Effects of chemical oxygen demand to sulfate ion ratio and concentrations

of chemical oxygen demand and sulfate ion in anaerobic digestion of sulfate

rich latex wastewater*
R.Y.B.Ananda , M.A.R.Hasanka , U.G.Y.S.Kawinda
Department of Chemical and Process Engineering
University of Moratuwa
Moratuwa, Sri Lanka
yasinthabimal@yahoo.com,mar.hasanka@gmail.com,ys.kawinda@gmail.com

Abstract
The objective of this research was to assess the ability of anaerobic digestion process to
treat sulfate rich wastewater at different chemical oxygen demands to sulfate
concentration ratios and also to assess the optimum chemical oxygen demand for
anaerobic digestion process of sulfate rich wastewater. Sulfate concentration reductions
by anaerobic sulfur reducing bacteria were measured in acclimation period and
continuous feeding period with two different feed concentrations. A reduction of 58.8%
sulfate was observed for 10:1 ratio in acclimation period and 48.5% reduction was
observed for 4:1 ratio in acclimation period. A reduction of 33.2% chemical oxygen
demand was observed for 10:1 ratio and 17.8% reduction was observed for 4:1 ratio in
acclimation period and first continuous feed period. In overall 10:1 ratio gives better
sulfate reduction and chemical oxygen demand of 5000 mg/L in that ratio gives further
better sulfate reduction.
Keywords: Anaerobic digestion; Sulfate; Sulfur reducing bacteria; Acclimation
period;Chemicaloxygendemand
To do the experiment we prepared
I. INTRODUCTION synthetic wastewater samples having
different : 4 2 ratios and
Generally sulfur rich wastewaters are 2
generated from industries such as gas different and 4 concentrations.
scrubbing, coal plants, metal plating and
latex industry. Here we considered the
II. MATERIALS AND
wastewater generated from latex industry.
The treatment method that we used in this METHODS
experiment was the anaerobic digestion Two glass bottles were taken as the two
process. In anaerobic treatment process of reactors to perform the anaerobic digestion
sulfur rich wastewater, waste streams process. The inner bottom of the bottle
consist of high concentrations of sulfate should be sealed by cement with a 10 cm
ions. The chemical oxygen demand of the diameter hole in the center to hold the
wastewater sample can have an impact on magnetic stirrer. Then the mouth of the
the efficiency of the anaerobic digestion bottle should be sealed with a polymer
process. Also the concentrations of both seal. Silicon sealer should apply over the
and 4 2 can have an effect on the seal to prevent air transferring to inside.
anaerobic digestion process. Therefore our The sealer consists of five SS tubes. One
objective was to find the connection and was for temperature sensor which was an
co-relation between and 4 2 open top sealed bottom one. Second one
concentrations of the waste water streams. was a shorter one opened in both sides to

[1]
collect the bio gas formed. Clear hose was factor for synthetic wastewater sample
connected at the top end of the tube. And having required value.
other end of the hose was put inside in a
water filled measuring cylinder to collect
the gas. The third tube was an opened one Graph of COD value Vs Dilution Factor
from both sides which was used to feeding 25000
and removing the wastewater from the
reactor. A short clear hose was connected 20000
y = 19.84x2 - 1303.x + 22300
to its outer end. The fourth and fifth tubes

COD (mg/L)
15000
were the inlet and outlet of the submerged
SS coil inside the reactor. It was used for 10000
the purpose of transferring hot water from
water bath through the reactor in order to 5000

keep the reactor temperature in the desired

0
level. The feeding tube should always keep 0 10 20 30
closed at the outer end by a hose clip Dilution Factor
except at feeding and removing of Fig. 2. Graph of COD value Vs Dilution Factor

wastewater. This was done in order to

prevent air go inside to the reactor.
TABLE 1. Dilution factors for samples with required COD
level
Required Corresponding
Sample COD Dilution Factor
10,000 mg/L 10
5,000 mg/L 18.3
4,000 mg/L 20
2,000 mg/L 25

