Journal of Scientific & Industrial Research

Vol. 65, October 2006, pp. 830-837

Aerobic and anaerobic treatment of fruit juice industry effluents
Emine Elmaslar Ozbas
1,
*, Nese Tufekci
1
, Gulsum Yilmaz
1
and Suleyman Ovez
2
1
Istanbul University, Faculty of Engineering, Environmental Engineering Department, Avcilar, 34320, Istanbul, Turkey
2
Istanbul Technical University, Faculty of Civil Eng, Environmental Engineering Department, Maslak, 34469, Istanbul, Turkey
Received 02 December 2005; accepted 05 June 2006
This study investigates biological treatment of fruit juice industry effluents in sequencing batch reactor (SBR), activated
sludge reactor (ASR) and anaerobic upflow sludge blanket reactor (UASB). At anaerobic biological treatability studies, seed
sludge was acclimated to the medium and 95% of COD removal was obtained within a few weeks. At the end of anaerobic
study, organic loading rate was increased to 5 kg COD/m
3
-day and the hydraulic retention time was decreased to 2.3 days.
At the aerobic biological treatability studies, 90-95% soluble COD removal was achieved for both wastewaters (sour cherry
and apple) in SBR and in ASR. In addition to aerobic biological treatability studies, microbiological investigation, and
kinetic and stociometric coefficients were determined. At the end of microbiological examination, fungi overwhelmingly
dominated the system.
Keywords: Activated sludge, Anaerobic up flow sludge blanket reactor (UASB), Fruit juice industry wastewater,
Sequencing batch reactor (SBR)
IPC Code: C02F3/12
Introduction
Wastewater effluents from the fruit juice industry
contain primarily high concentrations of organic
materials, which are occasionally discharged into the
municipal wastewater collection system and
processed in wastewater treatment plants along with
domestic wastewater. Major problems in the treatment
of raw effluents from the fruit juice industry are low
pH values, imbalance of nutrients, and the very
considerable fluctuations in the amount of effluent
and waste matter produced
1,2
. Sequencing batch
reactor (SBR) is successfully applied to the treatment
of strong wastewaters with effective organic carbon
and nutrient removal
3-5
. Anaerobic upflow sludge
blanket reactor (UASB) is a simple and easily
operated anaerobic system. In such systems, sufficient
refining efficiency can be achieved at low
temperatures, as well as high temperatures
6
.
This study deals with the anaerobic and aerobic
treatments of wastewater from fruit juice industry
before it is discharged into the municipal wastewater
treatment plant. In addition to treatability studies,
kinetic and stociometric coefficients (maximum
specific growth rate
m
and endogenous decay rate
b
H
) were determined for aerobic treatment systems.

Materials and Methods
UASB Reactor consisted of: total volume, 16.5 l;
internal diam, 12 cm; and height, 150 cm. Feeds were
prepared each day and pumped to the reactors using
variable speed peristaltic pumps. Anaerobic reactor is
thoroughly mixed due to influent input from the
bottom of tank and gas emission during treatment,
which also drag a portion of the biomass when rising.
Gas bubbles that are separated from the liquid and
solid phases at the bottom of the funnel, which is
placed to the precipitation section, leave the system
with the gas line. In the mean time, released sludge
particles return to the body. Similarly, some amount
of sludge, which is dragged with the hydraulic up-
flow, precipitates at the stable exterior part of the
funnel and returns to the body section. The treatment
efficiency is improved through efficient mixing and
contact of the effluent with biomass in the reactor.

Activated sludge reactor (ASR) and SBR were
operated with a hydraulic retention time (HRT) of
24 h, sludge age of 10 days and F/M ratio 0.5
(Table 1). After start-up period (24 h), SBR was
operated with a cycle time of 12 h.
___________
*Author for correspondence
Tel: +90 212 4737070/17732; Fax: +90 212 4737180
E-mail: elmaslar@istanbul.edu.tr
OZBAS et al: AEROBIC & ANAEROBIC TREATMENT OF FRUIT JUICE INDUSTRY EFFLUENTS


831

Analytical Methods
The reactors pH and temperature were controlled
continuously. The main parameters including COD,
alkalinity, total suspended solids (TSS), volatile
suspended solids (VSS) and sludge volume index
(SVI) were measured. All analyses were carried out in
accordance with Standard Methods
7
. Maximum
growth yield and endogenous decay rate were
determined by using respirometric method
2
.

