The present disclosure generally relates to the field of wastewater treatment, more
specifically the present disclosure relates to a microbial consortium for treating
industrial wastewater and a method for treatment thereof.
BACKGROUND OF THE INVENTION
Water is one of the essential natural resources without which life cannot exist.
However, Water- the natural resource is getting scarce due to more demand of
overpopulated planet. Unfortunately, the retained water resource has been getting
5 further contaminated due to industrialization and urbanization. Industries or
factories involved in a large amount of processes such as refineries, generation of
electricity, leather processing, rubber processing, foods processing, and so on,
produce abundant effluents. Such effluents involve a large quantity harmful
wastes- chemical, physical, radiochemical, etc. When such wastes are consumed
10 along with water by living beings, the wastes can be severely harmful thereto.
Even if such wastewater is left uncovered for a long time, the wastewater can be
harmful to the environment, and may be unfit for survival of living beings. For
example, the wastewater can be foul. In another example, the wastewater can emit
a large number of harmful gases, and chemicals to flora, fauna within the reservoir
15 of wastewater. Water can also be contaminated by any means of human use such
as domestic, industrial, commercial, or agricultural activities, and sewer inflow or
sewer infiltration. Ministry of Statistics and Programme Implementation reports
59.3% of the population (rural and urban) facing sewer disposal of wastewater
Around 80% of water in India is polluted including most of the river water.
20 Hindon river water of India is highly contaminated. Around 674-million-liter
sewage and industrial waste drains into Hindon river every day. The river’s water
has been assessed as not suitable for consumption, and parameters thereof have
been enlisted in Table 01. The Hindon river water is highly contaminated with
heavy metals such as lead, cadmium, chromium, and pesticide concentration
6
including endosulphan II Beta, Alpha beta, beta BHC, heptachlor, heptachlor,
epoxide, endosulphan, etc.
Apart from Hindon river, there has been many reservoirs of wastewater founded
by Industries which have contamination rate more than that of the treatment
5 thereof. Other examples include Indian Oil Corporation’s Mathura refinery- one
of the major reasons for polluting water of Yamuna River. Effluent discharged
from the refinery is responsible for the death of thousands of fishes in the Yamuna
River. Large-scale growth of water hyacinth is also found in the river due to the
effluent and waste products released by the refinery. Some petroleum
10 hydrocarbons are also mapped in the samples of Yamuna River which are
potentially carcinogenic in nature which makes it unfit and above all safety
parameters.
In the tanning industry also, each processing step leads to the creation of waste
and emissions, as seen by the breakdown of a typical leather tanning plant.
15 Tannery waste includes both solid and liquid waste, such as splits, shavings, and
trimmings, which contain fats, minerals, wastewater, and sludge. Leather
manufacturing pollutes the environment by releasing “heavy metal chromium into
water, high COD, gaseous contaminants (H2S, NH3), and ammonia (as N)
wastes”.
20 The natural rubber manufacturing industry is also one of the major contributors
for water pollution. Such industries run processes which involve water, electricity,
chemicals, and other resources, emitting a lot of waste and effluent. Rubber
manufacturing wastewater includes high levels of Biological oxygen demand
(BOD), Chemical oxygen demand (COD), and SS. Coagulation serum, field latex
25 coagulation, and skim latex coagulation. The effluent released after rubber
processing is acidic in nature and contains ammonia and nitrogen compounds in
high amounts along with sulphate.
There have been many conventional arts to treat industrial wastewater, for
example, chemical, physical, and so on. Some conventional methods are multi-
7
levelled processes which involve exposure of wastewater to different treatments at
different levels. Physical wastewater treatment methods like screening,
sedimentation, aeration, and skimming. Chemical treatment methods involve
exposure of the wastewater involves many chemicals like pH neutralizers, anti5 foaming agents, coagulants, flocculants, and so on. However, such methods are
very costly and time taking.
