MANOHARBHAI PATEL INSTITUTE OF ENGINEERING AND TECHNOLOGY

GONDIA FORWARDING LETTER

Forwarded herewith to the Rashtrasant Tukdoji Maharaj Univesity, Nagpur, and the dissertation

TREATMENT OF DOMASTIC WASTE WATER BY PYTOREMEDATION (LEMNA PLANT)

Submitted by- Sandeep P. Ajmire , in partial fulfillment of the award of the degree of Master of Technology in Environmental Engineering.

Prof. A. L. Nashine Head of department Dept. of Civil Engg. MIET , Gondia

Prof. Dr.S.S. Rathor Principal MIET Gondia

MANOHARBHAI PATEL INSTITUTE OF ENGINEERING AND TECHNOLOGY GONDIA

CERTIFICATE
This is to certify that dissertation entitled

TREATMENT OF DOMASTIC WASTE WATER BY PHYTOREMEDATION (LEMNA)

Submitted by Sandip P.Ajmire , in practical fulfillment of the requirement for the award of Degree of Master of Technology in Environmental Engineering to The Rashtrasant Tukdoji Maharaj University, Nagpur , is bonafide research work carried out under my supervision and guidance. The work embodied in this dissertation has not submitted previously for the award of any degree or diploma.

Prof . A.M. Deshpande Supervisor Dept. Of Civil Engineering MIET GONDIA

Prof.AL Nashine Head Of Department Dept. Of Civil Engineering MIET GONDIA

ACKNOWLEDGEMENT
I express my profound gratitude towards Prof. A.M. Deshpande ,Lecturer , Department of Civil Engineering. MIET Gondia, for this able guidance. I am extremely Grateful to Hon President Mr. Choudhari & CEO A.V. Dhoke, Municipal Council Gadchiroli . Mr .M.G. Nisal , Lab Asst . Environmental Engineering Lab MITE ,Gondia , without whose help the project might have been completed. Mr. S.P. Waghmare, Executive Engineer Jeewan Pradhikarn Gadchiroli & his technical and non technical staff , without whose help the project might have been completed. I express heartfelt thankful to Prof. Dr. S.S. Rathod , Principal & Prof .A.L.Nashine, H.O.D., Civil Engineering & Prof. P.E.Mishra Coordinate, PG Deptt. Of Environmentel Engg.,MIET, Gondia, for providing necessary facilities in the completion of this work and for his constant encouragement.

Sandeep P. Ajmire

CONTENTS
INTRODUCTION
1.1. General 1.2 . Polution Problem 1.3 . Standards of Disposal 1.4. Treatment methodology 1.5. Objective and scope of study

LITERATURE REVIEW
2.1. General 2.2. Characteristics of domestic waste water 2.3. Tretment Processes 2.4. Process selection criteria for treatment of various domestic waste water 2.5. Application of Phytoremedation to domestic waste water

PHYTOREMEDATION
3.1. History & back round 3.2. Definition & types of Phytoremedation 3.3.Introduction of Phytoremedation by Lemna 3.3.1 3.3.2 3.3.3 3.3.4 3.3.5 3.3.6 3.3.7 3.3.8 3.3.9 3.4. 3.5

PLANTS AND METHOD 4.1 4.2 4.3 4.5 4.6 4.7 4.8 4.9 4.9.1 4.9.2 4.9.3 4.9.4 4.9.5 4.9.6 4.9.7 4.10

OBSERVATIONS ,RESULTS,AND DISCUSSION

5.1 5.2 5.3 5.4 5.5 5.6 5.7

Reference:
PHOTOGRAPHS

INTRODUCTION 1.1. General

The population of glob is increasing, the problem of municipal & industrial waste tedious day by day. The legacy of rapid urbanization , industrialization ,fertilizer & pesticide use has resulted in major pollution problems in both terrestrial and aquatic environments. In developing countries is major problem to treat the polluted water from above sources. Chemical & mechanical menace are used for this purpose is expensive. In response , conventital, remediation systems based on high physical and chemical engineering approaches have been developed and applied to avert or restore polluted sites. Much as these conventional remediation systems are efficient, they are sparsely adopted because of some economical and technical limitations. Generally, the cost of establishment and running deter their use and meeting the demand particularly in countries with week economy. Logical this high cost technology can neither be applied justifiably where 1. The discharge is abruptly high for short time but the entire average load is relatively small.
2. The discharge is very low but long term (entire load is medium). 3. The discharge is continuously decreasing over a long duration.