Fig. 1. Configuration of the anaerobic reactor apparatus B. Preparation of activated

sludge sample
We had to prepare three types of samples
for the experiment. Those were, We did not need to specially prepare the
1. Synthetic wastewater sample. activated sludge sample since we could
2. Activated sludge sample containing obtain the sample directly from wastewater
anaerobic microorganisms. treatment facility. Here we obtained our
3. Nutrients samples for sludge sample from Lalan Factory
microorganism activity. situated at Mathugama, Sri Lanka.
i. Basal nutrients.
ii. Trace nutrients. C. Preparation of nutrients
A. Preparation of synthetic samples
wastewater sample
a. Basal nutrients
We had to prepare wastewater samples TABLE 2. Basal nutrients composition
with different values. So we prepared Chemical Concentration
the samples by diluting 1M, Acetic Acid in Substance
distilled water. We made a series of
4 174 /
samples with different dilutions and did a
2 4 28 /
testing and made a graph of (4 )2 4 28 /
value against the dilution factor. Using that
45 /
graph we could find the required dilution

[2]
 b. Trace nutrients
TABLE 3. Trace nutrients composition
Chemical Concentration
4 50 /
4 500 /
3 3 50 /
50 / period we ran the reactor with 30 day
2 (4 )3 2000 /
500 /
36 % 1 /
1 liter from each sample were prepared
and kept inside plastic bottles inside a
refrigerator.
D. Final synthetic wastewater
sample preparation
We added 60 ml of Basal nutrients and 10
ml of Trace nutrients for a 1L of synthetic
wastewater sample formerly prepared and
according to that formula, 2 liter from each
wastewater sample were prepared. The
prepared samples were collected into 2.5
liter plastic bottles and the bottles were
kept at refrigerator. Then we measured the
4 2 concentration in prepared samples
and found it was 1360 mg/L. Since Acetic
acid and Distilled water do not contain
4 2 ions the only possible way of
adding those from nutrient samples. Since
the 4 2 inhibition concentration is 1440
mg/L we did not increase the current
concentration value. Therefore adding of
2 4 was not needed to adjust the
4 2 concentration in the wastewater
sample.
E. Operation of the Anaerobic
Reactor
First we cleaned the reactor bottle well
with clean water and again added 2500 ml
of water and mark the 2500 ml level
outside the reactor with a cello tape and
removed the water inside the bottle. Then
in the first day of operation we added 400
ml of wastewater sample first and then
filtered sludge sample of 800 ml and
sealed the reactor with the plug and
silicone sealers. Their onwards in each
upcoming day we added 200 ml of
wastewater until the level of reactor
liquids came up to 2500 ml mark. That
period was considered as the Acclimation
Period. At the end of the acclimation
retention time. So their onwards we first
removed 83 ml from reactor and then
added 83 ml from wastewater sample. This
procedure was carried out daily and in
each day we measured the 4 2 ion
concentration in order to measure the
reduction of 4 2 ions from the
wastewater sample and in the final day we
measured the COD of the sample. The
COD sample was prepared by first normal
filtering and then micro filtering the
diluted original sample.
We had two reactor bottles and therefore
we first ran the reactors having
4 2 ratios of 10 1 and 4 1. Here in
10 1 ratio reactor we used 10000 mg/L
COD wastewater sample
having 4 2 concentration of 1360
mg/L. In 4 1 ratio reactor we used 4000
mg/L wastewater sample
2
having 4 concentration of 1360
mg/L. To have a 10 1 ratio we actually
needed to prepare waste water sample
having COD of 13600 mg/L. But we
measured the 10000 COD value before the
addition of nutrients. So we assumed that
COD of the sample increases in to 13600
mg/L COD after the addition of nutrients.
We made the same assumption on other
samples also. We named the 4 1 ratio
reactor as Reactor 1 and 10 1 ratio
reactor as Reactor 2. From the beginning
of the acclimation period to the end of the
reactors run we measured the daily
production of bio gas.
At the end of the run of reactors in above
concentrations we changed the and
4 2 concentrations in the feed.
Therefore their onwards we added feed to
the Reactor 1 with wastewater having
[3]
2000 mg/L COD and 680 mg/L 4 2
concentrations and for Reactor 2 we add
wastewater having 5000 mg/L COD and
680 mg/L 4 2 concentrations. The
collected biogas volume, 4 2
concentration and COD were measured.
F. Anaerobic Digestion Process
Here the main objective was to treat
