Bioprocess to control the turbidity of wastewater by the

microbial consortium Scenedesmus spp. and Chlorella

spp.
Sergio Hernán Mateus Osorio
Energy Engineering
smateus7@unab.edu.co

Autonomous University of Bucaramanga

RESUMEN 5. JUSTIFICATION
Se propone un proceso de biorremoción de la turbidez mediante The insoluble particles in suspension, colloidal or very fine that
un proceso de bioadsorción con filtros empaquetados a los cuales are found in certain types of water are those that cause turbidity
se les agrega un consorcio de algas verdes con Scenedesmus spp y making difficult the transmission of light and are very difficult to
Chlorella spp. Estas algas se cultivarán en fotorreactores aireados decant and filter. These substances can form deposits in water
y en presencia de luz. Las algas son fotosintéticas, transforman la pipes and equipment. In addition, it interferes with most processes
luz en energía química, asimilan carbono en forma de dióxido de to which the water can be destined. Adsorption with organisms
carbono CO2 y producen oxígeno O2. Los nutrientes del agua y las will improve water quality by retaining materials and providing
partículas en suspensión que producen la turbidez, serán su fuente oxygen by photosynthetic algae.
de alimento las cuales se fijarán en filtros empaquetados donde
realizarán la adsorción de las partículas en suspensión.
6. OBJECTIVES
ABSTRACT 6.1 General objective
A process of bioremediation of turbidity is proposed by means of Develop a bioprocess to control the turbidity of wastewater
a biosorption process with packed filters, to which a consortium of through the microbial consortium Scenedesmus spp. and Chlorella
green algae is added with Scenedesmus spp. and Chlorella spp. spp.
These algae will be grown in aerated photoreactors and in the 6.2 Specific objectives
presence of light. Algae are photosynthetic, transform light into  Characterizing wastewater
chemical energy, assimilate carbono in the form of CO2 carbon
 Adapt and identify Scenedesmus spp. and Chlorella
dioxide and produce oxygen O2. The nutrients of the water and
the particles in suspension that produce the turbidity, will be your spp. from mixed cultures.
source of food which will be fixed in packed filters where they  Prepare a microbial consortium between
will realize the adsorption of the particles in suspension. Scenedesmus spp. and Chlorella spp. in liquid
culture media.
1. Knowledge Area  Measure the turbidity of the wastewater using a
Biotechnology and Environment turbidimeter as a starting point for the bioprocess.
2. Keywords  Set up a photobioreactor to carry out the bioprocess
by inoculating increasing amounts of the microbial
Chlorella, Scenedesmus, turbidity, wastewater, microbial
consortium consortium to 100,000, 500,000, 1 million and 2
million of the consortium's microorganisms and let
3. Related them act for 1, 5, 10, 20 and 30 days with daily
ODSWater and Sanitation. turbidity measurements.
4. INTRODUCTION
In Colombia the processes of eutrophication of aquatic systems
have accelerated due to the urban development that the country
7. RESEARCH METHODOLOGY
has had, which implies a strong deterioration of these systems,
especially in the most populated areas such as cities. This problem Adaptation of algae to PHOTO BIOREACTOR
waste water
is being studied intensively in order to find the ecological value of
this type of ecosystem, which are fundamental for maintaining the
balance in a region. For this purpose, we want to use a microbial
Waste
consortium, which allows the monitoring of pollution and Consortium
water
provides tools for decision making about the use, recovery and
conservation of water resources. In this bioprocess we will make
the water turbidity decrease with these green algae that are going 8. Wastewater
METHODS AND TECHNIQUES
collection and
to be used in the crop. Turbidity measurements
USED characterization
8.1 Obtaining the waste water
El agua residual se recolecto del lavado de máquinas en una
empresa láctea donde se recolecta 12 L. Se refrigera hasta 8°C
hasta su utilización. Después de cierto tiempo de refrigeración se
utilizan frascos de 3 L y se les añade a 3 recipientes el agua
residual.
8.2 Characterization of wastewater
pH. The changes in acidity were observed by means of the pH
meter and the initial value of the water obtained was 11.45
Phosphates. By means of spectrometry an initial data (P04) 12
mg/L was obtained
Nitrogen. By means of spectrometry an initial data of 13mg/L was
obtained
Figure 1. Initial turbidimeter data
Turbidity. Turbidity was measured with the Merck TurbiQuant
1500IR. The procedure initially consisted of a calibration with
reference samples with 1000 NTU, 10 NTU and 0.02 NTU. Then
the wastewater sample is placed and the initial result is 432 NTU.
Figure 1.
8.3 Adaptation of algae to wastewater
Figure 2. Scenedesmus spp. and Chlorella spp.
To initiate the adaptation process of Scenedesmus spp. and
Chlorella spp. the wastewater is subjected to the pasteurization
process to remove microorganisms at 80°C for 5 minutes.
Four assemblies of 500 ml each were made with residual water
and algae broth with the concentrations respectively of: 25% and
75%, 50% and 50%, 75% and 25%, finally 100% residual water.
These assemblies were made for each type of algae, in which each
culture had a respective time for its growth that was 3 days for the
first, 3 days for the second, 4 days for the fourth and 10 days for
the last assembly.
8.4 Algae consortium photobioreactor and
turbidity measurements
It was counted per neubauer chamber for each algae in the
following concentrations of 100 thousand cells, 500 thousand
cells, 1 million cells and 2 million cells. From these
concentrations to 50% of each algae to form the consortium,
where each assembly with oxygen with respective times of
turbidity measurements at 5, 10, 20 and 30 days. Figure 3.
Figure 3. Assembly of photobioreactors with oxygenation
9. THEORETICAL REFERENCES
9.1 Scenedesmus spp.
Scenedesmus spp. algae line up in short rows of 4, 8 or 16 cells;
other times they alternate forming a tiny zigzag. It is a genus of
over one hundred species, all of which are common in freshwater
plankton. Many of them are very variable, making it difficult to
identify them [1].
The central cells of the Scenedesmus are usually straight-sided
while those located at the ends are slightly curved in the form of a
crescent, which often ends in two long appendages like a thorn
[4].
6.2 Chlorella spp.
It is a green unicellular algae with a spherical shape from 2 to 10
µm in diameter and does not have a flagellum. It is found in most
freshwater bodies. Chlorella spp. contains the green
photosynthesizing pigments of chlorophyll a and b in its
chloroplast.
Through photosynthesis it multiplies rapidly, requiring only
sunlight, carbon dioxide, water and small amounts of minerals. It
is used to manufacture nutritional supplements, anti-aging
cosmetic products, to stimulate the immune system and for heavy
metal detoxification.
6.3 Wastewater
Water is a natural resource used in various human activities. From
the use of this water in daily use, contaminated or waste water is
produced.
These are differentiated depending on their origin. For example, it
is called black water that is contaminated with feces or urine.
Sewage is water that comes from domestic use and as sewage that
passes through the sewer, but in reality any of these distinctions
are part of the term "wastewater" [6].
6.4 Turbidity pH 60 days
Turbidity is a measure of the degree to which water loses its
transparency due to the presence of suspended particles [11]. Inicial P1 P2 P3 P4

