GREENER JOURNAL OF BIOLOGICAL SCIENCES

ISSN: 2276-7762                      ICV: 5.99

Submitted:05/12/2017                 Accepted:08/12/2017                   Published:12/12/2017

Research Article (DOI: http://doi.org/10.15580/GJBS.2017.7.120517176)

Biological Control of Water Pollution at Challawa Industrial Area Kumbotso Local Government, Kano using Moringa oleifera Seed Extract

Asiya A¹ and *Ali M²

¹Department of Basic Studies, College of Basic and Remedial Studies Hassan Usman Katsina Polytechnic Katsina State.

²Microbiology Department, Kano University of Science and Technology Wudil.

¹Email: asiyaanas70 @gmail .com; Phone: +2348062570501

*Corresponding Author’s Email: alimuhd4real @gmail. com Phone: +2347032967252

ABSTRACT

Water purification and reduction of water borne diseases is a major public health goal. The study was aimed to utilize Moringa oleifera seeds extract for purification of ground water at Challawa industrial area Kumbotso Local Government Kano State, Nigeria.  Ground water was collected from four different locations in the area. All water samples were subjected to bacteriological and physiological analysis using standard laboratory methods. Extract from the seeds of Moringa oleifera was used in the process of purifying the water as antibacterial and anticoagulant agent. The result showed that 5 bacterial isolates (Escherichia coli, Salmonella typhi, Proteus mirabilis, Enterobacter spp and Klebsiella pneumoneae) were recovered and all were susceptible to Moringa oleifera seeds extract. The most susceptible isolate is E. coli with aveage zone of inhibition of 17.82 mm while Klebsiella pneumoneae is the least susceptible with average zone of inhibition of 13.00 mm. The turbidity assessment showed that at a concentration of 1000mg/L the Moringa seed suspension can reduce the turbidity of ground water from 4NTU to 1NTU after exposing for 90 minutes. Moringa, therefore, could be regarded as one of the remedy to reduce the incidence of water borne disease leading to high incidence of death especially in the developing world.

Keywords: Moringa oleifera; bacteria, physicochemical; Extract

INTRODUCTION

The ensuring of good quality drinking water is a basic factor in guaranteeing public health, the protection of the environment and sustainable development (Ranjini et al., 2010). Water of good drinking quality is of basic importance to human physiology and man’s continued existence depens very much on its availability (Lemikanra, 1999; FAO, 1997). A significant proportion of the world’s population use potable water for drinking, cooking, personal and home hygiene (WHO, 2004). Before water can be described as potable, it has to comply with certain physical, chemical and microbiological standards, which are designed to ensure that the water is potable and safe for drinking (Tebutt, 1983).

Potable water is defined as water that is free from disease producing microorganisms and chemical substances deleterious to health (Ihekoronye and Ngoddy, 1985). Water is the most common solvent for many substances and it rarely occurs in its pure nature. Water can be obtained from a number of sources, among which are streams, lakes, rivers, ponds, rain, springs and wells (Okonko et al., 2008). Drinking water has always been a major issue in many countries, especially in developing countries like Nigeria. In Nigeria, majority of the rural populace do not have access to potable water and therefore, depend on well, stream and river water for domestic use (Shittu et al., 2008). Water of good drinking quality is of basic importance to human physiology and man’s continued existence depends very much on its availability (Lamikanra, 1999; FAO, 1997). During passage through the ground, water dissolves minerals in rocks and collects suspended particulate matter, particularly those of organic sources as well as pathogenic microorganisms from faecal matters (Onuh and Isaac, 2009). In some areas, water sources are shared with the animals making the water dirty and contaminated. The quality and quantity of available water have implication on the health status of a community. Over 50,000 people die daily due to water borne diseases (Marque et al., 2003). About 2.3  billion  people  Worldwide  have  mortality  and  morbidity  associated  with water related ailment. Certain minerals are also toxic such as the heavy metals. Although, some of the heavy metals such as Zinc, manganese, nickel, and copper act as micro-nutrients at lower concentrations, but become toxic at higher concentrations.

Moringa oleifera Lam. (Moringa) is the most widely cultivated species of a monogeneric family, the Moringaceae. It is a tree native to India and cultivated in all sub-tropical areas. This tropical multipurpose fast growing tree is resistant to drought. In the present study, M. oleifera seeds were used in purification of ground water and evaluated for their efficiency in removing turbidity and microorganisms from the water sample.

MATERIALS AND METHODS

Sample Collection

Ground water samples with different proximity to industrial area collected from four (4) different locations within Challawa industrial area in Kumbotso Local Government Area Kano State, Nigeria. The water samples were collected in sterile bottles, transported to the water laboratory Department of Geography, Bayero University Kano for analysis. Microbiological analysis of water samples conducted at SLT Laboratory Kano state School of Technology, and was conducted as soon as possible after collection to avoid unpredictable changes in the microbial population.

