moringa oleifera seed as antibacterial agent in water treatment

8/12/2019 Moringa oleifera Seed as Antibacterial Agent in Water Treatment

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Paper for National Conference on Household Wake Trea tment Technology, at Hindustan College ofSc. And Tech. Farah, Mathura 24, July 2008

oringaol if r Seed as Antibacterial Agent in Water TreatmentJadhav Swapnali Mohan', Bipinraj N K2, Milind R Gidde3

AbstractWater extract of oringaoeifera seed, a natural anti-coagulant, was checked for its ability to reduce thetotal bacterial count in ground water. Using MPN method different concentration of the seed extract wastested for antibacterial potency. The results were compared with chlorine, which is the commonly useddisinfectant. MPN levels were decreased to greater extent with the treatment seed extract. Maximumreduction of 77.27 in MPN was found when the ground water was treated for 30 min. with 50mg/ml ofseed extract. An attempt was made to find out the MIC seed extract against E coli by TVC method andfound out that 12.5mg/ml concentration of seed extract could reduce >999 Ecoli count. The ability of theextract reduced when heated at 80c for 30 min. but normal conditions (30c) it act as an antibacterial agent.

This study highlights an environmental friendly method of ground water disinfection. Moringa is acommonly used vegetable and widely available in all part of India. Moreover the seed extract is alsoreported as an anti coagulant and used for water treatment in several parts of the world. These reportsstrongly suggest that Moringa seed extract can replace Chlorine in water treatment after standardization ofthe method.

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Key words - Water, Moringa seed extract ground water.

1- BVIEER, Bharati Vidyapeeth University, Pune2- Dept. of /cell and Molecular Biology, Rajiv Gandhi Institute of IT and Biotechnology, Bharati

Vidyapeeth University, Pune3- Professor, Dept. of Civil Engineering, Bharati Vidyapeeth University College of Engineering,

oDhanakawadi, Pune - 411043 (Maharashtra) Corresponding author E-mail: [email protected]

IntroductionWater is the unique molecule that preserves the life on the Earth. All life forms on the Earth depend onwater. About 70 of Earth is covered with water in that 97 is part of Oceans. Only a small percentage ofthe total water is fresh water, which is used by humans for drinking, farming and washing. Safe drinkingwater is a major requirement of any society. Ensuring people's access to safe drinking-water and adequatesanitation will improve the quality of life of millions of individuals. According to WHO report Safe waterfor better health One in every 10 diseases and 6 of all deaths globally are caused by unsafe water andimproper hygiene. In India alone, 1.03 crore people die annually of which, nearly 7.5 deaths are related to

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water, sanitation and hygiene. Diarrhoeal diseases make up a major part of this number causing 4.02 lakhdeaths. According to the report, 88 of cases of diarrhea worldwide are attributable to unsafe water.Diarrhoeal diseases like cholera, typhoid and dysentery caused mainly by the ingestion of pathogens resultin 1.5 million deaths each year. This large number of death due to diarrhea is mainly due to drinkingcontaminated water. Lack of adequate drainage system and distribution of clean drinking water are themain reasons of high mortality due to waterborne diseases in developing countries. Natural sources ofdrinking water are ponds, lakes, streams, rivers and rain. Although the pond water can be directlyconsumed it can transmit harmful pathogen. And it increases the risk for diseases in human as well asanimal body. Ground water is another important source of drinking water. Collected through well or tubeswells they are rich source of minerals, but due to industrial pollution ground water also gettingcontaminated with several heavy metals and non-metals. An average person may consume not more than 2to 3 lit water. However, the per capita consumption of water drawn from public supply is quite large.Untreated or partially treated domestic and industrial wastes released into the water sources affect thehealth of consumers and causes death by different pathogenic microorganisms.

