Microbiological aspects of aerobic thermophilic treatment of swine waste

A thermophilic strain (D2) identified as a Bacillus sp. was isolated from an aerobic digestor of swine waste after several months of operation at 55 degrees C. Aerobic thermophilic batch treatment of swine waste inoculated with strain D2 was studied in a 4-liter fixed-bed reactor. Stabilization of the waste was achieved in less than 30 h when the original chemical oxygen demand (COD) was between 15 and 20 g/liter or in less than 48 h when the COD was around 35 g/liter. When the COD was higher than 30 g/liter, the pH of the waste reached 9.2 to 9.5 during the treatment, and periodic adjustment of the pH to 8.5 was necessary to maintain the activity of the biofilm. In this reactor, ammoniacal nitrogen was completely eliminated by desorption in less than 72 h of incubation. The different packing materials used resulted in similar rates of degradation of organic matter. The thermophilic treatment was also efficient in the 75-liter digestor, and stabilization was achieved in approximately 50 h. A bank of 22 thermophilic bacterial strains originating from different environments and adapted to the thermophilic treatment of swine waste was established. This thermophilic treatment allows, in one step, rapid stabilization of the waste, elimination of the bad smell, and complete elimination of ammonia nitrogen by stripping.     

Efficiency of bacterial protein synthesis during anaerobic degradation of cattle waste

The rate of [15N]ammonia (15NH3) uptake or incorporation into bacterial cells was studied, using stirred, 3-liter benchtop digestors fed on a semicontinuous basis with cattle waste. The fermentations were carried out at 40 and 60 degrees C and at four different loading rates (3, 6, 9, and 12 g of volatile solids per liter of reactor volume per day). The rate of NH3-N incorporation for the period 1 to 5 h after feeding at the four different loading rates was 0.49, 0.83, 1.05, and 1.08 mg/liter per h in the mesophilic digestor and 0.68, 1.07, 1.17, and 1.21 mg/liter per h in the thermophilic digestor. Values were lower 7 to 21 h after feeding in both digestors and were related to the rate of fermentation or CH4 production. In the mesophilic digestors, the rate of bacterial cell production ranged from 3.97 to 8.72 mg of dry cells per liter per h, 1 to 5 h after feeding at the different loading rates. Corresponding values for the thermophilic digestors ranged from 5.46 to 9.77 mg of dry cells per liter per h. Cell yield values ranged from 2.3 to 3.1 mg of dry cells per mol of CH4 produced in the mesophilic and thermophilic digestors at the two lower loading rates. The values were higher (2.8 to 3.4) in the mesophilic digestors at the two higher loading rates because of the accumulation of propionate and a consequent reduction in CH4 production. Low cell yields such as those measured in this study are characteristic of low-specific-growth rates under energy-limited conditions.     

Efficiency of bacterial protein synthesis during anaerobic degradation of cattle waste.

The rate of [15N]ammonia (15NH3) uptake or incorporation into bacterial cells was studied, using stirred, 3-liter benchtop digestors fed on a semicontinuous basis with cattle waste. The fermentations were carried out at 40 and 60 degrees C and at four different loading rates (3, 6, 9, and 12 g of volatile solids per liter of reactor volume per day). The rate of NH3-N incorporation for the period 1 to 5 h after feeding at the four different loading rates was 0.49, 0.83, 1.05, and 1.08 mg/liter per h in the mesophilic digestor and 0.68, 1.07, 1.17, and 1.21 mg/liter per h in the thermophilic digestor. Values were lower 7 to 21 h after feeding in both digestors and were related to the rate of fermentation or CH4 production. In the mesophilic digestors, the rate of bacterial cell production ranged from 3.97 to 8.72 mg of dry cells per liter per h, 1 to 5 h after feeding at the different loading rates. Corresponding values for the thermophilic digestors ranged from 5.46 to 9.77 mg of dry cells per liter per h. Cell yield values ranged from 2.3 to 3.1 mg of dry cells per mol of CH4 produced in the mesophilic and thermophilic digestors at the two lower loading rates. The values were higher (2.8 to 3.4) in the mesophilic digestors at the two higher loading rates because of the accumulation of propionate and a consequent reduction in CH4 production. Low cell yields such as those measured in this study are characteristic of low-specific-growth rates under energy-limited conditions.     

