s.

Proceedings of the
First Annual Conference on Innovations
and Developments in Civil Engineering, ACIDIC-2014
MID) D©
©
ii
N/ TK, Surathkai, India.
19-20 May 2014
KM

MUNICIPAL WASTEWATER RECLAMATION BY MICROALGAE IN

PRODUCTIVE WAYS

Pragadeesh K S * , T Sundararajan2 M Velan 3, and R Saravanane4

Abstract : Water security and wastewater reclamation have been the focus for research
and development in developing countries like India, due to diverse landscapes and
unequal water availability and distribution. A whooping estimate of 151 BLD (Billion
Litres per Day) of wastewater is expected to be generated in India, by 2030. This picture is
very alarming necessitating the need to use water efficiently and exploring the possibilities
to reuse wastewater. Despite many strategies that are available for treatment, only one-
fourth of the generated wastewaters from various sources is treated, and reused. Of the
various potential sources, municipal sewage holds much promise for reuse as it is less
toxic and hence comparatively easier to treat and reuse. Microorganisms have a natural
tendency to break down complex substances and feed them , and hence advantageous for
treatment of wastewater. Among them, microalgae are still advantageous as they are
‘photoautotrophic’ . Further, they can intake almost 70% - 80% of the nutrients available
and generate biomass.
It has been reported that a number of studies have been carried out on the use of algae for
treating wastewaters from sources like municipal, piggery, dairy etc., having the potential
for reuse. In earlier reported studies, the potential of specific types such as: Chlorella sp.,
Botryococcus braunii, Dunaliella sp. have been investigated. But, there is a need to look
at the use of cultures based on a consortium of mutually beneficial microorganisms along
with microalga. This will make a more natural environment for large scale wastewater
application, which will be free from hinderence from other microbes. Further, studies
conducted so far, shows that introduction of C02 from lignite firings enhances biomass
generation.
In this paper, a comprehensive review of reclamation of wastewater from various
wastewater sources using microalgae, especially from municipal wastewater has been
highlighted, along with critical observations thereof. Further, preliminary results of a
laboratory-based batch study using microalgae for reclaiming water from municipal
wastewater has also been highlighted. It is found that the results indicate good nutrient
utilisation by microalgae along with native microbes and that exposure of the wastewater
wherein microalgae is grown, to the smoke generated by burning lignite has increased the
nutrient level. Further studies are in progress with focus on better treatment and higher
biomass production.

Keywords : water reuse; municipal wastewater; microalgae; CO2; biomass production

1 PG student, Department of Civil Engineering, Pondicherry Engineering College, Puducherry, India,

-
e mail:pragadeesh sekar @gmail .com
2 Professor, Department of Civil Engineering, Pondicherry Engineering College, Puducherry, India,
e-mail:saravananae@gmail.com; tsrajan5621@gmail.com
3 Chief Manager, Corporate Environment Cell (CEC), Neyveli Lignite Corporation ( NLC), Neyveli,
-
Tamilnadu, India, e mail: velanji@gmail.com

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INTRODUCTION
India’ s per capita availability of water is 1545 cubic meters ( cu.m. ) as per the 2011 census
( Ministry of Water Resources, India, 2012). The average annual per capita availability of
water was 1816 cu.m, during 2001 which had reduced to 1545 cu .m, during 2011 . The
projected demand for various uses upto the year 2050 is given in Table 1. Thus, water
security has become the primary concern, both for the public and Government , in India. In
fact, India is poised for a severe water and energy crisis, in a few decades from now.
Whereas, in well developed countries there are well thought out action plans in place to face
and tackle the above crisis, India seems to lack far behind in this context . In spite of good
potential in terms of availability of fresh water, assured supply of clean and safe drinking
water to all the people of India is still a distant dream, in India . In spite of the above stark
reality, many public-spirited organisations and individuals have realised the gravity of the
problem and are trying to help people to get access to water and use it efficiently. On the
other hand, researchers are concentrating on sourcing drinking water from various hitherto
untapped sources like: sea water, sewage, grey water, industrial effluents etc.

