE); (5) digested sludge (SLUDGE); and (6) SWINE mixed with SLUDGE (1:1) (SWINE/SLUDGE). The SC_OFMSW was separately and collected from university restaurant. The selected conditions were: 25% of inoculum, 30% of total solid and 55 degrees C of temperature, optimum in the thermophilic range. The six inoculum sources showed an initial start-up phase in the range between 2 and 4 days and the initial methane generation began over 10 days operational process. Results indicated that SLUDGE is the best inoculum source for anaerobic thermophilic digestion of the treatment of organic fraction of municipal solid waste at dry conditions (30%TS). Over 60 days operating period, it was confirmed that SLUDGE reactor can achieve 44.0%COD removal efficiency and 43.0%VS removal. In stabilization phase, SLUDGE reactor showed higher volumetric biogas generated of 78.9 mL/day (or 35.6 mLCH(4)/day) reaching a methane yield of 0.53 LCH(4)/gVS. Also, SWINE/SLUDGE and SWINE were good inoculums at these experimental conditions.

Real-time polymerase chain reaction diagnosis of leishmaniasis in Panama from both fresh and frozen tissue.

Skin biopsies stored in ethanol from 49 patients with suspected cutaneous leishmaniasis (CL) were tested in a real-time polymerase chain reaction (PCR) assay and compared with conventional diagnostic methods. With clinical diagnosis as the gold standard, PCR had a sensitivity of 96% (47/49) vs. 61% (30/49) for histopathology and 33% (16/49) for culture. In addition, DNA was extracted from 70 frozen smears of lesions from suspected cases of CL and tested with the same assay. In these samples, the PCR had a sensitivity of 61% (43/70) vs. 56% (39/70) for histopathology and 41% (29/70) for culture. In this study, real-time PCR offered a rapid diagnosis with an enhanced sensitivity over conventional methods. Although the yield of PCR diagnosis was lower when testing frozen smears, the assay still outperformed existing diagnostic modalities.

Influence of operational conditions on biofilm specific activity of an anaerobic fluidized bed reactor.

A key parameter in water and wastewater treatment technology is the biomass activity in terms of substrate removal ability. The effects of organic load rate and percentage of bed expansion on biofilm specific methanogenic activity were determined in an anaerobic fluidized bed reactor treating wine-distillery wastes in the thermophilic range (55 degrees C). The proposed activity tests are highly reproducible: an experiment with three identical tests has shown that the standard deviation with respect to the mean values is less than 3%. Specific tests are applied to measure the maximum methanogenic activities of the biomass carrier in lab-scale anaerobic biofilm reactors. These tests have been successfully applied for monitoring the support colonization process and the evolution of biofilm activity in reactors, anaerobic filter and fluidized bed, with different operating conditions. The results show a dependence between the percentage of bed expansion and the specific activity of methanogenic microbiote on biofilm. There is a relationship between the percentage of bed expansion, the sheer stress on the biofilm and the hydrodynamic conditions in the system. Initial biofilm detachment can be compensated with the increase of biomass and of its activity due to the reduction of the substrate diffusional limitations to the microorganism growth inside the support pores.

Kinetics of thermophilic anaerobes in fixed-bed reactors.

The main objective of this study is to estimate growth kinetic constants and the concentration of "active" attached biomass in two anaerobic thermophilic reactors which contain different initial sizes of immobilized anaerobic mixed cultures and decompose distillery wastewater. This paper studies the substrate decomposition in two lab-scale fixed-bed reactors operating at batch conditions with corrugated tubes as support media. It can be demonstrated that high micro-organisms-substrate ratios favor the degradation activity of the different anaerobic cultures, allowing the stable operation without lag-phases and giving better quality in effluent. The kinetic parameters obtained--maximum specific growth rates (mu(max)), non-biodegradable substrate (S(NB)) and "active or viable biomass" concentrations (X(V0))--were obtained by applying the Romero kinetic model [L.I. Romero, 1991. Desarrollo de un modelo matemático general para los procesos fermentativos, Cinética de la degradación anaerobia, Ph.D. Thesis, University of Cádiz (Spain), Serv. Pub. Univ. Cádiz], with COD as substrate and methane (CH4) as the main product of the anaerobic process. This method is suitable to calculate and to differentiate the main kinetic parameters of both the total anaerobic mixed culture and the methanogenic population. Comparison of experimental measured concentration of volatile attached solids (VS(att)) in both reactors with the estimated "active" biomass concentrations obtained by applying Romero kinetic model [L.I. Romero, 1991. Desarrollo de un modelo matemático general para los procesos fermentativos, Cinética de la degradación anaerobia, Ph.D. Thesis, University of Cádiz (Spain), Serv. Pub. Univ. Cádiz] shows that a large amount of inert matter is present in the fixed-bed reactor.

Measurement of microbial numbers and biomass contained in thermophilic anaerobic reactors.

