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FUNAI Journal of Science & Technology,

3(1), 2017, 48-66

BIOREMEDIATION OF INDUSTRIAL WASTE WATER WITH PSEUDOMONAS

AERUGINOSA ISOLATED FROM REFUSE SITE SOILS

O. F. Obidi, S.C. U. Nwachukwu and N. M. Jimah

Department of Microbiology, University of Lagos, Nigeria

(Received: 10 August, 2016; Revised: 22 May, 2017; Accepted: 26 May, 2017)

Abstract

Unregulated discharge of wastewater containing toxic compounds and heavy metals from

industries constitute a major environmental hazard in Nigeria. Pseudomonas aeruginosa isolated

from refuse site soil samples and identified by the Analytical Profile Index (API) 20NE was used

to bioremediate wastewater samples from three industries in Lagos, Nigeria. The samples were

inoculated with P. aeruginosa at 2.53 x 104

cfu/ml, optical density of 0.47 at 600 nm and

incubated at room temperature for 14 days. Physico-chemical parameters such as pH,

temperature, BOD5, COD, dissolved oxygen, nitrogen, sulphide, chloride, ammonia, sulphate, oil

and grease were determined at day 0 and 14 by standard methods for examination of water and

waste water. Heavy metals including Pb, Cr, Ni, Mn, Cd, Co, Cu, Zn and Fe were determined

using atomic absorption spectroscopy. At the end of the study period, a reduction of 100, 50-97,

74-94, 85-95, 42-87, 40-76 and 33-92% was observed for Pb, Cu, Ni, Mn, Co, Zn, Fe and Cr

respectively. BOD5 reduced from 66 - 21.28 and 16-10.28 mg/l while COD reduced from 120 -

25.9 and 33-19.4 in two of the three samples. The study authenticates the potentials of P.

aeruginosa in the degradation of polluted industrial wastewater.

Keywords: Pseudomonas aeruginosa, biodegradation, wastewater, refuse site

Bioremediation of Industrial Wastewater...

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FUNAI Journal of Science and Technology, 3(1), 2017 Page 49

1. Introduction

Industrial wastewater from mining,

metallurgical, electronic, electroplating

and metal finishing processes discharged

indiscriminately into rivers and lakes is

potentially toxic to human and aquatic life

(Israel et al., 2007). On many occasions,

industrial wastewater does not only drain

into rivers and lakes directly, but also

seeps into the ground, contaminating wells

and food chains. When heavy metals enter

food chain, they accumulate in high

concentrations in the human body, leading

to major health hazards (Babel and

Kurniawan, 2003). The level of heavy

metal accumulation in the environment has

continued to increase through constant

application of pesticides, fertilizers and

metal-contaminated sewage (Herland et

al., 2000). Heavy metals become

problematic in the environment because

they are relatively stable in nature,

compared to organic contaminants (Bruins

et al., 2000). The pollution of the

ecosystem by heavy metals is therefore a

real threat to the environment because

metals cannot be naturally degraded like

organic pollutants and so persist in the

ecosystem, having accumulated in

different parts of the food chain ((Igwe et

al., 2005; Smejkalova et al., 2003).

Subsequently, cost-effective and

environmentally friendly cleanup

technologies that can be applied for the

treatment of large volumes of wastewater

with various contaminants are in urgent

demand (Canstein et al., 1999), especially

in developing countries like Nigeria. In

advanced nations, such generated

municipal and industrial wastewaters are

subjected to special treatments to prevent

the toxic effects on surface water quality

(LaPara et al., 2000). Chemical and

mechanical methods have been employed

in the treatment of industrial wastewater.

These methods have been found to be

environmentally unfriendly and expensive

(Volesky, 2001; Volesky and Naja, 2007).

However, previous studies have shown the

use of biological treatment processes to be

more beneficial and advantageous

compared to chemical treatment processes,

due to their enhanced rate of

biodegradation and environmental

friendliness without negative implications

(Rozich and Colvin. 1997).

In many instances, heavy metals such as

Lead, Cadmium, Nickel, Copper, Iron,

Chromium, Manganese and Zinc, have

been implicated as the major toxic

pollutants in the biosphere as a result of

industrial, agricultural and domestic

activities (Srivastava et al., 2005). Lead is

a highly toxic metal. The effect of acute

lead poisoning and intoxication include

inattention, hallucinations, delusions, poor

memory, and irritability. Lead absorption


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in children may affect their development

and also results in behavioral problems,

nephropathy and plumbism (Dietrich et al.,

2001; Jacobs et al., 2003). Cadmium is

another heavy metal known to interfere

with metabolic processes in plants; and it

has potential to bioaccumulate in aquatic

organisms and enters the food chain.

Exposure to cadmium has been known to

affect the cardiovascular system, as well as

constitute a risk factor for insulin

resistance development (Bernhoft, 2013:

Chen et al., 2009; Satarug and Moore,

2012).