Table: 1 Projection of Total Water Requirement for Different Uses:

Uses 1997-98 Year 2010 Year 2025 Year 2050
Low High Low High Low High
Irrigation 524 543 557 561 611 628 807
Industries 30 42 43 55 62 90 111
Domestic 30 37 37 67 67 81 81
Power 9 18 19 31 33 63 70
Inland Navigation 0 7 7 10 10 15 15
Flood Control 0 0 0 0 0 0 0
Env. ( 1 ) Afforestation 0 0 0 0 0 0 0
Env.(2 ) Ecology 0 5 5 10 10 20 20
Evaporation Losses 36 42 42 50 50 76 76
Total 36 42 42 50 50 76 76
Source: “ Water in India: Situation and Prospects” , UNICEF 2013.

The best available option at the moment is to reuse wastewater for various purposes. As all
the purposes/ sectors need not require pure water, we can use treated water for reuse in
sectors like agriculture, industry, wasteland renaissance. Wastewater reclamation is the
treatment of wastewater to make it reusable. Further, it has to satisfy the basic requirements
like: using reliable treatment method, not compromising on public health, and public
acceptance for the reuse ( Asano and Bahri, 2011). Some of the applications of treated
wastewater are: irrigation, cooling towers, toilet flushing, indirect potable reuse, wetlands
augmentation, fire protection etc.

In India, out of the localised wastewaters generated, only 51% ( i.e. 8040 MLD), 32% ( 11553
MLD ) and 8% (i.e.233.7 MLD ) are alone treated in metropolitan cities, Class I cities, and
Class II Towns respectively, using available conventional treatment facilities (CPCB, 2009).
In India, sewage treatment plants (STPs) with integrated conventional treatment options like
physical, chemical and biological methods are adopted in a generic way. Though the above
processes satisfy the current norms set out by Government of India (Gol), they do not comply

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with the really acceptable limits. However, there are numerous newer treatment technologies
that are available since the last 2 decades, which include: constructed wetlands ( CWs), bio -
refineries, sprinkle/drip type applications ( Kaur et al., 2012). Based on the economic status of
the locations, a variety of wastewater reclamation techniques like primary treatment + ultra
filtration, primary treatment + ultra filtration + reverse osmosis etc. have been adopted for
complete reuse ( Kaur et al., 2012).

Biological methods have certain distinct advantages like: lower cost, eco -friendliness,
practicable etc. In recent days, people are very keen in extracting productive ways from
wastewater. Researchers have carried out treatment, by utilising varieties of specific micro -
organisms or mixed cultures of many microbes. Emphasis on ‘quality’ , rather than quantity of
treatment, has been increasing day-by-day along with studies on ascertaining feasibility of
product development from wastes.

Diverse microbial forms are available for treating wastewater with varied efficiencies and
abilities. For example, Superbugs, which are the result of genetic modifications, are popular
for degrading aromatic longer chain compounds. There are some other groups of microbes
which can yield high value by-products out of treatment making the processes profitable. In
more common terms, biomass itself is a good output for us to utilise. Biomass can be used for
the production of bio-oils, bio-alcohols, cosmetics, dye pigments etc. Among the
microorganisms, relatively more beneficial live forms are algae, in terms of productivity,
lesser toxicity, and photoautotrophic nature. Microalgae are especially advantageous to
choose as they have the ability for larger nutrient utilisation which is available in
wastewaters. This is as a result of their smaller size (larger surface area ).

