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Chemosphere 61 (2005) 1032–1041 www.elsevier.com/locate/chemosphere Microbial activity in a combined UASB–activated

Chemosphere 61 (2005) 1032–1041

Chemosphere 61 (2005) 1032–1041 www.elsevier.com/locate/chemosphere Microbial activity in a combined UASB–activated

www.elsevier.com/locate/chemosphere

Microbial activity in a combined UASB–activated sludge reactor system

J.S. Huang a, * , C.S. Wu b , C.M. Chen c

a Department of Environmental Engineering, Kun Shan University of Technology, Tainan County 710, Taiwan, ROC b Department of Resource and Environment, Leader University, Tainan City 709, Taiwan, ROC c Department of Environmental Engineering, National Cheng Kung University, Tainan City 701, Taiwan, ROC

Received 29 August 2004; received in revised form 3 March 2005; accepted 7 March 2005 Available online 26 April 2005

Abstract

A combined upflow anaerobic sludge bed–activated sludge (UASB–AS) reactor system with consistently wasting of excess biomass was used to treat suspended-solids pre-settled piggery wastewater (COD = 2000 mg l 1 , total Kjeldahl nitrogen TKN = 400 mg l 1 , suspended solids = 250–400 mg l 1 ). Thus, the activity of nitrogen-related microbial groups in each individual bioreactor was investigated. When the granules retention time (GRT) of 20–50 d in the UASB reactor, the solids retention time (SRT) of 10–25 d in the AS reactor and the recycle-to-influent ratio (R e ) of 1 were maintained, the combined system removed 95–97% of chemical oxygen demand (COD), 100% of TKN and 54–55% of total nitrogen (TN). Denitrification and methanogenesis occurred in the UASB reactor so that both biochemical pro- cesses contributed to most of the COD removal and, complete nitrification (most of the TKN removal) occurred in the AS reactor. Compact granules with good settling abilities developed in the UASB reactor, and rapid rates of granula- tion of break-up granules in the UASB reactor were confirmed by experiments. The activity of nitrifiers and denitrifiers (a n = 0.68–0.87; a dn = 0.55–0.70) and the calculated specific nitrification and denitrification rates (q n = 0.26–

0.47 mg NH

–N mg VSS 1 d 1 ) significantly increased with decreasing

SRT and GRT, respectively. Accordingly, the combined UASB–AS reactor system should be regarded a promising

alternative for the removal of organic carbon and nitrogen from piggery wastewater. 2005 Elsevier Ltd. All rights reserved.

þ

4

–N mg VSS 1 d 1 ; q dn = 0.046–0.076 mg NO

x

Keywords: Combined system; UASB; Activated sludge; Microbial activity; Piggery wastewater

1. Introduction

A separate-sludge system consisting of an anaerobic denitrifying/methanogenic reactor and an additional aerobic nitrifying reactor has been proven efficient in

* Corresponding author. Tel.: +886 6 2051331; fax: +886 6

2050540.

E-mail address: huangjsd@mail.ncku.edu.tw (J.S. Huang).

removing organic carbon and nitrogen from wastewater (Borzacconi et al., 1999; Bernet et al., 2000). Borzacconi et al. (1999) demonstrated that a denitrifying/methano- genic UASB reactor, combined with a nitrifying rotating biological contactor with effluent recycle, could success- fully remove organic carbon and nitrogen from sanitary landfill leachate. Bernet et al. (2000) found that a com- bined anaerobic–aerobic system (two sequencing batch reactors with effluent recycle) could effectively remove organic carbon and nitrogen from piggery wastewater.

0045-6535/$ - see front matter 2005 Elsevier Ltd. All rights reserved.

doi:10.1016/j.chemosphere.2005.03.027

J.S. Huang et al. / Chemosphere 61 (2005) 1032–1041

1033

In the separate-sludge system, appropriate conditions can be purposely established in each individual bio- reactor to favor denitrification/methanogenesis and nitrification (Chen and Lin, 1993; Lin and Chen, 1995; Hendriksen and Ahring, 1996; Borzacconi et al., 1999;