Bioremediation involves the use of

microorganisms, plants, or fungi to remove

or neutralize pollutants from a

contaminated site. It is a waste

management technique that is considered

safe, efficient and eco-friendly. It has high

potentials to replace conventional chemical

and mechanical techniques for the removal

of heavy metals and other pollutants

(Olukanni and Kokumo, 2013). The use of

microbial biomass as an adsorbing agent

for the removal and recovery of uranium

present in industrial effluents and mine

wastewater has been reported (Nakajima

and Sukaguchi, 1986). Previous studies

have also shown the success of biosorption

by fungi and agricultural by-products as an

alternative treatment option for wastewater

containing heavy metal (Igwe et al., 2005).

Certain algae, bacteria, yeast, fungi and

other microorganisms have been reported

to have ability to accumulate and adsorb

heavy metals on/within their cells (Mitani

and Misic, 1991; Vinita and Radhanath,

1992). In view of the beneficial properties

of such biological processes in detoxifying

the environment, this study sets out to

evaluate the bioremediation potentials of

Pseudomonas aeruginosa on industrial

wastewater samples obtained from

different sources.

2. Materials and methods

2.1.Sampling site and procedures

Soil samples for the isolation of

Pseudomonas aeruginosa were obtained

from five different refuse site locations at

the University of Lagos, Lagos - Nigeria.

Industrial wastewater samples used for the

study were collected aseptically in screw-

cap bottles from paint, Aluminum and

toothpaste manufacturing industries in

Lagos and were labeled A, B and C

respectively. The wastewater samples

which were typically exposed to high level

pollution, including pollution with organic

and inorganic compounds, were

immediately transferred to the laboratory

for analysis.

2.2. Microbiological analysis


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2.2.1. Cultivation and characterization

of organism

Pseudomonas aeruginosa was isolated

from refuse soil samples collected from the

study area by directly plating serially

diluted soil samples onto nutrient agar

(NA) medium (Oxoid) and Pseudomonas

Agar Base (PAB), using standard plate

technique. A 28 g portion of NA medium

was dissolved in 1L of distilled water. This

was vigorously agitated, melted with the

use of a water bath and then autoclaved at

121oC for 15 min. PAB was prepared and

used for isolation of Pseudomonas

aeruginosa as per the supplier’s protocols.

Inoculated plates were subsequently

incubated aerobically at 37oC for 24 hr.

Developed colonies were counted, purified

and characterized by conventional

biochemical tests (Bosshard et al., 2004).

Pseudomonas aeruginosa strain was

identified based on the analytical profile

index (API), using the (API) 20 NE kit

(bioMerieux, Marcy l’Etoile, France).

2.2.2. Preparation of bacterial

suspension

To prepare the inoculum, pure colonies of

P. aeruginosa were harvested from an

overnight culture plate. This was then

suspended in 100 ml of sterile nutrient

broth medium and incubated aerobically

for 24 h at 37oC. Subsequently, the

bacterial suspension was monitored by

setting the inoculum optical density at an

absorbance wavelength of 600nm (OD600)

at 0.47, corresponding to 2.53 x

104.CFU/ml.

2.2.3. Experimental set-up

The experiment was carried out for two

weeks at room temperature. The

physicochemical characteristics of the

industrial waste water samples were

determined at day 0 and at day 14. Two

liters of each industrial wastewater

samples were labelled as Sample A, B and

C respectively. A volume of three hundred

and fifty (350) mililitres (ml) from each

wastewater sample was inoculated with 5

ml of P. aeruginosa suspension in sterile

500 ml conical flasks as the experimental

samples and labelled as A1, B1 and C1.

These represented the different industrial

samples with the test organism at day 0.

Samples A2, B2, C2 represented industrial

samples with test organism at day 14. The

control samples had no bacterial

suspension and were labeled as AI

CONTROL, B1 CONTROL, C1

CONTROL. These were the different

industrial samples without test organism at

day 0. A2 CONTROL, B2 CONTROL, C2

CONTROL represented different industrial

samples without test organism at day 14.


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2.3. Determination of wastewater

physicochemical parameters

Wastewater samples from industries

typically comprise mixtures of organic and

inorganic substances; hence the following

physico-chemical parameters were assayed

at day 0 and day 14 for the experimental

and control samples, viz: pH, temperature,

BOD5, COD, Dissolved oxygen (DO),

nitrogen, sulphide, chloride, ammonia,

sulphate and oil and grease concentration.

They were determined by conventional

methods described by the American Public

Health Association (APHA, 1998). The

percentage degradation was calculated as

described by El-Bestaway et al., (2005).

2.3.1. Heavy metal analysis

Heavy metal analysis of wastewater

samples was done as described by Kulhari

et al. (2013), using Perking Elmer Analyst

200 flame Atomic Absorption

Spectrophotometer equipped with

acetylene gas. Appropriate standards of Fe,

Ni, Co, Cd, Cu, Pb, Cr, Mn and Zn were

used as reference analytes for the analyte

calibration and respective estimation of the

heavy metals. A standard stock solution

of 1000 ppm was diluted further to prepare

working standards solutions ranging from

1ppm to 8 ppm. Subsequently, a

calibration curve was obtained by a plot

between concentration (ppm) and

measured absorbance. The operating

parameters used in the determination of the

heavy metals are presented in Table 1

2.4. Statistical analysis

The analyses were performed with PASW

Statistics (version 18.0) of the SPSS

statistical package. One way Analysis of

Variance (ANOVA) was used to compare

the treated effluent samples from the

three companies studied with their

respective controls and the NESREA

Limits, to evaluate the inherent variability

based on selected parameters.

