Bioremediation of Arsenic(As) and lead(pb) by Aspergillus Niger

isolated from soil

Hajra Kharala, Sheeraz Ahmed Memona, Sajid Hussain Mangia,Rafi uz zaman Brohia, Abdul Aziz
Chan, Arsalan Ali Bughio.
a
Institute of Environmental Engineering and Management Mehran- UET Jamshoro, Sindh, Pakistan

Abstract

“Like other underdeveloped and developed countries such as Pakistan also relies on water as a main source for domestic,
agriculture and industrial purposes. And the main source is surface and groundwater. Heavy metals pollution in water
is mostly caused by rapid population growth and industrial development which is accumulated in water sources further
that is consumed through water and different food sources and deteriorates the heath. Among those heavy metals,
Arsenic is one of the major toxic heavy metal. This study focuses on the bioremediation of Arsenic (As) and Lead (pb)
through Aspergillus Niger, isolated from soil. Water samples were collected from most Arsenic rich areas and Isolated
Aspergillus Niger was employed and parameters were optimized and Removal efficiency was calculated. From this
study it observed that Aspergillus Niger is a potential Bioremediation tool for Heavy Metals.”

© 2022 Hajra Kharal, Sheeraz Ahmed Memon, Sajid Hussain Mangi, Rafi uz zaman Brohi, Abdul Aziz Chan, Arsalan Ali Bughio.“Selection and/or
peer-review under responsibility of Energy and Environmental Engineering Research Group (EEERG), Mehran University of Engineering and
Technology, Jamshoro, Pakistan.”

Keywords: Bioremediation; Aspergillus Niger;Ground water; Lead; Arsenic.

1. Introduction

Modern sciences has reached pinnacle of developments in result to which human civilization has advanced
unexpectedly. Rapid Increase in population has affected the nature of climate since the rapid urbanization and industrial
revolution has begun (Padhan et al, 2021). Among them heavy metals contaminated water is one of the most threatening
and challenging issue in this Era (Leong et al 2021). Heavy metal contamination either of water or soil ultimately
degrades economic value of soil. Once the soil or water is contaminated due to heavy metals amalgamated it leads to
degrade plant crops .Heavy metals contamination has come from natural as well as fertilizers ,pesticides including
industrial and municipal wastewater. Since the world has become a global village and new developments have been
introduced, besides that there is a dire need of such technologies to be introduced that are efficient, eco-friendly and cost
effective ones which can effectively degrade and remediate environmentally related pollutants in order to sustain healthy
environment (liu et al.,2019). Bioremediation is a branch of Biotechnology that employs the use of living organisms,
like microorganisms such as Bacteria, fungi and other microbes in the removal of contaminants from soil, water and
other environments (Cory Mitchell 2020). Bioremediation is a process which is used to treat contaminated soil and
water through microbial activity by altering Environmental conditions to stimulate and industry .Since microorganisms
are promising source as they possess ability to detoxify, degrade and totally remediation of organic and inorganic
toxicants. Aspergillus are saprophytic fungi that possess capability for its growth in aerobic environments. It is easily
found in oxygen rich environments. Aspergillus are feasible enough for multiple use including agriculture and industry.
Aspergillus possess oligotrophic characteristics which means it is capable to grow in less nutrient environments.
Aspergillus is used for recovery of contaminated soil and water .The focus of this study is on remediation of toxic
pollutants such as Arsenic and Lead through Aspergillus Niger isolated from soil.
 6th International Conference on
Energy, Environment and Sustainable Development 2022 (EESD 2022)
2. Material and Methodology

2.1. Sample Collection and adsorbent preparation

For Fungus isolation, soil samples were collected from Institute of Environmental Engineering and Management
MUET jamshoro.
Potato dextrose agar was used as culture media.
The Aspergillus Niger Biomass was prepared on Potato Dextrose Broth media. Potato Dextrose broth was prepared by
taking Potato Dextrose Agar 840 mg and diluted in 30 ml of distilled water .Broth was prepared. After that 2 gm of
Glucose was added and 70 ml of distilled water was added in order to make 100 ml solution .And the flasks were
autoclaved with proper covering of aluminum foils .The solution was autoclaved at 121 C for 15 mints .Later the flasks
were opened and under proper contamination free environment in laminar hood with proper safety and the fungus
isolated was inoculated in flasks .And the flasks were mixed on a rotary shaker for 3 to 4 days for at least 4 five hours a
day on 150 Rpm and 30 C. After 3 to 4 days of rotation the thick bed biomass was developed and was used for
Biosorption experiment.

(a) (b)
Fig. 1. Adsorbent preparation from Aspergillus Niger (a) Isolated Aspergillus Niger and (b) Adsorbent prepared.

