Palmitic acid was the main long chain fatty acids (LCFA) that accumulated onto the anaerobic sludge when oleic acid was fed to an EGSB reactor. The conversion between oleic and palmitic acid was linked to the biological activity. When palmitic acid was fed to an EGSB reactor it represented also the main LCFA that accumulated onto the sludge. The way of palmitic acid accumulation was different in the oleic and in the palmitic acid fed reactors. When oleic acid was fed, the biomass-associated LCFA (83% as palmitic acid) were mainly adsorbed and entrapped in the sludge that became "encapsulated" by an LCFA layer. However, when palmitic acid was fed, the biomass-associated LCFA (the totality as palmitic acid) was mainly precipitated in white spots like precipitates in between the sludge, which remained "non-encapsulated." The two sludges were compared in terms of the specific methanogenic activity (SMA) in the presence of acetate, propionate, butyrate, and H(2)CO(2), before and after the mineralization of similar amounts of biomass-associated LCFA (4.6 and 5.2 g COD-LCFA/g of volatile suspended solids (VSS), for the oleic and palmitic acid fed sludge, respectively). The "non-encapsulated," sludge exhibited a considerable initial methanogenic activity on all the tested substrates, with the single exception of butyrate. However, with the "encapsulated" sludge only methane production from ethanol and H(2)/CO(2) was detected, after a lag phase of about 50 h. After mineralization of the biomass-associated LCFA, both sludges exhibited activities of similar order of magnitude in the presence of the same individual substrates and significantly higher than before. The results evidenced that LCFA accumulation onto the sludge can create a physical barrier and hinder the transfer of substrates and products, inducing a delay on the initial methane production. Whatever the mechanism, metabolic or physical, that is behind this inhibition, it is reversible, being eliminated after the depletion of the biomass-associated LCFA.

Long-chain fatty acids (LCFA) associated with anaerobic sludge by mechanisms of precipitation, adsorption, or entrapment can be biodegraded to methane. The mineralization kinetics of biomass-associated LCFA were established according to an inhibition model based on Haldane's enzymatic inhibition kinetics. A value around 1,000 mg COD-LCFAÁg VSS À1 was obtained for the optimal specific LCFA content that allowed the maximal mineralization rate. For sludge with specific LCFA contents of 2,838 F 63 and 4,571 F 257 mg COD-LCFAÁg VSS À1 , the specific methanogenic activities in the presence of acetate, butyrate, and H 2 /CO 2 were significantly enhanced after the mineralization of the biomass-associated LCFA. For sludge with a specific LCFA content near the optimal value defined by the kinetic model, the effect of adding VFA to the medium was studied during the mineralization of the biomass-associated LCFA. Different patterns were obtained for each individual substrate. Acetate and butyrate were preferentially consumed by the consortium, but in the case of propionate no evidence of a sequential consumption pattern could be withdrawn. It was concluded that LCFA do not exert a bactericidal neither a permanent toxic effect toward the anaerobic consortia. A discussion is addressed to the relative roles of a reversible inhibitory effect and a transport limitation effect imposed by the LCFA surrounding the cells. B

Oleic acid toxicity and biodegradability were followed during long-term operation of two similar anaerobic fixed-bed units. When treating an oleate based effluent, the sludge from the bioreactor that was acclimated with lipids during the first operation period, showed a higher tolerance to oleic acid toxicity (IC50 = 137 mg/l) compared with the sludge fed with a non-fat substrate (IC50 = 80 mg/l). This sludge showed also the highest biodegradation capacity of oleic acid, achieving maximum methane production rates between 33 and 46 mlCH4(STP)/gVS.day and maximum percentages of methanization between 85 and 98% for the range of concentrations between 500 and 900 mg oleate/l. When oleate was the sole carbon source fed to both digesters, the biomass became encapsulated with organic matter, possibly oleate or an intermediate of its degradation, e.g. stearate that was degraded at a maximum rate of 99 mlCH4(STP)/gVS.day. This suggests the possibility of using adsorption-degradation cycles for the treatment of LCFA based effluents. Both tolerance to toxicity and biodegradability of oleic acid were improved by acclimatization with lipids or oleate below a threshold concentration.

The aim of the present work was to study the maximum potential methane production in batch assays of sludge samples taken along the operation of two EGSB reactors (RI inoculated with granular sludge and RII inoculated with suspended sludge) fed with increasing oleic acid concentrations between 2 and 8 gCOD/l (HRT = 1 day). After removing the residual substrate, the sludge was incubated in batch vials without any added carbon source. A maximum methane production rate of 152+/-21 mlCH4(STP)/gVS.day was obtained for the suspended sludge taken on day 70, when oleate at a concentration of 2 g COD/l was fed with a co-substrate (50% COD). The maximum plateau achieved in the methane production curve was 1145+/-307 mlCH4(STP)/gVS, obtained for the suspended sludge taken on day 162, when oleate was fed as the sole carbon source at 6 g COD/I. The methanization rate of the adsorbed substrate was enhanced under stirring conditions and was inhibited by adding oleic acid. Extraction and GC analysis confirmed that the main adsorbed substrate was palmitate, and not oleate. Accumulated palmitate adsorbed onto the sludge and further beta-oxidation was inhibited when in the presence of oleic acid. If oleic acid was removed from the medium beta-oxidation proceeded with methane production. Suspended sludge was more efficient than granular sludge.

Fenton treatment (Fe(2+)/H(2)O(2)) and different ozone-based Advanced Oxidation Processes (AOPs) (O(3), O(3)/OH(-) and O(3)/H(2)O(2)) were evaluated as pre-treatment of a mature landfill leachate, in order to improve the biodegradability of its recalcitrant organic matter for subsequent biological treatment. With a two-fold diluted leachate, at optimised experimental conditions (initial pH 3, H(2)O(2) to Fe(2+) molar ratio of 3, Fe(2+) dosage of 4 mmol L(-1), and reaction time of 40 min) Fenton treatment removed about 46% of chemical oxygen demand (COD) and increased the five-day biochemical oxygen demand (BOD(5)) to COD ratio (BOD(5)/COD) from 0.01 to 0.15. The highest removal efficiency and biodegradability was achieved by ozone at higher pH values, solely or combined with H(2)O(2). These results confirm the enhanced production of hydroxyl radical under such conditions. After the application for 60 min of ozone at 5.6 g O(3)h(-1), initial pH 7, and 400 mg L(-1) of hydrogen peroxide, COD removal efficiency was 72% and BOD(5)/COD increased from 0.01 to 0.24. An estimation of the operating costs of the AOPs processes investigated revealed that Fe(2+)/H(2)O(2) was the most economical system (8.2 € m(-3)g(-1) of COD removed) to treat the landfill leachate. This economic study, however, should be treated with caution since it does not consider the initial investment, prices at plant scale, maintenance and labour costs.