Metabolic Activity of Fatty Acid-Oxidizing Bacteria and the Contribution of Acetate, Propionate, Butyrate, and CO(2) to Methanogenesis in Cattle Waste at 40 and 60 degrees C

The quantitative contribution of fatty acids and CO(2) to methanogenesis was studied by using stirred, 3-liter bench-top digestors fed on a semicontinuous basis with cattle waste. The fermentations were carried out at 40 and 60 degrees C under identical loading conditions (6 g of volatile solids per liter of reactor volume per day, 10-day retention time). In the thermophilic digestor, acetate turnover increased from a prefeeding level of 16 muM/min to a peak (49 muM/min) 1 h after feeding and then gradually decreased. Acetate turnover in the mesophilic digestor increased from 15 to 40 muM/min. Propionate turnover ranged from 2 to 5.2 and 1.5 to 4.5 muM/min in the thermophilic and mesophilic digestors, respectively. Butyrate turnover (0.7 to 1.2 muM/min) was similar in both digestors. The proportion of CH(4) produced via the methyl group of acetate varied with time after feeding and ranged from 72 to 75% in the mesophilic digestor and 75 to 86% in the thermophilic digestor. The contribution from CO(2) reduction was 24 to 29% and 19 to 27%, respectively. Propionate and butyrate turnover accounted for 20% of the total CH(4) produced. Acetate synthesis from CO(2) was greatest shortly after feeding and was higher in the thermophilic digestor (0.5 to 2.4 muM/min) than the mesophilic digestor (0.3 to 0.5 muM/min). Counts of fatty acid-degrading bacteria were related to their turnover activity.     

Metabolic Activity of Fatty Acid-Oxidizing Bacteria and the Contribution of Acetate, Propionate, Butyrate, and CO2 to Methanogenesis in Cattle Waste at 40 and 60°C

The quantitative contribution of fatty acids and CO₂ to methanogenesis was studied by using stirred, 3-liter bench-top digestors fed on a semicontinuous basis with cattle waste. The fermentations were carried out at 40 and 60°C under identical loading conditions (6 g of volatile solids per liter of reactor volume per day, 10-day retention time). In the thermophilic digestor, acetate turnover increased from a prefeeding level of 16 μM/min to a peak (49 μM/min) 1 h after feeding and then gradually decreased. Acetate turnover in the mesophilic digestor increased from 15 to 40 μM/min. Propionate turnover ranged from 2 to 5.2 and 1.5 to 4.5 μM/min in the thermophilic and mesophilic digestors, respectively. Butyrate turnover (0.7 to 1.2 μM/min) was similar in both digestors. The proportion of CH₄ produced via the methyl group of acetate varied with time after feeding and ranged from 72 to 75% in the mesophilic digestor and 75 to 86% in the thermophilic digestor. The contribution from CO₂ reduction was 24 to 29% and 19 to 27%, respectively. Propionate and butyrate turnover accounted for 20% of the total CH₄ produced. Acetate synthesis from CO₂ was greatest shortly after feeding and was higher in the thermophilic digestor (0.5 to 2.4 μM/min) than the mesophilic digestor (0.3 to 0.5 μM/min). Counts of fatty acid-degrading bacteria were related to their turnover activity.     

Anaerobic digestion of cattle waste at mesophilic and thermophilic temperatures

Methanogenesis was studied using stirred, bench-top fermentors of 3-1 working volume fed on a semi-continuous basis with waste obtained from cattle fed a high grain, finishing diet. Digestion was carried out at 40 and 60°C. CH₄ production was 11.8, 18.3, 61.9 and 84.5% higher in the thermophilic than the mesophilic digestor at the 3, 6, 9 and 12 g volatile solids (VS) l^–1 reactor volume loading rates, respectively. When compared on an energetic basis CH₄ production was 7.4, 18.3, 72.9 and 107.3 kJ day higher in the thermophilic than the mesophilic digestor. CH₄ production decreased more rapidly with each increase in VS loading rate and decrease in retention time (RT) in the mesophilic than the thermophilic digestor. When expressed as l g^–1 VS fed or as kJ kJ^–1 fed, the amount of CH₄ was 49% less at the highest compared to the lowest loading rate in the mesophilic digestor. In the thermophilic digestor the decrease was only 16%. Propionate accumulated in the mesophilic digestor at the two highest loading rates, reaching concentrations of about 50 mM, but were only about 13 mM in the thermophilic digestor. Isobutyrate, isovalerate plus 2-methylbutyrate, and valerate also accumulated at the higher loading rates.     

