Treatment of Domestic Waste Water Using Natural Flocculants

Int. J. LifeSc. Bt & Pharm. Res. 2012 Narmadha D and Mary Selvam Kavitha V J, 2012

Narmadha D1* and Mary Selvam Kavitha V J1

*Corresponding Author: Narmadha D,  [email protected]

Wastewater generated from wastewater treatment plant contains odors some non pathogens, fecal Coliforms, fungus etc., and chemicals such as phenols, nitrate, chloride etc., in addition to usual constituents such as urine and faeces. The present investigation was hence made in wastewater treatment plant situated in Kallaraimedu of Kodaikanal. Presently the waste water is passing through an underground system to the wastewater plant. In this study analyze the Physio-chemical parameters and biological parameters of septic water before and after treatment with horse and cow dung; and portable water. Isolate and identify the microbes present in horse and cow dung by using biochemical tests.

Keywords: Waste water, microbial treatment of waste, Flocculants

INTRODUCTION such as urea, fruit sugars, soluble proteins, drugs, Wastewater can be defined as the flow of use pharmaceuticals, etc.; Inorganic particles such water discharged from homes, businesses, as sand, grit, metal particles, ceramics, etc.; industries, commercial activities and institutions Soluble inorganic material such as ammonia, which are directed to treatment plants by a road-salt, sea-salt, cyanide, hydrogen sulfide, carefully designed and engineered network of thiocyanates, thiosulfates, etc.; Animals such as pipes. This wastewater is further categorized and protozoa, insects, arthropods, small fish, etc.; defined according to its sources of origin as 1. Macro-solids such as sanitary napkins, nappies/ domestic or sanitary wastewater and 2. Industrial diapers, condoms, needles, children’s toys, dead wastewater animals or plants, etc.; Gases such as hydrogen Pathogens present in waste water include sulfide, carbon dioxide, methane, etc.; Emulsions bacteria, viruses, prions and parasitic worms and such as paints, adhesives, mayonnaise, hair Non-pathogenic bacteria. Organic particles such colorants, emulsified oils, etc.; Toxins such as as faeces, hairs, food, vomit, paper fibers, plant pesticides, poisons, herbicides, etc., are the material, humus, etc.; Soluble organic material constituents of waste water.

1 Department of Biotechnology, Mother Teresa Women's University, Kodaikanal.

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SEWAGE TREATMENT requires the use of an ozone generator, which By definition, treatment process means a series decontaminates the water as Ozone bubbles of actions or changes. Treatment facilities percolate through the tank. incorporate numerous processes which in Treated wastewater can be reused as drinking combination achieve the desired water quality water, in industry (cooling towers), in artificial objectives. These processes involve the recharge of aquifers, in agriculture and in the separation, removal and disposal of pollutants rehabilitation of natural ecosystems. In order to present in the wastewater. The treatment of achieve ecological wastewater treatment, wastewater is accomplished by four basic ecological engineering, a closed-loop treatment methods or techniques; physical, mechanical, system is recommended. Many present day biological and chemical. Most wastewater is systems are a “disposal-based linear system”. treated in industrial-scale wastewater treatment The traditional linear treatment systems must be plants (WWTPs) which may include physical, transformed into the cyclical treatment to promote chemical and biological treatment processes. the conservation of water and nutrient resources. However, the use of septic tanks and other On- Using organic waste nutrient cycles, from point- Site Sewage Facilities (OSSF) is widespread in of-generation to point-of-production, closes the rural areas, serving up to one quarter of the homes resource loop and provides an approach for the in the U.S. The most important mechanical management of valuable wastewater resources. aerobic treatment system is the activated sludge Chemical treatment methods enhance the process, based on the maintenance and efficiency of other process operations and provide reconciliation of a complex biomass composed specialized treatment as a result of their addition of micro-organisms able to absorb and adsorb at various treatment stages. The commonly used the organic matter carried in the wastewater. physiochemical processes that require chemical Anaerobic processes are widely applied in the action precipitation, chemical oxidation, carbon treatment of industrial wastewaters and biological adsorption, ultra filtration, reverse osmosis, sludge. Some wastewater may be highly treated electro dialysis, volatilization and gas stripping. and reused as reclaimed water. In some waste They are either very pollutant specific, or more waters ecological approaches using reed bed expensive than dumping and using more water. systems such as constructed wetlands may be The action of bacteria and other micro- appropriate. Modern systems include tertiary organisms are biological methods of treatment, treatment by micro filtration or synthetic which play a vital role in the removal of pollutants membranes. After membrane filtration, the treated which cannot be effectively achieved by other wastewater is indistinguishable from waters of means. the natural origin of drinking quality. Nitrates can be removed from wastewater by microbial SEPTIC TANK TREATMENT denitrification, for which a small amount of BY BACTERIA methanol is typically added to provide the bacteria with a source of carbon. Ozone Waste Water Although there are many methods used, there Treatment is also growing in popularity, and are numerous concerns raised regarding the

