HELIYON ISSN: 2405-8440

https://www.journals.elsevier.com/heliyon

Experimental evaluation of the natural effect of Moringa Oleifera to

reduce the turbidity and bacterial colonies present in water
Nasly A. Monedero Jaramilloa, Natalia Álvarez Lozanob, Miguel A. Argotea, Daniel F. Peñaa and
Ruthber Rodríguez Serrezuelaa
"a Industrial Engineering Program, University Corporation of Huila, Neiva, Colombia"
"b Department of Modern Languages, University Corporation of Huila, Neiva, Colombia"

ARTICLE INFO ABSTRACT

Article history: The present investigation evaluated the efficiency of moringa seeds as a coagulant and bactericide to
Received 23 December 18 counteract contamination and bacterial colonies as a possible alternative for the purification of water for
Received in revised form 20 January 19 human consumption. The research was carried out under the methodological context of experimental design
Accepted 24 February 19 with descriptive - correlational scope which allowed the evaluation of different concentrations of moringa
extract in the untreated water with low turbidity from the Las Ceibas River in the Municipality of Neiva.
Keywords: Jar-tests were used as well as total coliform colony counts, E. coli and sensory analysis. The study allowed
Bactericide, natural coagulant, bacterial defining the concentration of 4,5ppm aqueous extract of moringa as the best treatment for low turbidity
colonies, aqueous extract, Moringa waters, achieving up to 70% turbidity removal. The evaluation of the flocculent effectiveness of the Moringa
Oleifera, turbidity. Oleifera allowed ratifying its effect as a flocculent. However, it does not remove the turbidity to values
acceptable for human consumption. Likewise, the extract did not prove to be effective in the removal of all
of the total coliforms and E. coli. On the other hand, the sensory analysis with samples of drinking water
allowed determining anomalous odors and flavors present in the water by the use of Moringa

1. Introduction

This document describes the study carried out on the flocculant and bactericidal properties of the Moringa Oleífera
(MO) plant in contaminated water as a possible alternative for water treatment in rural areas. Considering that
28% of the Colombian rural population faces a critical situation due to the lack of aqueduct, thousands of people
have a hard time trying to consume water from wells and rivers exposing themselves to diseases and infections
by consuming non-potable water [1]. In addition, in the world-wide spectrum there are studies on MO carried out
in different countries. On the other hand, in Colombia, in particular in the Department of Huila, little is known
about the properties of MO.

Water safety and quality are fundamental to human development and well-being. Providing access to safe water
is one of the most effective instruments in promoting health and reducing poverty. For this reason, the international
authority on public health and water quality, WHO leads global efforts to prevent transmission of waterborne
diseases. This is achieved by promoting health-based regulations to governments and working with partners to

* Ruthber Rodriguez Serrezuela. Tel.: +57 8670772.

Electronic Engineer. Master in Industrial Control Engineering. Doctoral student in Biomedicine.
E-mail address: ruthber.rodriguez@corhuila.edu.co
University Corporation of Huila
2 Heliyon ISSN: 2405-8440

promote effective risk management practices to water suppliers, communities and households [2]. There are
several alternatives to disinfect and purify water that can be applied on a small scale at home, such as: boiling
water, chemical disinfection with chlorine, iodine or sodium permanganate and filtration [3]. However, the
disinfection of water as an essential factor is usually not enough to control the diseases related to it and its
sanitation [3]. Bearing in mind that crystalline water has a turbidity below 1 NTU, and the water becomes visible
turbid at 4 NTU and above, and that this is well above the expected levels in well-maintained and treated surface
water supplies, and in most groundwater supplies [4]. For this reason, to ensure water safety, water treatment
plants in cities and municipalities must comply with the provisions of Colombian legislation [5], which establish
the characteristics, control and monitoring of water quality for human consumption. As reference for this study,
the allowed values of turbidity, total coliforms and E. coli corresponding to 2NTU (Nephelometric turbidity units)
and 0CFU (Colony formation units).

Previous studies propose an alternative treatment of raw water to Moringa Oleifera affirming a positive effect in
the reduction of turbidity and elimination of bacterial colonies [6]. In their investigation in the Sinú River [7],
Feria, Bermúdez, & Estrada postulated the MO as an excellent alternative for water purification in rural areas
given turbidity values of raw water between 200NTU and 360 NTU. Similarly, it has been proven through the
collection and analysis of samples from containers for water storage treated with MO seed of MO that this helps
to a better sedimentation [8].