Bernet et al., 2000). Accordingly, denitrification/metha- nogenesis proceeds in the anaerobic reactor to utilize

the inflow organics and the recycled NO

from the aerobic reactor and, nitrification proceeds in the aerobic reactor at a low concentration level of organic materials. Although the mechanism of granulation in upflow anaerobic sludge bed (UASB) reactors is not well under- stood, some influencing factors, including wastewater constituents (Grotenhuis et al., 1991), growth environ- ments (Schimidt and Ahring, 1996), and superficial veloc- ity (u s ) (Tay and Yang, 1996; Huang et al., 2003; Jih et al., 2003) have been claimed. In recent studies, granulation in UASB denitrifying reactors has been observed (Borzac- coni et al., 1999; Rouse et al., 1999; Bhatti et al., 2001; Eiroa et al., 2004). Bhatti et al. (2001) found that the UASB reactor (with periodically wasting of excess bio- mass granules) could successfully remove nitrate from soft ground water. Nonetheless, attempts to carry out denitrification and/or methanogenesis in UASB reactors with consistently wasting of excess biomass granules were not found in the literature. Such an operation mode for UASB reactors may be regarded one of the measures that can be taken to improve the activity of microbial groups in UASB reactors; other measures include the elevation of operating temperature (Lin and Yang, 1991; Chou et al., 2004) and the maintenance of favorable growth environments (Grady et al., 1999). If a UASB reactor with wasting of biomass granules is to be adopted, a sludge granulation experiment should be conducted to ensure granulation in the UASB reactor is rapid enough. In this work, a laboratory-scale combined UASB– activated sludge (AS) reactor system with consistently wasting of excess biomass granules was used to treat sus- pended-solids pre-settled piggery wastewater. Thus, the activity of nitrogen-related microbial groups in each individual bioreactor was investigated. The biogas con- tents and biogas production rate over the UASB reactor and the alkalinity change over the AS reactor were mon- itored. The batch reactors, which were provided with growth environments similar to the AS and UASB reac- tors, were used to study nitrification and denitrification kinetics, respectively. A reliable method was also used to estimate the variations in the activity of nitrifiers and denitrifiers with different solids retention times (SRTs) and granules retention times (GRTs) of the AS and UASB reactors, respectively. Moreover in this article, the specific nitrification and denitrification rates together with relative percentage enhancement of micro- bial activity in each individual bioreactor are presented as well.

3

and NO

2

2. Materials and methods

2.1. Piggery wastewater

Liquid wastewater generated in pig farms in Taiwan generally contained fairly high concentrations of sus- pended solids (2000–3000 mg SS l 1 ). Thus, it has been

difficult to evaluate the performance and associated nitrification and denitrification kinetics of biological processes on suspended solids removal. In engineering practice, piggery wastewater is suggested to remove most of suspended solids by using the dissolved air flo- tation process prior to biological treatment. Accord- ingly, piggery wastewater after being shipped to the laboratory was first sieved through 1 mm diameter mesh and then placed in a cooling room (4 ± 1 C) for 6 h to settle most of the suspended solids. Thereafter, the supernatant was stored in the cooling room before use. After settling, the chemical oxygen demand (COD) and total Kjeldahl nitrogen (TKN) of the supernatant were 1600–2700 and 320–416 mg l 1 , respectively. To maintain stable influent water quality (COD =

2000 mg l 1 , TKN = 400 mg l 1 ), piggery wastewater

used in this work was either diluted with tap water or supplemented with methanol and ammonium chloride.

Other wastewater characteristics included pH (7.5–7.6), alkalinity (1250–1550 mg CaCO 3 l 1 ), organic nitrogen (40–80 mg l 1 ), total phosphorus (90–130 mg l 1 ), and

suspended solids (250–400

mg l 1 ). The substrate-feed

tank was placed in a water bath at 8 ± 2 C.

2.2. Experimental system

A schematic diagram of the combined UASB–AS reactor system is presented in Fig. 1.

2.2.1. UASB reactor

The UASB reactor, with dimensions of 6.0 (length) · 6.0 (width) · 105 cm (height) and six-equal distance sampling ports along the reactor height, had an active liquid volume of 3.78 l. The dimensions of the gas– liquid–solids separator of the UASB reactor were 6.0 (length) · 6.0 (width) · 18 cm (height). The UASB reactor was seeded with 2.0 l of anaerobic sludge, which was obtained from the anaerobic reactors of a local pig-

gery wastewater treatment plant and a laboratory single sludge (nitrification–denitrification) system. The temper- ature was maintained at 30 ± 1 C by circulating hot water in the outer-water jacket.

2.2.2. AS reactor

The AS reactor consisted of an aeration tank and a set- tler with active liquid volumes of 6 and 6 l, respectively. The AS reactor was seeded with aerobic sludge, which was obtained from the activated sludge reactors of a local piggery wastewater treatment plant and a laboratory

1034

J.S. Huang et al. / Chemosphere 61 (2005) 1032–1041

1. Thermistor

Biogas 2. UASB reactor 3. Outer-water jacket 4. Gas-liquid-solids separator 5. Internal recycle 4 6.
Biogas
2. UASB reactor
3. Outer-water jacket
4. Gas-liquid-solids separator
5. Internal recycle
4
6. Sampling port
7. Excess biomass
8. Activated sludge reactor
9. Automatic heating rod
10. Settler
11. Sludge return
3
2
3
12. Effluent holding tank
13. Effluent recycle
7
9
5
5
10
Effluent
8
6
12
7
11
Thermometer
1
13
Influent

Fig. 1. Schematic diagram of the combined UASB–AS reactor system.

single sludge (nitrification–denitrification) system. To maintain the dissolved oxygen (DO) concentrations at 3–4 mg l 1 and the temperature at 30 ± 1 C, the AS reac- tor was thoroughly aerated with a diffuser and installed with a completely submersible automatic heating rod (Visi-Therm, Aquarium System, Italy), respectively.

