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International Journal of Trend in Scientific Research and Development (IJTSRD)
Volume 4 Issue 4, June 2020 Available Online: www.ijtsrd.com e-ISSN: 2456 – 6470
@ IJTSRD | Unique Paper ID – IJTSRD31019 | Volume – 4 | Issue – 4 | May-June 2020 Page 491
Water Treatment and Purification using
Moringa Oleifera Seed Extract
Areeba Ansari1, Malika Ahuja2
1Student, 2Lecturar,
1,2B.N.N. College of Arts, Science and Commerce, Bhiwandi, Maharashtra, India
ABSTRACT
Water is the most abundant chemical and important natural resource.Various
concentrations of water at given place contributes to water quality. The
suitability of water and its specific use are evaluated by examining its quality
parameters. The adverse health effects have been observed in developing
countries due to drinking contaminated water. The natural resources have
serious threat due to development and urbanization in countries. People are
made to use low quality water because of high cost of treated water which
results in exposing them to waterborne diseases. The seed extract of Moringa
oleifera is used for purification of drinking and wastewater duetopresenceof
soluble cationic coagulant. It has capability toreducetheturbidityfromwater.
In the present study, the collected water samples wereexaminedwithvarious
physical, chemical and biological parameters. Obtained values of each
parameters were compared with standard values set by World Health
Organization.
KEYWORDS: Moringa oleifera, Physico-chemical, Bacteriological, Parameters,
Seed extract, Purification
How to cite this paper: Areeba Ansari |
Malika Ahuja "Water Treatment and
Purification using Moringa Oleifera Seed
Extract" Publishedin
International Journal
of Trend in Scientific
Research and
Development
(ijtsrd), ISSN: 2456-
6470, Volume-4 |
Issue-4, June 2020,
pp.491-494, URL:
www.ijtsrd.com/papers/ijtsrd31019.pdf
Copyright © 2020 by author(s) and
International Journal ofTrendinScientific
Research and Development Journal. This
is an Open Access article distributed
under the terms of
the Creative
CommonsAttribution
License (CC BY 4.0)
(http://creativecommons.org/licenses/by
/4.0)
INTRODUCTION
Water is an essential natural resource, it is the source of
sustenance for all living organisms, ecological system, food
production and economic development(Saxena et.al.,2011).
Sustenance of human health and influence of life clean and
safe drinking water is major requirement. The vital concern
of developing countries throughout the worldisprovisionof
good quality, clean, purified and safe water (Khan et.al.,
2013). Most widely circulated and distributed abundant
substance is water. In total there is 1400 million billionliters
of water out of which only 3% is fresh water and 1% is
available for portable use (Hujare et.al., 2008).Waterisused
for various activities of human life like drinking,cookingand
agricultural purposes as well as regulation of human body
such as nutrient transport, thermal regulation, digestion,
metabolic activities and flush of toxic waste (Kumar et.al.,
2013, Abera et.al., 2011). As water is directly connected to
human welfare its quality checking and maintaining is vital
concern to mankind. Food security, health and hygiene
mismanagement of water resources has strong socio-
economic repercussions (Khalid et.al., 2007). For use of
water in different areas in have requirements, composition
and purity is to be analyzed on regular basis to confirm
suitability for its use ( Saxena et.al., 2011). According to
study of world health organization inadequate sanitation,
polluted water or unavailability of water are the main
reasons of about 80% of all the sickness, illness anddiseases
(Vagarali et.al., 2011). The pathogens presenting serious
threat to disease if found in waterincludeSalmonellaspecies,
Shigella species, Escherichia coli, Vibrio cholerae, Yersinia
enterolitica, Cmpylobacter species. There is requirement of
systemic water quality monitoring for environmental and
public health (Kumar et.al., 2013). The water treatment
procedures of many emerging countries involves
coagulation, flocculation, sedimentation and disinfection
which are expensive because of high cost involved for
clarification and assessing thechemical coagulantsincluding
alum (Sunita et.al., 2014). The high cost associated with
water treatment leads to use of readily available source for
people which is of low quality expose them to water borne
diseases (Vrushali et.al., 2018). Moringa oleifera is found to
be a viable coagulant for chemical replacement. The seed
extract of Moringa oleifera have water soluble cationic
protein which act as primary coagulant in process of water
clarification treatment having the ability to reduce the
turbidity (Md Saduzaman et.al.,2013).
MATERIALS AND METHODS
Collection of water sample
The two water samples were collected in clean and surface
sterilized bottles from local area of Bhiwandi, Thane,
Maharashtra.
Collection of Moringa oleifera seeds
Moringa oleifera seeds were separated from fresh
drumsticks, the seed were dried in sunlight for few daysand
powdered using a mixer grinder for further experimental
purposes.