Thus conventional remediation approaches are best for circumstances of high pollutants discharge like in industrial mining and domestic waste water. Recently , it is evident that durability restoration and long term contamination control in conventional remediation is questionable because

in the long run the pollution problem is only is suspended or transferring from one site to another. The efficiency of duckweed (Lemna gibba L.) as an alternative cost effective natural biological tool in wastewater treatment in general and eliminating concentrations of both nutrients and soluble salts was examined in an outdoor aquatic systems. Duckweed plants were inoculated into primary treated sewage water systems (from the collector tank) for aquatic treatment over eight days retention time period under local outdoor natural conditions. Samples were taken below duckweed cover after every two days to assess the plants efficiency in purifying sewage water from different pollutants and to examine its effect on both phytoplankton and total and fecal coliform bacteria. The Lemnaceae family consists of four genera to most other plants, does (Lemna, Spirodela, Wolffia & Wolffiella) and 37 species have been identified so far. Compared duckweed has low fiber content (about 5%), since it these, wastewater treatment was not require structural tissue to support leaves and stems. Of

applications of Lemna gibba L (duckweed) in

found to be very effective in the removal of nutrients, soluble salts, organic matter, heavy metals and in eliminating suspended solids, algal abundance and total and fecal coliform densities. macrophyte belonging Outdoor Duckweed is a floating aquatic to the botanical family Lemnaceae, which can be

found world-wide on the surface of nutrient rich fresh and brackish waters. experiments to evaluate the performance of the duckweed as a purifier of domestic wastewater in shallow mini-ponds (20 & 30 cm deep) showed that quality of resultant secondary effluents met irrigation reuse criteria. Wastewater ammonia was converted into a protein rich biomass, which could be used for animal feed or as soil fertilizer. The economic

benefit of the biomass by-product reduced wastewater expenditures to approx. US$ 0.05 per treated m3 of wastewater, which was in the range of conventional treatment in oxidation ponds. The present study was concerned with decreasing pollution of municipal waste waster up to degree Standards of Disposal as per

National pollution control board.

1.2 . Pollution Problem The quality of municipal wastewater of stagnant/ slow velocity may create problem of high epidemics of malaria & other water born diseases. The efficiency was tested by measuring some of physicochemical characteristics of the control and plant treatments after each seven days. The experiment lasted for a month, and recorded the rates of reduction. The highest rates of reduction were for heavy metals, accounting 99.8%, 99.6%, 98.7% and 72% for Copper, Cadmium, Lead and Zinc, respectively, followed by Turbidity and Nitrate recorded 64% and 57.1% respectively. The percentage reduction of BOD5 was 49.6 %, while for COD 32.7%, either the rest of the physicochemical characteristics as follows: 48.9% for Soluble Solids and 43% of Oils and Grease, 41% of Total Alkalinity, 40% for Phenols, 39.1% for Sulfide, 38% of Suspended Solids and 30% of Phosphate. Lower rates of reduction were recorded of each Temperature (17.2%), pH and Sulfate (13.4%). The results showed that this aquatic plant can be successfully used for wastewater pollutants removal

Refinery wastewater is producing in a huge quantity in most the cities of the country that contain a diverse range of pollutants including Heavy Metals ,Oil and Grease ,Phenols, Sulphide, Sulphate ,Nitrate ,Phospate, Dissolved Solids, Suspended Solids, COD, BOD, which Its disposal and treatment has become a challenge for the municipalities; Many of the municipalities in growing cities neither have proper disposal system nor have any treatment facility due to higher cost and in such a situation refinery wastewater is discharge in to aquatic bodies like river, ponds and lakes, where it is posing a serious threat to the water quality and become a big environmental problem throughout the development of the petrochemical industry [1] [2]. Physical, chemical, and biological technologies have been developed to treat oily waste water and restore environmental quality; However their costs are high and most of them are difficult to use under field conditions, hence in such a condition there is an urgent need to study natural, simple, and cost-effective techniques for control pollution from industrial effluents and treating such wastewater, such as phytoremedation [1] [3]. Viewing this fact Phytoremediation was assumed to be very useful, as it is an innovative, eco-friendly and efficient technology in which natural properties of plant is used in engineered system to remediate hazardous wastes through physical, chemical, and biological processes from wastewater and sewage [4] [2]. Phytoremedation is the utilization of plants accumulation capabilities to remove contamination from water, soil and air, the capacity of aquatic plants to remove pollutants from water is well documented [5]. The recent application of phytoremediation technology by duckweed in wastewater treatment and management is quite interesting and revealing. Phytoremediation systems by duckweed are one of the options that have been widely applied for combined handling of wastewater with the nutrients used for poultry and aqua-cultural projects [6] [7].

