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Industrial effluents being polluted in nature need proper management before disposing off, as it may save our economical crops & hence life



Environmental Pollution is a hot major issue globe wide. It needs to be solved on priorities basis. Dyes are used in textile, dyeing, printing, cosmetic, leather industries etc. Most of the industries are releasing their waste directly into the water systems without treatment which is threatening aquatic life seriously. When such water is used for irrigation purpose, than crops are contaminated, which ultimately disturb the whole food chain. So, there is an utmost need to treat industrial effluent before its disposal into main water body. Industrial polluted water is used for irrigation purpose which is contaminating our crops. The attention would be focused to work on degradation/detoxification of industrial wastes using microbes, enzymatic profiles; toxicity evaluation etc. Our experimental strategy involves to irrigate the multiple varieties of Solanum lycopersicum (tomato) with normal water, with industrial effluents (polluted untreated water from three different sources) and with treated polluted water, followed by determination of different parameters like soil properties, germination, growth parameters, photosynthetic attributes, oxidants, antioxidants (enzymatic, non-enzymatic), microbial contamination of soils, protein profiling, qualitative & quantitative aspects of yield, heavy metal contents determination, estimation/localization of industrial effluents under study in root shoot leaf and fruit part. 

What actions do you propose?


The whole experimental plan will be comprised of following parts

Part I

1.      Optimization of different experimental parameters for degradation/detoxification of industrial wastes using white rot fungi (WRF)

2.      To study the ligninolytic enzymes profile of WRF responsible decolorization as well as de-toxification of industrial effluents

3.      Spectral analysis of metabolites of treated industrial effluents

4.      Toxicity estimation of treated industrial effluents

Part II

Multiple varieties of Solanum lycopersicum (tomato) will be grown in pots and will be irrigated as follow

a-      Control- irrigation with normal water

b-       T1- Irrigation with industrial effluents (polluted untreated water from three different sources)

             T2- irrigation with treated polluted water

Different parameters like soil properties, germination, growth parameters, photosynthetic attributes, oxidants, antioxidants (enzymatic, non-enzymatic), microbial contamination of soils, protein profiling, qualitative & quantitative aspects of yield, estimation/localization of industrial effluents under study in root shoot leaf and fruit part, will be taken into account.


A.       Scientific/technical methodology (give details):


1. Degradation/Mineralization of Industrial effluents using white rot fungi

Different strains of fungi will be grown on Kirk’s medium slants for a period of one week at optimum pH and temperature, purified and stored at 4°C.

1.1 Preparation of biodecolorizable mixture

Biodecolorizable mixture (for each effluent) will be prepared using Kirk’s nutrient medium. The triplicate industrial effluent containing flasks will be inoculated with respective fungi and incubated at 32 oC for 10 days. The flasks having basal medium and industrial effluent will serve as control. Sample (1mL) will be withdrawn from flasks (after 24 hours) and centrifuged at 1200rpm for 10 min to check absorbance at λmax. Supernants obtained after centrifugation (1200rpm for 10 min) will be run though spectrometer (Model T60U). The absorbance values will be used to calculate degree of decolorization. The uninoculated medium having industrial effluent will serve as blank. The results will be represented as an average of triplicates [10].

The fungal strain exhibiting best decolorization will be chosen for development/optimization of the biodecolorization process.

1.2 Optimization of biodecolorization process

Microbial progression and their enzyme assembly are the key role models responsible for decolorization and mineralization of effluents involving dyestuffs. Literature study revealed the reliance of fungal enzymes on specific growth parameters for their maximum working efficiency. Superlative growth factors are pH, temperature, inoculum’s size and effect of different additives [11]. (Ganesh et al., 1999). The process will be optimized for chosen fungi showing maximum degradation (in screening experiment) of effluents under study. Growth parameters will be optimized by varying one parameter, retaining others unvarying. 

1.3       Enzyme assays

Ligninolytic enzymes (Laccase, Lignin peroxidase, manganese peroxidase) profiles will be studied using standard methodologies [12,13,14].

1.4       Evaluation of treated industrial effluents through quality assurance   parameters

      It will be done by measuring BOD, COD, TSS and TOC of treated industrial effluents [15].

1.5       Metabolite study and Toxicity estimation

      Metabolite study of treated industrial effluents will be done by spectral techniques, Toxicity estimation will be carried out following the standard protocol [16].


PART II          

Multiple varieties of Solanum lycopersicum (tomato) will be grown in pots and will be irrigated as follow

c-      Control- irrigation with normal water

d-      T1- Irrigation with industrial effluents (polluted untreated water)

             T2- irrigation with treated polluted water

In order to evaluate the effects of industrial wastes as discharged on the microbiological soil properties as well as crop production. The plants will be grown in a vertical manner, having nylon cords placed between plants collar and iron wires settled lengthwise in the course of the plant rows, and fastened to the topmost of the net house, from the ground.