MICROALGAE AND THE SCOPE FORWARD

Microalgae used for bio-treatment of liquid wastes, are photo synthetic microorganisms
consuming organic matter in sewage and metabolise the nutrients with the help of solar
energy, preventing eutrophication. Moreover, the algal growth factors are unfavourable for
the growth of coliform bacteria. Microalgae are believed to remove heavy metals from
wastewater. Many studies showed that algae production can be done using various
wastewaters like: piggery, municipal, steel facility etc. (Aravinthan et al., 2014). Having the
producer property, microalgae can uptake CCh from external sources like power plants and
other burning processes etc. This makes the platform for abating man made greenhouse gases
( GHGs) especially, CCE ( the giant among them) in a natural way. Possibility of harvesting
many value-added products from algae increases the scope of microalgae culturing,
development and commercialisation. There are numbers of methods suggested for the
cultivation of microalgae associated with wastewater treatment . Some of them include:
hyper-concentrated cultures, immobilised cell systems, dialysis cultures, photo bioreactors,
stabilisation ponds ( Abdel- Raouf et al., 2012). There are certain physical, chemical and biotic
factors that can strongly influence microalgal growth. Some of them are: pH, light intensity,
turbidity/colour of the wastewater, nutrient composition, other chemical substances,
temperature, dissolved oxygen, geometry of the bioreactor, dominant species in microbial
consortia etc.

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ADVANTAGES WITH MUNICIPAL SEWAGE

Municipal sewage has the benefit of relatively lesser nutrient concentration when compared
to other wastewaters. This holds good for growing desired microbe by using it as the
substrate medium. Corrosion of culturing equipments is lesser in using municipal sewage and
less toxic to most of the microalgal species.

EARLIER RESEARCH HIGHLIGHTS

The most critical step in microalgal culture is to choose a particular microbial group that can
perform better. Various microalgal species have been reported to yield high biomass contents
with greater nutrient uptake rates. The most prominent among them are: Chlorella sp. ,
Dimeliella sp. , Spirulina sp., Chlamydomonas sp. etc. These are generally found in
wastewaters in all locations. For cultivation of microalgae along with CO 2 input, CO2 tolerant
species need to be analysed and chosen carelfully. Previously reported microalgal species
with more CO2 tolerance are: Cyanidium caldarium, Scenedesmus sp., Chlorococcum
littorale , Euglena gracilis, Chlorella sp. , Dimeliella tertiolecta, Chlamydomonas sp.,
Nannochloris sp., Tetraselmis sp . etc. (Ono and Cuello, 2010). Outdoor culture of
Chaetoceros muelleri and Dunaliella sp. during winter and spring showed higher growth
rates and final densities of both species were found in the alternative system independently of
the season ( Dorame et al 2010). It was also shown that 14 algae strains from the genus of
Chlorella, Haematococcus, Scenedesmus, Chlamydomonas, and Chloroccum were able to
grow on centrate (Li et al., 2011 ).

Growth characteristics of B. braunii 765 cultured with 2-20% CO2 aeration showed that the
strain could grow well without any obvious inhibition under all tested CO 2 concentrations
with an aeration rate of 0.2 vvm, even without any culture pH adjustment (Ge et al., 2011).
Feasibility study of CO2 sequestration using Chlorella sorokiniana showed that 5% CO2 (v/v)
in air was the most suitable concentration for the growth of this organism (Kumar and Das,
2012).

It is noted that microalgae contains more lipid content in their cell composition. This fact also
opens the door for choosing the microalgae with higher biomass productivity (proportionate
for lipid synthesis). Botryococcus braunii grown from secondary treated wastewater was
reported to yield a biomass of 345.6 mg/L/day with 17.85% lipid content ; Chlamydomonas
reinhardtii grown using centrate yielded 2000 mg/L/day of biomass with lipid content of
25.25%; primary treated wastewater with CO2 injection resulted the yield of 270.7 mg/L/day
biomass containing 9% cellular lipids from a mix of Chlorella sp . , Micractinium sp .,
Actinastrum sp . (Pittman et al., 2011).

Even though the above reported studies show the potential of wide range of microalgal
species, one should not be canned away by the above, as application for a particular
wastewater is always a puzzle. Hence, there is a need to identify the native microalgal species
and mutually benefiting microbes that ease nutrient uptake by algae by breakdown complex
organic matters into simpler forms. It is always better to work with clusters (i.e. mixed
consortia of microbial genera ) rather than with isolated one. This above fact is also true for
wastewater applications. This could be understood by considering the costs involved in

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isolated growth processes and their maintenance from inhibition/contamination by other

microorganisms. Hence, we need to look initially for locally dominant microalga plus
bacterial consortium before carrying out further work in wastewater reclamation studies.