High-cell-density cyclic fed-batch fermentation of a poly(3-hydroxybutyrate)-accumulating thermophile, Chelatococcus sp. strain MW10

Different fermentation strategies were employed for the cultivation of a new poly(3-hydroxybutyrate)-accumulating thermophilic bacterium, Chelatococcus sp. strain MW10, with the aim of achieving high-cell-density (HCD) growth and high poly(3-hydroxybutyrate) [poly(3HB)] productivity. Enhanced cultivation was achieved by a cyclic fed-batch fermentation (CFBF) technique (42-liter scale). Maximal poly(3HB) productivity was obtained during the second cycle [16.8 ± 4.2 g poly(3HB)/liter]. At the end of CFBF (265 h), an HCD of up to 115.0 ± 4.3 g cell dry weight/liter was achieved.     

Methanogenesis in a Thermophilic (58 degrees C) Anaerobic Digestor: Methanothrix sp. as an Important Aceticlastic Methanogen

Aceticlastic methanogens and other microbial groups were enumerated in a 58 degrees C laboratory-scale (3 liter) anaerobic digestor which was fed air-classified municipal refuse, a lignocellulosic waste (loading rate = 1.8 to 2.7 g of volatile solids per liter per day; retention time = 10 days). Two weeks after start-up, Methanosarcina sp. was present in high numbers (10 to 10 CFU/ml) and autofluorescent Methanosarcina-like clumps were abundant in sludge examined by using epifluorescence microscopy. After about 4 months of digestor operation, numbers of Methanosarcina sp. dropped 2 to 3 orders of magnitude and large numbers (most probable number = 10 to 10/ml) of a thermophilic aceticlastic methanogen morphologically resembing Methanothrix sp. were found. Methanothrix sp. had apparently displaced Methanosarcina sp. as the dominant aceticlastic methanogen in the digestor. During the period when Methanothrix sp. was apparently dominant, acetate concentrations varied between 0.3 and 1.5 mumol/ml during the daily feeding cycle, and acetate was the precursor of 63 to 66% of the methane produced during peak digestor methanogenesis. The apparent K(m) value obtained for methanogenesis from acetate, 0.3 mumol/ml, indicated that the aceticlastic methanogens were nearly saturated for substrate during most of the digestor cycle. CO(2)-reducing methanogens were capable of methanogenesis at rates more than 12 times greater than those usually found in the digestor. Added propionate (4.5 mumol/ml) was metabolized slowly by the digestor populations and slightly inhibited methanogenesis. Added n-butyrate, isobutyrate, or n-valerate (4.5 mumol/ml each) were broken down within 24 h. Isobutyrate was oxidized to acetate, a novel reaction possibly involving isomerization to n-butyrate. The rapid growth rate and versatile metabolism of Methanosarcina sp. make it a likely organism to be involved in start-up, whereas the low K(m) value of Methanothrix sp. for acetate may cause it to be favored in stable digestors operated with long retention times.     

Mesophilic and thermophilic biotreatment of BTEX-polluted air in reactors

This study compares the removal of a mixture of benzene, toluene, ethylbenzene, and all three xylene isomers (BTEX) in mesophilic and thermophilic (50 degrees C) bioreactors. In the mesophilic reactor fungi became dominant after long-term operation, while bacteria dominated in the thermophilic unit. Microbial acclimation was achieved by exposing the biofilters to initial BTEX loads of 2-15 g m(-3) h(-1), at an empty bed residence time of 96 s. After adaptation, the elimination capacities ranged from 3 to 188 g m(-3) h(-1), depending on the inlet load, for the mesophilic biofilter with removal efficiencies reaching 96%. On the other hand, in the thermophilic reactor the average removal efficiency was 83% with a maximum elimination capacity of 218 g m(-3) h(-1). There was a clear positive relationship between temperature gradients as well as CO(2) production and elimination capacities across the biofilters. The gas phase was sampled at different depths along the reactors observing that the percentage pollutant removal in each section was strongly dependant on the load applied. The fate of individual alkylbenzene compounds was checked, showing the unusually high biodegradation rate of benzene at high loads under thermophilic conditions (100%) compared to its very low removal in the mesophilic reactor at such load (<10%). Such difference was less pronounced for the other pollutants. After 210 days of operation, the dry biomass content for the mesophilic and thermophilic reactors were 0.300 and 0.114 g g(-1) (support), respectively, reaching higher removals under thermophilic conditions with a lower biomass accumulation, that is, lower pressure drop.     

