WASTEWATER TREATMENT USING MICROALGAE AZNA BINTI ABDUL AZIZ THESIS SUBMITTED TO FULLFILL THE REQUIREMENT FOR MASTER OF BIOTECHNOLOGY FACULTY OF SCIENCE UNIVERSITY OF MALAYA KUALA LUMPUR JANUARY 2013 Abstract Three species of selected microalgae isolated from Tasik Taman Jaya, Oscillatoria limnosa, Scenedesmus quadricauda and Pediastrum duplex, were cultured in Chu no. 10 defined medium. After the microalgae reached their optimum growth they were grown in wastewater to observe their capability to remove inorganic nutrients in wastewater. The microalgae showed steady increase in cell density for the duration of laboratory observation of eighteen days. As the cell density of the microalgae increased, both nitrate and phosphate concentration in wastewater decreased. This study showed that all the three species of microalgae have the ability to reduce the nutrients. Among them, Pediastrum duplex was most efficient in reducing inorganic nutrients in wastewater. It reduced 60% and 75% of the total nutrients of wastewater from Station A and Station B respectively. By using this biological wastewater treatment, less chemical and physical treatment is needed to improve wastewater quality. The biologically treated wastewater can then be discharged into the reservoirs, rivers or streams. ii Abstrak Tiga spesies mikroalga yang diasingkan dari Tasik Taman Jaya iaitu Oscillatoria limnosa, Scenedesmus quadricauda dan Pediastrum duplex telah dikulturkan di dalam media pertumbuhan Chu no. 10 di dalam makmal. Setelah mikroalga tersebut mencapai pertumbuhan yang optima, ia telah membiak di dalam air kumbahan, seterusnya menunjukkan keupayaannya untuk menyingkirkan nutrien tak organik dalam air sisa. Selepas 18 hari pemerhatian makmal dilakukan, ia menunjukkan peningkatan yang agak stabil dari sudut kepadatanya. Ketika kepadatan sel mengalami peningkatan, kandungan nitrat dan fosfat di dalam air sisa menunjukkan bacaan yang menurun. Dapatan dari kajian menunjukkan bahawa Pediastrum duplex ialah sel yang paling berkesan untuk mengurangkan kandungan nutrien tak organik di antara ketiga-tiga spesies mikroalga ini. Hasil dari bacaan makmal ke atas Stesen A dan B, ia menunjukkan bahawa sebanyak 60% dan 75% jumlah kandungan nutrien air sisa telah dikurangkan kepekatannya. Sehubungan itu, dengan mengaplikasikan kaedah rawatan secara biologi ke atas air sisa ia juga mampu mengurangkan kebergantungan ke atas kaedah rawatan secara kimia dan fizikal untuk memastikan kualiti air terjamin. Seterusnya, apabila proses rawatan secara biologi dilaksanakan maka penyaliran air sisa ke dalam kawasan reserviour dan sungai dapat disalirkan. iii ACKNOWLEDGEMENT In the name of Allah, The Most Gracious, Most Merciful. Praise is to Allah, The Cherisher and Sustainers of the Worlds for without Him nothing is possible. This dissertation would not have been possible without the assistance, support and guidance from a number of people. Thus, I would like to express my deepest appreciation and gratitude to those individuals. First of all, I would like to give my thankful to my supervisor Prof. Datin Dr. Aishah Salleh for her helpful and give me ideas to build up my project. Thanks are also to my lab mate at MUCED; Kak Nadia, Kak Zan, Sarini and Diana as they help me in my lab work and identification of the microalgae. Nevertheless, to Saiful, the lab assistant, in helping me providing the sampling apparatus and lab apparatus. To my beloved family (abah, mak, angah, cik and adik) thank you so much for supporting me during my studies and during my research. Nothing is more precious than a family love. And also to my husband, Mohd Hanafia thank you for everything as you never gives up supporting me and always gives a motivational advice to me……… AZNA BINTI ABDUL AZIZ SGF 040026 2012/2013 iv List of Contents Abstracts ii Abstrak iii Acknowledgement iv List of Content v List of Figures viii List of Tables ix List of Plate xi List of Abbreviation xii List of Symbols xiii List of Appendices xiv Chapter 1 1 :Introduction 1 1.1 : Objectives 5 Chapter 2 2 : Literature review 6 2.1 : Water Pollution 6 2.2 : Eutrophication 11 2.3 : Phytoplankton 15 2.3.1) Growth Factors for Phytoplankton 20 2.4 : Microalgae and Environment 25 2.5 : Application of microalgae in wastewater using various way of treatment 28 v Chapter 3 3 : Methodology 31 3.1 : Tasik Taman Jaya and collection of water sample 31 3.2 : Microalgae identification 35 3.3 : Water Quality 35 a) Nitrate 36 b) Phosphate 36 c) Silica 36 3.3.1 : Water quality data analysis 37 3.4 : Culture Preparation 37 3.4.1) Media 37 3.4.2) Isolation 38 3.4.3) Culture Method 39 3.5 : Nutrients Removal 39 3.6 : Growth 40 Chapter 4 4 : Results and Discussion 41 4.1 : Species of microalgae at Tasik Taman Jaya 41 4.2 : Water quality analysis 42 4.2.1) Nitrate Removal 47 4.2.2) Phosphate Removal 48 4.2.3) Silica Removal 49 vi 4.3 : Growth 50 4.4 : Relation between growth of wastewater and nutrient removal 53 Chapter 5 5 : Conclusion 66 References 67 vii List of Figures Figure 4.1 : Growth of Oscillatoria limnosa in water sample from Station 51 A and Station B during the experiment period Figure 4.2 : Growth of Scenedesmus quadricauda in water sample from 52 Station A and Station B during the experiment period Figure 4.3 : Growth of Pediastrum duplex in water sample from Station A 53 and Station B during the experiment period Figure 4.4 : Nutrients reduction vs. growth of Oscillatoria limnosa in 55 water sample from Station A Figure 4.5 : Nutrients reduction vs. growth of Oscillatoria limnosa in 56 water sample from Station A Figure 4.6 : Nutrients reduction vs. growth of Pediastrum duplex in water 58 sample from Station B Figure 4.7 : Nutrients reduction vs. growth of Pediastrum duplex in water 59 sample from Station A Figure 4.8 : Nutrients reduction vs. growth of Scenedesmus quadricauda 61 in water sample from Station B Figure 4.9 : Nutrients reduction vs. growth of Scenedesmus quadricauda 62 in water sample from Station A viii List of Tables Table 2.1 : Type of pollutants 7 Table 2.2 : The physical characteristics of wastewater 11 Table 2.3 : The general characteristic of oligotrophic and eutrophic lake 12 Table 2.4 : Type of nutrition found in the algae 17 Table 4.1 : Microalgae composition in Tasik Taman Jaya 42 Table 4.2 : Nutrients concentration of water collected from Station A and 43 inoculated with Oscillatoria limnosa Table 4.3 : Nutrients concentration of water collected from Station A and 43 inoculated with Scenedesmus quadricauda Table 4.4 : Nutrients concentration of water collected from Station A and 43 inoculated with Pediastrum duplex Table 4.5 : Nutrients concentration of water collected from Station A 44 without inoculated Table 4.6 : Nutrients concentration of water collected from Station B and 44 inoculated with Oscillatoria limnosa Table 4.7 : Nutrients concentration of water collected from Station B and 44 inoculated with Scenedesmus quadricauda Table 4.8 : Nutrients concentration of water collected from Station B and 45 inoculated with Pediastrum duplex Table 4.9 : Nutrients concentration of water collected from B without 45 inoculated Table 4.10 : Amount of nitrate in water sample from Station A 45 Table 4.11 : Amount of phosphate in water sample from Station A 46 Table 4.12 : Amount of silica in water sample from Station A 46 ix Table 4.13 : Amount of nitrate in water sample from Station B 46 Table 4.14 : Amount of phosphate in water sample from Station B 47 Table 4.15 : Amount of silica in water sample from Station B 47 x