Wastewater Characterization
As fruit juice wastewater is poor in nutrients
(Table 2), NH
4
Cl and Na
2
PO
4
were added to influent
to improve C:N:P in anaerobic studies
(C/N/P=300/5/1) and in aerobic studies
(C/N/P=100/5/1). Besides, in anaerobic studies,
system was fed with NaHCO
3
in order to provide
enough alkalinity in the reactor and to buffer the CO
2

and volatile acids. Temperature (35-37C) and pH
(6.5-7.8) within the reactor were maintained in
anaerobic studies. pH was maintained around 7.0 by
NaOH addition during aerobic studies. Reactors were
inoculated with the sludge taken from Pasabahce
Tekel Raki Factory.

Results and Discussion
Anaerobic Treatability Studies
During the start-up period, main parameters in
reactor were: Organic loading rate (OLR), 1.40 kg
COD/m
3
/day; HRT, 5.7 days; VSS concentration,
7700 mg/l; and pH, 7-8. In this period, the seed sludge
was acclimated to the medium and 95% of COD
removal was obtained within a few weeks (Fig. 1).
This period was observed for 73 days with no
problems.
After the start up period, OLR was increased
to 3.47 COD/m
3
/day and HRT was decreased to
2.3 days. In the first few days, the efficiency dropped
due to instability of pH and increase in OLR
was observed. Buffering the feeding effluent with
NaHCO
3
resulted in 90% of COD removal in
the system within 10 days. During this period,
OLR was kept at 3.37-5 kg COD/m
3
/day for 30 days
and 90% of COD removal was obtained on the
average. The effluent COD value was kept between
600-800 mg/l except for the over loadings (Fig. 1).
During the study, biogas production in the UASB
reactor was about 0.395 m
3
CH
4
/kg COD removed
and the average CO
2
ratio in the biogas was 20%
(Table 3).
Table 1Characteristics of activated sludge and
sequencing batch reactors
Characteristics Activated sludge
reactor
Sequencing batch
reactor

Liquid volume, l 4 4
Internal diam (), cm 12 14
Height, cm 80 64.5
Sludge age, day 10 10
F/M (F= COD;
M=MLVSS) ratio
0.5 0.5

Table 2Characterization of wastewaters
Parameters COD
mg/l

TKN
mg/l
TP
mg/l

Sourcherry juice
wastewater
1000-8000 3.3-55 0.104-10
Sourcherry juice
concentrate
1.000.000 3000 95
Apple juice
wastewater
1600-2500 73-114 0.63-0.98
Apple juice
concentrate
124.800 5694 49.14
Table 3Biogas production in the UASB reactor
Parameter Min. Max. Average St. deviation

Volatile loading rate,
kgCOD/m
3
day
2.3 21.1 5.7 2.2
Biogas production,
m
3
/day
880 11.000 5.500 1.980



Fig. 1COD removal efficiency values determined in UASB
reactor for fruit juice effluent. COD
i
: Influent COD; COD
e
:
Effluent COD
J SCI IND RES VOL 65 OCTOBER 2006


832
Aerobic Treatability Studies
Studies with Sour Cherry Juice Wastewater
Reactors were operated using sour cherry juice
wastewater during 133 days with HRT of 24 h and
80-96% COD removal. Aeration and settling period
used in ASR were approx 23 h and 30 min,
respectively. The durations for filling, aeration,
settling and withdraw phases were 45 min, 22 h and
15 min, 30 min and 30 min, respectively, for SBR.
After the start-up period, ASR was operated at:
HRT, 24 h; aeration period, 23 h; and settling period,
30 min. After 45
th
day, SBR reactor was operated with
a cycle time of 12 h. During second stage, the
durations of filling, aeration, settling and withdraw
phases for SBR were 45 min, 10 h and 15 min, 30 min
and 30 min, respectively. After start-up period,
average COD removal were achieved for ASR and
SBR as 92% (Table 4) and 93% (Table 5),
respectively. MLSS and MLVSS decreased after day
50, because a little amount of microorganism was
removed from the reactor to maintain the sufficient
settling (Table 4). When OLR changed, COD removal
values decreased because of insufficient aeration. In
SBR, when operating cycle was changed as 12 h,
microorganism concentration increased rapidly. SVI
values varied for ASR (30-90 ml/g) and for SBR
(30-80 ml/g).