In relation to this, the study carried out in the swamp of Malambo (Atlántico) by Meza, Riaños, Mercado, Olivero,
& Jurado [9] evaluated and compared the coagulating power of the Moringa Oleifera seed with the chemical
substance most used worldwide in the processes of water clarification, that is, aluminum sulphate. This study
showed that MO is an alternative for partially replace aluminum sulfate since this decreases the turbidity by 96%,
while the seeds of MO reduce this same parameter by 64%.

In addition to this, Sandoval & Laines conducted a study in which the coagulation efficiency was compared among
three types of solutions obtained from Moringa Oleifera seeds and aluminum sulphate by jar tests [10]. They
concluded that the water treatment with aluminum sulfate gave the highest values of turbidity removal (95.60%)
and color (98.32%), followed by the treatment of the MO solution in sodium chloride. Also, the study showed
that the treatments with MO did not change the chemical properties of the treated water, and that the turbidity
elimination efficiency was greater when using solutions with sodium chloride (92.03%) and seawater (90.72%)
than when using distilled water as solvent (56.02%) because the presence of ions promotes a greater separation
of soluble proteins, which are responsible for coagulation.

On the other hand, some authors highlight the bactericidal or antimicrobial properties of the seed in the treatment
of raw water, as described by Mangale, Chonde, Jadhav, & Raut in their study [11], where they managed to reduce
the UFC in a positive sample for fecal coliform, which was treated with a high dose of 150 mg / l of seed MO.
However, the water intended for consumption must present 0CFU.

Along the same lines, Kabore, Savadogo, Rosillon, Traore, & Dianou recorded in their study [12] a high reduction
in the load of microorganisms in water treated with MO seed: 99.92% for fecal coliforms, 99.92 % for E. coli and
100% for fecal Streptococcus, although the addition of organic matter from the seed can cause the growth of
bacteria in the treated water.

The effectiveness of Moringa Oleifera as an effective coagulant and flocculant for water treatment has also been
evaluated through nanotechnology. T. R. T. Santos, et al. [13] carried out an investigation using MO as a natural
coagulant functionalized with magnetic nanoparticles of iron oxide. They stated that: "the MO coagulant could
effectively eliminate 90% of the turbidity, 85% of the apparent color and 50% of the compounds with absorption
at UV254nm, in surface water under the influence of an external magnetic field in 30 minutes".

A study by Ashenafi Delelegn, Samuel Sahile and Azamal Husen has suggested that MO seeds have potential as
antibacterial compounds against pathogens and their ability to block or evade mechanisms of resistance, could
improve the treatment and eradication of microbial strains. Therefore, plant seed extracts could be used in the
treatment of infectious diseases caused by microbes. These extracts could probably be promising natural
antibacterial agents with potential applications to control the bacteria that cause bacterial diseases transmitted by
water [14].

Moringa oleifera Lam. Seeds, (often called horseradish tree) are widely used as an alternative water treatment
 Heliyon ISSN: 2405-8440 3

method in developing countries. A suspension of triturated seed reduces turbidity and improves water quality,
making it more suitable for human consumption [15].

Finally, the efficiency of moringa seeds in the removal of turbidity varies depending on the initial turbidity levels
of the water samples since the samples with initial turbidity of 70, 250 and 400NTU suffered, respectively, greater
reductions or equal to 89.5, 95.5 and 97.5%. This is stated in a study by Muniz, G.L, Duarte, F.V., & Oliveira
S.B. [16], where a linear relationship was observed between levels of initial turbidity of the water by the best
dosage of seeds.

In this regard, with the aim of supporting this reality, the coagulant and antibacterial effects of Moringa oleifera
seed extract were evaluated, which are used not only to eliminate suspended matter, but also to condition surface
water for human consumption [17]. Allowing to identify if the levels of turbidity and bacterial colonies present in
raw water decreased or not, and to what extent their effect was more efficient, in accordance with the Colombian
legislation corresponding to the treatment of water for human consumption

1. Materials and Methodology

The study is quantitative due to its sequential and measurable nature since this type of research uses data collection
to test hypotheses based on numerical measurement and statistical analysis [18]. Likewise, the scope was set as
descriptive - correlational due to the fact that the results obtained in the water purification treatments carried out
with the different concentrations of Moringa Oleifera were processed to specify their properties and
characteristics [18], and determine the best treatment in order to show the relationship between the results of
variables when comparing them [19].