The experiment will set down in a RCBD manner with three irrigation treatments, each will be repeated thrice.


2.1  Growth Parameters

The following germination parameters will be calculated during experimentation.

  • Germination rate, Germination percentage (G%), Germination Energy, Germination Index, Mean Germination Time (MET), Days to 50% Emergence (E50), Coefficient of Uniformity of emergence (CUE), Growth rate.

2.2  Soil chemical analysis

The soil EC and pH will be evaluated on 1:2 (w/v) and 1:2.5 (w/v) aqueous soil extracts, respectively. The accessible phosphorus will be characterized using standard protocol [17], TOC by titration [18], soluble NO3–N and NH4–N by standard procedure [19]. Soil physic-chemical parameters and soil microbiological attributes will be used altogether, for multivariate analysis, to find out the effects of the two soil measurements (i.e., GW, TW) on the dynamics of the bacterial communities.

2.3 Yield and fruit qualitative analysis

During the gathering of ripened crop, the sellable and toss out fruit will be considered and weighted, to judge the various constituents of the tomato yield. The following parameters will be considered: yield/plant, mean diameter, fruit skin off and pulp steadiness, soluble solids proportion of the flesh, measurable acidity [20], dry matter content [21], a*/b* ratio (CI) [22], and all inorganic Heavy metal contents in fruit.


2.4 Microbiological analysis

The GW and TW samples will be analysed E. coli and fecal Enterococci enumeration, according to methodology UNI EN ISO 19250:2013. The bacteriological analysis will be carried out by the spread plate method.

2.5  Protein profiling

 Protein profiling in the high-input manner is a most practicable method that permits establishment of reference informations for cells and tissues and performance of comparative proteomics. Protein will be extracted from the tomato plant, followed by 2D gel electrophoresis, in-gel digestion and identification of soluble proteins by MALDI-MS) and nano-liquid chromatography (nano-LC)-MS/MS. It will accompany to the identification rate of well-separated protein spots from a gel.

ROS and Anti-oxidant activity determination

Antioxidant activity will involve in the determination of various parameters i.e., reduction potential, H2O2, Ascorbic acid, MDA, total phenolic and flavonoid content, SOD, POD, Catalase, determination of DPPH scavenging activity etc.

Who will take these actions?

Dr. Shumaila Kiran (Principal Investigator)

Where will these actions be taken?

Department of Applied Chemistry, Government College University, Faisalabad, Pakistan

What are other key benefits?

                                i.            To study the degradation and mineralization of industrial effluents to reduce its toxicity or possibly de-toxify to be used safely for irrigation purposes

                              ii.            To study the effects of treated agro-industrial wastewater on qualitative aspects of tomato crop, microbiological contamination of tomato fruit, heavy metal contents, the microbiological soil properties, protein profiling, antioxidant activities etc.

It would cause the improvement of quality of commercial crops like tomato in terms of toxicity reduction caused by industrial effluents used for irrigation purpose.

Other Key Benefits

1.      To give awareness to public sector

2.      Strictly banned the industries to directly dispose of the toxic effluent in the outlet

3.      Providing adopted treatment technology  to industrialists and applying legislation rules

4.      Giving awareness to farmers to use treated water for irrigation purposes to avoid health hazards

What are the proposal’s costs?


A.      Salaries and Honorarium

i.                     Principal Investigator (PI) = 0.120465

ii.                   Studentships = 0.9 M

Total = 1.020465M

B.      Permanent Equipment

i.                     Refrigerator (-20 °C) = 0.2 M

ii.                    Colorimeter (Germany) = 0.145 M

iii.                  Orbital Shaker (Temperature controlled, 96 Flasks capacity) = 0.175 M

iv.                  Wooden Hood (Germany) = 0.1 M

v.                   Incubator (Germany) = 0.35 M

vi.                  Oven Temperature controlled (Germany)  = 0.15

vii.                Digital Balance (0.0001g capacity) = 0.1 M

Total = 1.22 M

C.      Expendable Supplies

i.                     Chemicals = 0.15 M

ii.                    Glassware = 0.20 M

iii.                  Solvents = 0.8 M

iv.                  Biological material = 0.4 M

Total = 1.55 M

D.      Local Travel

i.                     TA/DA for for sample collection = 0.025 M

ii.                    TA/DA for conferences/ seminars for paper presentations = 0.045 M

iii.                  spectrophotometric analysis = 0.1 M

Total = 0.17 M

E.      Others (Literature, documentation, information, online literature search, contingencies, postage, etc.)

i.                     Journal publication fee / Online material = 0.05 M

ii.                    Stationary/Contingency  = 0.03 M


Total = 0.08 M

F.       Miscellaneous

i.                     Audit Fee = 0.01 M

ii.                   Accountant Fee = 0.01 M

Total = 0.02 M

G.      Indirect cost (University overheads)

15% of total direct cost to meet office support and utilities etc. of ORIC = 0.64657 M