Production of PHA from starchy wastewater via organic acids

Polyhydroxyalkanoate (PHA) was produced from a starchy wastewater in a two-step process of microbial acidogenesis and acid polymerization. The starchy organic waste was first digested in a thermophilic upflow anaerobic sludge blanket (UASB) reactor to form acetic (60-80%), propionic (10-30%) and butyric (5-40%) acids. The total volatile fatty acids reached 4000 mg l(-1) at a chemical oxygen demand (COD) loading rate of 25-35 g l(-1) day(-1). A carbon balance indicates that up to 43% of the organic carbon in the starchy waste went to the organic acids and the rest to biogas, volatile suspended solids and residual sludge accumulated in the reactor. The acid composition profile was affected by COD loading rate: a medium rate around 9 g l(-1) day(-1) gave a high propionic acid content (29% wt) and a high rate around 26 g l(-1) day(-1) led to a high butyric acid content (34% wt). The acids in the effluent solution after microfiltration were utilized and polymerized into PHA by bacterium Alcaligenes eutrophus in a second reactor. Fifty grams of PHA was produced from 100 g total organic carbon (TOC) utilized, a yield of 28% based on TOC, which is comparable with 55 g PHA per 100 g TOC of pure butyric and propionic acids used. PHA formation from individual acids was further investigated in a semi-batch reactor with three acid feeding rates. With a limited nitrogen source (80-100 mg NH(3) per liter), the active biomass of A. eutrophus, not including the accumulated PHA in cells, was maintained at a constant level (8-9 g l(-1)) while PHA content in the cell mass increased continuously in 45 h; 48% PHA with butyric acid and 53% PHA with propionic acid, respectively. Polyhydroxybutyrate was formed from butyric acid and poly(hydroxybutyrate-hydroxyvalerate) formed from propionic acid with 38% hydroxyvalerate.     

The effect of pH on the efficiency of an SBR processing piggery wastewater

To treat piggery wastewater efficiently, the hydrolysis of urea (mainly derived from swine urine) in piggery wastewater with the change of sewage pH must be considered. Using activated sludge, piggery wastewater was treated in a sequencing batch reactor (SBR), and the effects of influent pH on SBR processing efficiency, sludge settle ability, and sludge activity were investigated. The results showed that a high influent pH value contributed to the improvement of the removal rate of ammonia nitrogen and reduction of the chemical oxygen demand (COD). When the influent pH was between 9.0 and 9.5, the removal rate of ammonia nitrogen was higher than 90%, and the reduction of COD from its original value was 80%. The influent pH had a greater influence on sludge concentration and sludge activity. When the influent pH increased from 7.0 to 9.5, the sludge concentration increased from 2,350 to 3,947 mg/L in the reactor, and the activities of ammonium oxidizing bacteria (AOB) and nitrite oxidizing bacteria (NOB) first increased and then decreased. When the influent pH was 9.0 and 8.0, the maximum values (0.48 g O₂/(g MLSS/day) and 0.080 g O₂/(g MLSS/day)) were reached, and the sludge settling ratio was nearly steady between 20 and 35% in each reactor.     

Key physiology of anaerobic ammonium oxidation.

The physiology of anaerobic ammonium oxidizing (anammox) aggregates grown in a sequencing batch reactor was investigated quantitatively. The physiological pH and temperature ranges were 6.7 to 8.3 and 20 to 43 degrees C, respectively. The affinity constants for the substrates ammonium and nitrite were each less than 0.1 mg of nitrogen per liter. The anammox process was completely inhibited by nitrite concentrations higher than 0.1 g of nitrogen per liter. Addition of trace amounts of either of the anammox intermediates (1. 4 mg of nitrogen per liter of hydrazine or 0.7 mg of nitrogen per liter of hydroxylamine) restored activity completely.     

Key Physiology of Anaerobic Ammonium Oxidation

The physiology of anaerobic ammonium oxidizing (anammox) aggregates grown in a sequencing batch reactor was investigated quantitatively. The physiological pH and temperature ranges were 6.7 to 8.3 and 20 to 43°C, respectively. The affinity constants for the substrates ammonium and nitrite were each less than 0.1 mg of nitrogen per liter. The anammox process was completely inhibited by nitrite concentrations higher than 0.1 g of nitrogen per liter. Addition of trace amounts of either of the anammox intermediates (1.4 mg of nitrogen per liter of hydrazine or 0.7 mg of nitrogen per liter of hydroxylamine) restored activity completely.     