sulfate ions by sulfate reducing anaerobic
bacteria living in the activated sludge.
Mainly anaerobic digestion process
consists of four stages. First stage is
Hydrolysis where complex organic
molecules are broken down to simple
sugars, fatty acids and amino acids.
Second stage is Acetogenesis which
results in further breakdown of remaining
components by acedogenic bacteria. Third
step is Acetogenesis where further
digestion occurs by acetogenic bacteria
making acetic acid. Fourth step is
Methanogenesis where methane is
produced by methanogens. We assumed
our reactor was in the acetogenesis stage
and therefore prepared our synthetic
wastewater samples using acetic acid.
G. Operational Conditions
According to the literature the suitable
optimum temperature for anaerobic
bacteria metabolism is 350C. To provide
that temperature we had to use temperature
controller device and hot water bath which
circulate hot water through the SS coil
inside the reactor. Due to lack of facilities
we did not prepared temperature control
set up. Since the room temperature was
also around 330C we assumed that the
temperature in the reactor was in the
optimum value.
Again in each feeding day we have to
check the pH of the reactor wastewater.
Anaerobic bacteria need the pH between
6.5-7.5 values for better metabolism.
Therefore we had to adjust the pH by
adding KOH base solution and acetic acid,
acidic solution in each day.
[4]
H. Possible Obstacles and
Remedies
a. Sulfate inhibition
According to the literature sulfate
inhibition concentration is 1440 mg/L.
since the wastewater feed streams
consisted of 1360 mg/L and 680 mg/L
sulfate concentrations respectively we
could come in to conclusion that the
reactors were safe enough to avoid sulfate
inhibition.
b. Sulfide inhibition
The pH of the reactor was always kept at
slightly alkaline ( 7.9 0.1) value which
limits the concentration of inhibitory
hydrogen sulfide. This allowed sulfate
reducing bacteria to predominate
throughout the experiment. Reduced
sulfate concentrations results in formation
of 2 which becomes inhibitory.
According to literature Methanogenic
microorganisms become inhibited at
: 4 2 ratios less than 7. And ratios
> 10 a large portion of 2 removed from
liquid phase reducing the inhibiting effect.
The inhibitory level for sulfide is 1500
mg/L. Since sulfate maximum
concentration is 1360 mg/L the maximum
expected sulfide concentration is 453.3
mg/L( since 4 2 : 2 ratio is 3:1
(96:32)) so we could come in to
conclusion that reactors were safe enough
to avoid sulfide inhibition.
c. Ammonia inhibition
The inhibitory level of ammonium ion is
3000 mg/L. The only way of adding
ammonium ion to the reactor was through
the addition of basal nutrients. According
to calculation the ammonium ion
concentration in the reactor was 1855
mg/L. So we could not expect ammonium
inhibition in the reactor.
 d. Too much high or low pH IV. RESULTS
values A. Acclimation period of Reactor
1
The optimum pH range for
microorganisms lies between values, (6.5- TABLE 4. Acclimation period data and results for reactor 1
Date Sludge Waste Total Daily Bio
7.5). So we daily adjusted the pH to suit Volume Water Volume Gas
the required value by adding KOH base or Added Volume in the Production
Added Reactor
Acetic Acid acidic compound.
2016/01/11 800 ml 400 ml 1200 ml
e. Uncontrolled Reactor 2016/01/12 200 ml 1400 ml
2016/01/13 200 ml 1600 ml
Temperature 2016/01/14 320 ml 1920 ml 10 ml
2016/01/15 8.75 ml
2016/01/16 8.75 ml
The optimum temperature for mesophilic 2016/01/17 8.75 ml
bacteria is (30-35)0C and we have to 2016/01/18 200 ml 2120 ml 8.75 ml
2016/01/19 200 ml 2320 ml 10 ml
control the temperature of the reactor to 2016/01/20 180 ml 2500 ml 15 ml
suit that temperature level. But due to lack 2016/01/21
2016/01/22 3.5 ml
of knowledge we could not prepare the 2016/01/23 3.5 ml
control unit. But room temperature also in 2016/01/24 3.5 ml
2016/01/25 3.5 ml
a value between (30-35)0C and therefore 2016/01/26
we considered uncontrolled reactor 2016/01/27
temperature would not yield severe
problems.
B. Running Period of Reactor 1
with First Feed Concentration
III. METHOD OF ANALYSIS
TABLE 5. Reactor 1 continuous feeding period 1 results
Throughout the experiment we took three Date Concentration Daily COD
[ ] Value
parameter measurements from reactor Before
Feeding
After
Feeding Reduction
samples that were 4 2 concentration, 2016/01/11 1360 mg/L 1360 mg/L 4000 mg/L
value and Bio gas volume 2016/01/25 700 mg/L 722 mg/L
measurement. 4 2 concentration was 2016/01/26 111 mg/L

measured by sulfate meter where two 2016/01/27 500 mg/L 529 mg/L 111 mg/L
2016/01/28 300 mg/L 336 mg/L 229 mg/L
diluted 10 ml samples were taken and
2016/01/29 600 mg/L
sulfate tablet was added to one sample.
2016/02/01 3288 mg/L
Then first use the blank sample and then
the other sample to get the direct reading
of the 4 2 concentration. values
were measured by using typical COD test.
C. Running Period of Reactor 1
Here reactor sample was micro filtrated with Second Feed
with 0.45m filter and then diluted sample Concentration
volume of 2.5 ml was added to the test TABLE 6. Reactor 1 continuous feeding period 2 results
tube with 0.167 M, 2 2 7 1.5 ml and Date Concentration Daily COD
[ ] Value
3M, 2 4 3.5 ml and placed inside the Before After
Reduction
Feeding Feeding
digester for two hours and titrated with
standard ferrous Ammonium sulfate 2016/02/08
2016/02/09 200 mg/L 216 mg/L 110 mg/L
solution with ferroin indicator. The bio gas
2016/02/10
were collected in a water filled up side 2016/02/11
down measuring cylinder placed in a water 2016/02/12 533 mg/L 537 mg/L 6882 mg/L
bath where the clear hose connected to the
bio gas tube from one end and other end
was placed in the measuring cylinder.
[5]
 D. Acclimation Period of Reactor G. Interpretation of Results
2
Acclimation Period Bio Gas Production
TABLE 7. Acclimation period data and results for reactor 2
Date Sludge Waste Total Daily Bio 80
Volume Water Volume Gas
Added Volume in the Production 70

Daily Bio Gas Production (ml)

Added Reactor 60
2016/01/07 800 ml 400 ml 1200 ml Reactor 2
2016/01/08 200 ml 1400 ml 60 ml 50
Reactor 1
2016/01/09 66.7 ml
2016/01/10 66.7 ml 40
2016/01/11 200 ml 1600 ml 66.7 ml
2016/01/12 200 ml 1800 ml 40 ml
30
2016/01/13 300 ml 2100 ml 20
2016/01/14 200 ml 2300 ml 35 ml
2016/01/15 10
2016/01/16
2016/01/17 0
2016/01/18 200 ml 2500 ml 0 10 20 30
2016/01/19 47 ml Day
2016/01/20 Fig. 3. Acclimation period bio gas production
2016/01/21 25 ml
2016/01/22 5 ml
2016/01/23 5 ml
2016/01/24 5 ml
2016/01/25 5 ml Daily Bio Gas Production in Reactor 2 for
2016/01/26 Continuos Feeding Period
2016/01/27 17 ml
250