There are greener alternatives such as Activated sludge process (ASP) utilizing
living bacterial flocs to degrade organic matter of the sewage and industrial waste
in aerated bioreactors. Such a process involves microorganisms digesting
10 biodegradable foods such as proteins, carbohydrates, fats, and other compounds
from wastewater. The ASP process mainly involves bacteria (95%), protozoa
(4%), and metazoa (1%). The bacteria can directly consume soluble organic
material, while solid particles consume through adsorption, and absorption. In
1953, Oswald suggested mutual interaction of algae and bacteria biomass to
15 mutually interact and degrade the wastes. However, bacteria require at least 0.1-
0.3 mg/L of oxygen to survive. Optimum pH between 7 and 7.5 and warm
temperature are required to enhance growth of bacteria. The bacteria may require
other basic nutrients such as carbon, nitrogen, phosphorus, trace amounts of
sodium, potassium, magnesium, and iron. Protozoa is a single cell microorganism
20 which is efficient in removal of amine containing pollutant only through ion
trapping mechanism. They can also be involved in removal of non-flocculent
bacteria and very small floc, therefore depending upon the bacteria. However, the
aforementioned microorganisms involved in the ASP process require nutrients to
grow.
25 As suggested by Oswald, microalgae can also play a good role in treatment of
industrial wastewater. Microalgal are the photosynthetic microorganisms that
convert water, sunlight, and CO2 to algal biomass. The microalgae makes own
food thereof. Hence, the microalgae can be the better option for treatment of
industrial wastewater. There are many microalgae which have been used in
30 treatment of industrial wastewater. For example, Chlorella protothecoides var.
8
acidicola, Ankistrodesmus, Scenedesmus, Euglena, Chlamydomonas, Oscillatoria,
Micractinium and Golenkinia, Scenedesmus, Nitzschia, Navicula and
Stigeoclonium , Galdieria sulphuraria, also denoted as Cyanidium caldarium.
The microalgae has tendency to grow on wastewater, however absorption capacity
5 thereof to eat nutrients from wastewater is nil. Hence, as the microalgae absorbs
enough nutrients from the wastewater to full capacity thereof, the microalgae
stops absorbing the nutrients therefrom further, thereby stopping the treatment
thereof thereafter. There has been research eliciting role of fungal in treating
pesticides from the wastewater, however overall treatment of the wastewater is
10 unknown through microbial consortia still.
Therefore, in light of the foregoing discussion, there exists a need for enhancing
capacity of microalgae to absorb nutrients from industrial wastewater, thereby
treating thereto at commercial scale at low cost without requiring any equipments.
SUMMARY OF THE INVENTION
15 In one embodiment of the present invention, a microbial consortium is disclosed.
The consortium includes an inoculation of a filamentous fungus, and at least one
green microalgae deployed into industrial wastewater. The filamentous fungi is
Aspergillus niger and the microalgae is Chlorella vuglaris. Spores of each of the
consortium are of size in the range of 1.0E5/L to 1.2E9/L. The consortium further
20 includes nanoparticles/nanomaterials, and/or combinations thereof. The
consortium is deployed into industrial wastewater in forms selected from a group
consisting of a sachet, a powder, a liquid, a spray, fumes, a gas, and so on.
In another embodiment of the present invention, a method for treating industrial
wastewater is disclosed. The method includes cultivating growth of each of
25 Aspergillus niger and Chlorella vulgaris separately by inoculating at least 10% of
seed cultures thereof with a culture medium, and shaking thereof. The method
further involves placing each of the cultures separately at 25+2
oC under a
continuous cool-white, fluorescence light illumination at 100µmol/(m2s).
Thereafter, the method involves mixing inoculation of spores of Aspergillus niger
30 of size in the range of 1.0E5/L to 1.29E9/L into the culture medium of Chlorella
9
vulgaris, forming a complex pellet. Finally, the method involves deploying at
least 100 L of the pellet in 100 mL of the wastewater.