Studies with Apple Juice Wastewater
Reactors were fed with apple juice wastewater. In
this period, ASR was operated with a cycle time of
24 h, and SBR with a cycle time of 12 h. The
durations of filling, aeration, settling and withdraw
phases for SBR were 45 min, 10 h and 15 min, 30 min
and 30 min, respectively. After 35 days, the influent
COD was increased from 1600 mg/l to 2750 mg/l.
COD removal efficiencies were 90% or higher until
45 days of operation. After initial COD increased,
COD removal decreased from 90% to 70-75% during
15 days in both reactors (Tables 6 and 7). Because of
this, operation cycle time was increased (24 h) and
VSS concentration was decreased (4000 mg/l) in
SBR. Beginning from 65
th
day, COD removal values
of both reactors reached 85% and above. COD
removal values reached 90% in the 90
th
day (Tables 6
and 7). SVI values were recorded as 30 ml/g in ASR
and 43 ml/g in SBR on the 36
th
day and remained at
the same level until the 45
th
day. When initial COD
values were increased, sludge settling was worst until
100
th
day. Beginning from 100
th
day, sludge settling
Table 4Main parameters of sour cherry juice for ASR
Time
days
COD
i

mg/l
COD
e

mg/l
COD removal
efficiency, %
TSS
mg/l
MLVSS
mg/l

13 2875 5890 5330
14 1145 45 96
20 1200 90 92 4520 4150
26 1400 70 94
34 1260 135 89 3600 3590
39 750 125 83
42 950 60 94
43 950 145 85 3540 3370
49 990 85 92
50 3940 3600
56 1145 95 92
74 1000 130 86 2760 2610
75 1310 95 92
76 1310 180 86
83 1085 130 88 2990 2810
90 1190 90 92
97 1500 85 92 4320 4160
104 1350 45 97 2390 2310
111 2990 2740
116 1030 45 96
120 2550 2420
123 1600 75 95
133 1100 50 95 3080 2870
Average 1257 93.42105 91.368 3547.5 3330
Standard deviation

432.3023 38.47 4.119 999.17 890.372

OZBAS et al: AEROBIC & ANAEROBIC TREATMENT OF FRUIT JUICE INDUSTRY EFFLUENTS


833
Table 5Main parameters of sour cherry juice for SBR
Time
days
COD
i

mg/l
COD
e
mg/l
COD removal
efficiency, %
TSS
mg/l
MLVSS
mg/l

13 2875 305 89 2400 2310
14 1145 55 95
20 1200 70 94 2720 2550
26 1400 70 95
34 1260 75 94 3440 3030
39 750 145 81
42 950 80 92
43 950 105 89 3860 3440
49 990 90 91
50 3620 3360
56 1145 50 96
74 1000 130 86 3310 3130
75 1310 110 91
76 1310 160 88
83 1085 70 93 3830 3580
90 1190 105 91
97 1500 50 95 5070 4800
104 1350 45 97 3360 3240
111 4130 3840
116 1030 35 97
120 5850 5620
123 1600 40 98
133 1100 65 94 4960 4750
Average 1257 92.75 92.3 3879.167 3637.5
Standard deviation

432.3 60.94 4.193 996.415 972.776
Table 6Main parameters of apple juice for ASR
Time
days
COD
i

mg/l
COD
e

mg/l
COD removal
efficiency, %
TSS
mg/l
MLVSS
mg/l

3 1650 55 97 2790 2490
7 1625 90 94
8 1625 60 96
10 1600 90 94
15 1600 95 94
16 1585 105 93
17 1600 105 93 3560 3440
22 1600 55 97
23 1600 70 96
31 1600 208 87
36 1900 195 90 6610 6450
39 2735 400 84
43
45 2740 1440 48
49 5920 5800
51 1980 1960
52 1755 1130 60 2530 2440
56
58 2900 360 87
59 4180 3910
61
62 2970 720 76 3470 3250
65 2500 320 87
69 2160 2040
(Contd)
J SCI IND RES VOL 65 OCTOBER 2006


834

Table 6Main parameters of apple juice for ASRContd
Time
days
COD
i

mg/l
COD
e

mg/l
COD removal
efficiency, %
TSS
mg/l
MLVSS
mg/l

72 2650 405 85
78 2635 300 88
84 2500 240 90 2520
91 3400 2990
97 3340 3100
98 2750 216 92 3270 3025
116 2470 130 95
117 4220 3980
125 2560 110 96 4550 4277
131 2500 144 94 5290 5025
139 2500 125 95 5830 5538
Average 2166 286.72 88.32 3860 3732.188
Standard deviation

