Bioresource Technology 96 (2005) 17301736

Poultry slaughter wastewater treatment with an up-ow anaerobic sludge blanket (UASB) reactor
vez P. a, R. Castillo L. a, L. Dendooven b, E.M. Escamilla-Silva C. Cha
a b

a,*

a Qu mica, Instituto Tecnolo gico de Celaya, Ave. Tecnolo gico y Antonio Garc a Cubas S/N, 38010 Celaya, Gto. Me xico Departamento de Ingenier a y Bioingenier a, Avenida Instituto Polite cnico Nacional 2508, C.P. 07000, Me xico D.F., Me xico CINVESTAV-IPN, Departamento de Biotecnolog Received 9 December 2002; received in revised form 27 August 2004; accepted 28 August 2004 Available online 4 June 2005

Abstract Removal of organic material from poultry slaughter wastewater as determined by changes in biological oxygen demand (BOD5) was investigated by adding three dierent types of inoculum combining cow manure, yeast extract or hydraulic residence time as variables with response vector of reduction of BOD5. In a 3-l reactors, a 95% removal of BOD5 from poultry slaughter wastewater was obtained with organic loading rates up to 31 kg BOD5 m3 d1 without loss of stability. This 95% removal was obtained between 25 and 39 C with a hydraulic residence time between 3.5 and 4.5 h. The growth of the consortium of micro-organisms in the reactor followed a rst-order kinetic with a constant specic growth rate of 0.054 h1. It was concluded that an inoculum from cow manure added with nutrients and yeast extract allowed a 95% removal of BOD5 from poultry slaughter wastewater at ambient temperatures within a hydraulic residence time of 4 h, sharply reducing possible environmental hazards. 2004 Published by Elsevier Ltd.
Keywords: Poultry slaughter wastewater; Up-ow anaerobic sludge blanket (USAB) reactor; Biological oxygen demand (BOD5); Hydraulic residence time and cow manure inoculum

1. Introduction One of the most important applications of biotechnology is the treatment of industrial and municipal wastewater to reduce environmental pollution (Lettinga et al., 1980). Euents from industrial poultry, porcine, or bovine slaughterhouses containing lipids, proteins, blood, and other organic material, might cause environmental damage if discharged untreated in rivers and creeks. Processing a chicken for human consumption requires 1012 l of water so the overall water consumption in a poultry processing plant is considerable. Sixty percent of the water is converted into wastewater with pH between 6.1 and 7.1, a biological oxygen demand
Corresponding author. Tel.: +52 461 61 175 75x152; fax: +52 461 61 177 44. E-mail address: eleazar@iqcelaya.itc.mx (E.M. Escamilla-Silva). 0960-8524/$ - see front matter 2004 Published by Elsevier Ltd. doi:10.1016/j.biortech.2004.08.017
*

(BOD) between 4500 and 12,000 mg l1 and a large percentage of solids, mostly clotted blood (more than 40% in volume), with a high fat content (Mercado, 1995). The rest of the wastewater is lost in the process through run-o. Most poultry wastewater is treated physicochemically, requiring large quantities of chemicals and energy to dry the euent and generating 20 g of sludge per litre of water. Deposition of the sludge is dicult, thus limiting the use of this technique. A better option to reduce the generated biosolids might be an anaerobic digestion using up-ow anaerobic sludge blanket reactors (UASB) (Speece, 1983; Young and Dahab, 1983; Young, 1991). In the USAB process, anaerobic bacteria convert organic material into methane, carbon dioxide, and biomass while purifying the wastewater (Del Nery et al., 2001). USAB systems are known for their high volumetric treatment rates, good CH4 productivity, and low

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sludge production, which makes the process economically and technologically attractive (Del Pozo et al., 2000). Since 1982, the applicability of UASB systems for the direct treatment of sewage has been tested (Lettinga et al., 1980; Lettinga and Pol, 1986). Investigations in Brazil (Souza, 1986), Indonesia (National Institute for Public Health et al., 1988), India (Siddiqi, 1990) and Colombia (Schellinkhout et al., 1985) showed that a BOD reduction of 75% is possible under tropical conditions with somewhat lower percentages in colder areas (Vieira and Souza, 1986). Because of the demonstrated capacity of UASB approaches for both domestic and industrial wastes, we investigated, in a preliminary study, the performance of a UASB treating poultry slaughter wastewater. Specically, we examined the eect of dierent types of inoculum on reactor performance as determined by reduction in BOD5. A full factorial experimental design was applied considering temperature, type of inoculum and hydraulic residence time as principal variables with response vector of reduction of BOD5.

