Bioresource Technology 85 (2002) 235241

Kinetics of anaerobic treatment of slaughterhouse wastewater in batch and upow anaerobic sludge blanket reactor
s Rodr Jesu guez-Mart nez *, Ivan Rodr guez-Garza, Estaban Pedraza-Flores, Nagamani Balagurusamy, Gerardo Sosa-Santillan, Yolanda Garza-Garc a
Department of Biotechnology, Chemistry Faculty, Universidad Autonoma de Coahuila, Blvd. V. Carranza and J. Ca rdenas V. Saltillo, Coahuila, ZC 25280, Mexico Received 3 July 2001; received in revised form 4 June 2002; accepted 5 June 2002

Abstract The kinetics of anaerobic treatment of slaughterhouse wastewater in batch and upow anaerobic sludge blanket (UASB) reactors was investigated. Dierent concentrations of organic matter in slaughterhouse wastewater did not change the rst order kinetics of the reaction. In batch digesters, methane and nitrogen production stopped after 3040, 2030 h, respectively, and in UASB reactors it was terminated after 3040 days. The constant of velocity was 3.93 and 0.23 h1 respectively, for methane and nitrogen production. The yield coecient, Yp was 343 and 349 ml CH4 per g of chemical oxygen demand at standard temperature and pressure conditions for batch reactors and UASB reactor, respectively. 2002 Elsevier Science Ltd. All rights reserved.
Keywords: Slaughterhouse wastewater; Anaerobic treatment; Kinetics denitrication; Methanogenesis

1. Introduction Anaerobic treatment is widely employed for treatment of most of the industrial wastewaters containing high concentrations of soluble organic matter (Jeris, 1983). The advantages of anaerobic treatment are widely reported by many workers (Fiestas, 1984; Olthof and Oleszkiewick, 1982). Conventional anaerobic treatment of slaughterhouse wastewater has also been reported by several workers (Rodr guez-Mart nez et al., 1997; Borja et al., 1995; Polprasert et al., 1992; Sayed et al., 1984, 1987). Borja et al. (1993) investigated the inuence of dierent supports on the kinetics of anaerobic purication of slaughterhouse wastewater. The kinetics of various processes of anaerobic digestion, viz., denitrication, sulfate reduction and methanogenesis in batch reactors as well as upow anaerobic sludge blanket (UASB) reactors fed with slaughterhouse wastewater is not yet fully investigated.

As nitrate and sulfate are present in considerable quantities in slaughterhouse wastewater, it is important to assess the inuence of these substrates on biomethanation of this wastewater. It is well known that presence of sulphate is inhibitory to methanogenesis in marine and fresh water sediments (Balderston and Payne, 1976) and in anaerobic digesters (Lawrence et al., 1966). The toxicity of sulde or free H2 S produced by microbial reduction of sulphate and competition for electron donors between sulphate-reducing bacteria (SRB) and methanogens are considered to be two important factors responsible for this inhibitory eect (Kristjansson et al., 1982; Kroiss and Wabnegg, 1983). The aim of this present work was to carry out a kinetic study on various processes that occur during anaerobic treatment of slaughterhouse wastewater in batch and UASB reactors.

2. Methods 2.1. Wastewater and inoculum

* Corresponding author. Tel.: +52-844-4100-722; fax: +52-844-4155752. E-mail address: jrodrigu@alpha1.sal.uadec.mx (J. Rodr guezMart nez).

The wastewater used in this study was collected from the municipal slaughterhouse of Saltillo, Coahuila (Mexico). The wastewater production was approximately

0960-8524/02/$ - see front matter 2002 Elsevier Science Ltd. All rights reserved. PII: S 0 9 6 0 - 8 5 2 4 ( 0 2 ) 0 0 1 4 1 - 4

