Biokaasun tuottaminen ja hyödyntäminen Lappeenrannassa

Lappeenrannassa kerätään ja hyödynnetään tällä hetkellä kaatopaikkakaasua 0,3 milj.m3 vuodessa. Biokaasua voitaisiin tuottaa Lappeenrannassa mädättämällä bioperäisiä jätteitä ja biokaasuntuotantoa varten kasvatettuja energiakasveja. Biokaasuntuotantoon soveltuvia jätteitä ovat erilliskerätty biojäte, jätevedenpuhdistamon jätevesiliete, puutarhajäte, lietelannat ja oljet. Kesannolla olevilla peltoaloilla voitaisiin kasvattaa ruokohelpeä. Biokaasun tuotantoon soveltuvia materiaaleja voitaisiin kerätä 143 000 t/a ja kasvattaa 68 000 t/a. Työssä tarkastellaan vaihtoehtoa, jossa mädätetään vain puhdistamoliete, sekä useita materiaaleja mädättävää yhteismädättämöä, johon liittyen tutkitaan kolmea eri vaihtoehtoa: kunnallisen jätteen mädätystä, kaiken jätteen mädätystä ja jätteen sekä energiakasvien mädätystä. Paras sijoituspaikka mädättämölle olisi jätevedenpuhdistamon läheisyydessä. Jätemateriaalista saataisiin kaasua enintään 12 milj. m3 ja energiakasveista enintään 16 milj. m3. Kaasusta voitaisiin tuottaa energiaa CHP-laitoksessa enintään 184 GWh. Mikäli biokaasun tuotannolla halutaan ensisijaisesti vähentää kasvi-huonekaasupäästöjä, kannattaa kaasu jalostaa ajoneuvopolttoaineeksi. Jalostettu kaasu on mahdollista myös syöttää maakaasuverkostoon. Suurimmat tulot on mahdollista saavuttaa yhdistetyssä sähkön- ja lämmöntuotannossa, mikäli biokaasulle suunniteltu syöttötariffi toteutuu. Muussa tapauksessa suurimmat tulot saadaan jalostamalla biokaasua ajoneuvojen polttoaineeksi.

Simultaneous determination of Cd and Pb in antibiotics used in sugar-cane fermentation process by GFAAS

A method has been developed for the simultaneous determination of Cd and Pb in antibiotics used in sugar-cane fermentation by GFAAS. The integrated platform of transversely heated graphite atomizer was treated with tungsten to form a coating of tungsten carbide. Six samples of commercial solid antibiotics were analyzed by injecting 20 µL of digested samples into the pretreated graphite platform with co-injection of 5 µL of 1000 mg L-1 Pd as chemical modifier. Samples were mineralized in a closed-vessel microwave-assisted acid-digestion system using nitric acid plus hydrogen peroxide. The pyrolysis and atomization temperatures of the heating program of the atomizer were selected as 600°C and 2200°C, respectively. The calculated characteristic mass for Cd and Pb was 1.6 pg and 42 pg, respectively. Limits of detection (LOD) based on integrated absorbance were 0.02 µg L-1 Cd and 0.7 µg L-1 Pb and the relative standard deviations (n = 10) for Cd and Pb were 5.7% and 8.0%, respectively. The recoveries of Cd and Pb added to the digested samples varied from 91% to 125% (Cd) and 80% to 112% (Pb).

Direct determination of calcium in milk by atomic absorption spectrometry using flow-injection analysis

A flow-injection system with sample and reagent addition by the synchronous merging zones approach for calcium determination in milk by flame AAS is proposed. Main parameters were optimized using a factorial design with central point. The optimum conditions were 2.5% (m/v) for La concentration, 8 mL min-1 for the carrier flow-rate, 20 cm for coiled reactor and 250 ìL for sample volume. Different sample preparation procedures were evaluated such as dilution in water or acid and microwave-assisted decomposition using concentrated or diluted acids. The optimized flow system was applied to determine Ca in eleven commercial milk samples and two standard reference materials diluted in water. Similar calcium levels were encountered comparing the results obtained by the proposed method (dilution in water) with those obtained using microwave-oven digestion. Results obtained in two standard reference materials were in agreement at 95% confidence level with those certified. Recoveries of spiked samples were in the 93% - 116% range. Relative standard deviation (n = 12) was < 5.4% and the sample throughput was 150 measurements per hour, corresponding to a consumption of 250 µL of sample and 6.25 mg La per determination.