2.3. Combined system operations

To ensure acclimatization of anaerobic and aerobic seed sludge in each individual bioreactor, the combined UASB–AS system was continuously fed with piggery wastewater with an inflow rate of 6 l d 1 , a recycle-to- influent ratio (R e ) of 1, and the COD and TKN concen- trations of 500 and 100 mg l 1 for one month. Then the combined system was gradually fed with piggery waste- water with increasing COD and TKN concentrations shown in run 1 (Table 1) for two more months. To begin granulation of the sludge, the u s of the UASB reactor was gradually increased from 0.069 to 2.0 m h 1 by con- trolling the inflow rate of 6 l d 1 and the step-wise increasing internal-recycling flow rate. Granulation in

the UASB reactor was allowed to proceed for two months. The GRT of the UASB reactor was controlled by wasting a designated amount of biomass granules

(= sludge-bed volume/GRT) daily from the lower-, mid- dle-, and upper-part of the sludge-bed zone. Each of a total of four test runs of the combined system was con- ducted (Table 1) and continuously operated for at least

three months. The COD, TKN, NO

–N

concentrations in the effluent of each individual bioreac- tor were monitored. Steady state was assumed to be reached after test results of the four chemical parameters were within 10% deviation for four consecutive samples (sampling twice weekly).

2

–N, and NO

3

2.4. Determination of granule size

When the combined system reached steady state, approximately 5–10 ml of biomass granules were ran- domly removed from the lower-, middle-, and upper- part of the sludge-bed zone of the UASB reactor. Then the granule diameter was determined using the method of image analysis (Chou et al., 2004).

Influent COD = 2000 mg l 1 ; influent TKN = 400 mg l 1 ; influent alkalinity in test runs 1, 2, 3, and 4 were 1250, 1250, 1450, and 1550 mg CaCO 3 l 1 , respectively. The inflow rate

Alk a

330

190

200

205

TN

184

182

180

183

NO 3 –N

1 . e ), and hydraulic retention times of the UASB and AS reactors were maintained at 6 l d 1 , 1, 15.1 h, and 24.0 h, respectively.

182

180

183

183

NO 2 –N

ND

ND

0.2

0.6

TKN

0.2

0.2

0.1

0.1

AS-effluent (mg l 1 )

COD

102

85

65

58

1680

1420

1350

1550

Alk a

TN

184

190

187

188

NO 3 –N

ND

ND

ND

ND

Table 1 Operating conditions and treatment performance of the combined UASB–AS reactor system

NO 2 –N

ND

ND

ND

ND

UASB-effluent (mg l 1 )

TKN

184

190

187

188

COD

254

182

180

205

8.2

10.7

12.5

7.1

AS

Biomass

UASB

37.6

31.6

26.5

43.3

(g)

VSS l 1 ) mean)

1180

1790

2080

1370

AS

i (weighted

UASB

9200

15 440

11 100

12 900

X (mg

ratio 3 l (R

in mg CaCO

AS

(SRT)

10

20

15

25

recycle-to-influent

UASB

GRT

(d)

50

40

20

25

a Expressed

Test run

(Q),

3
4

1
2

J.S. Huang et al. / Chemosphere 61 (2005) 1032–1041

1035

2.5. Determination of biokinetic constants

Prior to determining the biokinetic constant of nitri- fiers (k n ), the enrichment of nitrifiers in a 5-liter batch reactor proceeded. First, biomass removed from the AS reactor was loaded into the batch reactor. Then the batch reactor was added into 1.19 g NH 4 Cl, 1 g KH 2 PO 4 , 8 g NaHCO 3 , and adequate amounts of trace metals (together with tap water to make an active liquid volume of 5 l) twice a day, and maintained at the DO concentrations of 3–4 mg l 1 . To keep an SRT of the batch reactor at 25 d (i.e., close to the SRT of the AS reactor in run 1), 0.2 l of mixed liquor was wasted daily prior to decanting of the supernatant. The above procedures were repeated for three months. To determine k n , an adequate amount of enrichment culture (nitrifiers) was taken from the previous batch reactor and loaded into a 1-liter batch reactor. The batch reactor was then added into 0.38 g NH 4 Cl,