Regarding the variables of the investigation, they were established according to the object of study: the
independent variable was defined as the amount of MO to purify the water while the dependent variables were the
degree of water decontamination and the concentration of moringa used.

The research design, which was specified as an experiment, was divided into two stages: the preliminary test and
the definitive test. For the definition of the sample of the optimal concentrations in ppm of MO, eighteen (18) tests
were performed in the preliminary stage with concentrations ranging from 6ppm to 36ppm in order to treat water
from the Las Ceibas River. For this test, the water had a turbidity of 9.85NTU. From the results obtained, the best
four (4) turbidity reduction results were chosen, which were subjected to microbiological analysis in the definitive
test.

Within this framework, the best MO concentration result was diluted in clean water and subjected to a flavor
evaluation with a random sample, to whom the relevant methodology was explained to so that it would contribute
to obtaining an acceptable analysis criterion. The processing of the results was carried out through the statistical
analysis of The Analysis of Variance (ANOVA) test which has long been an important tool for researchers
conducting studies on multiple experimental groups, and one or more control groups and Tukey data. Tukey’s
multiple comparison analysis method evaluates each experimental group against each control group [20].

Seed treatment
The Moringa seeds are morphologically round, protected with a brown seed coat containing three white wings
[21], or cotyledon that was removed to manually obtain a yellowish or beige-white walnut (endosperm). These
nuts were reduced in size in an electric mill [22], where fine granules were obtained and easily agglomerated
due to their high fat content.

The material obtained was separated into two portions, one of which was preserved in the same state as that
obtained from the mill and the other portion was subjected to a fat extraction process through Soxhlet equipment,
which worked with n-hexane as solvent. According to studies, the extraction of solvents is of great commercial
importance for the chemical and biochemical industries, since it is often the most efficient method of separating
valuable products from complex raw materials or reaction products. [23].

After this process, the samples were taken (with grease and fat-free) to an oven at 105°C for 24 hours to completely
4 Heliyon ISSN: 2405-8440

eliminate moisture. The result obtained was stored in a desiccator during the time of the tests so that it would be
preserved without moisture.

Extraction of the coagulant

A balanced solution of 5g of moringa powder and 5g of NaCl in 500ml of distilled water was prepared to mix it
homogeneously in a 500ml graduated balloon. This mixture was centrifuged at 3000rpm to separate the
sedimented particles generated by the powder and the aqueous extract. Subsequently, it was filtered in vacuum
equipment using filter paper discs. After this, the result was ready to be used in water treatment tests.|

Jar-tests
The objective of carrying out the jar test was to establish the optimum proportion of moringa coagulant and to
identify its effectiveness in reducing turbidity (UNT) of treated water to eliminate impurities and bacteria fulfilling
the quality guidelines that satisfy < 5 units of nephelometric turbidity (NTU) for the requirement of potable water
according to the World Health Organization [24]. The tests were carried out in a JTP-6P jars test equipment that
had 6 rotating blades and an equal number of 1000ml samples with a rapid cycle of 100rpm for 1 minute and a
slow mixture of 40rpm of 15 minutes with a settling time of 30 minutes. Subsequently turbidity was measured
with a Hach tabletop turbidimeter.

Microbiological tests
Samples of water treated with moringa were tested in the laboratory Diagnosticamos - Neiva, where the total
coliform count and E. coli were carried out. Subsequently, they were evaluated in accordance with Colombian
regulations, specifically Resolution 2115 of 2007, which mentions the minimum parameters of drinking water
quality [5].

Organoleptic perception
A cupping test was performed to identify whether the water treated with moringa coagulant has any residual
sensory-like effect perceptible by the human sense. For this reason, three (3) different samples of water were
subjected to organoleptic evaluation. The first one had a concentration of 4.5ppm of moringa in its natural state
(with its fats and oils), the second one had a concentration of 4.5ppm of moringa totally degreased and the last
corresponded to a sample of drinking water.

The population studied was the students of the Corporación Universitaria del Huila 'Corhuila', estimated at 4,700
in 2017. The procedure to calculate the sample size was for a finite population, which turned out to be 146 people.
They were applied a test in order to know their perceptions. In the test, three (3) phases were evaluated: visual,
olfactory and gustatory.