A simple method for the spectrophotometric determination of cephalosporins in pharmaceuticals using variamine blue

A simple spectrophotometric method for the determination of cefotaxime, ceftriaxone, cefadroxil and cephalexin with variamine blue is presented. The determination is based on the hydrolysis of β-lactam ring of cephalosporins with sodium hydroxide which subsequently reacts with iodate to liberate iodine in acidic medium. The liberated iodine oxidizes variamine blue to violet colored species of maximum absorption at 556 nm. The absorbance is measured within the pH range of 4.0-4.2. Beer's law is obeyed in the range of 0.5-5.8 µg mL-1, 0.2-7.0 µg mL-1, 0.2-5.0 µg mL-1 and 0.5-8.5 µg mL-1 for cefotaxime, ceftriaxone, cefadroxil and cephalexin respectively. The analytical parameters were optimized and the method is successfully applied for the determination of cefotaxime, ceftriaxone, cefadroxil and cephalexin in pharmaceuticals.

Epäpuhtauksien poistaminen prosessivesistä

Kemira Chemicals Oy in Äetsä produces sodium chlorate as its main product. It is produced with electrolysis in electrolyte cells. During the manufacturing process impurities, out of which the largest one is iron, accumulate in the cells. These impurities are removed in cell wash with hydrochloric acid liquid, after which the wash water is precipitated with sodium hydroxide and sodium carbonate, and filtered with filter press. After the treatment the wash water is recycled back to the manufacturing process. The aim of this thesis was primarily to improve the treatment of wash water in order to remove the impurities with low costs. This would result in more impurity-free water and in sufficient capacity of impurity removal. The second aim was to maintain the chromium in the treated wash water because it forms a diaphragm of chromium hydroxide to cathode which prevents the flow of anions to cathode. The literature part investigates properties, use and manufacturing of sodium chlorate, electrolyte cell and its wash technique, and impurities of wash water. The beginning of the applied part investigates alternatives of separation methods which could be used to improve the treatment of wash water. In the experiments an optimum pH for the precipitation of wash water was determined, and a research of the use of sodium hydroxide, sodium carbonate, calcium hydroxide and calcium chloride as a precipitant was carried out. Also a suitable flocculant and a filter cloth for the treatment of wash water were determined. Finally, process changes were introduced, partly by applying the current equipment, and the costs and savings were calculated.

Sensitive spectrophotometric determination of lamotrigine in bulk drug and pharmaceutical formulations using bromocresol green

Two new, simple, rapid and reproducible spectrophotometric methods have been developed for the determination of lamotrigine (LMT) both in pure form and in its tablets. The first method (method A) is based on the formation of a colored ion-pair complex (1:1 drug/dye) of LMT with bromocresol green (BCG) at pH 5.02±0.01 and extraction of the complex into dichloromethane followed by the measurement of the yellow ion-pair complex at 410 nm. In the second (method B), the drug-dye ion-pair complex was dissolved in ethanolic potassium hydroxide and the resulting base form of the dye was measured at 620 nm. Beer's law was obeyed in the concentration range of 1.5-15 µg mL-1 and 0.5-5.0 µg mL-1 for method A and method B, respectively, and the corresponding molar absorptivity values are 1.6932 x 10(4) and 3.748 x 10(4) L mol-1cm-1. The Sandell sensitivity values are 0.0151 and 0.0068 µg cm-2 for method A and method B, respectively. The stoichiometry of the ion-pair complex formed between the dug and dye (1:1) was determined by Job's continuous variations method and the stability constant of the complex was also calculated. The proposed methods were applied successfully for the determination of drug in commercial tablets.

Development of a molecular method for detection and identification of Xanthomonas campestris pv. viticola