BRIEF DESCRIPTION OF THE DRAWINGS
Other objects, features, and advantages of the embodiment will be apparent from
5 the following description when read with reference to the accompanying
drawings. In the drawings, wherein like reference numerals denote corresponding
parts throughout the several views:
Referring to FIG. 1, illustrated is a figure depicting a pellet combination of algae
and fungi of a microbial consortium, in accordance with a present embodiment;
10 Referring to FIG. 2, illustrated is a flowchart depicting a method (100) for treating
industrial wastewater through a microbial consortium, in accordance with the
present embodiment;
Table
Referring to Table 01, enlists latest water quality parameters of the Hindon river;
15 and
Referring to Table 02, enlists latest water quality parameters of the Hindon river
being improved after the Hindon river wastewater gets exposure to the microbial
consortium.
DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS
20 The embodiments herein and the various features and advantageous details thereof
are explained more fully with reference to the non-limiting embodiments that are
illustrated in the accompanying drawings and detailed in the following
description. Descriptions of well-known components and processing techniques
are omitted so as to not unnecessarily obscure the embodiments herein. The
25 examples used herein are intended merely to facilitate an understanding of ways
in which the embodiments herein may be practiced and to further enable those of
10
skill in the art to practice the embodiments herein. Accordingly, the examples
should not be construed as limiting the scope of the embodiments herein.
The present embodiment discloses a microbial consortium for the treatment of
industrial wastewater. The consortium includes an inoculation of a filamentous
5 fungi, and at least one green microalga. In the embodiment, the filamentous fungi
is Aspergillus niger and the microalga is Chlorella vuglaris. As the fungi and
algae are mixed together, the fungi and algae together makes a complex pellet as
shown in Fig. 1. In the mixture, spores of each microbe of the microbial
consortium are of size in the range of 1.0E5/L to 1.2E9/L. When the Aspergillus
10 niger and Chlorella vuglaris mix together to form the complex pellet, and
deployed into the wastewater, the Chlorella vuglaris starts eating all the nutrients
contained in the wastewater. As the Chlorella vuglaris absorbs the nutrients, the
Aspergillus niger absorbs all the nutrients absorbed by the algae. Hence, as the
Chlorella vuglaris absorbs all the nutrients from the wastewater thereby treating
15 thereto, the Aspergillus niger absorbs all the nutrients from the Chlorella vuglaris.
As the wastewater is treated, the Chlorella vuglaris may not have nutrients to
absorb, however the Chlorella vuglaris starts making nutrients spontaneously
which may be further absorbed by the Aspergillus niger. In the embodiment, as
the purified water gets more wastes, the Chlorella vuglaris again starts absorbing
20 the nutrients, thereby treating the wastewater instantly. Hence, the Aspergillus
niger enhances the absorption capacity of the Chlorella vuglaris. Chlorella
vuglaris is the only type of microalgae which is well suited to make a complex
with Aspergillus niger.
Chlorella vulgaris is a unicellular species of green microalga in the Division
25 Chlorophyta. C. vulgaris is a green eukaryotic microalga in the genus Chlorella.
The C. Vulgaris is well known for high protein rich source, and is consumed as a
dietary supplement. Hence, the algal strain when introduced into the water, the
water may be fit for direct consumption by the living beings.
Aspergillus niger is a fungus and one of the common species of the genus
30 Aspergillus. It causes a disease called black mold on certain fruits and vegetables
11
such as grapes, apricots, onions, and peanuts, and is commonly found as
contaminant of food. The fungal strain is a haploid filamentous fungus known for
the involvement thereof in many industrial processes such as production of citric
acid and extracellular enzymes.
5 The consortium further includes nanoparticles/nanomaterials, and/or combinations
thereof. Examples of the nanomaterials may include such as but are not limited to
titania, silica dioxide, fullerenes, liposomes, vesicles, nanodroplets, and so on.
The consortium is deployed into industrial wastewater in forms selected from a
group consisting of a sachet, a powder, a liquid, a spray, fumes, a gas, and so on.