0.2 g KH 2 PO 4 , 1.6 g NaHCO 3 , and adequate amounts of trace metals, and maintained at the DO concentration of 3–4 mg l 1 . In a mixing state, the initial biomass con- centration in the batch reactor was measured, while the

NH

–N concentration remaining in the solution was

measured for every 20 min. Thereafter, linear regression was applied to search for k n which would fit in experi- mental data. Prior to determining the biokinetic constant of denit- rifiers (k dn ), the enrichment of denitrifiers in a 5-liter batch reactor proceeded. First, biomass granules (re- moved from the UASB reactor) together with a few glass beads were loaded into a 500-ml serum vial, followed by placing the serum vial on a shaker (30 ± 1 C; 80 rpm) for two hours to break up biomass granules. The size distribution of the obtained dispersed sludge was deter- mined using a laser particle size distribution analyzer (Horiba, model LA-920, Japan). More than 86% of the particles were found within the size range of 0.26– 200 lm. Thereafter, the dispersed sludge was loaded into

the 5-liter batch reactor installed with a mixing device (approximately 80 rpm) and equipped with a cover but provided with a gas vent to strip off oxygen using nitro- gen gas. The batch reactor was added into 7 g KNO 3 and piggery wastewater (approximately 700 mg COD) twice a day. To maintain an SRT of the batch reactor at 50 d (i.e., close to the GRT of the UASB reactor in run 1), 0.1 l of mixed liquor was wasted daily prior to decanting of the supernatant. The above procedures were repeated for five months and biogas was monitored to ensure that no methane production occurred in the batch reactor.

To determine k dn , an adequate amount of enrichment culture (denitrifiers) was taken from the previous batch reactor and loaded into a 500-ml serum vial. The serum vial was then placed on a shaker (30 ± 1 C; 80 rpm) and

added into 0.3 g KNO 3 and piggery wastewater to make

þ

4

1036

J.S. Huang et al. / Chemosphere 61 (2005) 1032–1041

an active liquid volume of 500 ml. In an agitation state, the initial biomass concentration in the serum vial was

–N concentra-

measured, while the NO

tions remaining in the solution were measured for every 30 min. Thereafter, linear regression was applied to

search for k dn .

3

–N and NO

2

2.6. Determination of activity of nitrogen-related microbial groups

When the combined system reached steady state, bio- mass was removed from the AS reactor and loaded into a 1-liter batch reactor. The batch reactor was thoroughly aerated with a diffuser and added into the effluent of the UASB reactor. At a temperature of 30 C and in an agi-

tation state, the initial biomass concentration (X B,i ) in

–N

the batch reactor was measured, while the NH

concentration remaining in the solution was measured for every 30 min. Thereafter, the initial rate method (Jih et al., 2003; Chou et al., 2004) was used to determine the mass fraction of nitrifiers (f n ) in the AS reactor in run 1 (SRT = 25 d) and the activity of nitrifiers

(a n ) in the AS reactor in runs 1–4, as shown in Eqs.

(1)–(3):

þ

4

k

0

i

¼ S B; 1 S B;þ1

2X B;i Dt

ð1Þ

f i ¼ k k 0 i

i

ðfor run 1; SRT ¼ 25 d and GRT ¼ 50 dÞ

ð2Þ

ð3Þ

a i ¼ k k i

where i denotes n and dn ; k i and k i 0 represent the maxi-

–N reduction)

rate of the enrichment- and mixed-culture, respectively;

S B, 1 , S B,0 and S B,+1 represent the bulk concentration

of NH

ples; X B,i represents biomass concentration in the batch reactor; and Dt represents the time interval. Similarly, biomass granules were removed from the UASB reactor and loaded into a 250-ml serum vial, fol- lowed by placing the serum vial on a shaker (30 ± 1 C; 80 rpm) for 2 h to break up granules. The serum vial was added into a mixture of piggery wastewater and the effluent of the AS reactor with a proportion just the same as the recycle-to-influent ratio (R e ) of 1. In an agi-

tation state, the initial biomass concentration (X B,i ) in

3 –N and

NO

2 –N concentrations remaining in the solution were

the serum vial was measured, while the NO

x NÞ in the initial three consecutive sam-

mum specific NH

0

i

ðfor runs 14Þ

þ

4

–N oxidation (NO

x

4

NðNO

þ

measured for every 20 min. Thereafter, the initial rate method was used to determine the mass fraction of den- itrifiers (f dn ) in the UASB reactor in run 1 (GRT = 50 d) and the activity of denitrifiers (a dn ) in the UASB reactor

in runs 1–4, as shown in Eqs. (1)–(3).