This article can be downloaded from http://www.ijlbpr.com/currentissue.php 207 Int. J. LifeSc. Bt & Pharm. Res. 2012 Narmadha D and Mary Selvam Kavitha V J, 2012 presence of constituents including heavy metals, and identified by using selective and differential pathogens and other toxic substances. This media. MR-VP medium (Methyl Red Voges requires the selection of the correct disposal Proskauer), nutrient agar, nutrient broth, tryptic method focusing on efficient and environmentally soy agar, starch agar, Mannitol salt agar, Simmon safe disposal. New technologies are being Citrate agar, lactose Broth were used. The pour produced to assist in the treatment and disposal plate technique was used for bacterial of sewage sludge, conforming to strict enumeration and determines the bacterial count environmental regulations. One of these new in a milliliter or gram of a specimen. After technologies being proposed is the use of incubation, colonies appearing on the agar are Effective Microorganisms (EM). counted. Each colony represents one colony forming unit (CFU). The CFU/ml or CFU/g is then Bacteria are living organisms, part of the calculated. Gram staining was used to distinguish earth’s biological system. In a septic system between gram positive and gram negative bacteria such as Bacillus subtilis aid in digesting bacteria. waste matter inspects the system approximately every three years. The sludge is pumped when EXPERIMENTAL DESIGN necessary to maintain a proper operating system to provide a healthy environment for bacteria to Sample Preparation achieve optimum population levels and proper Two litres of septic water samples were collected operation within the septic tank. from water treatment plant, Kallaraimedu, Kodaikanal and dispensed into 4 conical flasks. The septic tank processes convert the organic nitrogen compounds into ammonium; however The volume of sample in each conical flask was no conversion of ammonium to nitrate is expected 500ml. The first conical flask with the septic water to occur in the septic tank. Since in the septic sample was not treated Septic water untreated tank phosphorus concentration is high (SWUT). 5g of Horse dung was weighed and wastewater coliform bacteria are not removed added into a conical flask containing 500ml of the because reduction of ammonium to nitrate does sample. This was named as septic water treated not occur. Flocculant1 (SWTS 1). 5g of Cow dung was weighed and added into a conical flask containing MATERIALS AND METHODS 500ml of the sample. This was named as septic Flocculant Collection and Analysis of water treated Flocculant2 (SWTS 2). The known Flocculant Bactizyme was used as an internal positive Flocculant was collected from Kodaikanal area control it was weighed 5g and added to the between Jan-Feb. 2011. Horse dung was used sample (SWTS 3). The 500ml of distilled water as Flocculant1 and cow dung was used as with no dung was kept as Negative control Flocculant2. (SWNC). Potable water was used as Standard (SWSS). Isolation and Identification of Microbes Present in Horse and Cow Dung Analysis of Septic Water Microbes present in the flocculants were isolated The collected samples were subjected to