2.2. Metal biosorption experiment

After the experiments carried out having different fungal isolates with metal concentrations, the mixtures were filtered
through whatman filter paper .And the filtrate was analysed through atomic absorption spectrophotometer for metal
concentrations In order to find out metal concentrations through atomic absorption spectrophotometer the metal solution
having metal concentrations were diluted for proper results.
The 1 mL of metal solutions of concentration 100ppm and 150pm and 200 were diluted up to 250ml and metal 250, 300,
350 and, 400 in volumetric flask with 500 ml of distilled water. The samples were stored in test tubes and were analysed
further in atomic absorption spectrophotometer.

2.3. Batch biosorption experiment

Batch study was conducted separately to evaluate the effects of operational parameters such as adsorbent contact time
and adsorbent concentration for arsenic and lead removal.
Samples richen with arsenic concentration were collected from different regions of Sindh having arsenic concentration
between 35 to 100 µg/L. The contact time for metals removal was adjusted from 01 hour to 4 hours to obtain optimum
contact time for removal of metals in water. And the dosages of adsorbent were selected from 1g, 2g, 3g, 4g and 5g. The
concentration of heavy metals was determined with atomic adsorption spectrophotometer. All the experiments were
conducted in 125 ml Erlenmeyer flasks on obituary shaker at (100 rpm) at room temperature (30 ºC ±2).

3. Results and discussion

 6th International Conference on
Energy, Environment and Sustainable Development 2022 (EESD 2022)
In this study Aspergillus Niger was isolated from soil and its biosorption capacity was studied against metals such as
Arsenic and Lead. Adsorbent were prepared by isolation of Aspergillus Niger and its bioremediation for Arsenic and
Lead were analyzed on different parameters such as adsorbent dosage and adsorbent contact time.

3.1. Effects of adsorbent dosages on bioremediation of arsenic

Various adsorbent dosages from Aspergillus Niger were used for bioremediation of arsenic from water. The effects of
adsorbent dosage were varied from 1g to 5g. From the experimental analysis the optimum dosage for bioremediation of
arsenic was observed as 3g as it yield 82 % removal efficiency of arsenic from water.

Effects of adsorbent dosages on

bioremediation of Arsenic
100
80
60
40
20
0
1 2 3 4 5

Dosage (Grams) Arsenic Removal Efficiency %

Fig. 2. Effects of adsorbent dosages on bioremediation of Arsenic

3.2. Effects of adsorbent dosages on bioremediation of Lead

Various adsorbent dosages from Aspergillus Niger were used for bioremediation of Lead from water. The effects of
adsorbent dosage were varied from 1g to 5g. From the experimental analysis the optimum dosage for bioremediation of
arsenic was observed as 4g as it yield 79 % removal efficiency of arsenic from water.

Fig. 3. Effects of adsorbent dosages on bioremediation of Lead

3.3. Effects of adsorbent contact time for bioremediation of Arsenic

 6th International Conference on
Energy, Environment and Sustainable Development 2022 (EESD 2022)
Influence of adsorbent contact time for arsenic removal was investigated in the range of 1 to 6 hours. Maximum removal
efficiency of 82% for arsenic was observed within 3 hours of contact time, after which the system reached to the
equilibrium point.
100
90
80
70
60
50
40
30
20
10
0
1 2 3 4 5 6

Contact time (hours) Arsenic Removal Efficiency %

Fig. 4. Effects of adsorbent contact time on bioremediation of Arsenic

3.4. Effects of adsorbent contact time for bioremediation of Lead

Influence of adsorbent contact time for lead removal was investigated in the range of 1 to 6 hours. Maximum removal
efficiency of 79% for arsenic was observed within 3 hours of contact time.

100
80
60
40
20
0
1 2 3 4 5 6

Contact Time (hours) Lead Removal Efficiency %

Fig. 5. Effects of adsorbent contact time on bioremediation of Lead

4. Conclusion

From this study it is concluded that the Aspergillus Niger isolated from soil possessed potential in respect of
bioremediation of arsenic and lead from water. The high adsorbent capacity of Niger make it suitable for removal of
contaminants present in surface and groundwater. The maximum arsenic removal efficiency utilizing Aspergillus Niger
used as adsorbent was observed was 82 % by applying 3g of adsorbent dosage with the contact time of 3 hours. And the
maximum removal efficiency for Lead removal through adsorbent prepared from Aspergillus Niger was observed 79%
using 4g of adsorbent dosage and contact time of 3 hours.
 6th International Conference on
Energy, Environment and Sustainable Development 2022 (EESD 2022)
Acknowledgements

The authors highly acknowledge the Institute of Environmental Engineering & Management Mehran University of
Engineering & Technology, Jamshoro, Sindh, Pakistan, for its support to carry out this research work.