In order to develop a molecular method for detection and identification of Xanthomonas campestris pv. viticola (Xcv) the causal agent of grapevine bacterial canker, primers were designed based on the partial sequence of the hrpB gene. Primer pairs Xcv1F/Xcv3R and RST2/Xcv3R, which amplified 243- and 340-bp fragments, respectively, were tested for specificity and sensitivity in detecting DNA from Xcv. Amplification was positive with DNA from 44 Xcv strains and with DNA from four strains of X. campestris pv. mangiferaeindicae and five strains of X. axonopodis pv. passiflorae, with both primer pairs. However, the enzymatic digestion of PCR products could differentiate Xcv strains from the others. None of the primer pairs amplified DNA from grapevine, from 20 strains of nonpathogenic bacteria from grape leaves and 10 strains from six representative genera of plant pathogenic bacteria. Sensitivity of primers Xcv1F/Xcv3R and RST2/Xcv3R was 10 pg and 1 pg of purified Xcv DNA, respectively. Detection limit of primers RST2/Xcv3R was 10(4) CFU/ml, but this limit could be lowered to 10² CFU/ml with a second round of amplification using the internal primer Xcv1F. Presence of Xcv in tissues of grapevine petioles previously inoculated with Xcv could not be detected by PCR using macerated extract added directly in the reaction. However, amplification was positive with the introduction of an agar plating step prior to PCR. Xcv could be detected in 1 µl of the plate wash and from a cell suspension obtained from a single colony. Bacterium identity was confirmed by RFLP analysis of the RST2/Xcv3R amplification products digested with Hae III.

Mädätysjäännöksen tuotteistamismahdollisuudet Kymenlaaksossa

Biokaasun tuotantoa ollaan selvästi lisäämässä Suomessa. Biokaasutuksen kokonaishyödyn kannalta on olennaista, että mädätyksen lopputuote eli mädätysjäännös saadaan lannoitekäyttöön. Tämän työn tavoitteena oli selvittää Kymenlaakson Jäte Oy:n mahdollisuuksia tuotteis-taa Kymen Bioenergia Oy:n yhteismädätyslaitoksen mädätysjäännöstä. Työssä keskityttiin hyötykäyttövaihtoehdoista lannoitekäyttöön maanviljelyssä sekä tilanteeseen jossa mädätyslaitos käsittelee sekä puhdistamolietettä että biojätettä ja mädätysjäännös kuivataan mekaanisesti. Mekaanisesti kuivatun mädätysjäännöksen ensisijaiset tuotteistamisvaihtoehdot maanviljelyyn ovat joko jäännös sellaisenaan tai termisesti kuivattuna ja rakeistettuna, eli kuivarakeena. Mäkikylän laitoksen mädätysjäännöksen arvo peltolannoitteena on syyskuun 2010 keinolannoit-teiden hintaan vertaamalla sellaisenaan noin 1–20 €/t ja kuivarakeena noin 2–60 €/t. Arvo riippuu siitä, miten tuotteiden typpeä ja fosforia huomioidaan kasveille käyttökelpoiseksi. Täl-lä hetkellä käyttökelpoisin tapa on ympäristötuen puhdistamolietetuotteita koskevien ehtojen mukaisesti ottaa huomioon vesiliukoinen typpi ja 40 % kokonaisfosforista. Tällöin mädätys-jäännöksen arvo on noin 6 €/t ja kuivarakeen n. 18 €/t. Käytön kannalta kuivarae on helpompi vaihtoehto ja alueen viljelijät ovat heille tehdyn kyselyn mukaan varsin kiinnostuneita kuivarakeesta lannoitteena. Muista tuotteistusvaihtoehdoista termisesti kuivaamalla mädätysjäännöksen tehollinen lämpö-arvo saapumistilassa on noin 10 MJ/kg. Vastaava arvo jyrsinturpeen kesäkuun 2010 hinnan mukaan on noin 30 €/t. Tuotteen soveltuvuus polttoon tulee silti varmistaa. Termisesti kuiva-tulla mädätysjäännöksellä on tuotteistamismahdollisuuksia hieman laajemmin kuin kompostoidulla. Kompostoidun mädätysjäännöksen tuotteistamisen lähtökohta on lähinnä viherrakentaminen. Maanviljelykäyttöä ajatellen mädätysjäännöstä ei välttämättä tarvitse kompostoida.