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Chlorination is the most widely used method for water treatment. However, chlorination has numerousdisadvantages such as production of trihalomethanes and chlorinated hydrocarbons which are consideredhealth hazards. It combines with inorganic material in water to form chloride salts, and with organic_material in water to form chlorinated organic chemicals. These chlorinated compounds are less likely todegrade and can cause the same hazards as that of chlorine. Chlorination also produce large amount ofsludge which is again an environmental pollutant. Due to these demerits of chlorine and high expense ofother physical method there is a requirement of safe and cheap water treatmentmethod.

Naturally occurring coagulants and disinfectants can be an alternative to chemical and physical watertreatment method. There are several biological agents, mainly plants, reported for its coagulating activityespecially removal of microorganisms. Coagulation effect ofMO seed on turbid water has been reported bya number of researchers John, 1988; Sutherland et aI., 1990; Nkhata, 2001; Muyibi and Evison, 1995).Seeds of the oringa o eifera found to be highly effective in removing suspended particle from water withmedium to high levels of turbidity ECHO, 17391Durance Rd., North Ft. Myers FL 33917, USA). Thesestudies propose that oringa o eifera seed powder can be an alternative to chemical and physical method.However, effectiveness of waste water treatment also depends on the removal of pathogenicmicroorganisms that cause water born diseases. The present study shows activity of oringa o eiferaseedpowder as an antimicrobial agent.

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2. Materials and Methods2.1 Preparation of Moringa ol lf r seed extractThe Moringa oleifera seed of dry pod were used for the extraction. The seed were removed from the pod,and stored at room temperature. The winged seed cover was shelled just before the extraction. The kernelwas ground to a fine powder by using a mixer and 5gm of the seed powder was mixed with 100 ml ofdistilled water. The suspension was stirred using a Cyclo Mixer for 10 min. The solution was then filteredthrough a muslin cloth. The filtered solution was used for disinfection experiment immediately.

2.2 MOST PROBABLE NUMBER (MPN) METHODMost probable number (MPN) is a routine microbiological method to find out the water quality. Thismethod checks the presence of coli forms in the water by using MacConky s broth. Presence of coli formsis an indirect proof of the possible occurrence of dangerous pathogen, since coli form contamination occursthrough fecal matters. In this experiment MPN of the ground water treated with Moringa seed extract waschecked and compared with non-treated water. MPN of the treated and non-treated water has performed infOUTeplicates. In each experiment 5 ml of seed extract (50 mg/ ml) has mixed with the ground water andchecked for MPN at 0 hr. and after 30 min. of incubation. Ground water sample without any treatment andsterilized distilled water with 5Omg/ml seed powder were used as controls.

2.3 Effect of seed extract on Bacterial growthThe effect of seed extract on bacterial growth was studied using E coli as test organism. In this experimentE coli (1 1 a was inoculated in nutrient broth containing different concentration (2.5, 5, 10, 50 mg) ofseed extract. The growth of the bacteria was checked by measuring optical density and compared with thecontrol flasks inoculated with E coli

2.4 MIC (Minimum Inhibitory Concentration)Minimum inhibitory concentration (MlC) is the lowest concentration of an antimicrobial that will inhibitthe growth of a microorganism. Minimum inhibitory concentrations are important to find out the efficiencyof an antimicrobial agent. MIC of the seed extract was found out using double dilution method. Initialconcentration of 50 mg/ml was diluted to 25, 12.5, 6.25, 3.12, 1.56 and 0.78 mg/ml in nutrient broth. Allthese tubes were then inoculated with 0.02 ml of 1 l E coli suspension and incubated for 18 hr.

2.5 Comparison between Chlorine and MoleiferaChlorine is the most widely used chemical for water treatment. It can also inhibit microbial growth in thewater. The ability of Moringa seed extract to inhibit the growth of bacteria was compared with that ofchlorine. For this experiment ability of 5 seed extract to inhibit the bacterial growth was compared withchlorine demand of the ground water (5mgIL). The comparison was performed by MPN method.