This article can be downloaded from http://www.ijlbpr.com/currentissue.php 208 Int. J. LifeSc. Bt & Pharm. Res. 2012 Narmadha D and Mary Selvam Kavitha V J, 2012 systemic analysis of the below stated Total Dissolved Solids physiochemical parameters, biological testing The total Dissolved Solids of the samples of the and microbial characteristics noted as Day 0. The treatment system was maintained at from SWUT analysis was continued for 20 consecutive days. from 0.01g. The same levels were maintained in The samples were tested for the following standard, controls in all the day of observation. physiochemical parameters pH, Total Suspended TDS was contributed predominantly by chlorides, Solids (TSS), Total Dissolved Solids (TDS), Sulphates, phosphates, Mg, Ca, Na, K etc. High Total Solids (TS), Chloride, Nitrate. The septic level of salts could adversely affect the biological water treated samples (using Flocculant 1 and treatment process. Some of them are reported Flocculant 2) were serially diluted and microbial to remain unchanged during the treatment analysis was done for bacteria - Total viable count process (Vishnu et al., 2008). (TVC) Presumptive test, confirmed test, Chloride completed test. Faecal coliform (FC) was tested and the number of colonies on the EMB agar Chloride content gradually decreased from Day plates were counted and recorded as CFU/ml. 0 to Day 20. In Day 0 untreated septic sample Staphylococcus was also tested (Cauppuccino, chloride content is 295.13mg/l and in day 20 it is 1992). Total fungal count (TFC) was performed 200.22mg/l. This is due to the inoculation of the as per the procedure outlined in the Microbiology Flocculants which act on compounds present in Laboratory Manual (Cappuccino, 1992). The the sample. The increasing value of chloride fungal colonies were counted and recorded as might be due to the precipitation of chlorides of CFU/ml. Calcium and Aluminium phosphate on hydrous aluminium oxide and the presence of more RESULTS AND DISCUSSION suspended solids (Ngtez et al., 1999; Rashed and Soltan, 2002). pH The sample from SWUT remained acidic. Nitrate However the change of pH in septic tank was Nitrate content gradually decreased from Day 0 observed to be very rapid in treatment time from to Day 20. Due to low hydraulic retention time day 0 to 20 days. The water slowly moved towards (HRT) in the septic tank, ammonium oxidizing neutral pH. Low pH in elevated temperature, bacteria could not get sufficient time to produce polymerization and condensation reactions take effects. Active by passing of the waste water in place resulting in the reduction of soluble and septic tank resulted in high concentrations of NH insoluble materials. Reduction of pH is due to 3 and HNO (Manipura and Burgess, 2008). chlorine because it degrades the volatile fatty 2 acids (Baloch and Akunna., 2003 and Mohammed Nitrogen is considered as one of the most et al., 2008). important nutrient causing water pollution and removal of compounds having nitrogen is Total Suspended Solids considered as the major activity in waste water The total Suspended Solids in the samples of the treatment (Koren et al., 2000 and Kasia et al., treatment system decreased from 0.50g to 0.03g 2005). Nitrogen was reduced much when treated septic tank from Day0 to Day 20.

This article can be downloaded from http://www.ijlbpr.com/currentissue.php 209 Int. J. LifeSc. Bt & Pharm. Res. 2012 Narmadha D and Mary Selvam Kavitha V J, 2012 with SWTS 1, SWTS 2 and SWTS3. Decrease Identification or increase in nitrogen content is due to bacteria The group of cultures were grown in differential which requires carbon source. These microbes and specialized media for the growth of specific use NO3-N as the terminal electron acceptor in microbe. The cultures identified as the absence of oxygen (Zhao et al., 1999). The Flocculant collected was used for isolation of organic nitrogen of waste water in the present microbes. The culture F 1A (Pseudomonas sp), investigation was observed to be increasing and F 1B (Bacillus subtilis), F 1C (Enterococcus sp), gradually decreases due to the conversion of F 1F (Aspergillus sp) , F 2A (Bacillus subtilis), F organic bound nitrogen in to an ammonium 2B (Streptococcus sp), F 2C (Lactobacillus sp), compound. and F 2F (Candida sp) are non pathogenic Microbiological Parameters for Potential naturally occurring microbes present in horse Faecal Pathogens dung and cow dung and are antibiotic resistant Faecal Coliform that destroy the majority of human pathogenic The Higher Faecal Coliform rate was noticed in organisms in waste water through biological Day 0 and it was gradually decreased from Day1 competition (Gerba et al.,1980; Ward et al.,1984 to Day 20. and Placha et al., 2008). The data obtained from the existing treatment system of waste water Staphylococcus indicate that the treated wastewater match with On day 0, 2-3 colonies were noticed and after positive control and standard. treatment effectively from Day 1 to Day 20 no The main species involved in effective waste staphylococcal colonies were obtained. There is water treatment include Lactic acid bacteria – no fermentation in lactose broth and no color Lactobacillus plantarum, L. casei, Streptoccus change from Day 0 to Day. lactis; Photosynthetic bacteria – Rhodop- Fungal Count seudomonas palustrus, Rhodobacter spaeroides; The total Viable Count of bacterial colonies Yeasts – Saccharomyces cerevisiae, Candida present in the sample. The viable count of fungi utilis Actinomycetes – Streptomyces albus, S. shows reduction in treatment from Day 1 to Day griseus; Fermenting fungi – Aspergillus oryzae, 20. The fungal colonies were identified based on Mucor hiemalis (Diver, 2001). The basis for using conidial structure - Candida Sp, Aspergillus, these EM species of microorganisms is that they Mucor. contain various organic acids due to the presence Identification and isolation of microbes of lactic acid bacteria, which secrete organic from flocculants acids, enzymes, antioxidants, and metallic Chelates (Higa and Chinen, 1998). This study Flocculant collected was used for isolation of microbes. The cultures F 1A, F 1B, F 1C and F has identified microbes in Flocculants 1 are 1F are horse dung isolates and F 2A, F 2B, F 2C Pseudomonas sp., Bacillus subtilis, Enterococcus and F 2F are cow dung isolates were identified sp., Aspergillus niger., and in Flocculant 2 are by colony morphology, growth characteristics and Bacillus subtilis, streptococcus sp., Lactobacillus biochemical tests. sp., Candida sp.