3. Results

3.1. Culture and characterization of

Pseudomonas aeruginosa

Pseudomonas aeruginosa was isolated

from refuse site soils and identified on the

basis of phenotypic characterization, using

the API 20NE kits. The total heterotrophic

bacterial population present in the refuse

site soil samples ranged from 1.86 x 104 to

2.86 x 106

cfu/g on NA, and between 1.2 x

104

and 1.0 x 105

cfu/g on PAB plate. The

microscopic and macroscopic examination

revealed slender, Gram-negative bacilli,

motile by one or two polar flagella,

arranged singly, in pairs or short chains.

Colonies were large, 2-3 mm in diameter,

smooth, translucent, irregularly round and

emit a characteristic fruity odour.


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3.2. Wastewater composition

The various components of the wastewater

samples were determined at the beginning

and at the end of the biodegradation study

to evaluate the efficiency of P. aeruginosa

in bioremediation of the samples. The

results showed that heavy metals such as

Pb, Ni, Mn, Cd, Cu, Zn, Fe, Cr and Fe

were present at concentrations ranging

from 0.0031-0.678 mg/l in the three

industrial wastewater samples. Amongst

the metals detected, Fe had the highest

concentration of 0.62, 0.67 and 0.40mg/l

in samples A, B and C respectively. The

BOD and COD ranged from 16-66 and 33-

120mg/l for all the samples. The

occurrence of oil and grease was observed

in all the three samples, ranging from 0.9

to 9.50mg/l. Tables 2-4 show the

composition of the wastewater samples

from the three industries.

3.3. Biodegradation of wastewater

using P. aeruginosa

At the end of the study period, BOD had a

decrease of 68%, and 35%, for sample A

and B respectively, compared to the

control samples. COD had a decrease of 78

and 47% for samples A and B respectively.

The BOD5 and COD values of samples A

and B thus fell within the acceptable limits

of the National Environmental Standards

and Regulation Enforcement Agency

(NESREA) and World Health

Organization (WHO) after the remediation

studies. However, in sample C, which had

a high concentration (0.314 mg/l) of Cd

before remediation, there was no decrease

in both BOD5 and COD. The

physicochemical parameters of the

wastewater samples, the controls, as well

as the NESREA and WHO limits are

shown in Tables 2-4. The dissolved

oxygen was 2.0 and 4.0mg/l in samples A

and B but 5.3 in sample C. This shows that

organic components that may result in

depletion of oxygen are less in sample C.

After the biodegradation study, DO was

observed to increase to 7.8 and 13.5 in

samples A and B, indicating that organic

matter that attract oxygen for

biodegradation has decreased over the

period. Hence, oxygen level increased. The

BOD and COD of the samples before the

remediation ranged from 16-66 and 33-

120mg/l in all the samples. These show

high levels of pollution when compared

with the WHO and NESREA limits of 10-

20mg/l and 40mg/l for BOD and COD

respectively. However, after the

remediation, BOD ranged from 10.28-

21.28 and COD from19.40-25.90 in

samples A and B respectively. Wastewater

sample A had the highest sulphate

concentration of 145mg/l, which decreased

appreciably to 19.78mg/l after the

remediation process. The sulphate

concentration in samples B and C also


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decreased from 45mg/l to 34.61 and

from17.30 to 9.06mg/l respectively. It is

noteworthy however, to observe that

sulphide was not degraded by P.

aeruginosa in all the samples. The

concentration of sulphide increased from

0.50-3.56 and 1.44-5.40mg/l in samples A

and C respectively. The sulphide

concentration however, remained static at

0.44mg/l in sample B. These values were

all far higher than the NESREA acceptable

limit of 0.2. However, the control samples

had values of 0.88, 0.36 and 4.16mg/l,

which suggests that indigenous microbes

in the control wastewater may be

degrading sulphide better than P.

aeruginosa. Wastewater sample B was

brown in colour and alkaline (pH 9.87)

before the biodegradation study. This is

higher than the NESREA acceptable limit

of 6-9. However, it could not achieve the

colourless nature after the bioremediation

process, but only became light brown with

pH reduced to an acceptable 8.85 value.

Samples A and C had pH of 7.62 and 7.12,

which were also reduced to 7.46 and 6.10

respectively after the biodegradation study.

These samples, which were colourless

before the experiment, retained their

colourless nature after the experiment.

3.4. Heavy metal concentration in

wastewater

The concentrations of heavy metals in the

different wastewater samples before and

after bioremediation are shown in Tables

2-4. Generally, all the metals show

appreciable reduction after the

biodegradation process. Mn reduced

considerably with 85.7%, 74.2% and 94%

for samples A, B and C respectively. An

appreciable reduction of 100%, 50% and

100% was achieved for Ni in samples A, B

and C, compared to the control samples.

