Review of the effect of Moringa oleifera seeds on reducing water

turbidity and pathogens

Nick Kelley








SCSC 658
Dr. Jacqueline Peterson
April 25, 2013
Review of M. oleifera seeds 2


Introduction
This is a review of two research papers that studied the effects of Moringa oleifera seeds
on the reductions of water turbidity and pathogen concentrations, specifically in the context of its
use in impoverished or third-world communities. These papers are the Use of Moringa oleifera
seed extracts to reduce helminth egg numbers and turbidity in irrigation water
19
; and
Bioremediation of turbid surface water using seed extract from Moringa oleifera Lam.
(Drumstick) Tree
11
. A third paper was also used as a supporting supplement, providing some
comparative information regarding various water parameters and their effects on the coagulation
properties of M. oleifera
18
, primarily referenced in the discussions about the former two studies.

Description of M. oleifera
Lea
11
provides an overview of the description of the M. oleifera tree, which is native to
northern India but is also widespread through Africa, Asia, and Latin America. The tree is easily
cultivated in tropical environments
4
, providing a convenient resource in parts of the world where
millions of people have access only to water of very poor quality. For example, the most
common source of irrigation water used in Ghana is derived from dug-outs, streams, and
stormwater that is typically polluted with untreated wastewater
19
. This is a scenario that is most
likely replicated in many other places around the world, especially along the tropical and sub-
tropical belts.
The M. oleifera tree provides two distinct benefits: exceptional coagulation and
flocculation properties, and a rich source of nutrients. As for the latter, almost every part of M.
oleifera is consumed as food, except for the trunk and roots
11
. The pods of the tree are typically
the most valuable part, while the seeds and flowers can also be eaten. The leaves are apparently
Review of M. oleifera seeds 3


high in nutrients, containing [] more Vitamin A than carrots, more calcium than milk, more
iron than spinach, more Vitamin C than oranges, and more potassium than bananas []
11
as
well as protein levels that rival eggs and milk.
In the context of water, however, M. oleiferas most important characteristic is the
coagulating property of its seeds. In the two main studies under review in this paper, Lea
11

found a reduction in turbidity of 80.0 99.5% using M. oleifera seeds, while Sengupta et al.
19

saw a reduction of 85 96%.

Action of Coagulation/Flocculation
The action of coagulation caused by M. oleifera is derived from proteins found within the
seeds
11
. These proteins are water-soluble organic polymers known as polyelectrolytes
13
. While
it is agreed that suspended particles have a negative charge and that the polyelectrolytes have a
positive charge, the exact method of how the polyelectrolytes bind suspended matter and form
floc is still not exactly known, though several explanations have been provided. One possible
mechanism is that the positive polyelectrolytes bind to the negative particles, neutralizing their
surface charge and resulting in reduced electrostatic repulsion, allowing the particles to
flocculate. Another explanation is that the polyelectrolytes adsorb to parts of the particle surface,
resulting in differently charged particle surface areas that attract oppositely charged surface areas
of other particles and form floc 1,2,8
15
.

Comparison of Coagulation Methods
The main link between the studies presented by Lea, and Sengupta et al. is that of the
coagulation effect of M. oleifera, along with consideration of its practical use in third-world or
Review of M. oleifera seeds 4


impoverished communities. Sengupta focused his study in the country of Ghana and the
application of M. oleifera for treating water from wastewater-polluted sources typically used for
irrigation of food crops. Turbidity levels of irrigation water used in Ghana may be as high as 791
NTU (nephelometric turbidity units) during dry seasons, which is far above the 2 NTU
recommended by the US EPA
6
. Lea references Jahn
10
, stating that M. oleifera seeds can
increase clarity of water with initial turbidity as high as 10,000 NTU. Leas study focuses on
developing specific preparation and application protocols of M. oleifera seeds that can be easily
followed by members of poor rural communities. Both studies considered dosage rates of M.
oleifera (measured in mg/L or ml/L) against varying levels of water turbidity (measured in
NTU).
Preparation of M. oleifera seeds was approached slightly differently in both studies.
Sengupta took a more empirical approach; crushing the seeds into powder using a mortar, mixing
the powder in tap water to final concentrations of 3% and 5% w/v, stirring the mixture and
filtering through a sieve cloth, and utilizing the filtrate as the coagulating agent. Sengupta also
performed tests in both a lab setting and in the field in Ghana.
Conversely, Lea provides basic protocols for preparing the coagulating agent which are
more rudimentary (assumedly, since Sengupta does not list every step of his preparation process)
and applicable to the people who would actually use the seeds to treat their water. One protocol
is an adaptation of a similar procedure by Price
17
and includes extracting oil from the seeds
which can then be used in other applications, thus increasing the value of the seeds. This protocol
can result in either a dried seed cake which can be stored for later use, or fresh seed powder
which is then used immediately. Either product is used to create a paste which is then diluted and
Review of M. oleifera seeds 5


filtered to obtain the extract solution. Leas tests are performed in the field under real-world
conditions, though he also references other studies for result support/validation.
Concerning the classification of turbidity levels, Sengupta et al. used three different
categories of turbidity for his laboratory tests and one category for the field tests while Lea
references four different ranges. Senguptas lab waters included low turbidity, or tap water
having <50 NTU; medium turbidity, or wastewater having 50 150 NTU; and high turbidity, or
turbid water having >150 NTU. As for his field tests, Sengupta used local irrigation water taken
from an on-farm pond in Kumasi, Ghana; the turbidity of this irrigation water ranged 42 183
NTU. Leas categories included: low at <50 NTU; medium at 50 150 NTU; high at 150 250
NTU; and extreme at >250 NTU. Coincidentally, Senguptas ranges were derived from Leas
study (Lea
11
is cited in Senguptas paper).