Total = 0.64657 M

Grand Total (A + B + C + D + E + F+ G): 4.667035 M

Time line


1.       Activity-1

Sample collection from various industries + screening of fungal strains of white rot fungi

(From start- 04 Months)

2.       Activity-2

Optimization of physic-chemical parameters for biodegradation + study of ligninolytic enzymes profile

(5-8 Months)

3.       Activity-3

Quality assurance parameters study + metabolite study of treated industrial effluents + Toxicity estimation

(9-10 Months)

4.       Activity-4

Application of different treatments on crop under study + growth parameters analysis

(11-13 Months)

5.       Activity-5

Soil chemical analysis + Yield and fruit qualitative analysis + Microbiological analysis

(14-17 Months)

6.       Activity-6

Microbiological analysis + Protein profiling + antioxidant activity determination

(18-20 Months)

7.       Activity-7

Data compilation + statistical analysis

(21-22 Months)

8.       Activity-8

Final report writing


Related proposals



[1]. Irshad, A., Ali, S. and Jan, M.R. (1997). Physico-chemical studies of industrial effluents. Proc. NSMTCC  97 on Environ. Pollut.Feb 24-26, Islamabad, Pakistan.

[2]. Emongor, V., Nkegbe, E., Kealotswe, B., Koorapetse.,Sankwasa, S. and Keikanetswe,S. (2005). Pollution Indicators in Gaborone Industrial Effluent.J. Appl. Sci., 5(1): 147-150.

[3]. Khan, H.R. (2006). Assessment of SPWAC (SoilPlant-Water-Air Continuum) Quality within and around Dhaka City. Report submitted to the Director of the Centre for Advanced Studies and Research in Biotechnological Sciences, University of Dhaka, Bangladesh.Pollution indicators in Gaberone effluent.J. Appl. Sci., 5: 147-150.

[4]. McGrath, S.P., Zhao, F.J. and Lombi, E. 2002. Phytoremediation of metals, metalloids and radionuclides. Adv. in Agronomy, 75: 1-56.

[5]. Wafaa, M.S. (2001). The Effect of Industrial Effluents and Vasicular-ArbuscularMycorrhizae on Nutrient Distribution Concentration of Wheat and Bean Plants.Online J. Biol. Sci., 1 (8): 689-693.

[6]. Pedrero, F., Kalavrouziotis, I., Alarcón, J.J., Koukoulakis, P. and Asano, T.(2010). Use of treated municipal wastewater in irrigated agriculture—review of some practices in Spain and Greece Agric. Water Manage., 97: 1233–1241

[7]. Ahmed, M., Talukder, M.S.U. and Mojid, M.A.(1993). Quality of groundwater for irrigation in Muktagacha area. J. Institute of Engineers, Bangladesh, 21: 91-98.

[8]. FAO (2011). The State of the World's Land and Water Resources for Food and Agriculture.The Food and Agriculture Organization of the United Nations AndEarthscan (2011) ISBN 978-1-84971-326-9 (hdk).

[9]. Chen, J., Liu, Y., Ni, J., Wang, Y., Bai, Y., Shi, J., Gan, J., Wu, Z. and Wu, P. (2011). OsPHF1 regulates the plasma membrane localization of low- and high affinity inorganic phosphate transporters and determines inorganic phosphate uptake and translocation in rice. Plant Physiol., 157: 269–278

[10]. Asgher, M., Asad, M.T. and Legge, R.G. (2011). Enhanced lignin peroxidase synthesis by Phanerochaetechrysosporium solid state bio-processing of lignocellulosic substrate.World J. Microbiol.Biotechnol.,22: 449-453.

[11]. Ganesh, R., Boardman, G.D. and Mochelson, F. (1994). Fate of azo dyes in sludges. Water Res., 28: 1367-1372.

[12].  Warishi, H., Valli, K. and Gold, M.H. (1992). Manganese (II) oxidation by manganese peroxidase of Phanerochaetechrysosporium: Kinetic mechanism and role of chelators. J. Biol. Chem., 267: 23688-23695.

[13].   Wolfendon, B.S. and Wilson, R. L. (1982). Radical cationsas a reference chromogens in studies of one electron transfer reactions: pulse radiolysis studies of 2, 2- azinbis-(3-ethylbenzthiazoline-6-sulfonate). J. Chem. Soc. Perkin. Trans., 2: 805-812.

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