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56 Current Environmental Engineering, 2015, 2, 56-63
Biological Denitrification: Screening of Packing Material, Comparison of
Denitrification Rate by Pseudomonas aeruginosa and Pseudomonas stutzeri,
Application and Design of Bioreactor
Akshay Jakhete, Sayali Titre, Pritam Patil
*
and Jitendra B. Naik
Department of Chemical Engineering, University Institute of Chemical Technology, North Maharashtra University,
Jalgaon, Maharashtra, 425001, India
Abstract: Current studies explore the viability, applicability and comparison of Pseudomonas aeruginosa and
Pseudomonas stutzeri based biological denitrification of waters containing high concentration of nitrate and nitrite waste
originating from various agricultural and industrial sources. This process converts the readily soluble nitrates to harmless
nitrogen gas through a formation of series of intermediates. Experimental work involved use of varying concentration of
nitrates and nitrites with different packing conditions, substrates and were analyzed by UV-Vis spectrophotometer. The
parameters such as C/N ratio, temperature and pH were optimized for decreasing concentration of nitrates and nitrites.
Best packing materials for nitrate reduction proved to be the agro waste based materials which have potential to be used
for nitrate reduction of various waste water samples obtained around the city. Average nitrate reduction rates obtained for
agro based waste packing condition exhibited about 96.55% and 95% for Pseudomonas aeruginosa and Pseudomonas
stutzeri respectively. When applied for denitrification of waste water sample using the obtained reaction parameters,
average nitrate reduction obtained was 53.84% and 33.72% for Pseudomonas aeruginosa and Pseudomonas stutzeri
respectively. Based on the obtained observations and experimental results, general bioreactor for denitrification process
has been designed.
Keywords: Agro waste, denitrification, nitrates, nitrites, Pseudomonas aeruginosa, Pseudomonas stutzeri.
1. INTRODUCTION
The increasing contamination of water of nitrogenous
materials and the destruction of habitats endangers the
availability of water for human consumption and wildlife
development. In the country like India, this problem is
mainly due to the excessive use of agricultural fertilizers,
septic tank systems, animal waste disposal, explosives and
industrial effluents. The increasing nitrate concentration in
ground water is one of the most important and serious
environmental problems because it poses serious health risk
to humans due to the body´s reduction of nitrate to nitrite [1,
2]. The nitrite reacts with the hemoglobin in the blood and
thus oxygen transfer to cells is inhibited. This phenomenon
is called methaemoglobinemia or the blue baby syndrome [3-
5]. Water containing high NO
3
-N concentrations should be
avoided for consumption, since it increases the probability of
non-Hodgkin’s lymphoma and gastric cancer [6]. Also,
scientific evidence shows that nitrate and nitrite are likely to
cause mutagenesis and teratogenesis, miscarriage in pregnant
women, coronary cardiac diseases, and cancer of the ovaries
and growth of hypertrophy of the thyroid [7-9]. Nitrate and
Nitrite nitrogen are known to be toxic for animals and
aquatic life, like for fish, benthic fauna, plants, and
bacterioplankton. At lower concentrations, nitrate poisoning
have known to result in increasing cases of still birth
incalves, ovaries with cyst development, lower milk
production, reduced weight gains, and vitamin A deficiency
*Address correspondence to this author at the Department of Chemical
Engineering, University Institute of Chemical Technology, North
Maharashtra University, Jalgaon, India; E-mail: patilpritamb29@gmail.com
[10, 11]. This danger necessitates the removal of nitrate from
water reserves. The U.S. Environmental Protection Agency
and WHO has set 10 ppm of nitrate nitrogen as desirable
limit for human consumption [12, 13] whereas Indian
standard for nitrates has been set at 45 ppm, which is
desirable for human consumption [14]. Nitrates from
different waste can exhibit the characteristics of either point
or nonpoint source pollution. Point sources occur at or near
the actual waste facility involved and typically exhibit high
levels of nitrate or ammonia in a limited area. Nonpoint
sources are spread over large areas [10]. Increased nitrate
concentrations in groundwater have led to the shutdown of
wells and rendered aquifers unusable as water sources.
Hence nitrates removal is an important aspect of present day
wastewater treatment process [15]. Several techniques are
available for nitrate reduction such as reverse osmosis,
chemical denitration, biological denitrification along with
physiochemical methods each with its own advantages and
disadvantages [16-20]. Physiochemical treatments involve
processes such as ion exchange and reverse osmosis which
produces secondary concentrated waste and is not feasible
[21]. Another way to remediate nitrates is by thermal
denitration which requires a temperature of more than 950
0
C
and also produces large amount NOx. The best way to
remediate nitrates is to convert them into nitrogen gas which
is achieved by both chemical denitration and biological
denitrification [21]. Chemical denitration results in nitrate
reduction to nitrogen gas using suitable reducing agents in
presence of catalysts but is feasible only when nitrate
concentration is very high. For lower concentrations of
nitrate, biological denitrification is the most effective method
and is generally preferred for industrial and domestic sewage
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