Dosage and Coagulation Results
Both studies consider optimum M. oleifera dosages with regards to turbidity levels.
Starting with Senguptas results, which are more empirical than Leas, Sengupta compared
turbidity with dosage as well as turbidity over time. He found that dosage was highly dependent
on the type of water being treated. Using two M. oleifera coagulating agent concentrations of 3%
w/v and 5% w/v, different optimum dosages were revealed for each turbidity group. The
irrigation water had optimum dosages of 100 ml/L for 3% solution and 20 ml/L for 5% solution.
Turbid water (>150 NTU) saw maximum reduction with 8 ml/L of 5%, while wastewater (50
150 NTU) saw maximum reduction with 4 ml/L of 5% solution.
Leas dosages, similar to his protocols, are more practical in the context of real-world
use. He references Doerr
3
and notes that one shelled seed is equivalent to about 200 mg and that
Review of M. oleifera seeds 6


this is typically enough to treat 1 liter of very turbid water. With this in mind, Lea suggests the
following dosages: 1 seed/4 liters of low turbidity water (<50 NTU); 1 seed/2 liters of medium
turbidity water (50 150 NTU); 1 seed/liter of high turbidity water (150 250 NTU); and 2
seeds/liter for extreme turbidity water (>250 NTU). Both studies found that M. oleifera is not an
effective coagulant for low turbidity (<50 NTU) water
11
, and in the case of Sengupta, found that
M. oleifera actually increased the turbidity of tap water. This is probably due to the fact that M.
oleifera itself has some level of turbidity
19
and that it was added to water that has extremely low
turbidity to begin with.
Despite the doses suggested by Sengupta and Lea, both studies mention using the jar test
14,15
to assess coagulation efficiency of the M. oleifera extract to determine the best clarification
dose for a specific water source, as not all water is the same. Lea includes this test as a support
protocol due to its ease of use within a primitive household.
Focusing now on Senguptas study and the relationship of turbidity and time, he found
that turbidity did decrease naturally with time, however, turbidity decreased faster with the
inclusion of M. oleifera agent. Irrigation water saw an 85% decrease in turbidity one hour after
treatment. Turbid water saw a 96% decrease in turbidity one hour after being treated, compared
to an 82% reduction without treatment; and wastewater saw an 85% reduction in turbidity one
hour after treatment compared to 54% without treatment. Optimal settling times were also
calculated to be 120 minutes for turbid water and 90 minutes for wastewater, after which no
significant changes in turbidity were observed.

Comparison of Pathogen Studies
Review of M. oleifera seeds 7


Along with reductions in turbidity, Lea and Sengupta also saw reductions in certain
pathogenic elements, specifically coliform bacteria, and helminth eggs, respectively.
Sengupta looked at helminth eggs specifically because contaminated irrigation water used
in Ghana has shown concentrations of helminth eggs in the range of 5 10 eggs per liter of water
16,9
. This range is above the World Health Organizations (WHO) standard for wastewater used
for irrigation, which requires a concentration of 1 helminth egg per liter
23
. Helminth parasites
of concern to human health include the roundworm (Ascaris lumbricoides), whipworm
(Trichuris trichiura), and hookworms (Ancylostoma duodenale and Necator americanus)
5,7
.
Helminth eggs are also notorious for being highly resistant to environmental stresses
22
and
wastewater treatment systems in poor areas (if present) may not adequately reduce helminth egg
concentrations to safe levels. High levels of turbidity can also impede treatment for helminth
eggs, as the eggs tend to flocculate with and settle at the same velocity as suspended particles
20
.
Higher efficiencies of turbidity reduction using M. oleifera may also reduce helminth egg
concentrations.
Sengupta et al.
19
found that helminth eggs were reduced by 94% when irrigation water
was treated with M. oleifera, from 14.0 eggs per liter down to 0.8. Helminth eggs were found to
reduce exponentially in treated water, with an optimum settling time of 2 2.5 hours. In lab
experiments, removal of helminth eggs was significantly higher in all water types treated with M.
oleifera compared to untreated water. On average, 93% of Ascaris eggs were recovered in treated
water compared to 74% in untreated water. In treated water, 92% of Trichuris eggs were
recovered compared to 68% in untreated water. Sengupta also found a positive correlation
between turbidity and number of helminth eggs in water treated with M. oleifera.
Review of M. oleifera seeds 8