10 In another embodiment of the present invention, a method (100) for treating
industrial wastewater is disclosed as shown in Fig. 2. The method (100) involves a
number of steps, sequence thereof provided herein is exemplary for the persons
skilled in the art to understand the present invention. The method (100) includes
cultivating growth of each of Aspergillus niger by inoculating at least 10% of seed
15 cultures thereof with a culture medium, and shaking thereof at step 102A. The
method (100) includes cultivating growth of each of Chlorella vuglaris by
inoculating at least 10% of seed cultures thereof with a culture medium, and
shaking thereof at step 102B simultaneously. The culture medium is BG-11
medium. For example, the BG-11 medium includes K2HPO4•3H2O (0.04 g/L),
20 MgSO4•7H2O (0.075 g/L), CaCl2•2H2O (0.036 g/L), citric acid (0.006 g/L),
ferric ammonium citrate (0.006 g/L), EDTA (0.001 g/L), NaNO3 (1.5 g/L),
Na2CO3 (0.02 g/L), and trace metal mix A5 (1.0 ml). Trace metal mix A5
solution consisted of H3BO3 (2.86 g/L), MnCl2•4H2O (1.81 g/L), ZnSO4•7H2O
(0.222 g/L), NaMoO4•2H2O (0.39 g/L), CuSO4•5H2O (0.079 g/L), and
25 CoCl2•6H2O (0.05 g/L).
Further, the method (100) involves placing each of the cultures separately at
25+2
oC under a continuous cool-white, fluorescence light illumination at
100µmol/(m2
s) at step 104. The method (100) involves mixing inoculation of
spores of Aspergillus niger of size in the range of 1.0E5/L to 1.29E9/L into the
30 culture medium of Chlorella vulgaris, forming a complex pellet at step 106. The
method (100) involves filtering the pellets at step 108. Finally, the pellets are
12
deployed into the wastewater by deploying at least 100 L of the pellet in 100 mL
of the wastewater at step 110.
Experimental data
Algal inoculums preparation
5 The algal strain C. Vulgaris was isolated from local wastewater from Hindon
river, Ghaziabad, India. The C. vulgaris was able to grow on both the classic BG11 medium under light and the BG-11 medium, with 10g/L glucose in the dark
condition. Therefore, the C. vulgaris has both the autotrophic and the
heterotrophic pathways, and can be considered as a facultative heterotrophic
10 strain.
The seed culture of the algal strain was cultivated in BG-11 medium containing
the following chemicals: K2HPO4·3H2O (0.04 g/L), MgSO4·7H2O (0.075 g/L),
CaCl2·2H2O (0.036 g/L), citric acid (0.006 g/L), ferric ammonium citrate (0.006
g/L), EDTA (0.001 g/L), NaNO3 (1.5 g/L), Na2CO3 (0.02 g/L), and trace metal
15 mix A5 (1.0 ml). Trace metal mix A5 solution included H3BO3 (2.86 g/L),
MnCl2·4H2O (1.81 g/L), ZnSO4·7H2O (0.222 g/L), NaMoO4·2H2O (0.39 g/L),
CuSO4·5H2O (0.079 g/L), and CoCl2·6H2O (0.05 g/L) and pH was adjusted to
7.0 with 10 g/L of glucose. The enriched seed cultures were inoculated at 10 % (υ
inoculation/υ medium) on 100 mL liquid medium in 250-mL flasks placed on a
20 horizontal shaker (100-120 rpm). The culture was kept at 25 ± 2°C under the
continuous cool-white, fluorescent light illumination at 100 μmol/(m2·s).
Fungal inoculums preparation
The fungal strain Aspergillus niger was isolated from local Ghaziabad city of
India. The isolated fungal species was stored in slant medium (24 g/L potato
25 dextrose broth with 20 g/L agar). The spore suspension was counted by using
optical microscope. The experiments were carried out in 250-ml of flasks placed
on horizontal shaker (100-120 rpm) under identical temperature and lighting
conditions mentioned above using BG-11 medium with 10 g/L glucose.
Complex pellet formation
13
The initial inoculum sizes of fungal spores from 1.0E5/L to 1.2E9/L were added
to both culture medium and algae culture medium for fungi pellets and fungi–
algae complex pellets formation.
As shown in Table 02, the water quality parameters of the Hindon river water
5 improved as provided therein as the wastewater was exposed to the
aforementioned pellet complex. Hence, the microbial consortium treats the
wastewater continuously at commercial scale economically.