Preparation of Moringa oleifera Seed Suspensions

Moringa oleifera seeds used were collected from the compounding Laboratory in the Department Pharmaceutical Technology, Kano State Polytechnic. The seed wings were removed and the kernels were air dried for two weeks, then grounded into a fine powder using sterile pestle and mortar under laboratory condition. Two grams (2g) of the seeds powder were extracted using 200ml distilled water by blending for 30s. The resulting suspension was filtered through a muslin cloth and the filtrate was made up to (1000ml) to give a stock solution of approximately (2000mg/L). From the stock solution different concentrations (250, 500, 750, 100 mg/L) were prepared and used for turbidity assessments.

Preparation of Moringa oleifera Seed extract

The preparation of ethanol extract of the Moringa oleifera Seed was carried out according to Bengum (2014). 50g of the powdered seed was weight out and dissolved in 500ml of ethanol in a sterile conical flask and allowed to stand for three days with constant shaking. The mixture was filtered using Whatman filter paper and the extracts were evaporated to dryness using rotary evaporator. The solid residues obtained were measured and reconstituted in 10% DMSO at various concentrations and stored in the refrigerator at 4 ⁰C until used (Ali et al., 2017).

Physico-chemical Analysis

The physico-chemical tests included the determination of temperature, turbidity, odour, colour, total solid, total dissolved solid, total suspended solid, pH, conductivity, acidity, total hardness and chloride content using the methods of FAO (1997).

Microbiological analysis

Membrane filters (MF) method. The method described by Noble et al. (2003) was followed; 100 ml of each water sample was filtered through sterile membrane which retained the bacteria on its surface. The membrane was removed aseptically and placed on a Nutrient agar (NA) as a basal medium and MacConkey agar as a differential medium to determine enteric bacteria. All the plates were incubated at 37°C for 24 hrs. Coliform colonies (indicating faecal contamination) growing on the surface of the membrane were counted and recorded as Coliform density (total Coliform colonies per 100 ml) or colony forming unit (CFU).

Characterization and Identification of Bacterial Isolates

Presumptive colonies were confirmed by gram staining and biochemical (Indole, Methyl-red, Vougues Proskeaur, Citrate utilization and Oxidase) tests and each plate were graded as positive or negative. Bacteria isolates were identified and characterized according to Bergy’s manual of systemic determinative Bacteriology by Jolt et al. (1994)

Testing for the Antibacterial Activity

Agar diffusion method was performed following Ahmed and Beg (2001), with some modifications was used for Testing for the antibacterial activity of Moringa. 0.1 ml of the standardized bacterial stock suspension (0.5 Mac Farland) were mixed with 20 ml of sterile Mueller Hinton agar and poured into sterile Petri dish, the agar left at room temperature to dry. Four wells of 6mm in diameter were cut using sterile cork borer. Wells were filled with 0.1 ml of the different Moringa seeds extract concentrations (50, 100, 150 and 200 mg/ml), three replicates for each extract for each testing organisms (Escherichia coli, Salmonella typhi, Enterobacter spp, Proteus mirabilis and Klebsiella pneumoneae). The extracts left to diffuse for one hour, and then the plates were incubated at 37⁰ C for 24 hours. Ciprofloxacin (Micro lab limited) 75mg/ml, was used as control in this research. After incubation, the diameter of the zones of inhibition around each well were measured to the nearest millimeters along straight line i.e. 180° to each other and the mean of the readings were then calculated.

Turbidity Assessment

Turbidity measurement was performed following the method of Myuibi and Evison (1995). Moringa seeds concentrations (250, 500, 750 and 1000 mg/L) were used. Hanna instrument (H193703) was used to assess turbidity in treated and non-treated ground water. The instrument has been designed according to the ISO 7027 international standard, the turbidity measurement units is Nephelometric Turbidity Unit (NTU). The four different concentrations of Moringa seeds were used at different time's intervals to assess its ability to coagulant tap water. Turbidity readings were taken at three times at intervals of 30 minutes each. The pH of the water samples was measured before and after dissolving each concentrate in 1L of it

RESULTS

Bacteriological analysis of water

The bacterial isolates recovered from water a sample is presented in Table 1. The result showed that Escherichia coli, Salmonella typhi, Proteus mirabilis, Enterobacter spp and Klebsiella pneumoneae identified. Escherichia coli, Salmonella typhi, and Klebsiella pneumoneae were present in all the water samples

Antibacterial activity of Moringa seed extract

The antibacterial activity of Moringa seed extract against bacterial isolates recovered from the samples is presented in Table 2. Five different faecal coliform bacteria (Escherichia coli, Salmonella typhi, Enterobacter spp, and Klebsiella pneumoneae) were identified. The result showed that the extract is active against the isolates. Antibacterial activity is increasing with the increase in concentration of the extract. The most susceptible isolate is E. coli with average zone of inhibition of 17.82 mm while Klebsiella pneumoneae is the least susceptible with average zone of inhibition of 13.00 mm

Physicochemical analysis of water

The average physicochemical analysis of the water samples is presented in Table 3. Both physical and chemical properties of water are presented. All of the samples have turbidity of 4NTU except sample 2, which indicated that the samples have high turbidity. The Ph of the water samples ranges from 6.4 - 6.6. Both the water samples were colourless and odourless. 