Lemna minor L. known as common duckweed is a small, free floating aquatic plant fast growing, adapt easily to various aquatic conditions and play an important role in extraction and accumulation of pollutants from waters [8]. In particular, species of Lemna are reported to accumulate toxic metals and therefore are being used as experimental model systems to investigate heavy metal induced responses, Bioavaibility and bioaccumulation of various heavy metals in aquatic and wetland ecosystems is gaining tremendous significance globally [9]. Aquatic macrophytes take up metals from the water producing an internal concentration several fold greater than their surroundings. Many of the aquatic macrophytes found to be the potential scavengers of heavy metals from aquatic environment and are being used in wastewater renovation systems [10] [11]. Aquatic plants have shown their efficiency in absorbing nutrients from various sources of polluted water, [12] [13]. This study aimed to assessing the efficiency of duckweed (Lemna minor) in phytoremediate the pollutants of Basrah oil refinery wastewater. Pour dterminer la tolrance et la capacit de phytoaccumulation du cuivre (Cu) et du nickel (Ni) par une espce de lentilles deau, Lemna gibba L., les plantes sont exposes diffrentes concentrations de Cu et Ni (0,1 2,0 mg/L) dans une solution de Coc et Lesaint dilu 1/4. Le pH est maintenu constant 6,0 ( 0,1) et le flux de lumire est de 12 h/jour. Le cuivre et le nickel sont tolrs par L. gibba des concentrations 0,3 mg/L et 0,5 mg/L, respectivement. Cependant, la croissance des plantes diminue de 50% (I50) quand le milieu de culture contient 0,45 mg/L de Cu ou 0,75 mg/L de Ni. La plus faible concentration causant une inhibition complte (LCI) est de 0,5 et 1,0 mg/L respectivement en prsence de Cu et Ni. Les rsultats de lanalyse du mtal dans les tissus des plantes rvlent une grande accumulation de Cu et une faible accumulation de Ni dans les tissus vgtaux (pour la concentration ne causant aucune inhibition dans la croissance). Une diminution de la concentration de mtal dans leau est galement observe. On peut conclure que L. gibba peut tre un bon candidat pour lpuration des eaux contamines par le cuivre.

ABSTRACT To assess the tolerance and phytoaccumulation ability of the duckweed Lemna gibba L. to copper (Cu) and nickel (Ni), the aquatic plants were exposed to different concentrations of Cu and Ni (0.1 2.0 mg/L) in quarter Coc and Lesaint solution at pH = 6.0 ( 0.1) and under a daily regime of 12 h light. Copper and nickel were tolerated by L. gibba at concentrations 0.3 mg/L and 0.5 mg/L, respectively. However, plant growth decreased by 50% (I50) when the medium contained 0.45 mg/L of Cu or 0.75 mg/L of Ni. The observed LCI (lowest concentration causing complete inhibition) were 0.5 and 1.0 mg/L respectively in the presence of Cu and Ni. Results from metal analysis in plant tissues revealed a high accumulation of copper and a low accumulation of nickel within the plant (for concentrations causing no growth inhibition) and a corresponding decrease of metals in the water. The duckweed L. gibba L. could be a good candidate for the removal of low concentrations of copper from polluted water. Certain plants are able to accumulate in their tissues several metals without showing any signs of toxicity. This natural accumulation is related with the resistance which represents response of plants to metal stress conditions. According to Papazoglou et al. (2005), metal resistance can be based on either avoidance or tolerance mechanisms. Avoidance reflects the cell protection against the metal whereas tolerance is the cell capability to protect themselves against injury by the metal (Sabreen & sugiyama, 2008). Kanoun-Boul et al. (2008) reported in the tolerance of duckweed to copper that the release of organic anions might be involved in the protection of plants by chelating the metal ions in the rhizosphere to form non toxic complexes. However, according to Sabreen and Sugiyama (2008), tolerant populations can be characterized by a lower metal accumulation than the sensitive one.