As enlisted in Table 01, untreated surface water of Hindon Rivers has Dissolved oxygen
(DO) 2.5 mg/L, Biological oxygen demand (BOD) 51 mg/L, Chemical Oxygen demand
10 (COD) 334 mg/L, Chlorine (Cl) 22 mg/L, pH 8.4, Electrical conductivity (EC) 10047,
transparency 27 cm. Heavy metals (Lead-Pb, Cadmium-Cd, Cromium-Cr) and average
pesticide concentration (Endosulphan II Beta, Alpha BHC, Beta BHC, Heptachlor,
Heptocolr Epoxide -Endosulphan I Alpha, Aldrin, BHC Isomers, Endosulphan Sulphate)
is also high. However, after treatment with the microbial consortium, all parameters have
15 significantly reduced.
The foregoing descriptions of exemplary embodiments of the present disclosure
have been presented for purposes of illustration and description. They are not
intended to be exhaustive or to limit the disclosure to the precise forms disclosed,
and obviously many modifications and variations are possible in light of the above
20 teaching. The exemplary embodiments were chosen and described in order to best
explain the principles of the disclosure and its practical application, to thereby
enable others skilled in the art to best utilize the disclosure and various
embodiments with various modifications as are suited to the particular use
contemplated. It is understood that various omissions, substitutions of equivalents
25 are contemplated as circumstance may suggest or render expedient but is intended
to cover the application or implementation without departing from the spirit or
scope of the claims of the present disclosure.

WE CLAIM
1. A microbial consortium comprising an inoculation of a filamentous fungi, and
at least one green microalga deployed into industrial wastewater, wherein the
filamentous fungi is Aspergillus niger and the microalga is Chlorella vuglaris,
5 spores thereof are of size in the range of 1.0E5/L to 1.2E9/L.
2. The consortium as claimed in claim 1, wherein the consortium further
comprising nanoparticles/nanomaterials, and/or combinations thereof.
3. The consortium as claimed in claim 1, wherein the consortium is deployed
into industrial wastewater in forms selected from a group consisting of a
10 sachet, a powder, a liquid, a spray, fumes, a gas.
4. A method for treating industrial wastewater, the method comprising:
-cultivating growth of each of Aspergillus niger and Chlorella vulgaris
separately by inoculating at least 10% of seed cultures thereof with a
culture medium, and shaking thereof at step 102A;
15 -cultivating growth of each of Chlorella vulgaris separately by inoculating
at least 10% of seed cultures thereof with a culture medium, and shaking
thereof simultaneously at step 102B;
-placing each of the cultures separately at 25+2
oC under a continuous
cool-white, fluorescence light illumination at 100µmol/(m2
s) at step 104;
20 -mixing inoculation of spores of Aspergillus niger of size in the range of
1.0E5/L to 1.29E9/L into the culture medium of Chlorella vulgaris,
forming a complex pellet at step 106;
-filtering the pellets at step 108; and
-deploying at least 100 L of the filtered pellets in 100 mL of the
25 wastewater at step 110.
5. The method as claimed in claim 4, wherein the complex pellet deploys into the
wastewater directly.
6. The method as claimed in claim 4, wherein a nanoparticle coated complex
pellet deploys into the wastewater.
15
7. The method as claimed in claim 4, wherein the culture medium is BG-11
medium.
8. The method as claimed in claim 7, wherein the BG-11 medium includes
K2HPO4·3H2O (0.04 g/L), MgSO4·7H2O (0.075 g/L), CaCl2·2H2O (0.036
5 g/L), citric acid (0.006 g/L), ferric ammonium citrate (0.006 g/L), EDTA
(0.001 g/L), NaNO3 (1.5 g/L), Na2CO3 (0.02 g/L), and trace metal mix A5
(1.0 ml). Trace metal mix A5 solution consisted of H3BO3 (2.86 g/L),
MnCl2·4H2O (1.81 g/L), ZnSO4·7H2O (0.222 g/L), NaMoO4·2H2O (0.39
g/L), CuSO4·5H2O (0.079 g/L), and CoCl2·6H2O (0.05 g/L).