PRELIMINARY STUDIES

In our institute, research in the area of utilisation of microalgae for wastewater reclamations
and CO2 sequestration for biomass growth and energy recovery thereof has been initiated,
recently. Preliminary studies were carried out using the primary treated sewage with
microalgae already present in waste stabilisation ponds maintained by the Govt of ,

Puducherry, in their treatment facilities. Details of preliminary investigations carried out and
the results obtained thereof are presented here.

Two sets of microalgal growth were studied in various sewage dilutions. The first set of study
involved the observation of algal growth in water samples from waste stabilisation pond rich
in microalgal growth and which are diluted with tap water at different ratios, till their growth
ceases. For this study, plastic containers (7 nos.) of 3.5L (working volume) have been used .
Algal water samples were diluted with tap water at various percentages, like : 6.66%, 16.66%,
33.33%, 50%, 66.66%, 83.33% and 0% (control). Top of the containers were kept open to the
atmosphere and sunlight . Meanwhile, suspended matter-removed raw sewage was added to
each of the containers, in appropriate dilution proportions ( a total volume of 50mL each), in
order to maintain algal growth. At the end of this study, dilution ratio which yielded the
highest biomass content was noted. Parameters like pH, Electrical Conductivity (EC) were
monitored, for the entire growth period.

In the second set of study, the sewage samples were diluted / mixed with microalgae rich
water, in higher sewage proportions so as to ensure nutrient becomes not a limiting factor. In
this case also, the ratio with the highest biomass yield was the target of study. For this study,
Erlenmeyer flasks ( 4 nos.) of 0.5L (working volume) were used . Various dilutions of raw
sewage with algae water ( i.e 60%, 70%, 80%, 90% and 100% (control)) were maintained
with the top of the container open to atmosphere and sunlight . After observing visible
growths in the samples, they were exposed to the smoke emanating from burning lignite, to
understand the response of the samples in terms of: pH (shows the level of smoke dissolution,
by decreased value), Electrical Conductivity (EC). Total Phosphorus (TP) and Total Kjeldahl
Nitrogen ( TKN ) values were analysed to know the effect of exposing the samples to smoke.
The characteristics of the samples were observed for one week after exposing them to the
smoke.

The volume of algal and tap water and compositions for various dilutions are given in Table 2
and 3 for the first and second set of study. Variations of pH and EC over the period of
observation for the two sets of study are shown in Figs. 1 to 4. Further, variations of TS,
TKN and TP over the period of observation for the second set of study are shown in Fig. 5 , 6.

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Municipal wastewater reclamation by microalgae in productive ways

Table 2: Volumes & composition of containers in the first set

Algal water Tap water
Additional Raw
Container
Volume(L) % Volume(L) % sewage (mL)
Control 3.5 100 0 0 50
Cl 3.26 93.33 0.24 6.66 46.66
C2 2.92 83.33 0.58 16.66 41.66
C3 2.34 66.66 1.16 33.33 33.33
C4 1.75 50 1.75 50 25
C5 1.16 33.33 2.34 66.66 16.66
C6 0.58 16.66 2.92 83.33 8.33

Table 3: Volumes & composition of containers in the second set

Raw sewage Algal water
Container Volume (mL) % Volume ( mL) %
Control 500 100 0 0
SI 450 90 50 10
S2 400 80 100 20
S3 350 70 150 30
S4 300 60 200 40

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14/ Aug 03/Sep 23/Sep 13 /Oct 14/ Aug 03/Sep 23/Sep 13/Oct
Days Days

Fig. 1: Variation in pH Fig. 2: Variation in EC

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 Proceedings of ACIDIC-2014
Municipal wastewater reclamation by microalgae in productive ways

1.3
£
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Fig. 3: Variation in pH Fig. 4: Variation in EC

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Fig. 5: Variation in TKN Fig. 6: Variation in TP

INFERENCES

The results from the first study showed that nutrient is a limiting factor, where the sewage is
considered as the nutrient and biomass yield was seen highest in the undiluted control
showing that there is no need to dilute the algal inoculums. This shows that hyper-
concentration of organisms is a better option and it is highly recommended .