Cu also had a 100 % reduction in all the

samples tested. Sample A had extremely

higher concentrations of Pb and Cu

compared to the permissible limit of 0.01

and 0.05 defined by WHO (2006). Sample

B also had a higher Pb concentration,

while sample C had higher Cd

concentration than the permissible limits

of 0.01. BOD5 was higher in all samples,

while COD was higher than the WHO

permissible limit in samples A and C.

Tables 5-7 show the degradation

percentages of the samples as revealed by

changes in the physico-chemical

parameters and heavy metal concentrations

in samples A, B and C after the 14-day

study period. A striking degradation

percentage of 100% was observed for Pb

and Cu in all the samples. Cadmium was

not observed in sample A. At the end of

the two-week biodegradation period, the

concentration of cadmium in sample C

increased from 0.314 to 0.452 and 0.397 in

the experimental and control samples


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respectively. These values were far higher

than the NESREA limit of 0.1mg/l. All the

other metals had higher degradation

percentage in the experimental samples

than the control samples that was not

inoculated with P. aeruginosa. The

analysis results show a P-Value greater

than the significance value of 0.05. This

suggests that there was no overall

difference between all the samples studied,

given the parameters measured. Although,

the treatment impacted a

remarkable decrease in hazardous heavy

metals such as lead and copper. Similar

results were obtained for all companies

sampled. The oil and grease content in the

different wastewater samples also

decreased to acceptable limits of < 1mg/l

(Tables 5-7). Although, oil and grease

detected in the wastewater samples were

within the NESREA acceptable limit, the

biodegradation process resulted in a

further reduction from 9.50 - 0.45, 6.40 -

0.86 and 0.9 – 0.38 mg/l in samples A, B

and C respectively, which is far below the

NESREA limit of 10mg/l. This reduction

confirms the effectiveness of P.

aeruginosa in biodegradation studies,

especially where oil is a major pollutant.

Table 1. Operating parameters used in heavy metal determination using flame atomic absorption

spectroscopy

Operating Parameters Heavy metals

Pb Ni Cr Cu Cd Co Mn Fe Zn

Wavelength (nm) 283.3 232.0 357.9 324.7 228.8 240.7 279.5 248.3 213.9

Flame Gas A-Ac A-Ac A-Ac A-Ac A-Ac A-Ac A-Ac A-Ac A-Ac

A-Ac, Acetylene gas


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Table 2. Physicochemical parameter of sample A before and after inoculation with P. aeruginosa and

their control samples

Parameters A A1 A1

CONTROL

A2 A2

CONTROL

NESREA

LIMITS

WHO

LIMITS

Temperature(oC)

pH (mg/l)

DO (mg/l)

BOD5 (mg/l)

COD (mg/l)

Sulphate (mg/l)

Sulphide (mg/l)

Nitrogen (mg/l)

NH3 (mg/l)

Pb (mg/l)

Ni (mg/l)

Mn (mg/l)

Cd (mg/l)

Co (mg/l)

Cu (mg/l)

Zn (mg/l)

Fe (mg/l)

Cr (mg/l)

Oil/grease (mg/l)

Colour

27

7.62

2.0

66

120

145

0.50

7.10

1.60

0.072

0.019

0.082

ND

0.019

0.092

0.203

0.62

0.016

9.50

Colourless

27.1

8.04

3.0

53

105

64

0.30

5.50

1.0

ND

0.016

0.023

ND

0.012

ND

0.141

0.53

0.014

1.80

Colourless

27

8.66

4.3

14

112

80

0.1

2.3

0.20

0.038

0.017

0.072

ND

0.027

0.021

0.201

0.59

0.011

2.5

Colourless

28

7.46

7.8

21.28

25.9

19.78

3.56

4.55

1.37

ND

0.0061

0.012

ND

0.0029

ND

0.065

0.20

0.0063

0.45

Colourless

27.5

8.04

12.8

17.92

49.5

29.66

0.88

3.24

0.86

ND

0.012

0.031

ND

0.0036

0.0036

0.104

0.294

0.0125

1.14

Colourless

40

6-9

Not<2

20

40

100

0.2

10

0.5

0.1

0.1

1.0

0.1

0.5

1.0

0.5

2.0

0.01

10

Colourless

30

6.5-8

10

10

40

250

N/A

N/A

0.1

0.01

0.02

0.1

0.003

0.05

0.05

1.0

1.0

0.05

N/A

Colourless

A: Initial test of sample A; A1: sample A + organism at 0day; A1 CONTROL: Sample A without organism at

0day; A2: Sample A + organism at 2 Weeks (14days) A2 CONTROL: Sample A without organism at 14 days.