IJTSRD31019
International Journal of Trend in Scientific Research and Development (IJTSRD) @ www.ijtsrd.com eISSN: 2456-6470
@ IJTSRD | Unique Paper ID – IJTSRD31019 | Volume – 4 | Issue – 4 | May-June 2020 Page 492
Estimation of Alkalinity
The alkalinity of water sample is its quantitative capacity to
neutralize a strong acid to a designated pH caused by HCO3
and OH- ions in water. Before titration process
standardization of HCl was carried and end point was notes
from yellow to orange colour (x cm3). The first alkalinity
titration is phenolphthalein alkalinity which is carried using
50cm3 water sample with 0.1N Na2S2O3 solution and
1%phenolphthaleim as indicator titrated against standard
HCl solution. End point is determined when pink color just
disappear or fade(y cm3). Alkalinity resultswerereported in
terms of mg of CaCO3.
Estimation of Chemical Oxygen Demand (COD)
Organic substances present in water sample are oxidized by
K2Cr2O7 in acidic medium. The sample was heated in
presence of Ag2SO4 as catalyst, HgSO4 was used for removal
of chloride interference. The excess of K2Cr2O7 was titrated
back against ferrous alum solution. Amount of oxidisable
organic matter measured as oxygen equivalent is
proportional to K2Cr2O7 consumed. In two round bottom
flasks few porcelain pieces, 1g 20% HgCl2, 5cm3 K2Cr2O7
conc.H2SO4, 50 cm3 of 0.025N K2Cr2O7 and 5 cm3 of 1%
Ag2SO4 solutions was added. In last add 25 cm3 of distilled
and sample waters in respectivetubesandkeepforrefluxing
on boiling water bath for 90 minutes.Now,titratethesample
after cooling with 0.025M ferrous alum using indicator
0.025M ferroin. End point will be from green to red colour.
This method determined the COD value of 50 mg/dm3 or
more.
Estimation of Biological Oxygen Demand (BOD)
Biological oxygen demand is the amountofdissolvedoxygen
needed by aerobic biological organism to break down
organic matter present in given water sample at certain
temperature over a specific time period. Samples were
labelled as sample1 (Day 1&5) and sample2 (Day 1&5). To
both the samples 2.0 ml of MnSO4 and 2.0ml of alkali iodide
azide were added. Precipitate of MnSO4 was formed which
was dissolved by adding 2.0ml of H2SO4. It was titrated
against sodium thiosulphate using starch as indicator and
readings were recorded in terms of 0.0125N sodium
thiosulphate for titration. The BOD value was expressed in
milligrams of oxygen consumed per liter of sample on both
day 1 and day 5 for both blank and test respectively.
Calculation:
Amount of Na2S2O3 = Sample 1– Sample 2
BOD in mg/L = A*Normality of Na2S2O3*8*1000*dilution
factor / Amount of sample
Isolation and Identification of Bacteria
Selective and differential media was prepared as per
standard protocol and after sterilization agar plates were
prepared under aseptic conditions. Different water samples
collected; were streaked on different media.Theplateswere
incubated at 37⁰C for 24 hours to observe bacterial growth.
Biochemical tests were done to identifythemicroorganisms.
Most Probable Number (MPN)
Most probable number analysis is a statistical methodbased
on the random dispersion of microorganism per volumeina
given sample. For presumptive test 10ml water sample was
inoculated in three double strength Lauryl Tryptose Broth
(LTB) tubes. In these tubes, 0.1ml and 1ml of water sample
was added in three single strength LTB tubes. The tubes
were incubated at 37⁰C for 48 hours for gas production
observation. Tubes showing positive presumptive test (gas
production) were inoculated one loopful in sterile BGLB
tubes and incubate at 37⁰C for 48 hours and check for gas
production in BLGB tubes constitutes positive confirmed
test.
Determination of Initial Number of Organisms Present
in Water Sample
Firstly, 1.0ml of both water samples were poured in sterile
petri plates and then at about 45⁰C R2A agar media was
poured, as agar solidified the plateswerekeptforincubation
at 37⁰C for 5 days. Regularly observation was taken and
results were noted.
Addition of Moringa oleifera seed powder in water
samples
The water samples were treated in the addition of Moringa
oleifera seed powder at different concentrations of 0.2% to
2.4% gm of seed powder. This added water sample were
kept into purification for 10 days.
Determination of Final Number of Organisms Presentin
Water Sample
The treated and control water sample wereexaminedonthe
basis of appearance, taste, pH, odour and microbial
evaluation, total bacterial count were taken for each treated
sample and control by pour plate method. Both the samples
were filtered with whatsman filter paper and 1ml of treated
sample was added on R2A agar media plate and incubated
for 24 hours. The evaluation of bacterial count was observe
and compared to untreated water sample.