527.51 339.88 11.589 1383.97 1365.53
Table 7Main parameters of apple juice for SBR
Time
days
COD
i

mg/l
COD
e

mg/l
COD removal
efficiency, %
TSS
mg/l
MLVSS
mg/l

3 4610 4460
7 1650 55 97
8 1625 45 97
10 1625 70 96
15 1600 50 97
16 1600 115 93
17 1585 110 93 5040 4650
22 1600 80 95
23 1600 55 97
31 1600 80 95
36 1600 208 97 4710 4500
39 1900 184 90
43 2735 761 70 8320 8060
45 2775 640 76
49 2740 11280 10290
51 3780 3760
52 4400 4200
56 1755 675 75
58 3480 3340
59 2900 1340 54
61
62 4080 3880
65 2970 930
69 2500 380 85 3730 3560
72 5170 4900
78 2650 70 97
84 2635 375 86 4450
91 2500 380 85 3440 3220
97 3900 3600
98 4610 4240
116 2750 192 93
117 2470 217 91 5170 4940
125 6460 6201
131 2560 105 96 6560 6232
139 2500 128 95 5610 5330
Average 2177 301.875 89.13043 5200 4964.611
Standard deviation

537.542 335.842 10.943 1909.77 1799.33
OZBAS et al: AEROBIC & ANAEROBIC TREATMENT OF FRUIT JUICE INDUSTRY EFFLUENTS


835
begun to be good. SVI value was 80 ml/g in SBR and
50 ml/g in ASR in the 125
th
day
8
.

Determination of Kinetic Coefficients
Determination of Endogenous Decay Rate (b
H
)
A reactor (vol, 2 l; initial biomass conc, 2000 mg/l)
was started for the determination of b
H
.
Oxygen consumption rates (OCRs) were measured
during 10 days for sour cherry juice wastewater.
During 12 days, OCRs were measured for apple
juice wastewater. Then, a graphic was drawn
for each wastewater using these values (Figs 2a and
3a). Slope of the curve on graph is equal to b
H
, which
for sour cherry juice wastewater and apple juice
wastewater was 0.32 day
-1
and 0.13 day
-1
,
respectively.

Determination of Maximum Specific Growth Rate (
m
)
A reactor (2 l) with F/M=4 mgCOD/mgMLVSS
was prepared for each wastewater. OCRs were
measured during 1 h. A graphic was drawn using
these values for each wastewater (Figs 2b and 3b).
Slope of the curve on graph is equal to (
m
-b
H
). The

m
values were determined for sour cherry juice
wastewater and apple juice wastewater as 5.15 day
-1

and 6.18 day
-1
, respectively.
Microbiological Examination and Monitoring of Aerobic
Reactors Systems
Microbiological examination of apple juice
production wastewater treatability research in
both aerobic reactors was begun at 69
th
day and
continued for the following 20 days. Most interes-
ting observation was the overwhelming dominance of
fungi, Aspergillus spp. (Fig. 4). This does not support
the previous arguments that growth rates associated
with bacteria are higher than fungi
9
. These
filamentous fungi cells have septa and foot cell
belonging to Aspergillus genus. Abundance of fungi
filaments was classified
10,11
as excessive or
dominant. The number of the filaments was over 50
within each floc, almost completely covering them.
This situation was probably caused by wastewater
composition of fruit juice production and pH. Because
the content of wastewater of fruit juice production had
very rich fruit sugar and other carbohydrates. pH of
the system has decreased to 6-6.5 and even under 6
even though the system pH was everyday adjusted to
7-7.5. Slightly acidic pH values and high
carbohydrate concentration are very favorable to
fungi, so they can multiply faster than other
competitors and get advantage to dominate the
system. Fungi can tolerate acidic conditions and


Fig. 2Determination for sour cherry juice wastewater of:
a) Endogenous decay rate; b) Maximum specific growth rate



Fig. 3Determination for apple juice wastewater of:
a) Endogenous decay rate; b) Maximum specific growth rate

J SCI IND RES VOL 65 OCTOBER 2006


836
adverse environments better than bacteria. Another
important cause of fungi domination is nutrient,
especially nitrogen (N) and phosphorus (P) is that
bacteria can flourish and multiply even in very low
concentrations of N and P
9
. N and P concentrations in
wastewaters of fruit juice production are very low
(N: 3.3-114 mg/l; P: 0.104-0.98 mg/l). This kind of
wastewater composition can provide advantage
primarily to fungi. It can be said that fungi are good
and valuable microorganisms to treat this kind of
wastewaters. On the other hand, this can be a problem
in aeration tank for bulking and in final sedimentation
tank for settlement of solids. These problems can
cause process control and low effluent quality
problems
11
.
Filamentous fungi (Aspergillus spp.) has increased
rapidly and dominated to the systems after 78
th
day.
Abundance of filamentous fungi reached excessive
numbers and caused solid separation and bulking
problems in apple juice production wastewater
treatability research. This problem has been
determined by microscopic examinations of the
activated sludge and the result of SVI measurements
(>150 ml/g). Fungi domination has not caused
significant decrease in COD removal yield other than
some slight decline. Zoogleal floc structure of the
activated sludge was very good and appearance was
normal. During the microbiological examinations, a
few kinds of ciliated, attached and flagellated
protozoa, a filamentous bacterium species, a
filamentous fungus, and gram-negative and gram-
positive bacteria have been observed in floc structure.
An unidentified filamentous bacterium, which
contributed to floc macro-structure with 2-3
filaments/flock in normal times, has helped to
overcome settlement problem of the activated sludge.
Filamentous bacterium has caused settlement problem
when the filaments spread into bulk solution from the
floc, causing a decrease in density of the floc. It has
not caused settlement and bulking problems when it