Turbidity assessment

The turbidity assessment after using Moringa seed suspension is presented in Table 4. The result showed that at a concentration of 1000mg/L the Moringa seed suspension can reduce the turbidity of ground water from 4 to 1NTU after exposing for 90 minutes

DISCUSSION

The results of the study showed different Moringa oleifera extract were considered to be active against the tested isolates. All the isolates showed sensitivity towards four concentrations of M. oleifera seeds extract E. coli was inhibited significantly at 200 mg/ml (21 mm). Klebsiella pneumoneae is found to be less susceptible to the extract with average zone of inhibition of 13mm. According to Castillo et al. (1999) as a general rule, plant seed extract are considered active against both fungi and bacteria. Kaspar et al. (1999) also concluded that aqueous extract of Moringa seeds showed very strong and superior anti-microbial activity especially against Gram positive bacteria Staphylococcus aures and Bacillus subtilis. However, Sobsey, (2001) the anti-microbial activity of Moringa seed extract might be due to the presence of lipophilic compounds that might bind within or internal to cytoplasmic membrane when the zone of inhibition is greater than 6 mm.

 The results of turbidity assessment from the study indicated that the Seeds of M. oleifera reduced ground water turbidity in the range of 25-75 % at different concentrations after 1 hour 30 minutes. Turbidity of ground water tested was best reduced up to 1 NTU (75 %) after 1 hour 30 minutes application of Moringa seeds suspension at the concentration of 1000 mg/L. This result of this study was inconformity with the results obtained from previous researchers who documented 70-99 % turbidity removal by M. oleifera seeds as primary coagulant both for raw waters and synthetics turbid waters (Mark and Frederic, 2003). The seed kernels of moringa contain significant quantities of a series of low molecular weight, water-soluble proteins which, in solution, carry an overall positive charge. The proteins are considered to act similarly to synthetic, positively charged polymer coagulants. Amino acid analysis and sequencing of M. oleifera showed high contents of glutamine, arginine and  proline  as  well  as  total  of  other 60 residues (Genthe and Franck, 1999). The results of this study, confirms that the use of Moringa seeds as natural coagulants for ground water removes turbidity up to 75 %. Comparing with other results using synthetic coagulants the average most probable number (MPN) reductions in turbidity obtained with alum it was only 7.7%, (Venkobachar et al., 1994). Finally, M. oleifera is a non toxic (Rijal et al., 2000) i.e. it could be used as a safe natural organic polymer coagulant. However, an extract from moringa seeds is principally a low molecular proteins which is the main effective coagulant agent used for water treatment (Martins et al., 1997).

CONCLUSIONS

M. oleifera offers an alternative solution to the use of expensive chemical coagulants. The experiments with these natural coagulants gave filtered water turbidity less than or almost equal to 1NTU and thereby met the turbidity criteria for drinking water as per WHO guidelines. Turbidity of ground water tested was best reduced up to 1 NTU (75 %) after 90 minutes application of Moringa seeds suspension at the concentration of 1000 mg/L. At the concentration of 200 mg/ml the antimicrobial activity against important human pathogens was also significant. Moringa, therefore, could be regarded as one of the remedy to reduce the incidence of water borne disease causes leading to high incidence of death in the developing world.

COMPETING INTEREST

The authors declared that no competing interest exist

REFERENCES

Ahmed, I. and Beg, A.Z. (2001). Antimicrobial and phytochemical studies on 45 Indian Medicinal plants against multi-drug resistance human pathogens. J Ethnopharmacol74, 113-123

Ali, M., Aminu, F., and Ibrahim, I. S. (2017). In-vitro Assessment of Antibacterial Activity and Phytochemical Screening of Vitex doniana on Clinical Isolate of Salmonella typhi International Journal of Advanced Academic Research | Science, Technology & Engineering V 3(1) PP 9-16

Castillo, G., Duarte, R., Ruiz, Z. Marucic, M. T., Honorato, B. Mercado, R., Coloma, V., Lorca, V. Martins, T. M. and Dutka, B. J. (1999).“Evaluation of Disinfected and Untreated Drinking Water Supplies in Chile by the H2S Paper Strip Test,” Water Research, Vol. 11, No. 28, pp. 1765- 1770. DsOI:10.1016/0043-1354(94)90249-6