This paper describes the determination of the microbial population in terms of the number, biomass, and composition of single- and two-phase, laboratory-scale thermophilic (55 degrees C) anaerobic reactors under steady-state conditions. Epifluorescence microscopy with 4',6-diamidine-2-phenylindole (DAPI) as fluorochrome was used to determine the total number of microorganisms in the reactors, and autofluorescence microscopy was used to determine the total methanogenic bacteria populations. The results obtained by the direct count methods were compared to the quantity of biomass contained in the system, which was determined by volatile suspended solids. The concentration of acidogenic bacteria was estimated by subtraction of the autofluorescence results from those of the DAPI epifluorescence microscopy. The viable bacterial population was determined by plating techniques using an anaerobic chamber. The total bacterial and methanogenic populations of single-stage digesters increase when the hydraulic retention time decreases; nevertheless, the percentages of the principal bacterial groups (acidogenic and methanogenic) remain constant at 87% and 13%, respectively. In the two-stage reactors, the percentages of the acidogenic and methanogenic groups are 99% and 26% of the total population in the acidogenic and methanogenic reactors, respectively. In the single-stage reactors, biomass determinations can be used to estimate microbial concentrations, and vice versa, as there is a high positive correlation between microorganism concentration and biomass. The syntrophic relationship between the bacteria involved in the anaerobic process is a possible explanation for the low values of viable population obtained in the reactors studied. Nevertheless, there is a high correlation between direct counts by epifluorescence microscopy and viable plate counts for the combined system studied.

Differential expression of nitric oxide by dermal microvascular endothelial cells from patients with scleroderma.

Vascular abnormalities in scleroderma are fundamental to the pathogenesis of this disease. The objective of this study was to characterize dermal microvascular endothelial cells (DMEC) isolated from scleroderma patients with respect to growth and expression of the constitutive form of endothelial nitric oxide synthase (eNOS). DMEC from patients with both systemic sclerosis (SSc) and localized scleroderma (Loc Scl) contained small intact microvascular structures in contrast to single cell isolations obtained from control skin. Immunoaffinity selection on anti-PECAM-1 beads yielded pure populations of DMEC expressing normal markers. While the morphology and initial growth of SSc DMEC closely paralleled control cells, the growth of SSc DMEC decreased with time in culture (doubling time of 3 days vs. 5 days). Expression of ecNOS mRNA was reduced in both Loc Scl and SSc as shown by semi-quantitative RT-PCR (p < 0.001). Western blots showed variable but generally lower ecNOS protein levels and decreased levels of nitrogen oxides in media were found from both SSc and Loc Scl relative to control cells. The results indicate an intrinsic defect in the mechanism of nitric oxide production in DMEC isolated from scleroderma patients and suggest its possible involvement in the pathophysiology of scleroderma.

Biodegradation kinetics of surfactants in seawater.

In this paper, a general kinetic model for degradation processes of surfactants is proposed. The model equation is v = K2S2 + K1S + K0, where v is the substrate consumption rate in the biodegradation process, S is the surfactant concentration in the medium and K2, K1, and K0 are kinetic constants. From this general expression, different simplified equations can be obtained (where K0 = 0; K2 and K0 = 0; K2 = 0; K2 and K1 = 0), which are representative of the process for different operating conditions. This model was tested by measuring the degradation of two different surfactants (Sodium dodecyl benzene sulfonate, LAS; and Sodium dodecyl sulfate, DSNa) under two different temperatures (5 and 20 degrees C). Values predicted by the model are close to experimental data obtained.

Membrane-type matrix metalloproteinases in human dermal microvascular endothelial cells: expression and morphogenetic correlation.

Membrane-type matrix metalloproteinases (MT-MMP) activate the zymogen form of MMP-2/Gelatinase A on cell surfaces and are expressed in invasive tumors. We sought to identify and characterize MT-MMP in a non-malignant cell type that undergoes a physiologic and reversible invasive phenotype during angiogenesis. Human dermal microvascular endothelial cells (HDMEC) were isolated from neonatal tissue and purified by anti-CD31 (PECAM) affinity beads. MT-MMP-1 and -3 transcripts were amplified by reverse transcriptase-polymerase chain reaction and northern blots showed a single 4.5 kB mRNA for MT-MMP-1 that was modulated by angiogenic factors and phorbol ester. Immunoblotting of reduced cellular extracts with different MT-MMP-1 antibodies showed the presence of the 63-65 kDa and 57-60 kDa forms, as well as additional forms at lower molecular weights. HDMEC membranes extracted with Triton X114 were incubated with gelatin-sepharose purified MMP-2 and MMP-9 to show activation of proenzymes. Pre-incubation of HDMEC with anti-MT-MMP-1 antibodies decreased proMMP-2 conversion activity only. The movement of HDMEC and the formation of tubule-like structures in three-dimensional collagen gels was markedly delayed by preincubation with the same anti-MT-MMP-1 antibodies. These results demonstrate the presence of MT-MMP in cutaneous microvascular cells in vitro. Modulation of these cell surface proteinases by angiogenic factors, demonstration of multiple processed forms, and specific attenuation of HDMEC morphogenetic patterns in three-dimensional collagen gels implicate their potential roles in the formation of new blood vessels in the skin.