References

1. Bilal, M., & Iqbal, H. M. (2020). Microbial bioremediation as a robust process to mitigate pollutants of
environmental concern. Case Studies in Chemical and Environmental Engineering, 2, 100011.
2. Talukdar, D., Jasrotia, T., Sharma, R., Jaglan, S., Kumar, R., Vats, R., ... & Umar, A. (2020). Evaluation of novel
indigenous fungal consortium for enhanced bioremediation of heavy metals from contaminated sites. Environmental
Technology & Innovation, 20, 101050.
3. Singh, P., Singh, V. K., Singh, R., Borthakur, A., Madhav, S., Ahamad, A., ... & Mishra, P. K. (2020).
Bioremediation: a sustainable approach for management of environmental contaminants. In Abatement of environmental
pollutants (pp. 1-23). Elsevier.
4. Kaur, S., & Roy, A. (2021). Bioremediation of heavy metals from wastewater using nanomaterials. Environment,
Development and Sustainability, 23(7), 9617-9640.
5. Leong, Y. K., & Chang, J. S. (2020). Bioremediation of heavy metals using microalgae: recent advances and
mechanisms. Bioresource technology, 303, 122886.
6. Das, A., & Osborne, J. W. (2018). Bioremediation of heavy metals. In Nanotechnology, food security and water
treatment (pp. 277-311). Springer, Cham.
7. Dash, H. R., Mangwani, N., Chakraborty, J., Kumari, S., & Das, S. (2013). Marine bacteria: potential candidates
for enhanced bioremediation. Applied microbiology and biotechnology, 97(2), 561-571.
8. Gouma, S., Fragoeiro, S., Bastos, A. C., & Magan, N. (2014). Bacterial and fungal bioremediation strategies. In
Microbial biodegradation and bioremediation (pp. 301-323). Elsevier.
9. Hinchee, R. E., Means, J. L., & Burris, D. R. (1995). Bioremediation of Inorganics (No. CONF-950483-). Battelle
Press, Columbus, OH (United States).
10. Irshad, S., Xie, Z., Mehmood, S., Nawaz, A., Ditta, A., & Mahmood, Q. (2021). Insights into conventional and
recent technologies for arsenic bioremediation: a systematic review. Environmental Science and Pollution Research, 1-
23.
11. Jayan, N., & Bhatlu, M. L. D. (2021). Isolation and studies on zinc removal using microorganism from contaminated
soil. Materials Today: Proceedings, 44, 1892-1897.
12. Kang, C. H., Kwon, Y. J., & So, J. S. (2016). Bioremediation of heavy metals by using bacterial mixtures. Ecological
Engineering, 89, 64-69.
13. Kaur, S., & Roy, A. (2021). Bioremediation of heavy metals from wastewater using nanomaterials. Environment,
Development and Sustainability, 23(7), 9617-9640.
14. Kumar, N., Mahessar, A. A., Memon, S. A., Ansari, K., & Qureshi, A. L. (2020). Impact assessment of groundwater
quality using WQI and geospatial tools: A case study of Islamkot, Tharparkar, Pakistan. Engineering, Technology &
Applied Science Research, 10(1), 5288-5294
15. Mallikarjunaiah, S., Pattabhiramaiah, M., & Metikurki, B. (2020). Application of nanotechnology in the
bioremediation of heavy metals and wastewater management. In Nanotechnology for food, agriculture, and environment
(pp. 297-321). Springer, Cham.
16. Mishra, S., Lin, Z., Pang, S., Zhang, Y., Bhatt, P., & Chen, S. (2021). Biosurfactant is a powerful tool for the
bioremediation of heavy metals from contaminated soils. Journal of Hazardous Materials, 126253.
 6th International Conference on
Energy, Environment and Sustainable Development 2022 (EESD 2022)
17. Padhan, D., Rout, P. P., Kundu, R., Adhikary, S., & Padhi, P. P. (2021). Bioremediation of Heavy Metals and Other
Toxic Substances by Microorganisms. Soil Bioremediation: An Approach Towards Sustainable Technology, 285-329.
18. Sharma, P., & Kumar, S. (2021). Bioremediation of heavy metals from industrial effluents by endophytes and their
metabolic activity: Recent advances. Bioresource Technology, 125589.
19. Sher, S., Ghani, A., Sultan, S., & Rehman, A. (2021). Bacterial Strains Isolated from Heavy Metals Contaminated
Soil and Wastewater with Potential to Oxidize Arsenite. Environmental Processes, 8(1), 333-347.
20. Verma, S., & Kuila, A. (2019). Bioremediation of heavy metals by microbial process. Environmental Technology
& Innovation, 14, 100369.
21. Vidali, M. (2001). Bioremediation. an overview. Pure and applied chemistry, 73(7), 1163-1172
22. Volarić, A., Svirčev, Z., Tamindžija, D., & Radnović, D. (2021). Microbial bioremediation of heavy metals.
Hemijska industrija, 75(2), 103-115.
23. Yadav, K. K., Gupta, N., Kumar, V., & Singh, J. K. (2017). Bioremediation of heavy metals from contaminated
sites using potential species: a review. Indian J. Environ. Prot, 37(1), 65.