2. Methods 2.1. Wastewater Wastewater originated from a poultry slaughter com xico). pany (Bachoco S.A. de C.V.) in Celaya (Gto., Me It was sampled from the container where the euent was separated from the larger residues such as feathers, bones and meat every 6 h for two weeks. A total of 100 l was obtained per day, homogenized and analysed chemically and microbiologically (ALPHA AWWA WPCF, 1990). 2.2. Pre-treatment of poultry slaughter wastewater

(Rojas, 1988). Three days later 10 g yeast extract per litre was added to the mixture (Stronach et al., 1986). (B) Ten litre poultry slaughter wastewater taken from the equalization basin was added to a 15 l closed glass container, equipped with gassing out orice and left to stand. After ve days, 50% of the concentrated poultry slaughter wastewater was replaced with fresh poultry slaughter wastewater and three days later, 50 mg ferric chloride, 15 mg sodium molybdate, 20 mg cobalt chloride and 10 mg nickel chloride were added as part of a 1 l solution (Kennedy and Droste, 1991; Keemer and McCallion, 1989). (C) Ten litre poultry slaughter wastewater taken from the equalization basin was added to a 15 l closed glass container, equipped with gassing out orice and left to stand. After ve days, 50% of the concentrated poultry slaughter wastewater was replaced with fresh poultry slaughter wastewater. One litre of fresh waste water was amended with 5 g cow manure, 50 mg ferric chloride, 15 mg sodium molybdate, 20 mg cobalt chloride and 10 mg nickel chloride prior of being added to the mixture. Thirty-six hours later, a yeast extract solution (10 g yeast extract per litre of poultry slaughter wastewater) was added. 2.4. Reactor characteristics A tubular glass bioreactor with 85 cm height, 6.7 cm internal diameter and 9 cm external diameter (CRODE; Celaya, Mexico) and a 3-l working volume operating in continuous ow through mode was used. Three pH sensors were installed in the bioreactor while its temperature was controlled by circulating water trough its jacket with a peristaltic pump. Sub-samples were taken each 15 h and analysed for chemical oxygen demand (COD) and BOD5. 2.5. Experimental process in bioreactor

The slaughter wastewater collected from an equalization pond of 185.5 m3 at Bachoco S.A. de C.V. to minimize uctuations in wastewater characteristics thereby providing optimum conditions for subsequent treatment. Retention time in the equalization basin was between 12 and 24 h. 2.3. Sludge activation process and treatments Three dierent inocula were produced and tested for their suitability as follows: (A) Ten litre poultry slaughter wastewater taken from the equalization basin was added to a 15 l closed glass container, equipped with gassing out orice and left to stand. After ve days, 50% of the concentrated slaughter wastewater was replaced with fresh slaughter wastewater and cow manure was added at a rate of 5 g l1

A factorial experimental design L9 (34-1) in triplicate (Montgomery, 1991; Moen et al., 1991) was used to investigate eects of hydraulic residence time, temperature and inoculum type on decrease of organic material as determined by changes in BOD5 in poultry slaughter wastewater (Table 1). BOD5 was determined by measuring dissolved oxygen with an OD YSI instrument (model 50-B-ILL, USA) before and after an incubation at 20 C for ve days. The BOD5 was dened as (Standard Methods for the Examination of Water and wastewater, 1989) BOD; mg l1 BOD; mg l1 D1 D2 ; P D1 D2 B1 B2 f P 1 2

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Table 1 Experimental factorial design (33) to investigate the eect of hydraulic residence time, temperature and inoculum type on the poultry slaughter wastewater treatment with an up-ow anaerobic sludge blanket (UASB) reactor Experiment Hydraulic residence Time (h) Real 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27
a

Temperature (C) Codea 1 0 1 1 0 1 1 0 1 1 0 1 1 0 1 1 0 1 1 0 1 1 0 1 1 0 1 Real 25 25 25 32.5 32.5 32.5 40 40 40 25 25 25 32.5 32.5 32.5 40 40 40 25 25 25 32.5 32.5 32.5 40 40 40 Code 1 1 1 0 0 0 1 1 1 1 1 1 0 0 0 1 1 1 1 1 1 0 0 0 1 1 1

Inoculum type Real 1 1 1 1 1 1 1 1 1 3 3 3 3 3 3 3 3 3 2 2 2 2 2 2 2 2 2 Code 1 1 1 1 1 1 1 1 1 0 0 0 0 0 0 0 0 0 1 1 1 1 1 1 1 1 1

ing-up of the system to an industrial level will require an adjusted HRT. The inuent ow rate was steadily decreased while keeping the time intervals constant between successive increments. Time intervals were dened when stable concentrations of COD and VFAs value in the euent under each operating conditions were obtained. This step-by-step organic load increase allowed the biomass to adapt continuously.