Fig. 4Microphotos belonging to reactors

OZBAS et al: AEROBIC & ANAEROBIC TREATMENT OF FRUIT JUICE INDUSTRY EFFLUENTS


837
stayed in the floc structure. Fungi filaments have
increased their numbers overwhelmingly in 100
th
day
of operation, and all flocs have been completely
covered by fungi filaments and even flocs could not
be seen due to the abundance of filaments. Fungi
spores have also contributed to this structure. There
was no settlement or bulking problems in these
situations. Probably, the reason is that, eukaryotic
cells (fungi cells) are bigger, heavier and longer than
prokaryotic cells (filamentous bacteria), so they could
settle easily. Whenever filamentous bacteria have
increased their numbers and spread out from the flock
towards the bulk solution, settlement and bulking
problems have occurred.


Conclusions
In anaerobic treatment of fruit juice industry
wastewaters ( 90%) COD removal efficiencies were
obtained with an organic loading up to 5 kg
COD/m
3
/day. In aerobic treatability studies, high
COD removal ( 90%) was obtained at treatment
studies of each wastewater at cycle time of 12 h in
SBR. Sometimes, COD removal values decreased.
When OLRs were changing and if there was not
enough aeration, low COD removal values were
observed. There were not settling problems in aerobic
treatability studies. Sometimes, SVI values increased
depending on pH. The difference in the endogenous
decay coefficient and maximum specific growth rate
constant for apple juice and sour cherry juice
wastewaters was caused by the different easily
biodegradable fractions of wastewaters. At the end of
microbiological examination, fungi overwhelmingly
dominated the system. But, there were no settlement
or bulking problems in these situations.
Acknowledgements
This research was supported by the Research Fund
of Istanbul University (Project Number: BYP-
281/03112003).

References
1 Austerman-Haun U, Seyfried C F & Rosenwinkel K-H,
UASB reactor in the fruit juice industry, Water Sci &
Technol, 36 (1997) 407-414.
2 Dold P L, Ekama G A & Marais G V R, A General model for
the activated sludge process, Wat Technol, 12 (1980) 47-77.
3 Bortone G, Gemelli S, Rambaldi A & Tilche A, Nitrification,
denitrification and biological phosphate removal in
sequencing batch reactors treating piggery wastewaters,
Water Sci & Technol, 26 (1992) 977-985.
4 Bortone G, Malaspina F, Stante L & Tilche A, Biological
nitrogen and phosphorus removal in an anaerobic/anoxic
SBR with separated biofilm nitrification, Water Sci &
Technol, 30 (1994) 303-313.
5 Subramaniam K, Greenfield P F, Ho K M, Johns M R &
Keller J, Efficient biological nutrient removal in high
strength wastewater using combined anaerobic-sequencing
batch reactor treatment, Water Sci & Technol, 30 (1994)
315-321.
6 Dold P L, Sam-soon A, Palmer H I & Marais G R, Anaerobic
treatment of an apple processing wastewater, Water Sci &
Technol, 19 (1987) 237-247.
7 Standard Methods for the Examination of Water and
Wastewater (American Public Health Assoc, Washington,
USA) 1995.
8 Elmaslar E, Tufekci N & Ovez S, Aerobic treatability of fruit
juice industry effluents in sequencing batch and activated
sludge reactors, Frese Environ Bull, 13 (2004) 985-988.
9 Bitton G, Wastewater Microbiology (John Wiley& Sons,
New York, USA) 1994, 67-80.
10 Eikelboom D H, Filamentous organisms observed in bulking
activated sludge, Water Res, 9 (1975) 365-388.
11 Jenkins D, Richard M G & Daiger G T, Manual on The
Causes and Control of Activated Sludge Bulking and
Foaming, 2
nd
edn (Lewis Publisher, Michigan, USA) 1993,
43-55.