2.7. Chemical analyses

Nitrate and nitrite were determined using the chromo- tropic acid method and the colorimetric method, respec- tively (APHA, 1995). The close reflux, titrimetric method (APHA, 1995) was used to determine COD. N 2 and CH 4 were analyzed using a Shimadzu gas chromatographic column system with a 150 cm · 3 mm stainless-steel col- umn (packed with 45/60 Carboxen 1000), a thermal con- ductivity detector, and helium carrier gas. Injector, column, and detector temperatures were 130, 130, and 150 C, respectively. Biomass (expressed in VSS), alka-

linity, TKN, and NH

Standard Methods (APHA, 1995).

þ –N were measured according to

4

3. Results and discussion

3.1. Performance of combined UASB–AS

reactor system

The operating conditions and performance data of the combined UASB–AS reactor system treating piggery wastewater are presented in Tables 1 and 2. As shown in Table 1, when the influent TKN and COD concentra-

tions of 400 and 2000 mg l 1 , respectively, the hydraulic retention time (HRT) of 15.1 h and the GRT of 20–50 d

in the UASB reactor, the HRT of 24.0 h and the SRT of

10–25 d in the AS reactor, and the R e of 1 were main- tained, biomass measured in the UASB and AS reactors decreased (43.3–26.5 g; 12.5–7.1 g) with decreasing GRT and SRT, respectively. As shown in Table 2, the COD, TKN, and TN re- moval efficiencies of the combined system varied slightly (45–97%; 100%; 54–55%, respectively) with different

GRTs and SRTs. According to material-balance calcula- tions using the performance data over the combined sys- tem, the removal of COD primarily occurred in the UASB reactor (80–85%); the removal of TKN mainly took place the AS reactor (92–95%); and the removal

of TN primarily occurred in the UASB reactor (51–53%).

Although the influent COD/TKN ratio of as high as 5.0 was maintained, complete nitrification occurred in the AS reactor at low COD bulk concentrations of 58– 102 mg l 1 (Table 1); nitrate was generally the main

product of nitrification (Table 1). This was primarily be- cause synchronous complete denitrification (utilizing the

recycled NO

and methanogenesis occurred in the UASB reactor,

resulting in the utilization of most of inflow organics.

A relative lower TN removal efficiency (compared with

TKN removal efficiency) found in this work was because

in the combined system the AS reactor was placed after

the UASB reactor. Thus, a part of NO

erated in the AS reactor was discharged from the com-

bined system. Accordingly, the combined UASB–AS

3

and NO

2 as electron acceptor; Table 1)

3

and NO 2 gen-

J.S. Huang et al. / Chemosphere 61 (2005) 1032–1041

1037

Table 2 COD, TKN and TN removal efficiencies of each individual bioreactor and the combined UASB–AS reactor system

Test run

Removal (%)

 

COD

TKN

TN

UASB

AS

Combined system

UASB

AS

Combined system

UASB

AS

Combined system

1

85

12

97

7

93

100

52

2

54

2

85

12

97

5

95

100

51

4

55

3

84

12

96

8

92

100

53

2

55

4

80

15

95

6

94

100

52

2

54

reactor system was proven to be an effective alternative for the removal of organic carbon and nitrogen from piggery wastewater.

3.2. Granule characteristics in UASB reactor

As shown in Table 3, a shorter GRT resulted in smaller

granule size (d p ) and lower biomass concentration (X i ) (weighted mean) in the UASB reactor. Even at the shortest GRT of 20 d, granulation of sludge (i.e., good set- tling properties) was also observed in the UASB reactor, and the measured d p and X i were 1.05 mm and 9200 mg VSS l 1 (weighted mean), respectively. At a GRT range of 20 to 50 d, the average d p and X i in the lower-part of the sludge-bed zone was the largest (1.06– 1.90 mm; 9850–19 040 mg VSS l 1 ), compared with the

average

zone (1.03–1.63 mm; 9480–14860 mg VSS l 1 ) and the average d p and X i in the upper-part of the sludge-bed zone (1.06–1.13 mm; 8250–11 720 mg VSS l 1 ). This is a typi- cal case of the distribution of granule size and biomass along the sludge-bed height (H) of UASB reactors (Grot- enhuis et al., 1991; Jih et al., 2003; Chou and Huang, 2005). The specific gravity (sp gr) and microbial density (X f ) of the granule in the lower-part of the sludge-bed zone (1.06–1.07; 101 500–133 200 mg VSS l 1 ) tended to be greater than those in the middle-part and upper-part of the sludge-bed zone (1.02–1.06; 83 900–109 500 mg VSS l 1 ). The sp gr of granules primarily depends on the occurrence of inorganic precipitation (Schimidt and Ahr- ing, 1996). Typical sp gr of granules in UASB reactors ran- ged from 1.03 to 1.08 (Schimidt and Ahring, 1996; Jih et al., 2003). According to calculations, sludge-bed poros-