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100 m3 /day. Of the animals slaughtered, cattles were largest in numbers (about 90%) and the rest were swine (about 10%). The inoculum for the study was obtained from an UASB reactor that treats slaughterhouse wastewater. The pH of the inoculum was 7.3 and the total solids (TS), volatile solids (VS) and volatile suspended solids (VSS) content of the inoculum was 44 3, 27:5 2 and 21 3 g per liter respectively. 2.2. Reactors and the experimental set-up Anaerobic batch reactors (120 ml volume) with 40 ml working volume including 5 ml of granular sludge were used as batch reactors. Various COD (g l1 ) concentrations; 11.7, 9.3, 6.8 and 6.5 were prepared by diluting the slaughterhouse wastewater (12.82 g l1 ) with distilled water. The COD of the slaughterhouse wastewater was increased to 29 g l1 by adding blood to the wastewater. To one set of digesters having a COD concentration of 29 g l1 , sulphate was added at a level of 1.0 g l1 . Thirty-ve milliliters of mineral medium without carbon source was maintained as control. A UASB reactor [14 l; 1.70 m (height) 10:5 cm (inner diameter)] with a water jacket for temperature control was also used in this study. The UASB reactor was built of acrylic material. The inoculum was incubated in diluted slaughterhouse wastewater at 37 2 C for 30 days. After this incubation period, continuous feeding of the wastewater into the UASB reactor was initiated and the COD concentration was gradually increased to the original level of slaughterhouse wastewater (12.82 g l1 ). The pH was in the range of 7.18.2 during the treatment process. The characteristics of slaughterhouse wastewater used in the present study are given in Table 1. 2.3. Analysis Methane and molecular nitrogen were analyzed by capillary gas chromatograph (Varian 3400; Varian Analytical Instruments, TX, USA) equipped with a thermal conductivity detector and molecular sieve column
Table 1 Characteristics of the slaughterhouse wastewater Characteristics COD (g/l) pH Sulphate (g/l) Phosphate (g/l) VSS (g/l) TSS (g/l) VFA (acetic acid) (g/l) Alkalinity (CaCO3 ) (g/l) Grease and oils (g/l) Nitrogen (Kjeldahl) (g/l) Nitrates (mg/l) Inuent to UASB reactor 12.82 7.5 0.97 0.41 26.5 58.20 0.88 0.53 0.25 0.531 0.96 Euent from UASB reactor 1.43 8.19 0.24 0.25 3.55 5.66 0.325 0.58 0.12 0.150 0.21

(30 m 0:53 ID) at 50 C. Helium was used as the carrier gas and the ow rate was 30 ml/min. The volatile fatty acids (VFA) were identied and quantied using a capillary gas chromatograph (Varian 3300) equipped with ame ionization detector and DB-FFAP column. The column temperature was programmed from 150 to 220 C. Helium was used as the carrier gas and the ow rate was 30 ml/min. The following parameters viz., chemical oxygen demand (COD), pH, TS, VS, mineral solids (MS), total suspended solids (TSS), VSS, mineral suspended solids (MSS), VFA (in terms of acetic acid), alkalinity, total nitrogen (Kjeldahl), nitrates, phosphates, oils and greases were analyzed by following standard methods (APHA, 1990). Sulfate was determined by following the procedures described by Cottenie et al. (1982). 2.4. Statistical analyses All treatments were conducted in duplicate. The values reported in this work are an average of three samples for every replication maintained. The data were statistically analyzed for dierences in means at the 5% probability level by general linear models procedure in SAS institute (1996) and by following Steel and Torrie (1980). Gm and KA were calculated by using time series processor (TSP), International Program, Version 4.0 D, Stanford, California, USA (Borja et al., 1993).

3. Results and discussion 3.1. Variation of methane volume produced with time The methane produced per hour in the batch reactor was dependent on the strength of slaughterhouse wastewater (Table 2). The methanogenic activity and velocity of reaction increased with an increase in COD. But methanogenic activity was less in digesters fed with slaughterhouse wastewater containing concentrated blood, although the COD was higher (Table 2). The molecular nitrogen and methane produced at dierent concentrations of COD in batch digesters are presented in Figs. 1 and 2 respectively. The rate of methane formation in the UASB reactor is shown in Fig. 3. The following equation (Fiestas et al., 1990) was used to calculate the kinetics of the present study G Gm 1 eka t 1

where G is the volume of methane produced (ml) in time t (days); Gm is the maximum volume of methane accumulated at an innite digestion time and is the product of the initial substrate concentration (S0 ) and the yield coecient of the product (Yp ): Gm S0 Yp ; and ka is apparent kinetic constant that includes the biomass concentration (x): ka kx.

J. Rodr guez-Mart nez et al. / Bioresource Technology 85 (2002) 235241 Table 2 Denitrication and methanogenic activity in batch reactors fed with slaughterhouse wastewater COD (g/l) Denitrication Activity g=g of VSS=day 103 29 29 1 g sulphate 12.3 11.7 9.3 6.8 6.5 1:17 0:020a 0:986 0:024a 6:78 0:78b 5:89 0:58c 4:72 0:47d 3:66 0:38e 2:54 0:35f Velocity (V) g=l=day 102 2:51 0:019a 2:18 0:038a 14:93 1:101b 11:90 1:171c 10:1 1:28d 7:92 1:28e 5:44 0:88f Methanogenesis Activity CH4 g=g of VSS=day 102 1:37 0:14a 0:44 0:05b 2:78 0:031c 2:48 0:021c 2:28 0:05c 2:08 0:024a 2:05 0:024a Velocity (V) (g/l/day) 0:295 0:055a 0:0986 0:011b 0:613 0:041c 0:51 0:128c 0:49 0:02c 0:45 0:028a 0:44 0:028a

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Granular sludge, VSS (g/l) 21:4 1:497a 22:2 2:3a 22 2:16a 20:2 2:7a 21:4 2:4a 21:6 2:4a 21:4 2:4a

Dierent superscripts indicate signicant dierences in values of means (P < 0:05, Schees test).