2.30 3.30 4.30 2.30 3.30 4.30 2.30 3.30 4.30 2.30 3.30 4.30 2.30 3.30 4.30 2.30 3.30 4.30 2.30 3.30 4.30 2.30 3.30 4.30 2.30 3.30 4.30

3. Results and discussion Physicochemical analysis of poultry slaughter wastewater obtained daily for 15 days showed that most parameters were above permissible international discharge limits for wastewater (e.g. USEPA, 2002) (Table 2). The main contaminant in the wastewater was organic matter with BOD5s ranging between 4500 and 8700 mg l1, 1025 times larger than norms established by USEPA (2002). Organic material thus has to be reduced before the wastewater can be discharged in the drainage system or reused as irrigation water. Other pollutants such as fats, oils, and surfactants were also above norms
Table 2 Physicochemical characteristics of poultry slaughter wastewater measured daily for 15 days Characteristics pH at 25 C Electrolytic conductivity at 25 C (mS m1) Total solids dried at 103105 C (mg l1) Total volatile solids (mg l1) Total xed solids (mg l1) Total suspended solids dried at 103105 C (mg l1) Volatile suspended solids (mg l1) Fixed suspended solids (mg l1) Total dissolved solids (mg l1) Volatile dissolved solids (mg l1) Fixed dissolved solids (mg l1) Settable solids (mg l1) Oils and grease (mg l1) Biochemical oxygen demand (BOD5) (mg l1) Chemical oxygen demand (COD) (mg l1) Sulphates (mg l1) Total alkalinity (mg l1) Phenolphthalein alkalinity (mg l1) Methylene blue active substances (mg l1) Fluorides (mg l1) Total phosphorus (mg l1) Phosphate, P (mg l1) Ammonium, N (mg l1) Organic nitrogen (mg l1) Total nitrogen (mg l1) Minimum 6.1 86.1 1082 938 124 726 623 66 344 174 12 10 147 4524 5800 561 7.5 6.30 5.47 3.25 7.17 2.75 6 1.2 10.5 Maximum 7.1 14.7 4558 4402 1492 1462 1310 172 3600 3564 1324 33 666 8700 11,600 1496 12.1 11.70 11.21 15.50 12.74 7.81 95 22.5 11,150 Mean 6.6 11.7 2771 2199 572 938 821 124 1833 1378 455 20 306 5500 7333 1107 12.0 10.88 7.76 7.62 9.52 4.58 62 17.2 74.9

Code interpretation: 1: low level of the factor, 0: middle level of the factor, 1: high level of the factor.

with D1 = dissolved oxygen of diluted sample immediately after preparation (mg l1); D2 = dissolved oxygen of diluted sample after 5 d incubation at 20 C (mg l1); P = decimal volumetric fraction of sample used; B1 = dissolved oxygen of seed control before incubation (4 d); B2 = dissolved oxygen of seed control after incubation (mg l1) (4 d), and; F = ratio of seed in diluted sample to seed in seed control = (% seed in diluted sample)/ (% seed in seed control). The bioreactor was inoculated with 10% inoculum (0.3 l) types A, B or C. The response variable used was BOD5. All statistical analyses were done with Statistica (StatSoft, Inc., USA, 2000). 2.5.1. Start-up The reactor was started-up by continuous feeding at a ow rate of 1 l h1 corresponding to a hydraulic residence time (HRT) of 2.38 h. This high loading rate allowed a progressive adaptation of the biomass to the loading rate while preventing a washing out of the micro-organisms and the biosolids. However, a scal-

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12500 11500 COD 10500 BOD5

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COD and BOD5 (mg l )

9500 8500 7500 6500 5500 4500 3500 2500 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 Time (days)

Fig. 1. Concentrations of COD and BOD5 (mg l1) in poultry slaughter wastewater sampled daily for 15 days.