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Organic materials within wastewater originate tables and figures shown above that effect of from plants, animals or synthetic organic SWTS 1 and SWTS 2 have the best activity. compounds, and enter wastewater via a number Further studies need to reduce the treatment time. of routes including human wastes, detergents, The effectiveness of individual microbes identified and industrial sources (Taylor et al., 1997). In the in this study in waste water treatment should be current wastewater treatment process (either evaluated. municipal or domestic on-site) microorganisms play a significant role in the treatment of domestic REFERENCES sewage. Many different organisms live within the 1. A Subset of Naturally Isolated Bacillus wastewater itself, assisting in the breakdown of Strains Show Extreme www.ncbi.nlm. certain organic pollutants (Taylor et al., 1997). The nih.gov/pubmed/20626457 creation of an antioxidant environment by 2. An Introduction to bacterial Identification microbes assists in the enhancement of the solid- Page 1: General Principles this Page liquid separation, which is the foundation for Amplifies Material Discussed in our UW- cleaning water (Higa & Chinen 1998). Madison Microbiology 102 laboratory Research revealed that Aspergillus niger, courses. Trichoderma harzianum, Bacillus cereus and 3. Bsci424—Pathogenic Microbiology - Fall Bacillus subtilis isolated from cow dung can 2000 http://www.life.umd.edu/classroom/ reduce the growth of Sclerotium rolfsii, Fusarium bsci424/LabMaterialsMethods/Catalase oxysporum, Pythium aphanidermatum, Test.htm Helminthosporium maydis and Rhizoctonia solani 4. Cappuccino G J (1992), Microbiology- A with inhibitory zones of up to 58%. Furthermore, Laboratory Manual, 3rd Edition Benjamin, B. subtilis isolated from cow dung can enhance Cumming Publishing Company Inc., plant growth, sulfur oxidation, phosphorus Newyork. solubilisation and was found to produce industrial 5. Carl Lamanna (1940), “The taxonomy of the enzymes such as amylase and cellulase (Swain genus bacillus I. Modes of spore germina- and Ray, 2006). tion”, Journal of Bacteriology, pp. 340-360. CONCLUSION 6. G C Delzer and S W McKenzie (2003), SWTS 1 and SWTS 2 were found to be Potential “Five-Day Biochemical Oxygen Demand”, Flocculant for treating waste water. Both F 1 U.S. Geological Survey, TWRI Book 9 and F 2 were found to have Bacillus subtilis ChapterA7.2, pp. 1-21 Other microbes found were Pseudomonas sp., 7. How to test water purity/e-how.com www. Enterococcus sp., Aspergillus niger., Strepto- ehow.comhow_378_test_waterpurity... coccus sp., Lactobacillus sp. and Candida sp. This study concluded that the microbes present 8. Identification flow charts Bergey’s Manual in SWTS 1 and SWTS 2 are known to treat waste of Determinative Bacteriology All of the water. SWTS 1 have the best activity when Unknowns will fall into the Following Groups compared to SWTS 2. Hence we conclude from in Bergey’s Manual of Determinative

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