During the first 10 days, slight changes in conductivity were observed but no visible algal
growth in any of the containers. However, after the introduction of raw sewage, algal growth
was visible and increased proportionately with conductivity of the samples. Over a certain
period of time, conductivity was found to be high in control and also in the samples in lower
dilutions indicating the possible growth of wastewater algae. Also, fresh water algae were
seen in the highest dilution showing that nutrient has limited the growth of wastewater
microalga in treated water.

Results of the study 2 showed that the proportion with highest raw sewage among the
samples i.e. Si (refer Table 3) has yielded the maximum biomass. Also , ‘control’ specimen of

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Municipal wastewater reclamation by microalgae in productive ways

the study 2 showed the maximum biomass growth among all the samples of study 2. This
indicates that the microalgae also have good nutrient utilisation capacity along with other
microorganisms. These results also convey that competitive nutrient uptake was severe in
control, which resulted in highest biomass, and also that microalgal hyper -concentration in
containers may help to achieve a higher biomass yield when compared to the control
specimen. The changes in parameters are almost relatively similar to samples with decreasing
sewage concentrations.

After smoke exposure to all the containers in second set , following observations were made:
pH decreased abruptly in all samples most probably due to the dissolution of CO 2, SO2,
NO2 as a whole, in the medium.
EC increased immediately and dropped to lower values, possibly due to the death of
microbial species as a result of toxic effect .
TKN increased a little in each of the samples but not in considerable levels, which may be
due to low solubility of oxides of nitrogen in water.
TP levels have gone up, showing the possibility of nutrient increase. But, this may be
also due to particulates’ interference in spectrophotometric analysis.

These results show that sparging of smoke is a very necessary part before exposing to
reaction vessels. Also, threshold value of CO2 uptake by the microalgae and associated
microbes needs to be identified.

CONCLUDING REMARKS

The preliminary studies are encouraging and show that microalgae can be used to our
advantage for reclamation of wastewater. Nutrient concentration, CO 2 concentration and
microalgal concentration have been identified as a few among the important parameters,
which seem to influence the growth of biomass. Further detailed studies are in progress and
will be reported later.

ACKNOWLEDGEMENTS

The authors thank the Principal, Pondicherry Engineering College (PEC) for extending
support and cooperation for carrying out the work reported in this paper. Further, this formed
a part of a large study of a sponsored research project in progress in the Department of Civil
Engineering, PEC with Dr. R.Saravanane, as the Principal Investigator ( PI).

REFERENCES

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Asano Takashi and Bahri Akica, 2011. Global Challenges to Wastewater Reclamation and
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CPCB, 2009. Status of water supply, wastewater generation and treatment in Class -I Cities &
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Dorame Manuel Becerra, Jose Antonio Lopez-Elias, Luis R. Martinez-Cordova, 2010. An

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Municipal wastewater reclamation by microalgae in productive ways

alternative outdoor production system for the microalgae Chaetoceros muelleri and
Dunaliella sp . during winter and spring in Northwest Mexico Aquacultural Engineering,
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Ge Yarning, Junzhi Liu, Guangming Tian, 2011 . Growth characteristics of Botryococcus
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Kaur R, Wani SP, Singh AK and Lai K, 2012. Wastewater production, treatment and use in
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Kumar Kanhaiya and Das Debabrata, 2012. Growth characteristics of Chlorella sorokiniana
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Pittman, J .K., Dean, A.P., Osundeko, O., 2011. The potential of sustainable algal biofuel
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Raouf N. Abdel, Al-Homaidan A. A., Ibraheem I.B. M., 2012. Microalgae and wastewater
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