RESULTS AND DISCUSSION
Collection of Water Samples
Water samples from lake and tap were collected in sterile
plastic bottles to avoid additional contaminationduetonon-
sterile bottles. The bottles were transported immediately to
laboratory with appropriate care and stored at 4⁰C till
further processing.
Collection of Seed Powder
Moringa oleifera seeds were collected from fresh drumstick
plant, dried on sunlight for few days and then powdered
using grinder for further test.
Determination of pH in water samples
Digital pH meter is used to check the hydrogen ion
concentration in both water samples. The readings were
taken after 1 minute constant value (Kanase et.al., 2016).
The results are observed as given below in table no.1
Table no.1: Determination of pH in water samples
Water Samples pH
Sample 1 7.2
Sample 2 7.2
Drinking water with a pH between 6.5 and 8.5 is generally
considered satisfactory. The hydrogen ion (pH) in both the
samples is 7.2 which is within the safe limit set by WHO
standard and hence will not cause any harmful effect.
Determination of Alkalinity in water samples
The water samples were checked for alkalinity,measured by
content of hydroxide and carbonate ion present in samples.
International Journal of Trend in Scientific Research and Development (IJTSRD) @ www.ijtsrd.com eISSN: 2456-6470
@ IJTSRD | Unique Paper ID – IJTSRD31019 | Volume – 4 | Issue – 4 | May-June 2020 Page 493
It is titrated with standard HCl, result are displayed in table
no.2
Table no.2: Determination of alkalinity in water
samples
Water Samples Alkalinity (mg/L)
Sample 1 350
Sample 2 330
Acid neutralizing capacity is alkalinity of water due to
presence of carbonates and bicarbonates. The range set by
WHO is 500mg/L (Kanase et.al., 2016). Both the samples
showed alkalinity in range of 400mg/L.
Determination of Chemical Oxygen Demand in water
samples
For both the samples chemical oxygen demand was
determined by using dichromate refluction method. The
results are as displayed in table no.3
Table no.3: Determination of chemical oxygen demand
Water Samples COD (ppm)
Sample 1 200
Sample 2 719
Oxygen is important component as it is required for
degradation of organic matter hence COD is important
parameter. Both the samples showed a large variation in
chemical oxygen demand.
Determination of Biological Oxygen Demand in water
samples
Biological oxygen demand of both the samples were tested,
the maximum value as per WHO standard is 2.0mg/L for
drinking purpose. The results are shown in table no.4
Table no.4: Determination of biological oxygen
demand
Water
Samples
Day 1
(mg/L)
Day 5
(mg/L)
Results
(mg/L)
Sample 1 13.5 2.9 1160
Sample 2 15.0 3.2 1180
The tests showed wide range obtained under BOD test with
minimum value 2.9mg/L observed. No similar results were
obtained. Sample1 and sample2 showed BOD as 1160mg/L
and 1180ma/L respectively.
Determination of Most Probable Number(MPN) Method
Determination of most probable number for both the water
samples was done using multiple tube method. The results
are given in table no.5
Table no.5: Determination of most probable number
Water
Samples
10 ml 1.0ml 0.1ml
MPN Index
per 100ml
Sample 1 0 1 0 2
Sample 2 1 1 0 4
According to WHO standard for drinking waterqualitythere
should be < 1 MPN/100ml of total coliforms and no
Escherichia coli in 1ml of water (Olowe et.al., 2016)reported
MPN index in the range of 2-60 MPN/100ml. Both the water
samples used for this study showed MPN index 2 to 4
MPN/100ml.
Determination of Initial Number of Organisms in water
samples
Pour plate method was used to determine initial count of
organisms in untreated water sample, they were observed
and found that lake water(sample1)containlargenumberof
organism as compare to tap water(sample2).The results are
as shown in table no.6
Table no.6: Determination of initial number of
organisms
Water
Source
Physical
appearance
Number of
organisms (cfu/ml)
Sample 1 High yellow 1800
Sample 2 Yellow 1500
Isolation and Identification of Microorganisms
Microbiological and biochemical investigations werecarried
out of various water sample. Olowe et.al., 2016, Vagarali
et.al., 2011, Abera et.al., 2011 reported presence of
Escherichia coli in the range of 10% to 17.3%. The present
study of both water samples confirmed presence of
Escherichia coli.
Determination of Final Number of Organisms in water
samples
The water sample was treated with Moringa oleifera seed
extract thus the treated sample was checkedforreductionin
total bacterial count, it was observed that highest bacterial
reduction count was on 2.4% of plant extract by pour plate
method. The results are displayed in table no.7
Table no.7: Determination of final number of organisms
Water Samples Physical appearance pH 0.2% 0.6% 1.0% 2.2% 2.4% Control
Number of organisms (cfu/ml)
Sample1 Clear 7.0 1600 300 90 50 20 3800
Sample2 Clear 7.0 1100 900 400 50 10 2500
Reduction in total bacterial count was observed that 2.4%. Similar results with reduction in bacterial count were observed by
Yongabi, et.al., 2011.