Lea states that M. oleifera can also reduce bacterial concentrations as well, namely fecal
coliforms by 90.00 99.99%
12
within the first one to two hours of treatment. This is due mainly
to the fact that bacteria tend to adsorb to suspended particles; the bacteria settle out of the water
along with the sedimented sludge. However, Lea does not advise M. oleifera treatment to be the
sole treatment method used for turbid waters. He notes that M. oleifera treatment does not
guarantee 100% removal of pathogens and suggests that M. oleifera be used as a pretreatment to
reduce turbidity, allowing additional treatment for pathogens to become more effective. He lists
several support protocols that may be used to treat specifically for pathogens, including: biosand
filtration; disinfection by natural pH shift (using naturally acidic solutions to lower pH to kill
pathogens); disinfection by solar pasteurization; and disinfection by chlorination (though it
results in harmful disinfection byproducts when exposed to organic matter, so use of chlorine in
conjunction with M. oreifera is not recommended).

Discussion of Turbidity Results
Both studies
11,19
evaluated M. oleifera coagulation treatment on waters of varying
turbidity levels. Unfortunately, the two studies used different metrics for the amount of M.
oleifera agent used. Sengupta used a w/v (weight per volume) suspension measured as an initial
percentage of M. oleifera powder in tap water, which was then filtered and the filtrate dosage
measured in ml/L. Lea used the more direct metric of mg/L of M. oleifera powder or cake in
water. This complicates the comparison of optimum dosages suggested by the two studies as
some kind of conversion is needed to get the dosages in the same metric. To help provide an
extra reference, Pritchard et al.
18
claims a dose of 50 mg/L as a general optimum dosage across a
turbidity range of 40 200 NTU, however, he does note that optimum dose increases linearly as
Review of M. oleifera seeds 9


initial turbidity increases. This is compared to Leas suggested dosage of roughly 200 mg/L for
turbidity levels greater than 50 NTU (he also notes that M. oleifera is not effective at levels
below 50 NTU). One reason for this discrepancy may be the difference in conditions under
which the M. oleifera was applied; Pritchard studied M. oleifera under laboratory conditions
while Lea prepared and applied the agent in the field under real-world conditions. An additional
finding made by Pritchard is that M. oleifera coagulation is influenced by water temperature and
that the warmer the water, the more effective the coagulation. Water temperature was not
mentioned in Leas study, and so the water temperature may have differed between Leas and
Pritchards studies (and Senguptas study, as well).

Discussion of Pathogen Results
While these two studies did not focus on the same pathogens, they did demonstrate that
M. oleifera is not only an effective coagulating agent but can also be an effective disinfectant,
depending on the target pathogen. It should also be satisfying to note that both studies do not
recommend M. oleifera treatment as a sole treatment method for water. Both Sengupta and Lea
recommend additional treatment options, for example, sand filtration, if the water is going to be
used for drinking or irrigating food crops. And the fact that M. oleifera not only cleans
particulate from the water but some pathogens as well makes additional treatment and
disinfection that much easier.

Comments
It is very interesting to learn that natural plant products can achieve equivalent results in
water quality compared to anthropogenic compounds that may be costly and/or harmful to
Review of M. oleifera seeds 10


synthesize, as well as toxic to handle. These two studies do a wonderful job of demonstrating the
effectiveness of a naturally-derived and easily-obtainable resource for effectively treating a
common water issue (turbidity) that exists across the globe, and is an issue that millions of
people struggle with every day. I think these studies
11,18,19
provide a reliable and significant
body of work regarding the use of M. oleifera in water treatment. I did come across some
mentions of similar plant compounds being used to treat water issues, however, I dont believe
the research regarding those compounds was as extensive as M. oleifera, or their effects as well
known. Moringa oleifera has been used by many peoples around the world for hundreds of years;
its only just within the last couple of decades that it has really been scientifically studied. It is
also interesting to note that M. oleifera has been successfully tested full-scale as the primary
coagulant at a water treatment facility in Malawi
21
, demonstrating that naturally-derived
compounds and methods have the ability to function at the same level as industrially-produced
chemicals typically used in water and wastewater treatment.
I really enjoyed Leas approach to studying M. oleifera. Rather than studying its effects
solely in a laboratory setting using equipment and methods that are not really not applicable or
practical outside a lab, he established a real-world connection. By taking his study beyond the
lab, he generated a protocol that is applicable outside a lab and allows the actual people who use
this plant to better treat their own water. Hopefully that is knowledge that the people using M.
oleifera can maintain within their communities and pass down to future generations. I think there
needs to be more scientific study that takes the science out of the lab and into the lives of those
affected by water quality issues, particularly those living in developing and under-developed
nations. Its one thing to assess the effectiveness of a method in a lab using calibrated equipment
specifically designed for empirical uses, which is necessary to determine potential or theoretical
Review of M. oleifera seeds 11


effectiveness and health effects, etc. However, transferring that method to a real-world situation
in which one does not have access to such equipment and making it practical is very important if
we, as members of developed and industrialized nations, are to continue addressing the water
quality issues and the people of less developed communities.

Review of M. oleifera seeds 12


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