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Table 3: Physicochemical parameters of sample B before and after introduction with P. aeruginosa and their

control samples

Parameters B B1 B1

CONTROL

B2 B2

CONTROL

NESREA

LIMITS

WHO

LIMITS

Temperature(oC)

pH (mg/l)

DO (mg/l)

BOD5 (mg/l)

COD (mg/l)

Sulphate (mg/l)

Sulphide (mg/l)

Nitrogen (mg/l)

NH3 (mg/l)

Pb (mg/l)

Ni (mg/l)

Mn (mg/l)

Cd (mg/l)

Co (mg/l)

Cu (mg/l)

Zn (mg/l)

Fe (mg/l)

Cr (mg/l)

Oil and grease

(mg/l)

Colour

27.1

9.87

4.0

16

33

45

0.44

5.30

0.40

0.046

0.034

0.062

ND

0.042

0.021

0.128

0.678

0.025

6.40

Yellow

27.3

9.72

4.20

14

27

36

0.20

3.40

0.20

ND

0.031

0.047

0.157

0.018

ND

0.124

0.672

0.027

2.672

Yellow

27

9.72

4.1

15

26

36

0.4

5.12

0.3

0.039

0.034

0.057

ND

0.056

0.018

0.127

0.6.86

0.032

4.74

Yellow

27.4

8.85

13.5

10.28

19.40

34.61

0.44

2.98

0.63

0.0135

0.017

0.016

0.123

0.0060

0.0083

0.025

0.406

0.0167

0.86

Brown

27.8

9.27

16.9

13.08

24.8

42.02

0.36

2.46

0.42

0.0169

0.0198

0.037

N D

0.0088

0.0115

0.017

0.594

0.0187

1.74

Light brown

40

6-9

Not<2

20

40

100

0.2

10

0.5

0.1

0.1

1.0

0.1

0.5

1.0

0.5

2.0

0.01

10

Colourless

30

6.5-8

10

10

40

250

N/A

N/A

0.1

0.01

0.02

0.1

0.003

0.05

0.05

1.0

1.0

0.05

N/A

Colourless

B: Initial test; B1: sample B + organism at 0day; B1 CONTROL: Sample B without organism at 0day; B2:

Sample B + organism at 2 Weeks (14days); B2 CONTROL: Sample B without organism at 2 Weeks (14days).


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Table 4: Physicochemical parameters of sample C before and after introduction with P. aeruginosa and

their control samples

Parameters C C1 C1

CONTROL

C2 C2

CONTROL

NESREA

LIMITS

WHO

LIMITS

Temperature(oC)

pH

DO (mg/l)

BOD5 (mg/l)

COD (mg/l)

Sulphate (mg/l)

Sulphide (mg/l)

Nitrogen (mg/l)

NH3 (mg/l)

Pb (mg/l)

Ni (mg/l)

Mn (mg/l)

Cd (mg/l)

Co (mg/l)

Cu (mg/l)

Zn (mg/l)

Fe (mg/l)

Cr (mg/l)

Oil and grease

(mg/l)

Colour

27.6

7.12

5.3

24.32

52.71

17.30

1.44

7.40

1.46

0.0056

0.046

0.0283

0.314

0.0022

0.0031

0.172

0.402

0.042

0.9

Colourless

27.6

7.81

3.10

50

92

13

9.30

6.40

1.20

ND

0.021

0.018

0.451

0.014

ND

0.162

0.342

0.0021

0.342

Colour

27.8

7.2

5.3

53.4

93.7

16.26

10.6

7.2

1.38

0.0048

0.0375

0.0261

0.432

0.019

0.0027

0.17

0.326

0.0039

1.58

Slightly

coloured

27.3

6.10

4.7

34.88

63.4

9.06

5.40

4.77

1.95

ND

0.0015

0.0018

0.452

0.011

ND

0.099

0.0969

0.0021

0.38

Colourless

27.4

6.54

6.9

26.56

58.90

14.01

4.16

6.02

1.65

0.0034

0.003

0.0046

0.397

0.0017

0.0021

0.077

0.141

0.0031

1.07

Colourless

40

6-9

Not<2

20

40

100

0.2

10

0.5

0.1

0.1

1.0

0.1

0.5

1.0

0.5

2.0

0.01

10

Colourless

30

6.5-8

10

10

40

250

N/A

N/A

0.1

0.01

0.02

0.1

0.003

0.05

0.05

1.0

1.0

0.05

N/A

Colourless

C: Initial test; C1: sample C + organism at 0 day; C1 CONTROL: Sample C without organism at 0day; C2:

Sample C + organism at 2 Weeks (14days); C2 CONTROL: Sample without organism at 2 Weeks (14days).


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Table 5: Percentage (%) degradation of sample A based on measured parameters after 14 days study

period

Parameters A A2 A2 control % degradation

for sample with

P. aeruginosa

%

biodegradation

for control

Lead

Nickel

Manganese

Cadmium

Copper

Cobalt

Zinc

Iron

Chromium

BOD

COD

Sulphate

Nitrogen

Oil & grease

0.072

0.019

0.082

-

0.092

0.019

0.203

0.62

0.016

66

120

145

7.1

9.5

-

0.0061

0.012

-

-

0.0029

0.065

0.2

0.0063

21.28

25.9

19.78

4.55

0.45

0.018

0.012

0.031

-

0.0036

0.0036

0.104

0.294

0.0125

37.92

49.5

29.66

3.24

1.14

100

68

86

-

100

85

68

67

60

68

78

86

35.92

95.26

75

37

62

-

61

81

47

53

22

44

59

53

54.37

88

A: Sample from industry A; A2: Sample A + Pseudomonas aeruginosa at 2 Weeks (14days); A2 CONTROL:

Sample A without organism at 2 Weeks (14days)


with …

Obidi et al.