CONCLUSION
Water plays important role for sustenance of industries,
agriculture and human existence. The healthy water
ecosystem is depended on physico-chemical and
bacteriological characteristics. It is observed that watergets
polluted significantly due to human activities, seed of
Moringa oleifera were used for water purification. Alkalinity
and pH of both the samples were as per WHO standard, the
COD of sample 2 was not found to permissiblelimitwhereas
BOD of both the samples had unacceptable values. MPN
index indicates presence of greater number of bacteria and
was positive for Escherichia coli. The turbidity and bacterial
count was reduced by using Moringa oleifera seed extract,
International Journal of Trend in Scientific Research and Development (IJTSRD) @ www.ijtsrd.com eISSN: 2456-6470
@ IJTSRD | Unique Paper ID – IJTSRD31019 | Volume – 4 | Issue – 4 | May-June 2020 Page 494
2.4% of powder has ability to reduce the impurities from
water.The study can be used to determine fungal, algal, viral
count present in water samples. Moringa oleiferaisa natural
coagulant produces biodegradable environment friendly
sludge as compared to chemical coagulant. Properties of
Moringa oleifera like adsorbent, coagulant, dewatering and
desalting agent can be checked and used for removal of
heavy metals from water.
ACKNOWLEDGMENT
I would like to thank Miss Malika Ahuja for helping me in
doing research as well as for givinghermajorcontributionin
writing and proof reading the research article.
REFERENCES
[1] Abera S, Zenyinudin A, Kebede B, Deribew A, Ali S,
Zemene E (2011); Bacteriological analysis of drinking
water sources. Afri.Micro.Res.2011:5(18):2638-2641.
[2] Chaubey Soni and Mohan Kumar Patil (2015).
“Application of Artificial Neural Network in Assessing
Water Quality: A Case Study”, International Journal of
Current Research Vol.7, Issue01, pp.11403-11407,
India.
[3] Dhanaji Kanase G, Shagufta Shaikh A, Pramod Jagadale
N (2016); Physico-Chemical analysis of drinking water
samples of different places in Kadegaon Tahsil,
Maharashtra (India);Pelagia ResearchLibrary;7(6):41-
44.
[4] Hujare, M. S. (2008): Seasonal variation of physico-
chemical parameters in the perennial tank ofTalsande,
Maharashtra. Ecotoxical. Environ. Monitor. 18(3):233-
242
[5] Khalid Anwar, Amir Haider Malik, Amir Waseem,
Shazmeen Zahra and Ghulam Murtuza (2011);
Qualitative and quantitative analysis of drinking water
of different localities in Abbottabad district, Pakistan;
Internation journal of the physical science; Vol 6(33).pp
7480-7489.
[6] Khan Naeem, Sayef Tasleem Husain, Asif Khan, Kyong
Su Kim (2013); Physico-chemical investigation of
drinking water sources from Tehsil Lachi, Kohat;
American journal of research communication;Vol1(5):
170-190.
[7] Kumar Deepesh, Shurutikirti Malik,MolluMadan,Anita
Pandey, Ashish K, Asthana (2013); Bacteriological
analysis of water by MPN method in a tertiary care
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Metagud (2011);Bacteriological analysis of water,
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[10] Olowe Busayo Mutiat, Jacob O. Oluyege and Oladiran
Famurewa (2016); Prevalance of water borne disease
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[11] Saxena Nidhi, S.N.Misraand R.N.Shukla (2011);
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[12] Shimpi Vrushali V., Jhondle Pooja R., Gangurde Ritu B.,
Jadhav Tejal D., KapseVikashni Nand, Maataa Matakite,
Koshy Kanayathu and Sotheeswaran Subramanium
(2012);Water pirification using Moringa oleifera and
other locally available seeds in Fiji for heavy metal
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[13] Shireen S. (2018); Water purification using Moringa
oleifera (Drumstick) seed powder as a natural
coagulant; Recent Advances in Engineering Science and
Management ;89-83
[14] Thakur Sunita Singh, Chaubey Sonal (2014);
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oleifera and okra for treatment of turbid water;
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[15] Yahya Daniyan Safiya, Enemaduku Moses Abalaka and
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extract in water purification; IJRAP; 2(4): 1265-1271.