Food and Agriculture Organisation (FAO) (1997).Chemical analysis manual for food and water, 5^thEd,FAO rome, 1: 20-26

Genthe, B. and Franck, B. (1999). “A Tool for Assessing Microbial Quality in Small Community Water Supplies,” WRC Report Pages to Water Research Commission by Division of Water, Environment and Forestry Technology, CSIR, Stellenbosch, Vol. 33, No. 961/1/99

Ihekoronye, A.I. and Ngoddy, P.O. (1985).Integrated food science and technology for tropics.Oxford: Macmillan Press, pp. 95-195.

Jolt, J.G., Krieg, N.R., Sneath, P.H., Stanley, J.T. and Williams, S.T. (1994).Bergey’s manual of systematic bacteriology, 9th edn. Williams & Wilkins Co. Baltimore, Maryland, p786.

Kaspar, I. Guillen, D. Revelli, T. Meza, G. Velazquez, L. Mino de Kasper, C. Pozolli, C. Nunez and G. Zoulek, (1999). “Evaluation of a Simple Screening Test for the Quality of Drinking Water Systems,” Annals of Tropical Medicine and Parasitology, Vol. 3, No. 43, pp. 124-127.

Lamikanra, A. (1999). Essential microbiological for students and practitioners of pharmacy, medicine and microbiology, 2nd Edition, Amkra books, 406pp.

Mark D. S. and Frederic, K. P. (2003). “Evaluation of the H2S Method for Detection of Faecal Contamination of Drinking Water,” WHO/SDE/WSH/02.08 Geneva

Marque, S., Jacqmin-Gadda, H., Dartigues, J.F. and Commenges, D.(2003).Cardiovascular mortality and calcium and magnesium in drinking water: an ecological study in elderly people. European Journal of Epidemiology 18:305–309

Martins, M., Castillo and G. Dutka, B. J (1997). “Evaluation of Drinking Water Treatment Plant Efficiency in Microor- ganism Removal by the Coliphage, Total Coliform and H2S Paper Strip Test,” Water Science and Technology, Vol. 3, No. 35, pp. 403-407. doi:10.1016/S0273-1223(97)00293-X

Okonko,I.O., Adejoje, O.D., Ogunnusi, T.A., Fajobi, E. and Shittu, O.B. (2008). Microbiological and physicochemical analysis of different water samples used for domestic purposes in Abeokuta and Ojota, Lagos Nigeria. African Jounal of Biolechnology7(5) 617-6721.

Onuh, J.O. and Isaac, V.U. (2009).Physicochemical and microbiological quality of water sources from some major towns in Igala land. Nigeria food Journal27(22): 66.

Ranjini K; Roland P; John C. and Vincent R. (2010). Microbiological and physicochemical analysis of drinking water in George town. Nature and Science, 8 (8)261-265.

Rijal, G. K., Fujioka and R. S. and Ziel, C. A (2000). “Evaluation of the Hydrogen Sulfide Bacteria Test, a Simple Test to De- termine the Hygienic Quality of Drinking Water, Abstracts of    the General meeting of the American Society for Microbiology,” Poster Presentation (Unpublished), American Society of Microbiology, Washington DC

Shittu, O. B, Olaitan, J.O. and Amusa, T.S. (2008).Physico-chemical and bacteriological analyses of water used for drinking and swimming purposes in Abeokuta, Nigeria. African Journal of Biomedical Research 11: 285-290.

Sobsey, M. D. (2001). “Microbial Detection: Implications for Exposure, Health Effects, and Control,” In: G. F. Craun, Ed, Safety of Water Disinfection: Balancing Chemical and Microbial Risks, Intl Life Sciences Inst Pres, Washington DC, pp. 11-51.

Tebutt T.H.Y. (1983). Principles of Quality Control. England: Pergamon, p.232. US Environmental protection Agency (USEPA) (1995).Guidelines for water suppl.y7th edition. New York: WHO press, pp.144 -162

Venkobachar, C., Kumar, D., Taireja, K., Kumar, A. and Iyengar, I. (1994). “Assessment of Bacteriological Quality Using a Modified H2S Strip test,” Aqua (Ottoro), Vol. 6, No. 43 pp. 311-314.

World Health Organization (2004). Guidelines for drinking water quality.3rd edition, Switzerland: WHO press pp.16,89

Cite this Article: Asiya A and Ali M (2017). Biological Control of Water Pollution at Challawa Industrial Area Kumbotso Local Government, Kano using Moringa oleifera Seed Extract. Greener Journal of Biological Sciences, 7(7):063-068, http://doi.org/10.15580/GJBS.2017.7.120517176