-1

established by USEPA (2002). The mean BOD5:COD ratio was 0.75 as calculated using data in Fig. 1. A value above 0.6 normally indicates that a biological treatment is better to remove organic matter from the euent than a physicochemical one (Lettinga and Pol, 1991). To overcome many of the problems of batch reactor studies and simulate waste-treatment processes, the ow reactor is widely used for kinetic studies. In general terms, the macroscopic material balance around a ow reactor is rate of accumulation in reactor rate of input rate of output reaction rate. 3 If we consider a well-mixed reactor then the concentration in the reactor is equal to the concentration in the euent. The material balance for biomass and substrate are V dX Q0 X 0 Q0 X V dt rate of biomass formation;

where h = fresh residence time; S 0 BOD0 5 ; S = BOD5; Q0 = volumetric ow. In the case that the kinetics follow a rst-order in a completely mixed reactor then we have rate of substrate consumption k 0 S . Substituting (6) into (7) gives S0 S k 0 h. S 8 7

Fitting the removal of BOD5 with Eq. (8) gave k = 0.2914 h1 with correlation coecient R2 = 0.975 for 4 h of HRT (Table 5). The kinetic of BOD5 removal indicated that organic matter biodegradation depended on concentration of BOD5 through time and could thus be described by a rst-order kinetic (Fig. 2). Start-up times were 1828 days for inoculum A, 715 days for inoculum B and 2.55 days for inoculum C. The start-up time using the third inoculation method was less than those reported in the literature for similar

25.00 20.00
y = 0.2914x - 2.2009 R2 = 0.9749

and V dS Q0 S 0 Q0 S V dt rate of substrate consumption;

15.00 10.00 5.00

respectively. Since substrate is consumed, the rate of substrate consumption is inherently negative. If steadystate conditions are maintained then the material balance Eq. (5) becomes rate of substrate consumption Q S0 S 0 S 0 S h V

(S0-S)/S

Experimental

0.00 0 20 40 (h) 60 80

Fig. 2. Removal BOD5 kinetics in a UASB bioreactor for poultry slaughter wastewater.

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vez P. et al. / Bioresource Technology 96 (2005) 17301736 C. Cha Table 4 Statistical analysis of dierent factors used in the optimisation study for the removal of BOD5 from poultry slaughter wastewater in UASB bioreactor Source of variation A: hydraulic residence time B: temperature C: inoculum AB AC BC AA BB CC Sum of squares 1849 902 6253 7 309 43 21 2 6815 Degrees of freedom 1 1 1 1 1 1 1 1 1 F 104.7 51.1 754.0 0.4 17.5 2.4 1.2 0.1 385.7 P 0.0000 0.0000 0.0000 0.5577 0.0006 0.1381 0.2962 0.7675 0.0000

nchez et al., bioreactors (Souza, 1986; Noyola, 1992; Sa 1993). The low start-up time obtained with high cellular reproduction load indicated that the system could support more organic load and remove it eciently in a short time. Data of maximum BOD5 removal obtained in this laboratory experiment will facilitate scale-up and design of an industrial bioreactor. Largest removal of BOD5 was obtained in experiments 6, 9, 15 and 18 (Table 3). Analysis of variance (ANOVA) showed that process hydraulic residence time (A), temperature (B), inoculum type (C) and interaction C C signicantly aected removal of BOD5 with inoculum type being the most important (Table 4). Through an analysis of the surface response and the factorial graphics it was possible to select the value for each variable that would remove the largest amount of BOD5 (Figs. 3 and 4). The surface response curved showed that at high temperatures and middle hydraulic residence times, the removal of BOD5 was optimal (Fig. 4A). The best inoculum was type 3 with a hydraulic residence time of 4 h (Fig. 4B). The two other inoculum types were not as ecient to reduce BOD5 as type 3 nor was reducing or increasing hydraulic residence time. Inoculum type 3 at the highest temperatures resulted in the largest removal of BOD5 and again the two other
Table 3 Results of the full factorial experimental (33) design for the removal of BOD5 from poultry slaughter wastewater using an UASB bioreactor Experiment BOD5 removal (%) Replicate 1 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 39.85 62.00 76.40 55.33 72.25 90.18 65.42 85.11 92.29 75.20 80.48 90.90 78.43 86.40 92.45 83.26 89.96 94.31 20.23 26.52 32.32 27.31 34.91 38.06 30.38 42.65 47.95 Replicate 2 40.82 62.52 76.21 52.36 72.21 89.96 65.10 85.15 91.94 75.07 80.03 90.50 78.08 86.02 91.82 83.32 89.89 94.23 20.30 26.89 32.26 27.32 34.83 37.92 30.10 42.72 47.91 Replicate 3 39.84 62.18 75.88 51.98 72.21 89.86 65.12 84.98 91.77 75.11 79.97 90.21 77.59 85.91 91.73 83.10 89.87 93.63 20.06 26.88 31.75 27.36 35.25 37.78 30.14 42.68 47.90 Mean 40.18 62.23 76.34 52.22 72.22 90.00 65.21 85.08 92.00 75.12 80.16 90.53 78.04 86.11 92.00 83.23 89.91 94.05 20.20 26.76 32.11 27.33 35.00 37.92 30.20 42.68 47.97