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Water Treatment and Purification using Moringa Oleifera Seed Extract

  • 1. International Journal of Trend in Scientific Research and Development (IJTSRD) Volume 4 Issue 4, June 2020 Available Online: www.ijtsrd.com e-ISSN: 2456 – 6470 @ IJTSRD | Unique Paper ID – IJTSRD31019 | Volume – 4 | Issue – 4 | May-June 2020 Page 491 Water Treatment and Purification using Moringa Oleifera Seed Extract Areeba Ansari1, Malika Ahuja2 1Student, 2Lecturar, 1,2B.N.N. College of Arts, Science and Commerce, Bhiwandi, Maharashtra, India ABSTRACT Water is the most abundant chemical and important natural resource.Various concentrations of water at given place contributes to water quality. The suitability of water and its specific use are evaluated by examining its quality parameters. The adverse health effects have been observed in developing countries due to drinking contaminated water. The natural resources have serious threat due to development and urbanization in countries. People are made to use low quality water because of high cost of treated water which results in exposing them to waterborne diseases. The seed extract of Moringa oleifera is used for purification of drinking and wastewater duetopresenceof soluble cationic coagulant. It has capability toreducetheturbidityfromwater. In the present study, the collected water samples wereexaminedwithvarious physical, chemical and biological parameters. Obtained values of each parameters were compared with standard values set by World Health Organization. KEYWORDS: Moringa oleifera, Physico-chemical, Bacteriological, Parameters, Seed extract, Purification How to cite this paper: Areeba Ansari | Malika Ahuja "Water Treatment and Purification using Moringa Oleifera Seed Extract" Publishedin International Journal of Trend in Scientific Research and Development (ijtsrd), ISSN: 2456- 6470, Volume-4 | Issue-4, June 2020, pp.491-494, URL: www.ijtsrd.com/papers/ijtsrd31019.pdf Copyright © 2020 by author(s) and International Journal ofTrendinScientific Research and Development Journal. This is an Open Access article distributed under the terms of the Creative CommonsAttribution License (CC BY 4.0) (http://creativecommons.org/licenses/by /4.0) INTRODUCTION Water is an essential natural resource, it is the source of sustenance for all living organisms, ecological system, food production and economic development(Saxena et.al.,2011). Sustenance of human health and influence of life clean and safe drinking water is major requirement. The vital concern of developing countries throughout the worldisprovisionof good quality, clean, purified and safe water (Khan et.al., 2013). Most widely circulated and distributed abundant substance is water. In total there is 1400 million billionliters of water out of which only 3% is fresh water and 1% is available for portable use (Hujare et.al., 2008).Waterisused for various activities of human life like drinking,cookingand agricultural purposes as well as regulation of human body such as nutrient transport, thermal regulation, digestion, metabolic activities and flush of toxic waste (Kumar et.al., 2013, Abera et.al., 2011). As water is directly connected to human welfare its quality checking and maintaining is vital concern to mankind. Food security, health and hygiene mismanagement of water resources has strong socio- economic repercussions (Khalid et.al., 2007). For use of water in different areas in have requirements, composition and purity is to be analyzed on regular basis to confirm suitability for its use ( Saxena et.al., 2011). According to study of world health organization inadequate sanitation, polluted water or unavailability of water are the main reasons of about 80% of all the sickness, illness anddiseases (Vagarali et.al., 2011). The pathogens presenting serious threat to disease if found in waterincludeSalmonellaspecies, Shigella species, Escherichia coli, Vibrio cholerae, Yersinia enterolitica, Cmpylobacter species. There is requirement of systemic water quality monitoring for environmental and public health (Kumar et.al., 2013). The water treatment procedures of many emerging countries involves coagulation, flocculation, sedimentation and disinfection which are expensive because of high cost involved for clarification and assessing thechemical coagulantsincluding alum (Sunita et.al., 2014). The high cost associated with water treatment leads to use of readily available source for people which is of low quality expose them to water borne diseases (Vrushali et.al., 2018). Moringa oleifera is found to be a viable coagulant for chemical replacement. The seed extract of Moringa oleifera have water soluble cationic protein which act as primary coagulant in process of water clarification treatment having the ability to reduce the turbidity (Md Saduzaman et.al.,2013). MATERIALS AND METHODS Collection of water sample The two water samples were collected in clean and surface sterilized bottles from local area of Bhiwandi, Thane, Maharashtra. Collection of Moringa oleifera seeds Moringa oleifera seeds were separated from fresh drumsticks, the seed were dried in sunlight for few daysand powdered using a mixer grinder for further experimental purposes. IJTSRD31019