200
Bio Gas Volume (ml)

E. Running Period of Reactor 2 150 Reactor 2

with First Feed Concentration 100
TABLE 8. Reactor 2 continuous feeding period 1 results 50
Date Concentration Daily COD
[ ] Value
Before After
Reduction 0
Feeding Feeding
2016/01/07 1360 mg/L 1360 mg/L 10000 mg/L
Day
0 5 10 15
2016/01/25 561 mg/L 588 mg/L Fig. 4. Reactor 2 bio gas production in continuous feeding
2016/01/26 561 mg/L 588 mg/L 27 mg/L Period
2016/01/27 533 mg/L 561 mg/L 55 mg/L
2016/01/28 297 mg/L 333 mg/L 264 mg/L
2016/01/29 264 mg/L 69 mg/L Sulphate Concentration in the reactors
2016/02/01 6685 mg/L
1600
1400
Reactor 2
Sulphate concentration (mg/L)

F. Running Period of Reactor 2 1200 Reactor 1

with Second Feed 1000

Concentration 800

TABLE 9. Reactor 2 continuous feeding period 2 results 600

Date Concentration Daily COD
[ ] Value 400
Before After
Feeding Feeding Reduction
2016/02/08 400 mg/L 410 mg/L
200
2016/02/09 300 mg/L 313 mg/L 110 mg/L 0
2016/02/10
0 10 20 30 40
2016/02/11
Day
2016/02/12 433 mg/L 441 mg/L 2867 mg/L Fig. 5. Sulfate concentration in reactors

[6]
 Daily Sulphate Reduction for Continuous H. Acclimation Period Average
First Feeding
300
Daily Bio Gas Production
For Reactor 1,
250
Reactor 2

Daily Sulphate Reduction (mg/L)

Reactor 1 =

200
(10 + 8.75 4 + 10 + 15 + 3.5 4)
=
150
11
= 7.64
100 For Reactor 2,

50 (60 + 200 + 40 + 35 + 47 + 25 + 20 + 17)
=
13
0 = 34.15
0 10 20
Day I. Continuous Feeding Period
Fig. 6. Daily sulfate reduction for continuous feeding period Average Daily Bio Gas
Production
For Reactor 1,
COD Concentration in the Reactors
We were unable to measure the value due
12000 to leak in bio gas tube.
For Reactor 2,
10000
Reactor 2
Reactor 1
(295 + 116 + 205)
=
8000 11
COD Concentration (mg/L)

= 56
6000
J. 4 2 Concentration
Reduction in Acclimation
4000 Period (3 Weeks)
For Reactor 1,
2000
(1360 700) /
= 100%
1360 /
0
0 10 20 30 40 = 48.5 %
Day
For Reactor 2,
Fig. 7. COD concentration in reactors

[7]
 (1360 561) / Continuous Feeding Period of
= 100%
1360 / First Feed Concentration
= 58.8 % For Reactor 1,

K. Average daily 4 2 =
(4000 3288) /
100%
Concentration Reduction for 4000 /
Reactor 1 = 17.8 %
For the First Feed Concentration, For Reactor 2,
2
4 reduction (10000 6685) /
4 2 = 100%
= 10000 /

= 33.2 %
(111 + 111 + 229)/
=
3 N. Reduction in between
Continuous Feeding Period of
= 150.33 /
First Feed Concentration and
For the Second Feed Concentration, (By Second Feed Concentration
ignoring 29/01/2016 [ 4 2 ] reading)
For Reactor 1,
4 2 reducton
(110)/ (3288 6882) /
= = 100%
11 3288 /

= 10 / = 109.31 %

L. Average daily 4 2 There was an accumulation of

Concentration Reduction for instead of reduction of in the reactor
1. This may be due to too much low
Reactor 2 concentration of COD which inhibits the
For the First Feed Concentration, anaerobic digestion process. Again this
value may be due to experimental error.
4 2 reducton
(27 + 55 + 264 + 69)/ For Reactor 2,
=
4 (6685 2867) /
= 100%
= 103.75 / 6685 /