and X i in the middle-part of the sludge-bed

d p

ity [e = 1 (X i /X f )] ranged from 0.85 to 0.92 (weighted mean). Accordingly, compact granules could develop in the UASB reactor with fairly large sludge-bed porosity. Moreover, a granulation experiment was conducted after all the analytical work required for run 1 had been completed. First, all the biomass granules (i.e., granule sizes ranged from 0.25 to 2.0 mm) were removed from the UASB reactor and loaded into four 1-liter serum vials. The serum vials were then placed on a shaker (30 ± 1 C; 80 rpm) for over 6 h to break up biomass granules to the sizes ranging from 0.25 to 0.75 mm.

Thereafter, the break-up granules were re-loaded into the UASB reactor to proceed with the granulation experiment. As shown in Fig. 2, the average d p (sur- face-area basis) of the break-up granules in the upper- part, middle-part, and lower-part of the sludge-bed zone restored to close to the average d p of the original gran- ules approximately at the sixth-, ninth-, and tenth-day, respectively. From an engineering perspective, the UASB reactor having an advantage of rapid granulation rates (i.e., time-saving for the start-up of the bioreactor) should be regarded a promising alternative in treating piggery wastewater.

3.3. Biogas production in UASB reactor

Biogas production and percent contents of N 2 and CH 4 in the UASB reactor were measured. At a GRT range of 20–50 d, the biogas produced was mainly com- posed of N 2 (55.8–60.1%; mean = 57.4%) and CH 4 (37.5–41.6%; mean = 39.7%). The monitored biogas

production rate ranged from 1.41 to 1.55 l d 1 (mean = 1.51 l d 1 ). The abundance of nitrogen gas generated in the UASB reactor suggested that denitrification was the main biological process occurred in this reactor. The presence of methane in the biogas also indicated that methanogenesis occurred with denitrification so that both processes contributed to the removal of organic carbon (COD) from piggery wastewater.

3 þ

5 H 2 OÞ, the consumption of one

H þ þ e ! mole of NO

5

6

–N generates 0.5 mole of nitrogen gas. By

using material-balance calculations together with

NO

From the stoichiometry of nitrate reduction ð 5 NO

1

3

10 N 2 þ 3

1

3 –N data over the UASB reactor (Table 1), the calcu-

lated N 2 production rates ranged from 0.94 to 0.96 l d 1 (mean = 0.95), which showed fairly good agreement with the experimental data (0.85–0.88 l d 1 ). This suggested

that the biogas (N 2 ) data could be used for monitoring and control of the combined UASB–AS reactor system.

3.4. Alkalinity change in AS reactor

Nitrification in the AS reactor is accompanied by the consumption of alkalinity. According to the stoichio-

–N consumes

metry of ammonia nitrification, 1 g of NH

þ

4

Weighted

106 400

95 000

96 800

111 300

mean

90 600

86 200

83 900

108 700

Upper

96 300

96 100

96 700

109 500

Middle

X f (mg VSS l 1 )

103 300

101 500

133 200

115 000

Lower

Weighted

9200

15 440

11 100

12 900

mean

Upper

8300

8250

11 720

10 640

Middle

X i (mg VSS l 1 )

9480

11 340

14 860

11 600

Lower

9850

13 200

19 040

16 220

Weighted

mean

1.04

1.04

1.06

1.05

Upper

1.04

1.04

1.02

1.03

Granule sp gr (–)

Middle

1.04

1.04

1.06

1.05

Lower

1.06

1.06

1.07

1.07

Weighted

mean

1.34

1.57

1.05

1.35

Table 3 Granule characteristics of UASB reactor

Upper

1.06

1.07

1.08

1.13

Middle

1.49

1.48

1.63

1.03

d p (mm)

Lower

1.90

1.06

1.47

1.51

H
(cm)

80

79

78

81

Test

run

3
4

1
2

1038

J.S. Huang et al. / Chemosphere 61 (2005) 1032–1041

2.0 1.5 1.0 Original granules During granulation 0.5 Upper Upper Middle Middle Lower Lower 0.0
2.0
1.5
1.0
Original granules
During granulation
0.5
Upper
Upper
Middle
Middle
Lower
Lower
0.0
0 2
4
6
8
10
12
Granule diameter (mm)

Time (d)

Fig. 2. Time course of granulation of break-up granules along the sludge-bed height of UASB reactor.