  • 2. International Journal of Trend in Scientific Research and Development (IJTSRD) @ www.ijtsrd.com eISSN: 2456-6470 @ IJTSRD | Unique Paper ID – IJTSRD31019 | Volume – 4 | Issue – 4 | May-June 2020 Page 492 Estimation of Alkalinity The alkalinity of water sample is its quantitative capacity to neutralize a strong acid to a designated pH caused by HCO3 and OH- ions in water. Before titration process standardization of HCl was carried and end point was notes from yellow to orange colour (x cm3). The first alkalinity titration is phenolphthalein alkalinity which is carried using 50cm3 water sample with 0.1N Na2S2O3 solution and 1%phenolphthaleim as indicator titrated against standard HCl solution. End point is determined when pink color just disappear or fade(y cm3). Alkalinity resultswerereported in terms of mg of CaCO3. Estimation of Chemical Oxygen Demand (COD) Organic substances present in water sample are oxidized by K2Cr2O7 in acidic medium. The sample was heated in presence of Ag2SO4 as catalyst, HgSO4 was used for removal of chloride interference. The excess of K2Cr2O7 was titrated back against ferrous alum solution. Amount of oxidisable organic matter measured as oxygen equivalent is proportional to K2Cr2O7 consumed. In two round bottom flasks few porcelain pieces, 1g 20% HgCl2, 5cm3 K2Cr2O7 conc.H2SO4, 50 cm3 of 0.025N K2Cr2O7 and 5 cm3 of 1% Ag2SO4 solutions was added. In last add 25 cm3 of distilled and sample waters in respectivetubesandkeepforrefluxing on boiling water bath for 90 minutes.Now,titratethesample after cooling with 0.025M ferrous alum using indicator 0.025M ferroin. End point will be from green to red colour. This method determined the COD value of 50 mg/dm3 or more. Estimation of Biological Oxygen Demand (BOD) Biological oxygen demand is the amountofdissolvedoxygen needed by aerobic biological organism to break down organic matter present in given water sample at certain temperature over a specific time period. Samples were labelled as sample1 (Day 1&5) and sample2 (Day 1&5). To both the samples 2.0 ml of MnSO4 and 2.0ml of alkali iodide azide were added. Precipitate of MnSO4 was formed which was dissolved by adding 2.0ml of H2SO4. It was titrated against sodium thiosulphate using starch as indicator and readings were recorded in terms of 0.0125N sodium thiosulphate for titration. The BOD value was expressed in milligrams of oxygen consumed per liter of sample on both day 1 and day 5 for both blank and test respectively. Calculation: Amount of Na2S2O3 = Sample 1– Sample 2 BOD in mg/L = A*Normality of Na2S2O3*8*1000*dilution factor / Amount of sample Isolation and Identification of Bacteria Selective and differential media was prepared as per standard protocol and after sterilization agar plates were prepared under aseptic conditions. Different water samples collected; were streaked on different media.Theplateswere incubated at 37⁰C for 24 hours to observe bacterial growth. Biochemical tests were done to identifythemicroorganisms. Most Probable Number (MPN) Most probable number analysis is a statistical methodbased on the random dispersion of microorganism per volumeina given sample. For presumptive test 10ml water sample was inoculated in three double strength Lauryl Tryptose Broth (LTB) tubes. In these tubes, 0.1ml and 1ml of water sample was added in three single strength LTB tubes. The tubes were incubated at 37⁰C for 48 hours for gas production observation. Tubes showing positive presumptive test (gas production) were inoculated one loopful in sterile BGLB tubes and incubate at 37⁰C for 48 hours and check for gas production in BLGB tubes constitutes positive confirmed test. Determination of Initial Number of Organisms Present in Water Sample Firstly, 1.0ml of both water samples were poured in sterile petri plates and then at about 45⁰C R2A agar media was poured, as agar solidified the plateswerekeptforincubation at 37⁰C for 5 days. Regularly observation was taken and results were noted. Addition of Moringa oleifera seed powder in water samples The water samples were treated in the addition of Moringa oleifera seed powder at different concentrations of 0.2% to 2.4% gm of seed powder. This added water sample were kept into purification for 10 days. Determination of Final Number of Organisms Presentin Water Sample The treated and control water sample wereexaminedonthe basis of appearance, taste, pH, odour and microbial evaluation, total bacterial count were taken for each treated sample and control by pour plate method. Both the samples were filtered with whatsman filter paper and 1ml of treated sample was added on R2A agar media plate and incubated for 24 hours. The evaluation of bacterial count was observe and compared to untreated water sample. RESULTS AND DISCUSSION Collection of Water Samples Water samples from lake and tap were collected in sterile plastic bottles to avoid additional contaminationduetonon- sterile bottles. The bottles were transported immediately to laboratory with appropriate care and stored at 4⁰C till further processing. Collection of Seed Powder Moringa oleifera seeds were collected from fresh drumstick plant, dried on sunlight for few days and then powdered using grinder for further test. Determination of pH in water samples Digital pH meter is used to check the hydrogen ion concentration in both water samples. The readings were taken after 1 minute constant value (Kanase et.al., 2016). The results are observed as given below in table no.1 Table no.1: Determination of pH in water samples Water Samples pH Sample 1 7.2 Sample 2 7.2 Drinking water with a pH between 6.5 and 8.5 is generally considered satisfactory. The hydrogen ion (pH) in both the samples is 7.2 which is within the safe limit set by WHO standard and hence will not cause any harmful effect. Determination of Alkalinity in water samples The water samples were checked for alkalinity,measured by content of hydroxide and carbonate ion present in samples.