11.45
10.52

10.16
9.85

9.79
9.63

9.54

9.45

9.15

9.06
9.02
Measurement principle: If optical radiation is introduced into a

8.79

7.9
dispersion system, the dissolved solids transform it into another
type of energy and reduce its intensity. This effect is known as
absorption. The relationship between initial and final energy is
defined as turbidity [13].
100 mil 1 millon 2 millon D at o I n i c i al
Factory calibration: WTW online sensors are calibrated from the
factory using a multi-point method. This procedure is very stable, Graph 2. pH data in relation to time
so no re-calibration is required. Formazin, diluted to the required
concentration, is used as the calibration standard [13].

Cleaning system: There are some parameters that can have an

TURBIDITY MEASUREMENT
impact on the turbidity value and that are automatically
compensated taking into account the influence of a secondary Inicial Dia 5 Dia 10 Dia 20 Dia 30
light source, the color of the particles and the medium. The optical

432
path must always remain clean and for this purpose WTW uses an
exclusive ultrasonic system [13].

264
6.5 Photobioreactors

125
92.8

105

76.5
75.2

73.3
A bioreactor is a vessel or system commonly cylindrical, of

65.8
45.3

40.3
32.5

54
various sizes and are usually manufactured in stainless steel where
it is sought to maintain an adequate environment such as pH,
temperature, oxygen concentration etc., either to maintain a living 100 mil 1 millon 2 millon D at o I n i c i al
organism or biochemically active substances derived from such
organisms [16]. Graph 3. Turbidity data in relation to time

