The anaerobic process functions effectively over two temperature ranges: the mesophilic range of 85-100°F (29-38°C) and the thermophilic range of 120-135°F (49-57°C). Although the rates of reaction are much greater in the thermophilic range, the maintenance of higher temperatures is usually not economically justifiable.
Methane organisms function over a pH range of 6.6-7.6 with an optimum near pH 7.0. When the rate of acid formation exceeds the rate of breakdown to methane, a process unbalance results in which the pH decreases, gas production falls off, and the CO2 content increases . pH control is therefore essential to ensure a high rate of methane production. According to German literature, the tolerable pH range for anaerobic microorganisms is between 6.8 and 7.5. This means that the anaerobic biocenosis is very pH-specific .
With regard to the influence of initial concentration on anaerobic degradation, preliminary laboratory and pilot-scale experimentation on diluted olive oil mill effluents
(OME)  showed that the anaerobic contact process was able to provide high organic removal efficiency (80-85%) at 35°C and at an organic load lower than 4 kg COD/m3-day; however, in particular at high feed concentration, the process proved unstable due to the inhibitory effects of substances
such as polyphenols. Moreover, additions of alkalinity to neutralize acidity and ammonia to furnish nitrogen for cellular biosynthesis were required.
To overcome these difficulties and improve process efficiency and stability, there are basically two methods that may be adopted : (a) the treatment of combined OME and sewage sludge in contact bioreactors; and (b) operation with more diluted OME in highrate bioreactors (such as UASB reactors and fixed-bed filters).
In the first method, conventional digesters can be overloaded with concentrated soluble wastes such as OME, and still operate satisfactorily. Moreover, nutrients such as ammonia and buffers are provided by degradation of proteineous substances from sludge. On this basis, laboratory-scale experimentation  has shown that removal efficiencies of 65 and 37% in terms of COD and VSS, respectively, were obtained at 35°C and at an organic load of 4.2 kg COD/m3-day (66% from sewage sludge, 34% from OME). Higher OME additions led to process imbalance due to the inhibitory effects of polyphenols. This method, based on anaerobic contact digestion of combined OME and sewage sludge, seems to be suitable only for those locations where the polluting load due to the OME is lower than the domestic wastewater load. In this regard it is worth considering that during the olive oil milling season, OME pollution largely exceeds that from domestic wastewater .
With regard to the second method, based on the use of high-rate bioreactors, experimentation on UASB reactors [46,47] showed that COD removal efficiencies of about 70-75% were obtained at 37°C and at an organic load in the range 12-18 kg COD/m3-day by adopting a dilution ratio in the range of 1:8 to 1:5 (OME: tap water; diluted OME initial concentration in the range 11-19 g COD/L). Slightly less satisfactory results were obtained by using anaerobic filters filled with macroreticulated polyurethane foam .
It is important to note that immobilization of methanogenic bacteria may decrease the toxicity of phenolic compounds. Another pilot-scale anaerobic-aerobic treatment of OME mixed with settled domestic wastewater  produced a final COD concentration of about 160 mg/L, provided that a dilution ratio of 1:60 to 1:100 was adopted, corresponding to a COD load ratio equal to 3:1 for OME and domestic wastewater, respectively. This ratio is typical for those locations with a high density of olive oil mills. However, in addition to the high value required for the dilution ratio, the final effluent did not comply with legal requirements in terms of color and nitrogen .
The aforementioned data clearly show that in the treatment of OME, even when carried out with the use of most appropriate technology, that is, anaerobic digestion, it was difficult to reach the treatment efficiencies required by national regulations throughout the Mediterranean area. In particular, methanogenesis, which represents the limiting step in the anaerobic digestion of soluble compounds, is severely hindered by the inhibition caused by the buildup of volatile fatty acids (VFAs) and/or the presence of a high concentration of phenolic compounds and/or oleic acid in the OME. As for phenol, 1.25 g/L leads to 50% activity reduction of acetateutilizing methanogens . As for oleic acid, it is reported that 5 mM is toxic to methanogenic bacteria .
The reader may refer to the following Case Study V to better understand the mechanism of biodegradation of the main compounds contained in the OME in relation to pH, temperature, and initial concentration of effluents, and in particular the mutual coherence of the two successive partial stages occurring in anaerobic digestion of OME, acidogenesis, and methanogenesis.
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