  • 3. International Journal of Trend in Scientific Research and Development (IJTSRD) @ www.ijtsrd.com eISSN: 2456-6470 @ IJTSRD | Unique Paper ID – IJTSRD31019 | Volume – 4 | Issue – 4 | May-June 2020 Page 493 It is titrated with standard HCl, result are displayed in table no.2 Table no.2: Determination of alkalinity in water samples Water Samples Alkalinity (mg/L) Sample 1 350 Sample 2 330 Acid neutralizing capacity is alkalinity of water due to presence of carbonates and bicarbonates. The range set by WHO is 500mg/L (Kanase et.al., 2016). Both the samples showed alkalinity in range of 400mg/L. Determination of Chemical Oxygen Demand in water samples For both the samples chemical oxygen demand was determined by using dichromate refluction method. The results are as displayed in table no.3 Table no.3: Determination of chemical oxygen demand Water Samples COD (ppm) Sample 1 200 Sample 2 719 Oxygen is important component as it is required for degradation of organic matter hence COD is important parameter. Both the samples showed a large variation in chemical oxygen demand. Determination of Biological Oxygen Demand in water samples Biological oxygen demand of both the samples were tested, the maximum value as per WHO standard is 2.0mg/L for drinking purpose. The results are shown in table no.4 Table no.4: Determination of biological oxygen demand Water Samples Day 1 (mg/L) Day 5 (mg/L) Results (mg/L) Sample 1 13.5 2.9 1160 Sample 2 15.0 3.2 1180 The tests showed wide range obtained under BOD test with minimum value 2.9mg/L observed. No similar results were obtained. Sample1 and sample2 showed BOD as 1160mg/L and 1180ma/L respectively. Determination of Most Probable Number(MPN) Method Determination of most probable number for both the water samples was done using multiple tube method. The results are given in table no.5 Table no.5: Determination of most probable number Water Samples 10 ml 1.0ml 0.1ml MPN Index per 100ml Sample 1 0 1 0 2 Sample 2 1 1 0 4 According to WHO standard for drinking waterqualitythere should be < 1 MPN/100ml of total coliforms and no Escherichia coli in 1ml of water (Olowe et.al., 2016)reported MPN index in the range of 2-60 MPN/100ml. Both the water samples used for this study showed MPN index 2 to 4 MPN/100ml. Determination of Initial Number of Organisms in water samples Pour plate method was used to determine initial count of organisms in untreated water sample, they were observed and found that lake water(sample1)containlargenumberof organism as compare to tap water(sample2).The results are as shown in table no.6 Table no.6: Determination of initial number of organisms Water Source Physical appearance Number of organisms (cfu/ml) Sample 1 High yellow 1800 Sample 2 Yellow 1500 Isolation and Identification of Microorganisms Microbiological and biochemical investigations werecarried out of various water sample. Olowe et.al., 2016, Vagarali et.al., 2011, Abera et.al., 2011 reported presence of Escherichia coli in the range of 10% to 17.3%. The present study of both water samples confirmed presence of Escherichia coli. Determination of Final Number of Organisms in water samples The water sample was treated with Moringa oleifera seed extract thus the treated sample was checkedforreductionin total bacterial count, it was observed that highest bacterial reduction count was on 2.4% of plant extract by pour plate method. The results are displayed in table no.7 Table no.7: Determination of final number of organisms Water Samples Physical appearance pH 0.2% 0.6% 1.0% 2.2% 2.4% Control Number of organisms (cfu/ml) Sample1 Clear 7.0 1600 300 90 50 20 3800 Sample2 Clear 7.0 1100 900 400 50 10 2500 Reduction in total bacterial count was observed that 2.4%. Similar results with reduction in bacterial count were observed by Yongabi, et.al., 2011. CONCLUSION Water plays important role for sustenance of industries, agriculture and human existence. The healthy water ecosystem is depended on physico-chemical and bacteriological characteristics. It is observed that watergets polluted significantly due to human activities, seed of Moringa oleifera were used for water purification. Alkalinity and pH of both the samples were as per WHO standard, the COD of sample 2 was not found to permissiblelimitwhereas BOD of both the samples had unacceptable values. MPN index indicates presence of greater number of bacteria and was positive for Escherichia coli. The turbidity and bacterial count was reduced by using Moringa oleifera seed extract,