10. CHRONOGRAM Graph 3 shows a decrease in initial turbidity of 432 NTU at 10

Mon Activities days in all concentrations with values of 45,32,5 and 40,3 NTU,
t being the sample with 1 million cells at 10 days the most efficient.
1-2 Characterize the wastewater
3-4 Adapt Scenedesmus spp. and Chlorella spp. to
different concentrations of wastewater
5 Prepare a microbial consortium between Scenedesmus
TURBIDITY MEASUREMENT at 60
spp. and Chlorella spp. from the adaptations days
6-10 Make an assembly of a photobioreactor to carry out
the bioprocess inoculating increasing amounts of the
microbial consortium to 100 thousand, 500 thousand, P1 P2 P3 P4
one million and 2 million of the microorganisms of
8.24

the consortium
11- Measure the turbidity of the wastewater using a
12 turbidity meter as the starting point of the bioprocess
5.04

and leave it for 1, 5, 10, 20 and 30 days with daily

4.32
4.09

turbidity measurements.
3.94
4
2.77

2.61
2.42

2.8

2.09

11. RESULTS
1.69

Graph 1 shows the decrease of the initial pH to values of 8.5 to

8.9 from the 5th day to 30 days in all the assemblies.
100 mil 1 millon 2 millon
pH MEASUREMENT
Graph 4. Turbidity data in relation to time
11.45

Inicial Dia 5 Dia 10 Dia 20 Dia 30

8.99

8.99
8.94

8.89
8.85
8.82

8.81
8.72

8.67
8.64
8.58

In graph 4 it was a sample after 60 days to the last sample that was
8.9

made as in graph 3. Each photobioreactor decreased the turbidity

considerably to the initial sample that was analyzed. The 100,000-
cell sample was the most efficient.

100 mil 1 millon 2 millon D at o I n i c i al

Graph 1. pH data in relation to time
 11. https://www.definicionabc.com/medio-ambiente/aguas-
residuales.php
MEASURING PO4 AND N
12. https://www.lenntech.es/turbidez.htm
Nitrogeno Fosfatos 13. https://www.wtw.com/fileadmin/upload/Service/Download
s/Kataloge/Online/ES/Onl_034_039_Turbidez-y-Solidos-

27
disuelto_390-KB_ES-pdf.pdf
14. http://apuntesbiotecnologiageneral.blogspot.com/2014/05/b
13

iorreactores.html
12

8.3
15. http://www0.unsl.edu.ar/~organica/archivos/Documento
%20didactico.pdf
I n i c i al Fi n al 16. https://www.revistaciencia.amc.edu.mx/online/MicroAlgas.
pdf
Figure 5. P04 and N data in [mg/L]
12. CONCLUSION

This test shows the experimental results reported through a

process of removal of organic matter contained in this wastewater,
it was possible to make several samples in order to reduce
turbidity. Four cultures were made in order to know which would
be the most optimal, but there was an incident in the laboratory
and the project was finished with only three culture media, each
one has different concentrations to determine which would be the
most efficient with the consortium of Scenedesmus spp. and
Chlorella spp.

The final sample in each of the cultures has shown that water
quality can be improved by 100%, that by decanting this
wastewater it can be used for irrigation or for the same cleaning of
a company. Considering the potential of the algae in consortium,
plus the multiple uses given to its biomass, it is suggested that its
cultivation is economically and environmentally attractive.

13. PROJECT IDENTIFICATION

Name of Seedbed Buiatá
Project Tutor Dra. Graciela Chalela Alvarez
Investigation Group Biotechnology & Environment
Research Line Biotechnology
Presentation Date November 06th, 2020
14. REFERENCES
1. https://www.seaweed.es/productos/scenedesmus/
2. http://www.scielo.org.co/pdf/vitae/v15n1/v15n1a04.pdf
3. file:///C:/Users/biblioteca/Downloads/microalgas.cadiz.pdf
4. https://www.biodiversidadvirtual.org/micro/Scenedesmus-
ellipsoideus-img228.html
5. https://contenidos.ceibal.edu.uy/fichas_educativas/_pdf/cie
ncias-naturales/reino-protista/002-chlorella-vulgaris.pdf
6. https://www.ecured.cu/Chlorella_Vulgaris
7. http://www.exeedu.com/publishing.cl/av_cienc_ing/2012/
Vol3/Nro2/16-ACI1105-11-full.pdf
8. https://aprendeenlinea.udea.edu.co/revistas/index.php/actbi
o/article/view/330682/20786958
9. https://harmonia.la/ecologa/que_son_y_como_se_tratan_la
s_aguas_residuales
10. http://mimosa.pntic.mec.es/vgarci14/aguas_residuales.htm