7.14 g of alkalinity (expressed in CaCO 3 ). Thus, material-

balance calculations together with TKN data over the AS reactor (Table 1) were used to calculate the change in alkalinity and compared with the measured values. The ratios of DAlk cal /DAlk exp over the AS reactor ranged from 0.94 to 1.14 (mean = 1.04), showing good agreement between the predicted and measured values. This suggests that the alkalinity data could be used for monitoring and control of the combined UASB–AS reactor system.

3.5. Evaluation of kinetic parameters

Ammonia nitrification in the batch reactor (with enrichment culture originally taken from the AS reactor

and initial biomass and NH

–N concentrations of

1430 mg VSS l 1 and 97 mg l 1 , respectively) followed zero-order kinetics. By performing linear regression of

the experimental data, the estimated k n value was

0.71 mg NH

–N

mg VSS 1 d 1 (Shieh and LaMotta, 1979; Neufeld

et al., 1980; Dincer and Kargi, 2000).

(i.e., piggery wastewater was

added to supply carbon source) in the batch reactor

(with enrichment culture originally taken from the

3 –N concen-

trations of 1200 mg VSS l 1 and 79 mg l 1 , respectively)

followed zero-order kinetics. By performing linear regression of the experimental data, the estimated k dn

value was 0.33 mg NO

mated k dn value in the present study was close to that

of Dincer and Kargi (2000) who utilized glucose as

carbon source (k dn = 0.23 mg NO

–N mg VSS 1 d 1 . The esti-

UASB reactor and initial biomass and NO

þ

4

þ

4

–N mg VSS 1 d 1 . Typical values of k n

þ

reported in the literature were 0.70–1.15 mg NH

Denitrification of NO

x

4

x

x

–N mg VSS 1 d 1 ).

3.6. Activity of nitrogen-related microbial groups

When the combined system reached steady state, bio- mass was removed from the AS reactor and loaded into

batch reactors to determine the mass fraction of nitrifiers

d as

of 50 value.

RPE for q dn c (%)

0

+65

+15

+35

reference

d as GRT

q dn d (mg NH 4 þ –N mg VSS 1 d 1 )

the SRT of at 25 the

rate (q dn ) at determined

0.062

0.046

0.076

0.053

RPE for a dn c (%)

+22

0

+16

+27

rate (q n ) determined

Activity of nitrogen-related microbial groups and specific nitrification and denitrification rates in the combined UASB–AS reactor system

a dn (–)

0.64

0.70

0.67

0.55

denitrification

value. by using the mass fraction of denitrifiers (f dn ) of 0.55 together with the performance data of the UASB reactor.

Initial NO 3 –N (mg l 1 )

and nitrication

reactor.

specific

60

57

57

81

AS

Biomass conc. (mg VSS l 1 )

(a specic dn

of ) the

Batch test

data

UASB

(a n ) and

1790

3290

2530

3250

on denitrifiers

performance

of nitriers

RPE for q n a (%)

+54

0

+19

+81

activity

the

q n b (mg NH 4 þ –N mg VSS 1 d 1 )

activity

with

the the

together

selecting

of by selecting 0.68

0.40

0.26

0.47

0.31

RPE for a n a (%)

(f n ) by

calculated

0

+16

+25

calculated

+28

nitriers

a n (–)

0.87

0.79

0.85

0.68

was

fraction of was

enhancement)

Initial NH 4 þ –N (mg l 1 )

enhancement)

using the mass

60

117

55

108

percentage

by percentage

Biomass conc. (mg VSS l 1 )

Batch test

(relative

(relative

1000

940

910

920

Calculated

d Calculated

AS

reference

Test run

a c RPE

RPE

Table 4

b

4

1
2 3

J.S. Huang et al. / Chemosphere 61 (2005) 1032–1041

(f n ) and the activity of nitrifiers (a n ) in the AS reactor in runs 1–4. As shown in Table 4, the biomass and initial

NH

–N l 1 ,

respectively; and the estimated f n value was 0.68 [i.e., cal-

were 910–1000 mg VSS l 1 and 55–117 mg NH

–N concentrations measured in the batch tests

þ

4

þ

4

culated by dividing k n by k n (SRT = 25 d)]. This implied that nitrifiers appeared to be the predominant microbial groups in the AS reactor. Although the yield coefficient of aerobic heterotrophic bacteria was generally higher than that of aerobic autotrophic bacteria (nitrifiers) (Metcalf & Eddy, 2003), most of the inflow organics (COD) in the present study were consumed in the UASB