Table 5 Parameters obtained tting a rst-order kinetic to BOD5 removal in a UASB bioreactor for poultry slaughter wastewater with dierent hydraulic residence times Residence time (h) 3:00 3:30 4:00 4:30 4:00 (repetition) 4:30 (repetition)
a

Removal rate (h1) 0.307 0.310 0.291 0.364 0.373 0.306 (0.029)a (0.024) (0.028) (0.034) (0.027) (0.020)

Intercept 3.011 2.081 2.220 3.151 2.725 2.420 (1.257) (1.307) (1.192) (1.429) (1.164) (0.850)

Correlation coecient 0.905 0.915 0.975 0.910 0.949 0.951

Standard error of the estimate.

inoculum types were not as ecient to reduce BOD5 nor did decreasing the temperature (Fig. 4C). A conrmatory test with the optimal parameters was done and corroborated that a 2.1% increase in reduction of BOD5 was obtained with hydraulic residence time of 4 h, inoculum type 3 and temperature 35 C. The hydraulic residence time of 4 h was less than reported by Rao et al. (1999) for treatment of similar wastewater and inoculum type 3, never reported before, removed 9597% of BOD5. The best removal of BOD5 was obtained at 35 C. However, maintaining a bioreactor at 35 C is economically unsustainable considering that mean outside yearly temperatures in this part of Mexico, i.e. Celaya, is 20 C (http://inegi.gob.mx). The conrmation test was repeated at 25 C: a temperature, which would be obtained in the UASB bioreactor. Changes in temperature had only a minimum eect on BOD5 removal and at 25 C a 95.6% removal of BOD5 was obtained, within the operational range of anaerobic processes reported by Lettinga et al. (1979) (Table 6). It was concluded that type 3 inoculum derived from cow manure added with yeast extract and nutrients with a short hydraulic residence time of 4.5 h allowed a 95% removal of BOD5 at ambient temperatures sharply reducing possible contamination of surface water with poultry slaughter wastewater.

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Residence time (h)
90.000
1.

1735

Temperature (C)

INOCULUM

Desirability

65.573
.5

71.060

43.085

0.

15.110

0.000

0.902

-1

-1

-1

Fig. 3. Factorial graphics showing the response of hydraulic residence time, temperature and inoculum type investigated with an experimental design 33 for poultry slaughter wastewater treatment.

Fig. 4. Surface response curve for the determination of optimal parameters in the poultry slaughter wastewater treatment: (A) temperature (C) versus residence time (h), (B) residence time (h) versus inoculum type, and (C) inoculum type versus temperature (C).

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Table 6 Removal eciencies (%) and organic volumetric loading (kg BOD m3 d1) at dierent residence times (h) and temperature (C) Experiment 1 2 3 4 5 6 7 8 9 10 Residence time (h) 1.5 1.9 2.5 2.9 3.0 3.5 4.0 4.5 4.0 4.5 Initial temperature (C) 24.0 22.0 24.0 25.0 24.5 26.0 27.0 25.0 26.0 23.0 Final temperature (C) 25.5 23.5 25.5 26.5 26.0 27.5 28.5 26.6 27.5 24.7 Removal eciency (%) 40.00 65.01 71.43 79.00 87.00 95.01 95.00 95.56 95.01 95.00 Organic volumetric loading (kg BOD m3 d1) 86.6 64.4 52.8 44.3 42.8 53.8 47.1 26.0 30.8 28.7

Acknowledgements The research was funded by Consejo del Sistema n Tecnolo gica (COSNET) grant Nacional de Educacio xico). C.C.-P. and R.C.-L. received 647.95-P (Me grant-aided support from Consejo Nacional de Ciencia xico). y Tecnologa (CONACyT, Me

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