Table 6: Percentage (%) degradation of sample B based on measured parameters after 14 days study

period

Parameters B B2 B2 control % degradation

for sample

with P.

aeruginosa

% degradation for control

samples

Lead

Nickel

Manganese

Cadmium

Copper

Cobalt

Zinc

Iron

Chromium

BOD

COD

Sulphate

Nitrogen

Oil & grease

0.046

0.034

0.062

-

0.021

0.042

0.012

0.678

0.025

16

33

46

5.3

6.4

-

0.017

0.016

-

-

0.0060

0.017

0.406

0.0167

10.28

17.4

34.61

2.98

0.86

0.0169

0.0198

0.037

-

0.0115

0.0088

0.025

0.594

0.0887

13.08

24.8

42.02

2.46

1.74

100

50

74

-

100

86

87

40

33

35

47

25

43.77

86.56

63

41

40

-

45

79

81

12

25

18

25

9

53.58

72.81

B: Sample from industry B; B2: Sample B + Pseudomonas aeruginosa at 2 Weeks (14days); B2 CONTROL:

Sample B without Pseudomonas aeruginosa at 2 Weeks (14days)

Table 7: Percentage (%) degradation of sample C based on measured parameters after 14 days study

period

Parameters C C2 C2 control % degradation

for sample with

P. aeruginosa

%

biodegradation

for control

Lead

Nickel

Manganese

Cadmium

Copper

Cobalt

Zinc

0.0056

0.046

0.0283

-

0.0031

0.022

0.172

-

0.0015

0.0018

-

-

0.0011

0.099

0.0034

0.003

0.0046

-

0.0021

0.0017

0.12

100

97

94

-

100

95

42

39

93

84

-

32

92

30


with …

Obidi et al.


Iron

Chromium

BOD

COD

Sulphate

Nitrogen

Oil &grease

0.402

0.028

54

96

17.3

7.4

0.9

0.0969

0.0021

26.56

58.9

9.06

4.77

0.38

0.141

0.0031

34.88

63.4

14.01

6.02

1.07

76

92

50

39

48

35.54

57.78

65

89

35

33

19

18.65

-

C: Sample from industry C; C2: Sample C + Pseudomonas aeruginosa at 2 Weeks (14days); C2 CONTROL:

Sample C without Pseudomonas aeruginosa at 2 Weeks (14days)

4. Discussion

Bioremediation has been shown to play an

important role in the degradation of heavy

metals and other toxic contaminants in

wastewater samples in the environment

(Ibegbulam-Njoku and Achi, 2014;

Olawale, 2014). The result of the present

study reveals the efficacy of P. aeruginosa

in degrading heavy metals and other toxic

wastes in industrial effluents. In order to

gain some insight into the effective

degradative status of P. aeruginosa and the

safety level of the wastewater after

biodegradation, we correlated the

physicochemical parameters of the

experimental samples with NESREA and

WHO acceptable limits of wastewater

components. The reduction in toxicity of

the wastewater was corroborated by the

corresponding decrease in BOD5 and

COD. Sample C had a high concentration

of Cd (0.314mg/L) detected before

remediation. Cadmium probably was not

biodegradable by P. aeruginosa, hence

there was no decrease in both BOD5 and

COD in this sample. Dissolved oxygen

(DO) increased from 2.0 to 7.8 and 4.0 to

13.5 in samples A and B; while there was

a decrease in sample C, probably because

the organic components in the original

samples were lower and therefore

decomposed faster in these two samples.

The decomposition in the organic matter

will lead to an increase in oxygen since the

organic components attracting oxygen for

biodegradation and causing depletion in

oxygen has decreased as a result of the

bioremediation. Therefore, such

wastewater with DO > 2 is safe when

discharged into the environment. The

Dissolved oxygen in sample C, though

decreased slightly, was still within the

NESREA limits of > 2. A sharp reduction


with …

Obidi et al.


in sulphate was observed in all the

samples, which ranged from 145 to 19.78,

45 to 34.61 and 17.30 to 9.06mg/l for

samples A, B and C respectively. This

decrease can be attributed to the utilization

of sources of sulphur in the wastewater for

the production of amino acids such as

methionine and cysteine by the inoculated

P. aeruginosa. This also explains why

sulphide, a component of sulphur was

reduced at the end of the study period.

Nitrogen was observed to decrease from

7.10-4.55, 5.30-2.98 and 7.40 -4.77mg/l in

samples A, B and C respectively. A key

observation was the 100% reduction in Pb

and Co, which further suggested that P.

aeruginosa is an effective biodegrader.