Determination of sugars and organic acids in pulp samples by capillary electrophoresis with UV detection

This MSc work was done in the project of BIOMECON financed by Tekes. The prime target of the research was, to develop methods for separation and determination of carbohydrates (sugars), sugar acids and alcohols, and some other organic acids in hydrolyzed pulp samples by capillary electrophoresis (CE) using UV detection. Aspen, spruce, and birch pulps are commonly used for production of papers in Finland. Feedstock components in pulp predominantly consist of carbohydrates, organic acids, lignin, extractives, and proteins. Here in this study, pulps have been hydrolyzed in analytical chemistry laboratories of UPM Company and Lappeenranta University in order to convert them into sugars, acids, alcohols, and organic acids. Foremost objective of this study was to quantify and identify the main and by-products in the pulp samples. For the method development and optimization, increased precision in capillary electrophoresis was accomplished by calculating calibration data of 16 analytes such as D-(-)-fructose, D(+)-xylose, D(+)-mannose, D(+)-cellobiose, D-(+)-glucose, D-(+)-raffinose, D(-)-mannitol, sorbitol, rhamnose, sucrose, xylitol, galactose, maltose, arabinose, ribose, and, α-lactose monohydratesugars and 16 organic acids such as D-glucuronic, oxalic, acetic, propionic, formic, glycolic, malonic, maleic, citric, L-glutamic, tartaric, succinic, adipic, ascorbic, galacturonic, and glyoxylic acid. In carbohydrate and polyalcohol analyses, the experiments with CE coupled to direct UV detection and positive separation polarity was performed in 36 mM disodium hydrogen phosphate electrolyte solution. For acid analyses, CE coupled indirect UV detection, using negative polarity, and electrolyte solution made of 2,3 pyridinedicarboxylic acid, Ca2+ salt, Mg2+ salts, and myristyltrimethylammonium hydroxide in water was used. Under optimized conditions, limits of detection, relative standard deviations and correlation coefficients of each compound were measured. The optimized conditions were used for the identification and quantification of carbohydrates and acids produced by hydrolyses of pulp. The concentrations of the analytes varied between 1 mg – 0.138 g in liter hydrolysate.

Learning through role-playing games: an approach for active learning and teaching

This study evaluates the use of role-playing games (RPGs) as a methodological approach for teaching cellular biology, assessing student satisfaction, learning outcomes, and retention of acquired knowledge. First-year undergraduate medical students at two Brazilian public universities attended either an RPG-based class (RPG group) or a lecture (lecture-based group) on topics related to cellular biology. Pre- and post-RPG-based class questionnaires were compared to scores in regular exams and in an unannounced test one year later to assess students' attitudes and learning. From the 230 students that attended the RPG classes, 78.4% responded that the RPG-based classes were an effective tool for learning; 55.4% thought that such classes were better than lectures but did not replace them; and 81% responded that they would use this method. The lecture-based group achieved a higher grade in 1 of 14 regular exam questions. In the medium-term evaluation (one year later), the RPG group scored higher in 2 of 12 questions. RPG classes are thus quantitatively as effective as formal lectures, are well accepted by students, and may serve as educational tools, giving students the chance to learn actively and potentially retain the acquired knowledge more efficiently.

Recovery and refining of manganese as by-product from hydrometallurgical processes