Duckweed commonly refers to a group of floating, flowering plants of the family Lemnaceae. The different species (Lemna, Spirodela, Wolffia and Wolfiella) are worldwide distributed in freshwater and wetlands, ponds and some effluents are the most common sites to find duckweed. The plants are fast growing and adapt easily to various aquatic conditions. They are able to grow across a wide range of pH, from pH 3.5 to10.5 but survive best between pH 4.5 to 8.3 (Environnement Canada, 1999; Cayuela et al., 2007). The plants are found in temperate climates and serve as an important food source for various water birds and fish (Drost et al., 2007). Some studies indicate that duckweed plants are sensitive to toxicity. Other studies however, report that duckweed plants are tolerant to environmental toxicity (Wang, 1990). To assess the tolerance of the species L. gibba to heavy metals, plants were exposed to concentrations of copper and nickel higher than those used in medium cultures. Toxic effect of pollutant on duckweed is generally evaluated by phytotoxicity tests based on growth inhibition (Geoffroy et al., 2004). Copper and nickel were chosen as the metals for this study for a number of reasons. Their presence above trace levels in the environment is an indicator of industrial pollution. On the other hand, they are essential micronutrients for plants; copper is a structural and catalytic component of many proteins and enzymes involved in metabolic pathways (Teisseire & Vernet, 2000) and nickel has an important role in the urease and hydrogenase metabolism (Harish et al., 2008). However, when the concentration reaches a threshold value, these essential metals become first inhibitory and afterwards toxic. Copper is responsible for many alterations of the plant cell (respiration, photosynthesis, pigment synthesis and enzyme activity) (Teisseire & Vernet, 2000; Kanoun-Boul et al., 2009). Nickel inhibits

germination, chlorophyll production and proteins (Zhou et al., 2009) in plants; several animal experimental studies have shown an increased cancer incidence associated with chronic exposure to nickel. Each plant species has different resistance and tolerance levels to different contaminants (Kamal et al., 2004). Therefore, several studies have been performed to elucidate heavy metal toxicity to plants. In earlier study, we demonstrated that the duckweed L. gibba L. native to the north-east of Algeria tolerated Zn up to 18 mg/L and was effective in removing 60% of the metal pollutant from the nutrient medium (Khellaf & Zerdaoui, 2009). The present study investigates copper and nickel toxicity to duckweed to determine tolerance of this aquatic species to Cu and Ni. The study also investigates the potential accumulation of these two metals by the duckweed L. gibba L. The goal was to assess the possibility to use L. gibba L. for phytoremediation of Cu and Ni contamination in water.

1.3 . Standards of Disposal

1.4. Treatment methodology

1.5. Objective and scope of study

MATERIALS AND METHODS Preparation steps. Primary treated sewage water were transferred to the laboratory from the tertiary sewage water treatment plant after the preliminary sieving step to get rid of large suspended solids. The transferred water was immediately collected into two opaque tanks (as replicates) to prevent light entering except at the top (Parr et al., 2002), each tank with dimensions of 50 cm long, 35 cm wide and 25 cm deep and was filled with 25 L primary treated sewage water. Duckweed (Lemna gibba L.) plants ere collected from Ganabiet-Tersa drain (Fig. 1). The stock were cleaned by tap water then washed by distilled water inocula of Lemna plants were transferred to the water systems for aquatic treatment. The experiment was kept under outdoor local environmental conditions for eight days retention time. Water sampling. Subsurface (under duckweed mt) water samples for physico-chemical, biological and bacteriological parameters were collected in polyethylene bottles from all sides of each tank and then mixed. This procedure carried out every 2 days. Samples volume taken every two days for each of phytoplankton count and chlorophyll a determination was 100 mL. Parameters measured. Physico-chemical analyses (Table I) were carried out according to standard methods for e examination of water and wastewater (APHA, 1992). Field parameters (pH, conductivity & dissolved oxygen) were measured in situ using the multi-probe system (model Hydralab-Surveyor) and rechecked in laboratory using bench-top equipment to ensure data accuracy for biological parameters including total

coliform count and fecal coliform. count, phytoplankton identification and counting andchlorophyll a determination. Determination of duckweed growth rate. This was determined for fresh and dry weights. Samples of 20 cm2 area of Lemna plants were harvested periodically at the designated time periods (every 2 days) and filtered using filter papers then fresh weights were determined. These samples were then dried at 60oC for 48 h to a constant weight and then dry weights were calculated. Protien content. Duckweed organic nitrogen content was estimated at the beginning of the experiment and after 8 days retention time by using MicroKjeldahl method, then the obtained values were multiplied by 6.25 to obtain protein content values.