Bodour et al. (2003) also reported the

efficacy of P. aeruginosa in

bioremediation of hydrocarbon and metal

polluted sites. This biodegradative

potential of P. aeruginosa is attributable to

its ability to produce both

monorhamnolipid and a mixture of mono-

and dirhamnolipid. The monorhamnolipid

is less soluble, sorbs to the surface more

strongly, solubilizes hydrocarbon to a

greater extent, and is able to bind cationic

metals up to ten times more strongly

(Zhang et al., 1997). Pb, which has been

ranked second in the priority list for

hazardous substances in the 2003

Comprehensive Environmental Response

and Liability Act due to its high level of

toxicity (Wu et al., 2006) was drastically

reduced by the bioremediation process.

Other metals such as Chromium, Iron,

Zinc, Silver, Cobalt, Nickel and Silver

were all reduced to acceptable NESREA

limits. A similar study by Canstein et al.,

1999 reported a near complete degradation

of mercury using P. putida. The study

which achieved 90-98% removal of

mercury showed the efficiency of

microbial detoxification process. In

wastewater samples A, B and C, there was

100% removal of lead and copper. Sample

C also had 94-97% removal of Ni and Mn.

Altogether, the total removal of these

heavy metals, which are considered

common pollutants, highly toxic and

sometimes carcinogenic (Musyoka et al.,

2013), in industrial wastewater from

samples obtained from the three industries

ranged from 33-100%. Thus, a bioprocess

for wastewater cleanup using

Pseudomonas aeruginosa can be adapted

to wastewater generated from different

types of industry.

5. Conclusion

The bioremediation strategy used in this

study provided useful information on P.

aeruginosa’s active involvement in the

biodegradation of wastewater from various

industries, thereby reducing pollution

effect of industrial effluent in the

environment. While the role of other

http://scialert.net/fulltext/?doi=rjes.2014.405.421#1341878_ja

http://scialert.net/fulltext/?doi=rjes.2014.405.421#1341878_ja


with …

Obidi et al.


indigenous microorganism is not clearly

demonstrated, though may be associated

with bioremediation, we suggest the use of

a consortium of organism for better

remediation performance. Due to the fact

that most of the microorganisms that

degrade organic components such as

Pseudomonas sp are also susceptible to the

damaging effect of heavy metals,

application of a mixed consortium might

be worth investigating. This work revealed

the bioremediation potentials of P.

aeruginosa on various industrial

wastewater samples. Bioremediation

proved to be a cost-effective, easy and safe

method of removing heavy metals and

toxic contaminants in polluted industrial

effluents. The results of the study also

confirm that toxic heavy metal pollution of

the environment may be derived from

aluminum, paint and toothpaste

manufacturing industries. Therefore, they

are potential sources of hazardous

contamination of the environment through

wastewater discharge.

REFERENCES

American Public Health Association

(APHA), (1998). Standard Methods

for Examination of Water and Waste

water. American Public Health

Association. APHA AWWA-WEF,

20th

Ed. Washington, D.C.

Babel S and Kurniawan, TA, (2003). Low-

cost adsorbents for heavy metals

uptake from contaminated water: A

review. Journal of Hazardous

Materials, 977: 219-243.

Bernhoft RA, (2013). Cadmium toxicity

and treatment. The Science World

Journal,

Article ID: 394652, pp1-7.

Bodour AA, Drees KP and Maier RM,

(2003). Distribution of biosurfactants-

producing Bacteria in undisturbed and

contaminated arid southern soils,

Applied and Environmental

Microbiology, 69(6): 3280-3287.

Bosshard PP, Abel S, Altwegg M, Bottger

EC and Zbinden R, (2004).

Comparison of conventional and

molecular methods for identification

of aerobic catalase-negative gram-

positive cocci in the clinical

laboratory. Journal of Clinical

Microbiology, 42: 2065-2073.

Bruins MR, Kapil S, Oehme FW, (2000).

Microbial resistance to metals in the

environment.

Ecotoxicology and Environmental

Safety 45(3): 198–207.

Canstein HV, Li Y, Timmis KN, Deckwer

WD Wagner-Doebler I, (1999).

Removal of mercury from chloralkali

electrolysis wastewater by a mercury-

http://academic.research.microsoft.com/Author/22955357/h-f-von-canstein

http://academic.research.microsoft.com/Author/47194939/ying-jie-li

http://academic.research.microsoft.com/Author/23389180/kenneth-n-timmis

http://academic.research.microsoft.com/Author/45028525/i-wagner-doebler


with …

Obidi et al.


resistant Pseudomonas putida strain.

Applied and Environmental

Microbiology, (65)12: 5279-5284.

Chen YW, Yang CY, Huang CF, Hung

DZ, Leung YM and Liu SH, (2009).

Heavy

metals, islet function and diabetes

development, Islets, (1)3: 169–176.

Dietrich KN, Ris MD, Succop PA, Berger

OG and Bornschein RL, (2001).Early

exposure to lead and juvenile

delinquency Neurotoxicology and

Teratology 23 (6):511-518.

El-Bestaway E, El-Masry MH, and El-Adl

NE, (2005). The potentials of free

Gram-negative

bacteria for removing oil and grease

from contaminated industrial

effluents. World Journal of

Microbiology and Biotechnology, 21:

815–822.