The consumption of manganese is increasing, but huge amounts of manganese still end up in waste in hydrometallurgical processes. The recovery of manganese from multi-metal solutions at low concentrations may not be economical. In addition, poor iron control typically prevents the production of high purity manganese. Separation of iron from manganese can be done with chemical precipitation or solvent extraction methods. Combined carbonate precipitation with air oxidation is a feasible method to separate iron and manganese due to the fast kinetics, good controllability and economical reagents. In addition the leaching of manganese carbonate is easier and less acid consuming than that of hydroxide or sulfide precipitates. Selective iron removal with great efficiency from MnSO4 solution is achieved by combined oxygen or air oxidation and CaCO3 precipitation at pH > 5.8 and at a redox potential of > 200 mV. In order to avoid gypsum formation, soda ash should be used instead of limestone. In such case, however, extra attention needs to be paid on the reagents mole ratios in order to avoid manganese coprecipitation. After iron removal, pure MnSO4 solution was obtained by solvent extraction using organophosphorus reagents, di-(2-ethylhexyl)phosphoric acid (D2EHPA) and bis(2,4,4- trimethylpentyl)phosphinic acid (CYANEX 272). The Mn/Ca and Mn/Mg selectivities can be increased by decreasing the temperature from the commonly used temperatures (40 –60oC) to 5oC. The extraction order of D2EHPA (Ca before Mn) at low temperature remains unchanged but the lowering of temperature causes an increase in viscosity and slower phase separation. Of these regents, CYANEX 272 is selective for Mn over Ca and, therefore, it would be the better choice if there is Ca present in solution. A three-stage Mn extraction followed by a two-stage scrubbing and two-stage sulfuric acid stripping is an effective method of producing a very pure MnSO4 intermediate solution for further processing. From the intermediate MnSO4 some special Mn- products for ion exchange applications were synthesized and studied. Three types of octahedrally coordinated manganese oxide materials as an alternative final product for manganese were chosen for synthesis: layer structured Nabirnessite, tunnel structured Mg-todorokite and K-kryptomelane. As an alternative source of pure MnSO4 intermediate, kryptomelane was synthesized by using a synthetic hydrometallurgical tailings. The results show that the studied OMS materials adsorb selectively Cu, Ni, Cd and K in the presence of Ca and Mg. It was also found that the exchange rates were reasonably high due to the small particle dimensions. Materials are stable in the studied conditions and their maximum Cu uptake capacity was 1.3 mmol/g. Competitive uptake of metals and acid was studied using equilibrium, batch kinetic and fixed-bed measurements. The experimental data was correlated with a dynamic model, which also accounts for the dissolution of the framework manganese. Manganese oxide micro-crystals were also bound onto silica to prepare a composite material having a particle size large enough to be used in column separation experiments. The MnOx/SiO2 ratio was found to affect significantly the properties of the composite. The higher the ratio, the lower is the specific surface area, the pore volume and the pore size. On the other hand, higher amount of silica binder gives composites better mechanical properties. Birnesite and todorokite can be aggregated successfully with colloidal silica at pH 4 and with MnO2/SiO2 weight ratio of 0.7. The best gelation and drying temperature was 110oC and sufficiently strong composites were obtained by additional heat-treatment at 250oC for 2 h. The results show that silica–supported MnO2 materials can be utilized to separate copper from nickel and cadmium. The behavior of the composites can be explained reasonably well with the presented model and the parameters estimated from the data of the unsupported oxides. The metal uptake capacities of the prepared materials were quite small. For example, the final copper loading was 0.14 mmol/gMnO2. According to the results the special MnO2 materials are potential for a specific environmental application to uptake harmful metal ions.

Sewage Sludge Electro-Dewatering

Original sludge from wastewater treatment plants (WWTPs) usually has a poor dewaterability. Conventionally, mechanical dewatering methods are used to increase the dry solids (DS) content of the sludge. However, sludge dewatering is an important economic factor in the operation of WWTPs, high water content in the final sludge cake is commonly related to an increase in transport and disposal costs. Electro‐dewatering could be a potential technique to reduce the water content of the final sludge cake, but the parameters affecting the performance of electro‐dewatering and the quality of the resulting sludge cake, as well as removed water, are not sufficiently well known. In this research, non‐pressure and pressure‐driven experiments were set up to investigate the effect of various parameters and experimental strategies on electro‐dewatering. Migration behaviour of organic compounds and metals was also studied. Application of electrical field significantly improved the dewatering performance in comparison to experiments without electric field. Electro‐dewatering increased the DS content of the sludge from 15% to 40 % in non‐pressure applications and from 8% to 41% in pressure‐driven applications. DS contents were significantly higher than typically obtained with mechanical dewatering techniques in wastewater treatment plant. The better performance of the pressure‐driven dewatering was associated to a higher current density at the beginning and higher electric field strength later on in the experiments. The applied voltage was one of the major parameters affecting dewatering time, water removal rate and DS content of the sludge cake. By decreasing the sludge loading rate, higher electrical field strength was established between the electrodes, which has a positive effect on an increase in DS content of the final sludge cake. However interrupted voltage application had anegative impact on dewatering in this study, probably because the off‐times were too long. Other factors affecting dewatering performance were associated to the original sludge characteristics and sludge conditioning. Anaerobic digestion of the sludge with high pH buffering capacity, polymer addition and freeze/thaw conditioning had a positive impact on dewatering. The impact of pH on electro‐dewatering was related to the surface charge of the particles measured as zeta‐potential. One of the differences between electro‐dewatering and mechanical dewatering technologies is that electro‐dewatering actively removes ionic compounds from the sludge. In this study, dissolution and migration of organic compounds (such as shortchain fatty acids), macro metals (Na, K, Ca, Mg, Fe) and trace metals (Ni, Mn, Zn, Cr) was investigated. The migration of the metals depended on the fractionation and electrical field strength. These compounds may have both negative and positive impacts on the reuse and recycling of the sludge and removed water. Based on the experimental results of this study, electro‐dewatering process can be optimized in terms of dewatering time, desired DS content, power consumption and chemical usage.