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3. Result and Discussions3.1MPNMPN results showed that seed extract was highly effective in reducing the MPN. In the case of groundwater control the MPN was 110 and sterilized distilled water with seed powder showed and MPN of 23.Ground water treated with seed extract showed an MPN of 94 at 0 hr incubation while after 30 min. theMPN is found to be 23. This value was equal to that obtained from sterile distilled water (Table 1).

Table I: MPN of treated and untreated samples.SET Nos. Test (Raw water with Test (Raw water Sterile distilled

Ground water seed extract) at 0 hr. with seed extract) water with seedafter 30 mi extract

no 94 23 23 no 70 23 23m no 94 23 23

AVERAGE no 86 23 233.2 Effect of seed extract on Bacterial growth

Moringa seed powder extract clearly inhibited the growth of coli(Figure 1). After 8 hours ofincubation the growth of control flask increased to 1 OD while the OD of the bacteria in flaskcontaining seed extract remained same as that Ohr. same result was observed with allconcentrations of seed extract tested. These results indicate that oring seed extract can be usedas an excellent natural antibacterial agent in treating the ground water. These results were similarto the results observed in other earlier reports(Mougli Suarez,Marisa Haenni,Ste phaneCanarelli,Florian Fisch,Pierre Chodanowski,published.Sept.2005p.3847-3857)

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1 1:: - 1 8 5mgml seed extracti L with e.coIiI6 4 2o == ===== == == ==

3 4 5 6 7 8nilE IIin..Figure 1: growth ofbacteria in presence of 5 mg/ml seed extrac

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3.3MICTubes with 12.5 and 25 mg/ml seed extracts showed minimum optical density 0.01 aD) after 18hrsincubation. Only 300 viable celli m1were found in the plate when checked the TVC, whereas in the casecontrol the TVC was 106 ceillmi Table 2.). 12.5-mglml seed extraction reduced the growth by more 99 .La usanne, feverier 2004.)

Table 2: Total Viable CountTune for Concentration of seed powder Total viable oount/ mlincubation19hr 12.5mg 19hr Control asKcoIi 1

3.4 Comparison with chlorineChlorine at 4.5 mg/l concentration reduced the MPN to zero, whereas Moringa seed extract could reducethe MPN to 23. Chlorine is widely used for water treatment because it can treat any kind of water andhighly reactive. However due to its high reactivity it can cause severe environmental and health relatedproblems. The rural people are not aware of the hazardous of chemical disinfectant and use themindiscriminately to treat drinking water. Chlorine forms the HIM tri-halo methane), which is knowncarcinogen D.W.Page, J.A. van Leeuwen, K.M.Spark, M.Drikas, Withers, D.E.Mulcahy; WMer Research362002 4884-4892 .

Table 2. Comparison of seed extract and chlorine treatmentUntreated NO. Of tubes giving positive reactionground water 50flOml 50flml 50fO.lml MPN5 3 1 11Ground water NO. Of tubes giving positive reactiontreated with 50flOml Sof Iml 5ofO.lml MPNseed extract 5 235 )Ground water NO. Of tubes giving positive reactiontreated with 50flOml 50flml 50fO.lml MPNChlorine

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4. ConclusionThis project has indicated that groundwater can be treated considerably with or ingo leifer seed extract.In this study it was found that the MPN of groundwater was reduced from 110 to 23 7 9 1 reduction bythe treatment with seed extract. It can also inhibit the growth of coli and the MIC was found between12.5 to 25 mg/ml of seed powder. Although this figure is higher than the earlier report by standardizationof different parameters affecting the disinfection it can be made more effective as that of chlorine.Moreover o ring is widely grown in India for use as a vegetable so the seeds will be easily available fortreatment of water. This will help in the development of rural water treatment using or ing seed extract.In conclusion oring seed extract is proposed to be an effective environmental friendly alternative forchlorine in ground water treatment and can be used in rural ground water treatment.5. AcknowledgementThe authors are thankful to the heads of the institutions of Bharati Vidyapeeth University in which they areworking.

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moringa oleifera seed as antibacterial agent in water treatment

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