Herland BJ, Taylor D and Wither K,

(2000). The distribution of mercury

and other trace

metals in the sediments of the Mersey

Estuary over 25 years 1974- 1998.

Science of the Total Environment 253,

45-62.

Ibegbulam-Njoku, P.N. and Achi, O.K.

(2014). Biodegradation of palm oil

mill effluents. British Microbiology

Research Journal, 4 (12): 1440-1450.

Igwe JC, Ogunewe DN, Abia AA, (2005).

Competitive adsorption of Zn (11), Cd

(11) and

Pb (11) ions from aqueous and non-

aqueous solution by maize cob and

husk. African Journal of

Biotechnology 4 (10): 1113-1116.

Israel AU, Obot IB, Umoren SA, Ikpeme

V and Ebong GA, (2007). Effluents

and solid waste analysis in a

petrochemical company – a case study

of Eleme Petrochemical Company

Ltd., Port-Harcourt, Nigerian E

Journal of Chemistry 5(1): 74-80

Jacobs DE, Mielke H and Pavur N, (2003).

The high cost of improper removal of

lead-

based paint from housing: a case

report. Environmental Health

Perspectives, 111(2): 185-186.

Kulhari A, Sheorayan1 A, Bajar S, Sarkar

S, Chaudhury A and Kalia RK,

(2013). Investigation of heavy metals

in frequently utilized medicinal plants

collected from environmentally

diverse locations of north western

India. Springer Plus 2: 676, 1-9.

LaPara TM, Nakatsu CH, Pantea L and

Alleman JE, (2000). Phylogenetic

analysis of bacterial communities in

mesophilic and thermophylic

bioreactors treating pharmaceutical

wastewaters. Applied and

Environmental Microbiology, 66

(9):3951-3952.

Mitani T and Misic DM, (1991. Copper

http://academic.research.microsoft.com/Journal/1088/aem-applied-and-environmental-microbiology

http://academic.research.microsoft.com/Journal/1088/aem-applied-and-environmental-microbiology


with …

Obidi et al.


accumulation by Penicillium sp.

isolated from soil. Soil Science and

Plant Nutrition, (37) 2: 347-349.

Musyoka SM, Ngila JC and Mamba BB,

(2013). Remediation studies of trace

metals in natural and treated water

using surface modified biopolymer

nanofibers. Phy Chem Earth. 66: 45-

50.

Nakajima A and Sakaguchi T, (1986).

Removal and recovery of uranium by

polyvinyl polyacrylate. Journal of

Chemical Technology and

Biotechnology, 36: 281–289.

Olawale AM, (2014). Bioremediation of

Waste Water from an Industrial

Effluent System in Nigeria Using

Pseudomonas aeruginosa:

Effectiveness Tested on Albino Rats.

Journal of Petroleum and

Environmental Biotechnology, 5: 167.

Olukanni DO and Kokumo KO, (2013).

Efficiency Assessment of a

Constructed Wetland Using

Eichhornia Crassipes for Wastewater

Treatment. American Journal of

Engineering Research, 2 (12): 450-

454.

Rozich AF and Colvin RJ, (1997). Design

and operational considerations for

thermophilic

aerobic reactors treating high strength

wastes and sludges. In Proceedings of

the 52nd Purdue Industrial Waste

Conference. J. E. Alleman (ed.), Ann

Arbor Press, Ann. Arbor, Mich. pp.1-

6.

Satarug S and Moore MR, (2012).

Emerging roles of cadmium and heme

oxygenase in type-2

diabetes and cancer susceptibility. The

Tohoku Journal of Experimental

Medicine, 228(4): 267–288.

Smejkalova M, Mikanova O and Boruvka

L, (2003). Effect of heavy metal

concentration on

biological activity of soil

microorganisms. Plant Soil

Environment, 49: 321-326

Srivastava R, Kumar D and Gupta SK,

(2005). Municipal Sludge-induced

Phytotoxicity.

ATLA. 33: 501-508.

Vinita VP, and Radhanath, PD, (1992).

Biorecovery of zinc from industrial

effluent using

native microflora. International

Journal of Environmental Studies, 44:

251-257.

Volesky B, (2001). Detoxification of metal

bearing effluents: Biosorption for next

century. Hydrometallurgy 59 (2-3):

203-216.

Volesky B and Naja G, (2007).

Biosorption technology: starting up an

enterprise. International Journal of


with …

Obidi et al.


Technology Transfer and

Commercialization 6: 196–211. WHO

(2006). A compendium of standards

for wastewater re-use in eastern

Mediterranean region 145/193.

Pp.289.

Wu CH, Wood TK, Mulchandani A and

Chen W, (2006). Engineering plant-

microbe symbiosis for

rhizoremediation of heavy metals. Applied

and Environmental Microbiology,

72(2): 1129-1134.

Zhang Y, Maier WJ and Miller RM,

(1997). Effects of rhamnolipids on the

dissolution, bioavailability and

biodegradation of phenanthrene.

Environmental Science and

Technology, 31:2211-2217.

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