For the Second Feed Concentration, (By = 57.11 %

ignoring 29/01/2016 [ 4 2 ] reading)

4 2 reducton V. DISCUSSION
(110)/
=
1 A. Bio Gas Formation
= 110 / Hydrogen sulfide, Methane and Carbon
dioxide are the major gases produced in
M. Reduction in the anaerobic digestion process. The bio
Acclimation Period and gases formations occur as follows.

[8]

612 6 () 32 () + 34 () (1) makes slightly higher sulfate reduction

than 10,000 mg/L level feed.
4 2 () + 42 2 () + 22 () + 2 () (2)

In high levels the nutrients for

4 2 () + 3 2 () + 23 () (3)
anaerobic bacteria is higher and therefore
In acclimation period, Reactor 2 having microorganism metabolism occurs well, so
the digestion process occurs well. In
: 4 2 ratio of 10: 1 showed much
higher bio gas production compared to 10,000 level the 4 2
Reactor 1, having 4: 1 ratio. For reactor 2, concentration is 1360 mg/L and it is much
continuous feeding period bio gas near value to sulfate inhibition
production was even higher than the concentration which is 1440 mg/L. In
acclimation period. Though we were 5,000 level the 4 2 concentration
unable to measure the bio gas production is 680 mg/L and it is not that much near to
in continuous feeding period for Reactor 1 inhibition concentration. So that may be
we can say Reactor 2 had higher rate the reason for 5,000 mg/L feed giving
considering acclimation period values. slightly higher daily sulfate concentration
Also bio gas production had increased in reduction.
continuous period feeding period using
second feed. C. Level Reduction

Bio gas formation is an indicator of the In the acclimation period and continuous
anaerobic digestion efficiency. When feeding period with first feed
digestion occurs better the bio gas concentration, Reactor 2 showed higher
formation is high. So we can come into the reduction (33.2%) than the reactor 1
conclusion that : 4 2 ratio of 10: 1 (17.8%). In the continuous feeding period
is much better for anaerobic digestion of with second feed concentration, reactor 2
sulfate rich wastewater with compared to showed much better reduction and
4: 1 ratio. Again we can come into reactor 1 shows accumulation.
conclusion that level of 5,000 mg/L Therefore we can come into conclusion
is better for anaerobic digestion with that the : 4 2 ratio of 10: 1 is much
compared to 10,000 mg/L level. better for anaerobic digestion of sulfate
rich wastewater with compared to 4: 1
B. Sulfate Reduction ratio and again level of 5,000 mg/L is
better for anaerobic digestion with
In the acclimation period reactor 2 showed compared to 10,000 mg/L level.
58.5% reduction of initial sulfate
concentration and Reactor 1 showed
48.5% reduction. In continuous feeding VI. CONCLUSION
period, Reactor 1 showed very high (150
According to the overall results we can
mg/L) and very low(10 mg/L) daily sulfate
come into the conclusion that the optimum
reductions for first and second feed
: 4 2 ratio for anaerobic digestion
concentrations respectively while Reactor
2 showed relatively high values(104 mg/L process is 10: 1 and also the optimum
and 110 mg/L) for the same parameters. level for anaerobic digestion process
Too much high value for Reactor 1 may is around 5000 mg/L.
due to practical errors such as sample
filtration errors. So we can come into
ACKNOWLEDGMENT
conclusion that : 4 2 ratio of 10: 1
gives better sulfate reduction than 4: 1 We would like to thank our research
ratio. Also 5000 mg/L level feed supervisor Dr.P.G.Rathnasiri for the

[9]
continuous guidance given. Again we have
to thank M.Sc. student Mr. I.Samaratunga
for the guidance and support given. Also
we would like to thank laboratory
assistants, workers and all the students
who helped us in various ways to complete
this research.
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