𝑍 2 𝑝𝑞𝑁
𝑛= (1)
𝐸 2 (𝑁−1)+𝑍 2 𝑝𝑞
Where;
n = Sample number
Z = 95% confidence level indicator (1.96 z value table)
p = Positive variability (0.5 probability of success)
q = Negative variability (0.5 error probability)
N = Number of students of the Corporación universitaria del Huila ‘Corhuila’
E = Sampling error 8% (0.08)
1,962 ∗0,5∗0,5∗4700
𝑛= (2)
0,082 ∗(4700−1)+1,962 ∗0,05∗0,05

𝑛 = 145,45 ≈ 146 𝐶𝑜𝑟ℎ𝑢𝑖𝑙𝑎 𝑠𝑡𝑢𝑑𝑒𝑛𝑡𝑠 (3)

 Heliyon ISSN: 2405-8440 5

2. Results

A. Preliminary test
Tests with different concentrations of Moringa Oleifera coagulant allowed to identify the level of concentration
with higher yield.

Table I. Jar-test results

Turbidity of the water of the river 9,85UNT

Test 1 Test 2 Test 3 Test o 4
Moringa 6ppm 8ppm 30ppm 36ppm
concentration
Final water 6,45NTU 7,85NTU 9,28NTU 9,32NTU
turbidity
Source. Own elaboration
From the above information, high concentrations of Moringa extract were ruled out, since the reduction of
turbidity was less than 6%, values that were not very representative for the study.

When the concentration of 6ppm was the one with the best turbidity removal response, with a value of 34.5%, the
4 concentrations were defined for the definitive test, these being 4ppm, 4.5ppm, 5ppm and 6ppm.

B. Definitive test
The concentrations determined in the preliminary test were subjected to three (3) replicates, which showed the
following results.

Table 2. Turbidity (NTU) according to different concentrations of Moringa

Data Turbidity (NTU)
Moringa 1st Replicate 2nd Replicate 3rd Replicate Average % Removal
0ppm 20,8 20,8 20,8 20,80 0,00
4ppm 6,88 6,36 6,39 6,54 68,54
4,5ppm 7,05 5,95 5,71 6,24 70,02
5ppm 7,6 6,27 6,58 6,82 67,23
6ppm 7,36 7,48 7,17 7,34 64,73

Source. Own Elaboration

The highest percentage of turbidity reduction was 70% removal by the concentration of 4.5ppm at an initial
turbidity of the raw water of 20.8NTU.
6 Heliyon ISSN: 2405-8440

Figure 1. Removal of turbidity with different concentrations of Moringa

Removal of Turbidity
80.00 68.54 70.02 67.23 64.73
% of Removal 60.00
40.00
20.00
0.00
0.00
0.0 ppm 4.0 ppm 4.5 ppm 5.0 ppm 6.0 ppm
Moringa Extract

Source. Own Elaboration

In spite of the similarities between the turbidity removal values (See Figure 1), and the four Moringa coagulant
concentrations, the study emphasized in the result that the best behavior presented with respect to the proposed
objectives.

The microbiological analytical tests were performed by experts in a specialized laboratory. The results obtained
in jar-tests were subjected to colony count analyzes, specifically total coliforms and E. coli to evaluate the
bactericidal capacity of the MO coagulant on water.

Table 3. Results of microbiological analyzes

4ppm 4,5ppm 5ppm 6ppm

Turbidity (NTU) 4,74 3,44 4,8 5,44
Total C. (CFU) 920 540 1600 920
E. coli (CFU) 240 110 31 350

Source. Own Elaboration

In the analyzes, a decrease of the microorganisms present in the water was identified, but with an irregular
behavior and without obtaining a continuous pattern by parts of the different concentrations used, reason why it
is ruled out that MO has bactericidal action.

Table 4. Results of the organoleptic evaluation of swallowed water with Moringa with and without fat

Results of the uncertainty of the of the

qualities of drinking water
Quality Moringa Moringa Average
with fat no fat
Clean 76% 65% 71%
Odorless 100% 94% 97%
Tasteless 53% 47% 50%

Source. Own Elaboration

 Heliyon ISSN: 2405-8440 7

Figure 2. Percentage of cupping results of water treated with Moringa with and without fat

Cupping Results
97%
100%

Percentage (%)
80% 71%

60% 50%

40%
Clean Odorless Tasteless
Quality

Source: Own Elaboration

According to the data obtained (See Figure 2) from the organoleptic evaluation carried out on the two samples of
water treated with Moringa (with a concentration of 4.5ppm with and without fat), it can be deduced that, on
average, 71% of the population that evaluated the sample, determined that this was evidenced to be clean; In the
same way, 97% of the respondents said that no odor is perceived in the samples; and 50% of people deny to
perceive any taste in the samples evaluated.