Production of fungal biomass protein using microfungi from winery wastewater treatment

This study was carried out to investigate the production of fungal biomass protein (FBP) in treatment of winery wastewater using microfungi. Three fungal strains, Trichoderma viride WEBL0702, Aspergillus niger WEBL0901 and Aspergillus oryzae WEBL0401, were selected in terms of microbial capability for FBP production and COD reduction. T. viride appeared to be the best strain for FBP production due to high productivity and less nitrogen requirement. More than 5 g/L of fungal biomass was produced in shake fermentation using T. viride without nitrogen addition, and by A. oryzae and A. niger with addition of 0.5-1.0 g/L (NH4)2SO4. The FBP production process corresponded to 84-90% COD reduction of winery wastewater. Fungal biomass contained approximately 36% protein produced by two Aspergillus strains, while biomass produced by T. viride consisted of 19.8% protein. Kinetic study indicated that maximum fungal cell growth could be achieved in 24h for T. viride and 48 h for A. oryzae and A. niger. Current results indicated that it could be feasible to develop a biotechnological treatment process integrated with FBP production from the winery waste streams.     

Impact of leachate recirculation and recirculation volume on stabilization of municipal solid wastes in simulated anaerobic bioreactors

The effects of leachate recirculation and the recirculation rate on the anaerobic treatment of domestic solid waste was investigated in three simulated landfill anaerobic bioreactors. A single pass reactor was operated without leachate recirculation while the other two reactors were operated with leachate recirculation. The leachate recirculation rate was 9 l/day (13% of the reactor volume) in Reactor₉, while the recirculation rate was 21 l/day (30% of the reactor volume), in Reactor₂₁. pH, chemical oxygen demand (COD), volatile fatty acids (VFA), ammonium–nitrogen (NH₄–N) total and methane gas measurements in leachate samples were regularly monitored. After 220 days of anaerobic incubation, it was observed that the pH, COD, VFA concentrations, methane gas productions and methane percentages in Reactor₉ were better than the single pass reactor and Reactor₂₁. When the leachate recirculation rate was increased to three times a decrease in pH, and an increase in VFA and COD concentrations were observed in Reactor₂₁. The COD values were measured as 47 000, 39 000 and 52 000 mg/l while the VFA concentrations were 15 000, 13 000 and 21 000 mg/l, respectively, in single pass, Reactor₉ and Reactor₂₁ after 220 days of anaerobic incubation. The values of pH were 5.89, 6.44 and 6.16, respectively, after anaerobic incubation. The mean methane percentages of single pass reactor, Reactor₉ and Reactor₂₁ were 30, 50 and 40%, respectively, after 50 days of incubation. Leachate recirculation reduced the waste stabilization time and was effective in enhancing methane gas production and improving leachate. However, leachate recirculation was not effective in removing ammonia from the leachate. The amounts of COD recovered by methane were 62.9, 162.3 and 94.6 g for single pass, Reactor₉ and Reactor₂₁, respectively, at the end of 220 days of anaerobic incubation.     

Biodegradation of p-nitrophenol in an aqueous waste stream by immobilized bacteria.

Microbiological analyses of activated sludge reactors after repeated exposure to 100 mg of p-nitrophenol (PNP) per liter resulted in the isolation of three Pseudomonas species able to utilize PNP as a sole source of carbon and energy. Cell suspensions of the three Pseudomonas sp., designated PNP1, PNP2, and PNP3, mineralized 70, 60, and 45% of a 70-mg/liter dose of PNP in 24, 48, and 96 h, respectively. Mass-balance analyses of PNP residues for all three cultures showed that undegraded PNP was less than 1% (less than 50 micrograms); volatile metabolites, less than 1%; cell residues, 8.4 to 14.9%; and water-soluble metabolites, 1.2 to 6.7%. A mixed culture of all three PNP-degrading Pseudomonas sp. was immobilized by adsorption onto diatomaceous earth biocarrier in a 1.75-liter Plexiglas column. The column was aerated and exposed to a synthetic waste stream containing 629 to 2,513 mg of PNP per liter at flow rates of 2 to 15 ml/min. Chemical loading studies showed that the threshold concentration for acute toxicity of PNP to the immobilized bacteria was 2,100 to 2,500 mg/liter. Further studies at PNP concentrations of 1,200 to 1,800 mg/liter showed that greater than 99 and 91 to 99% removal of PNP was achieved by immobilized bacteria at flow rates of 10 and 12 ml/min, respectively. These values represent hydraulic retention times of 48 to 58 min and PNP removal rates of 0.99 to 1.1 mg/h per g of biocarrier at 25 degrees C under optimal conditions. This study shows the successful use of immobilized bacteria technology to remove high concentrations of PNP from aqueous waste streams.     