reactor, causing nitrifiers to be the predominant micro- bial groups in the AS reactor (at low COD concentration levels). The obtained a n and relative percentage enhance- ment (RPE; a n determined at the SRT of 25 d was selected as reference value) values were 0.68–0.87 and 0–(+28)%, respectively; and a shorter SRT resulted in higher a n and RPE values, implying that the activity of nitrifiers was increased with decreasing SRT. Moreover, the mass fraction of denitrifiers (f dn ) and the activity of denitrifiers (a dn ) in the UASB reactor in runs 1–4 were determined using batch reactors. As shown

in Table 4, the biomass and initial NO

measured in the batch tests were 1790–3290 mg VSS l 1

and 57–81 mg NO

3 –N concentrations

0

3 –N l 1 , respectively; and the esti-

mated f dn value was 0.55 [i.e., calculated by dividing k 0 by k dn (GRT = 50 d)]. This revealed that denitrifiers ap-

peared to be the predominant microbial groups in the UASB reactor. The obtained a dn and RPE (i.e., a dn deter- mined at the GRT of 50 d was selected as reference value) values were 0.55–0.70 and 0–(+27)%, respectively; and a shorter GRT gave higher a dn and RPE values, showing that the activity of denitrifiers in the UASB reactor was increased with decreasing GRT.

dn

3.7. Specific nitrification and denitrification rates

If the f n and f dn values were multiplied by the bio- mass measured in the AS and UASB reactors, together with the use of performance data shown in Tables 1 and 2, the specific nitrification and denitrification rates

(q n ; q dn ) could be calculated, respectively (Table 4). The calculated q n and q dn values (0.26–0.47 mg NH

4

–N mg VSS 1

N mg VSS 1 d 1 ;

d 1 ) and the RPE values for q n and q dn [0–(+81)%; 0–(+65)%] increased with decreasing SRT (of the AS reactor) and GRT (of the UASB reactor). Thus, a shorter SRT resulted in higher activity of nitrifiers in

the AS reactor, while a shorter GRT gave higher activity

of denitrifiers in the UASB reactor.

The q n values of the present study were relatively lower than those of Campos et al. (1999) (q n = 0.5–

0.7 mg NH

nitrification in an AS reactor at high ammonia loading

–N mg VSS 1 d 1 ), because they conducted

þ

0.046–0.076 mg NO

x

þ

4

rates of up to 7.5 kg NH

þ

4

–N m 3 d 1 and without the

1039

1040

J.S. Huang et al. / Chemosphere 61 (2005) 1032–1041

addition of organic carbon. In addition, transportation of ammonia from the bulk liquid to the nitrifier cells could be hindered by the crowded heterotroph cells (Ha- nake et al., 1990), which were actually present in our activated sludge reactor. The q dn values of the present study were generally higher than those of Nyberg et al. (1992), and Lemmer et al. (1997) (q n = 0.014–

0.052 mg NO

–N mg VSS 1 d 1 ), who conducted deni-

trification in fixed-film reactors. In other words, the activity of denitrifiers in the biomass granule was rela- tively higher than that of denitrifiers in the biofilm.

x

4. Conclusions

The results of the present study showed that the com- bined UASB–AS reactor system (with consistently wast- ing of excess biomass and at the recycle-to-influent ratios (R e ) of 1) achieved efficient removal of COD (95–97%), TKN (100%) and TN (54–55%) from suspended-solids pre-settled piggery wastewater. Denitrification and meth- anogenesis occurred in the UASB reactor, contributing to most of the COD removal for a GRT range of 20–50 d and, complete nitrification (most of the TKN removed) occurred in the AS reactor. Compact granules with good settling abilities developed in the UASB reactor, yielding average granule sizes (d p ) of 1.05–1.57 mm, granule spe- cific gravity of 1.04–1.06, biomass concentration (X i ) of

9200–15 440 mg VSS l 1 , microbial densities (X f ) of 95 000–111 300 mg VSS l 1 , and sludge-bed porosity (e) of 0.85–0.92. The estimated mass fractions of nitrifiers (f n ) and denitrifiers (f dn ) were 0.68 and 0.55, respectively, reflecting the fact that nitrifiers and denitrifiers were the predominant microbial groups in the AS and UASB reac- tors, respectively. Specific nitrification and denitrification

mg

VSS 1 d 1 ; 0.046–0.076 mg NO

rates significantly increased (0.26–0.47 mg NH

þ

4

x

mg VSS 1 d 1 , respec-

tively) with decreasing SRT (25–10 d) and GRT (50–20 d), respectively. Consequently, the combined UASB–AS reactor system should be regarded a promis- ing alternative for the removal of organic carbon and nitrogen from piggery wastewater.

Acknowledgment

Financial support of this research from Grant Num- ber COA 90-AST-1.5.2-AID-U1 (1) from the Council of Agriculture of the Republic of China is greatly appreciated.

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