Lannoitetehtaan prosessivesien käsittelyvaihtoehtojen soveltuvuuden arviointi

Phosphorus and nitrogen cause eutrophication of water bodies, causing severe damage to the ecosystem. Eutrophication of the waters causes oxygen depletion, which in turn increases fish mortality, releasing toxins in waters. The released toxins can cause damage to animals and humans, which is the reason in many countries to set emission limits for waste water. Nutrients exist naturally, but due to human activities there is high nutrient leaching to water bodies. Human activity is one of the main reasons to the eutrophication. The aim of this thesis was to estimate the suitability of different water treatment options for Yara Finland’s fertilizer plant’s process waters in Siilinjärvi. The fertilizer plant process waters are high concentrate and especially nitrogen concentrations are high, which bring challenge to the treatment. At the theoretical part was investigated conventional and as well advanced wastewater treatment methods like reverse osmosis, adsorption and ion exchange. Beside different treatment methods corporate environmental requirements, responsibility and strategies were researched. At the empirical part of the thesis the goal was to find out possibil-ities to intensify the efficiency of purification at lamella clarifier with chemical precipitation. In addition possibility to use already existing chemical purifying plant for process waters was estimated. As a result of the research Yara has a possibility to intensify lamella clarifier’s action by addi-tion of calcium hydroxide and thus to obtain the phosphorus and fluorine to precipitate out of the water. But in practice this would be too expensive. It is possible to eliminate nitrogen compounds by adsorption or ammonia stripping, both methods requires additional testing. It is possible to process waters in chemical purifying plant, if ammonium nitrogen has been reduced before. Reverse osmosis is possible to exploit for the phosphoric acid plant’s waters.

Quali/quantitative characterization of biogas produced in batch digesters supplied with six distinct substrates

The present research was carried out at the DER/FCAV, São Paulo State University - Brazil, with the purpose of analyzing the quantity and quality of biogas produced by six types of substrates. The substrates used in the anaerobic digestion were characterized as: 1- Litter of broiler (LB); 2- Grinded broiler litter (GBL); 3- Broiler litter with wood shavings (BLWS); 4- Grinded broiler litter with wood shavings (GBLWS); 5- Broiler litter with peanut hulls (BLPH); 6- Grinded broiler litter with peanut hull (GBLPH). It was concluded from the collected data that: the (GBL) substrate showed a superior biogas accumulated production in relation to the other substrates, while the BLWS presented an inferior accumulated production; the grinded substrates showed higher quantities of accumulated biogas in relation to the non-grinded substrates, except for GBLPH, with 20.9 m³ inferior than BLPH; the period of maximum biogas production started from 45 to 60 days, declining after 120 days; at 57 days after filling up the digesters the biogas produced had levels of CH4 superior than 53%, and from 99 days all of them produced biogas with levels of CH4 superior than 70%; LB and GBL presented higher concentrations of CH4 in the biogas.

Performance of UASB reactors in two stages under different HRT and OLR treating residual waters of swine farming

In this study it was evaluated the effects of hydraulic retention time (HRT) and Organic Loading Rate (OLR) on the performance of UASB (Upflow Anaerobic Sludge Blanket) reactors in two stages treating residual waters of swine farming. The system consisted of two UASB reactors in pilot scale, installed in series, with volumes of 908 and 188 L, for the first and second stages (R1 and R2), respectively. The HRT applied in the system of anaerobic treatment in two stages (R1 + R2) was of 19.3, 29.0 and 57.9 h. The OLR applied in the R1 ranged from 5.5 to 40.1 kg CODtotal (m³ d)-1. The average removal efficiencies of chemical oxygen demand (COD) and total suspended solids (TSS) ranged, respectively, from 66.3 to 88.2% and 62.5 to 89.3% in the R1, and from 85.5 to 95.5% and 76.4 to 96.1% in the system (R1 + R2). The volumetric production of methane in the system (R1 + R2) ranged from 0.295 to 0.721 m³CH4 (m³ reactor d)-1. It was found that the OLR applied were not limiting to obtain high efficiencies of CODtotal and TSS removal and methane production. The inclusion of the UASB reactor in the second stage contributed to increase the efficiencies of CODtotal and TSS removal, especially, when the treatment system was submitted to the lowest HRT and the highest OLR.