Fig. 1. Kinetics of methane formation from slaughterhouse wastewater at dierent concentrations of COD; n 5.

Eq. (1) is in agreement with the nding empirically established by Roediger (Edeline, 1980). Almost constant biomass concentration (x) in the digesters (Table 2) assisted in comparison and interpretation of the results. Using the same parameters as indicated in Eq. (1) for the methane and nitrogen production, it is demonstrated that the reactions are of rst order kinetic model (Figs. 4 and 5) (Winkler, 1986; Edeline, 1980; McCarty and Mosey, 1991). The mean values of ka (with limit of condence of 95%) for the methane and nitrogen formation was 3:93 0:05 and 0:23 0:01 per day and hour respectively, for batch reactors. At a higher con-

centration of COD (12.3 g COD/l), Eq. (1) did not exhibit a pattern characteristic of a linear model. This indicated that the formation velocity of methane can be inuenced by dierent concentrations of COD (Figs. 6 and 7). Conversely molecular nitrogen appeared rst among all the electron acceptors present in the system becoming detectable usually during the rst 10 h of incubation (Figs. 2 and 8). This is in accordance with the earlier reports of Westermann and Ahring (1987) and Lens et al. (1998) that nitrates if present are rst used as electron acceptors during the anaerobic digestion. Afterwards, H2 S formation was observed

Fig. 2. Kinetics of molecular nitrogen formation from slaughterhouse wastewater at dierent concentrations of COD; n 5.

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Fig. 3. Kinetic of methane formation in UASB rector; OLR: 1.73.0 g COD/l/day; n 5.

Fig. 4. Kinetic formation of methane as a function: ln V0 ln K n lnGm ; where V0 is initial velocity of methane and Gm is accumulated methane; n 5.

Fig. 5. Kinetic formation of N2 as a function: ln V0 ln K n lnGm ; where V0 is initial velocity of N2 and Gm is accumulated nitrogen; n 5.

indicating the sequence of sulphate reduction after the disappearance of nitrate ions (Fig. 8). Sulphate reduction reduced methane production either as a consequence of competition, due to their higher anity of the hydrogenase system or due to the inhibition of the activity of methanogenic bacteria to the presence of hydrogen sulde (Westermann and Ahring, 1987). For kinetics of the methanogenic reaction in the UASB reactors, methane formation under dierent organic

loading rates (OLR) coincided with the batch reactors having COD concentrations of 612 g l1 . If the OLR is higher than this level, the removal eciency of UASB reactor decreased. The formation of methane as a function of organic loading rate is within the rank of between 1.7 and 3.0 (Fig. 9). From the relationship between OLR and hydraulic retention time (HRT), it was observed that with the OLR higher than 4 g COD/l per day, methane formation decreased as in the batch

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Fig. 6. Inuence of Gm (accumulated methane) on initial formation velocity of methane; n 5.

Fig. 7. Inuence of initial concentration of COD on initial consumption velocity of COD and their relationship to CH4 formation; n 5.

Fig. 8. Kinetic formation of N2 , H2 S and CH4 during anaerobic treatment of slaughterhouse wastewater (29 g COD/l) with 1 g of sulphate; n 5.

reactor. After this stage the linearity of the reaction disappeared (Fig. 10). 3.2. Yield coecient The yield coecient Yp at standard temperature and pressure (STP) was determined from the volume of

methane produced and the initial and nal COD. As can be seen from the results, the methane produced in the batch reactors was proportional up to the concentration of 12.3 g/l of COD (Fig. 2). In the case of the UASB reactor, the yield coecient was constant. By tting G and COD uptake value pairs to a straight line, the yield coecient under STP conditions was found

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Fig. 9. Methane (g/day) as function of OLR (g of COD/l/day) in UASB reactor; n 5.

Fig. 10. Biogas formation and OLR as a function of HRT in UASB reactor; n 5.

to be 343 5 and 349 5 ml CH4 under STP conditions per g COD for the batch and UASB reactors, respectively. The results are in conformation with the earlier report of Borja et al. (1993). 4. Conclusion Based on these studies, it appears that the reactions of anaerobic treatment of slaughterhouse wastewater in this study agree with the rst order kinetic model. The mean values of ka (with limit of condence of 95%) for the methane and nitrogen formation was 3:93 0:05 and 0:23 0:01 per day and hour, respectively, in batch reactors. At higher concentrations of COD (12.3 g COD/l), there was no linearity, which showed that the formation velocity of methane can be inuenced by dierent concentrations of COD. The yield coecient of methane at STP conditions was found to be 343 5 and 349 5 ml CH4 for the batch and UASB reactors, respectively. Acknowledgements This work was supported by Consejo Nacional de Ciencia y Tecnolog a (CONACYT), Mexico.

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