  • 4. International Journal of Trend in Scientific Research and Development (IJTSRD) @ www.ijtsrd.com eISSN: 2456-6470 @ IJTSRD | Unique Paper ID – IJTSRD31019 | Volume – 4 | Issue – 4 | May-June 2020 Page 494 2.4% of powder has ability to reduce the impurities from water.The study can be used to determine fungal, algal, viral count present in water samples. Moringa oleiferaisa natural coagulant produces biodegradable environment friendly sludge as compared to chemical coagulant. Properties of Moringa oleifera like adsorbent, coagulant, dewatering and desalting agent can be checked and used for removal of heavy metals from water. ACKNOWLEDGMENT I would like to thank Miss Malika Ahuja for helping me in doing research as well as for givinghermajorcontributionin writing and proof reading the research article. REFERENCES [1] Abera S, Zenyinudin A, Kebede B, Deribew A, Ali S, Zemene E (2011); Bacteriological analysis of drinking water sources. Afri.Micro.Res.2011:5(18):2638-2641. [2] Chaubey Soni and Mohan Kumar Patil (2015). “Application of Artificial Neural Network in Assessing Water Quality: A Case Study”, International Journal of Current Research Vol.7, Issue01, pp.11403-11407, India. [3] Dhanaji Kanase G, Shagufta Shaikh A, Pramod Jagadale N (2016); Physico-Chemical analysis of drinking water samples of different places in Kadegaon Tahsil, Maharashtra (India);Pelagia ResearchLibrary;7(6):41- 44. [4] Hujare, M. S. (2008): Seasonal variation of physico- chemical parameters in the perennial tank ofTalsande, Maharashtra. Ecotoxical. Environ. Monitor. 18(3):233- 242 [5] Khalid Anwar, Amir Haider Malik, Amir Waseem, Shazmeen Zahra and Ghulam Murtuza (2011); Qualitative and quantitative analysis of drinking water of different localities in Abbottabad district, Pakistan; Internation journal of the physical science; Vol 6(33).pp 7480-7489. [6] Khan Naeem, Sayef Tasleem Husain, Asif Khan, Kyong Su Kim (2013); Physico-chemical investigation of drinking water sources from Tehsil Lachi, Kohat; American journal of research communication;Vol1(5): 170-190. [7] Kumar Deepesh, Shurutikirti Malik,MolluMadan,Anita Pandey, Ashish K, Asthana (2013); Bacteriological analysis of water by MPN method in a tertiary care hospital and adjoining area western up, India. IOSR Journal of Environmental science toxicology and food technology 4(14). [8] Md saduzzaman, S. Sharmila, Jayanthi rebebeca L.; Domestic waste water treatment using leaf extract of Moringa oleifera; Research journal of pharmaceutical, biological and chemical; 4(2): 833-839 [9] Mnjula A, Shamkar G, Karadesai Preeti, Sharada Metagud (2011);Bacteriological analysis of water, Journal of Bioscience and Technology 2(1):220-222. [10] Olowe Busayo Mutiat, Jacob O. Oluyege and Oladiran Famurewa (2016); Prevalance of water borne disease and microbial assessment of drinking water qualoty in Ado-Ekiti and its environs; British microbiology research journal; 12(2): 1-13. [11] Saxena Nidhi, S.N.Misraand R.N.Shukla (2011); Physico-chemical and Bacteriological analysisofwater quality under different environmental condition; Journal of chemistry and pharmaceutical research; 3(2):162-167. [12] Shimpi Vrushali V., Jhondle Pooja R., Gangurde Ritu B., Jadhav Tejal D., KapseVikashni Nand, Maataa Matakite, Koshy Kanayathu and Sotheeswaran Subramanium (2012);Water pirification using Moringa oleifera and other locally available seeds in Fiji for heavy metal removal; International journal of applied science and technology ;2(5): 125-129 [13] Shireen S. (2018); Water purification using Moringa oleifera (Drumstick) seed powder as a natural coagulant; Recent Advances in Engineering Science and Management ;89-83 [14] Thakur Sunita Singh, Chaubey Sonal (2014); Assessment of coagulation efficiency of Moringa oleifera and okra for treatment of turbid water; Archives of applied science and research; 6(2): 24-30. [15] Yahya Daniyan Safiya, Enemaduku Moses Abalaka and Euro E.O. (2011); The use of Moringa oleifera seed extract in water purification; IJRAP; 2(4): 1265-1271. [16] Yongabi K. A., Lewis D. M. andvHarris L. P. (2011); Application of phytodisinfectants in water purification in rural Cameroon; African journal of microbiology research; 5(6): 628-635.