Biodegradation of p-nitrophenol in an aqueous waste stream by immobilized bacteria

Microbiological analyses of activated sludge reactors after repeated exposure to 100 mg of p-nitrophenol (PNP) per liter resulted in the isolation of three Pseudomonas species able to utilize PNP as a sole source of carbon and energy. Cell suspensions of the three Pseudomonas sp., designated PNP1, PNP2, and PNP3, mineralized 70, 60, and 45% of a 70-mg/liter dose of PNP in 24, 48, and 96 h, respectively. Mass-balance analyses of PNP residues for all three cultures showed that undegraded PNP was less than 1% (less than 50 micrograms); volatile metabolites, less than 1%; cell residues, 8.4 to 14.9%; and water-soluble metabolites, 1.2 to 6.7%. A mixed culture of all three PNP-degrading Pseudomonas sp. was immobilized by adsorption onto diatomaceous earth biocarrier in a 1.75-liter Plexiglas column. The column was aerated and exposed to a synthetic waste stream containing 629 to 2,513 mg of PNP per liter at flow rates of 2 to 15 ml/min. Chemical loading studies showed that the threshold concentration for acute toxicity of PNP to the immobilized bacteria was 2,100 to 2,500 mg/liter. Further studies at PNP concentrations of 1,200 to 1,800 mg/liter showed that greater than 99 and 91 to 99% removal of PNP was achieved by immobilized bacteria at flow rates of 10 and 12 ml/min, respectively. These values represent hydraulic retention times of 48 to 58 min and PNP removal rates of 0.99 to 1.1 mg/h per g of biocarrier at 25 degrees C under optimal conditions. This study shows the successful use of immobilized bacteria technology to remove high concentrations of PNP from aqueous waste streams.     

Liquefaction and gasification during anaerobic digestion of coffee waste by two-phase methane fermentation with slurry-state liquefaction

In order to improve the gas evolution rate during anaerobic digestion of coffee waste by two-phase methane fermentation with slurry-state liquefaction, the liquefaction and gasification processes were separately investigated. In the liquefaction process (including the acidification process), treatment at a pH above 6 had no major effects on the liquefaction and acidification rates. However, the VFA production rates were 880 and 320 mg/l·d during mesophilic (37°C) and thermophilic (53°C) liquefaction, respectively. Mesophilic conditions were superior to thermophilic conditions in the liquefaction. With respect to the gasification process, a high TOC volumetric loading rate of 21 g/l·d was achieved during thermophilic gasification. However, the mesophilic gasification did not yield stable data, even at a low TOC volumetric loading rate of 2 g/l·d. The gas yield was 1.7 l/g TOC consumed during thermophilic gasification. The thermophilic liquefaction and thermophilic gasification reactors were connected in series and a two-phase experiment was conducted with the reactors at various volumetric ratios. The maximum gas evolution rate of 1.43 l/l·d was achieved with a combination of a gasification reactor with a 0.45l working volume and liquefaction reactor with a 2l working volume. This rate was 1.7 times higher than the rate obtained in a previous study.     

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).     

Non-Sterilized Fermentative Production of Polymer-Grade L-Lactic Acid by a Newly Isolated Thermophilic Strain Bacillus sp. 2–6

Background

The demand for lactic acid has been increasing considerably because of its use as a monomer for the synthesis of polylactic acid (PLA), which is a promising and environment-friendly alternative to plastics derived from petrochemicals. Optically pure l-lactic acid is essential for polymerization of PLA. The high fermentation cost of l-lactic acid is another limitation for PLA polymers to compete with conventional plastics.

Methodology/Principal Findings

A Bacillus sp. strain 2–6 for production of l-lactic acid was isolated at 55°C from soil samples. Its thermophilic characteristic made it a good lactic acid producer because optically pure l-lactic acid could be produced by this strain under open condition without sterilization. In 5-liter batch fermentation of Bacillus sp. 2–6, 118.0 g/liter of l-lactic acid with an optical purity of 99.4% was obtained from 121.3 g/liter of glucose. The yield was 97.3% and the average productivity was 4.37 g/liter/h. The maximum l-lactic acid concentration of 182.0 g/liter was obtained from 30-liter fed-batch fermentation with an average productivity of 3.03 g/liter/h and product optical purity of 99.4%.

Conclusions/Significance

With the newly isolated Bacillus sp. strain 2–6, high concentration of optically pure l-lactic acid could be produced efficiently in open fermentation without sterilization, which would lead to a new cost-effective method for polymer-grade l-lactic acid production from renewable resources.