As can be seen, the results of the evaluation of the organoleptic qualities of the water treated with moringa, allowed
confirming the acceptable level of use of the seed as coagulant in the concentrations indicated and thus rule out
rejection by potential users, to identify differences with the drinking water without the addition of Moringa
oleifera.

3. Conclusions
The jars-test allowed to determine the optimal coagulant dose to reduce turbidity present in the water. This showed
a percentage of removal of around 70%, which according to the Tukey statistical analysis shows a significant
difference in the flocculant effect, product of the use of different concentrations of Moringa. It was observed that
the best results obtained, in terms of a decrease in turbidity, are achieved with a concentration of 4.5ppm of
aqueous Moringa extract.

The study confirms the flocculating power of moringa on the turbidity present in the water. In addition, as the
study conducted by Asrafuzzaman, Fakhruddin and Alamgir Hossain [25], it is concluded that moringa seed
coagulants work better with high and turbid water, compared to medium or low water, since the same aqueous
solution of Moringa presents its own turbidity. Because of what is required by Colombian regulations, Resolution
2115 of 2007, regarding the treatment of drinking water or for human consumption is a maximum turbidity of
2NTU, this value is not achieved with the use of moringa as a flocculant.

Likewise, it is identified that the natural Moringa coagulant does not decant the totality of the colloidal dispersion
present in the water. Although, it was possible to greatly reduce the colonies of microorganisms, this result is not
attributed to the use of Moringa as a bactericide, because it does not eliminate 100% of the microorganisms present
in the water, as required by the Colombian standard. Then, that decrease of bacterial colonies can be attributed to
the Moringa's flocculating power, which, when decanting the colloidal suspension of the water, can drag part of
the microorganisms that are adhered to these particles.

The flocculant effectiveness of Moringa Oleifera is not competent in comparison with the efficient effect provided
by chemicals such as aluminum sulfate, ferric chloride, calcium carbonate or synthetic organic polymers, which
are, according to, the most commonly used chemicals in the coagulation-flocculation processes [9].

In relation to the previous implications, once it has been identified that the natural coagulant of Moringa
8 Heliyon ISSN: 2405-8440

effectively works as a flocculant, by decreasing the turbidity levels of contaminated water in a good percentage,
prevents water quality to be compromised by the presence of infectious agents as an essential product for life
according to the World Health Organization [26]. Therefore, it is recommended that after filtering the treated
water, substances such as chlorine or sodium hypochlorite can be used in controlled quantities to disinfect water,
and thus, achieve a safe purification.

In the organoleptic acceptance test applied to the student community of Corporación Universitaria del Huila
‘Corhuila’, a sample of drinking water treated with the natural Moringa coagulant including its essential oils and
another with degreased Moringa solution were submitted to judgment. After the analysis of the results, it was
observed that on average: a percentage of 70.5% of the community asserts that the samples are colorless, 97% of
the respondents assure that the samples are odorless and 50% of the people claim that the water consumed is
tasteless. To this last characteristic it is emphasized that the water treated with Moringa without degreasing
treatment, a percentage of 24% of the community argues to perceive a bitter taste in the water, this in response to
the sensory perception in front of the presence of essential oils typical of the seed.

4. Declarations
Data Availability Statement
All the data type used to support the findings of this study are available from the corresponding author upon
request.

Author contribution statement

Ruthber Rodriguez Serrezuela, Natalia Alvarez Lozano, Miguel A. Argote and Daniel F. Peña: Conceived and
designed the analysis; Analyzed and interpreted the data.
Ruthber Rodriguez Serrezuela, Nasly A. Monedero Jaramillo: Conceived and designed the analysis; Analyzed
and interpreted the data, wrote the paper.

Funding statement
This research was financed by the University Corporation of Huila – Corhuila

Interest statement
The authors declare the following conflict of interests: Ruthber Rodriguez Serrezuela, Nasly A. Monedero
Jaramillo and Natalia Alvarez Lozano are professors of University Corporation of Huila - CORHUILA. The
authors Miguel A. Argote and Daniel F. Peña are graduates of Industrial Engineering of the University
Corporation of Huila – CORHUILA

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