Peanut shell as natural adsorbent of wastewater treatment containing copper II sulfate (CuSO4) and its toxicological effects on oreochromis niloticus

THAI NGUYEN UNIVERSITY UNIVERSITY OF AGRICULTURE AND FORESTRY RIZKY GUSTIANI VIDYA PEANUT SHELL AS NATURAL ADSORBENT OF WASTEWATER TREATMENT CONTAINING COPPER-(II)-SULFATE (CuSO4) AND ITS TOXICOLOGICAL EFFECTS ON OREOCHROMIS NILOTICUS BACHELOR THESIS Study Mode Major Faculty Batch : Full-time : Environmental Science and Management : Advanced Education Program : 2014-2019 Thai Nguyen, 23/09/2019 Thai Nguyen University of Agriculture and Forestry Degree Program Bachelor of Environmental Science and Management Student name Rizky Gustiani Vidya Student ID DTN1454290074 Thesis Title Peanut Shell as Natural Adsorbent of Wastewater Treatment Containing Copper-II- Sulfate (CuSO4) and Its Toxicological Effects on Oreochromis niloticus Supervisor (s) Prof Nguyen The Hung Dr Ho Ngoc Son Supervisors’ Signatures Abstract: High levels of water pollution can destroy aquatic life Pollutants are taken up by plants and animals and can enter the human body, resulting in many health problems The situation is worse in countries where people not have access to potable water, and in many instances polluted water is used as a source of drinking water The present study has been conducted to determine peanut shell as natural adsorbent of wastewater treatment containing copper-(II)sulfate (CuSO4) and its toxicological effects on Nile Tilapia (Oreochromis niloticus) This study reported that the removal of copper-(II)- sulfate (CuSO4) from aqueous solution could be using pure and physically pretreated biomass from Arachis hypogea (peanut shell) which this material was indigenous, easily available, surplus by-products for biosorption studies The preliminary toxicity test was carried out with different concentration treatments to determine LC50 and it found to be 21.91 mg/L Adsorbent experiments used fish as bioindicators of heavy metal adsorption using concentration variations with treatments and replications The used concentrations of CuSO4 are 21.91 mg/L without adsorbent; 17.53 mg/L with adsorbent; 19.72 mg/L with adsorbent; 21.91 mg/L with adsorbent; 24.10 mg/L with adsorbent; 26.29 mg/L with adsorbent The results showed that Nile Tilapia mortality was still found in treatments using adsorbents accompanied by changes in fish behavior such as loss of equilibrium, change in body color, irregular i swimming activity, rapid jerk movement and aggressiveness before the fish died The adsorption capacity of peanut shell for adsorbent adsorption examination was effective during the contact time between pollutants and adsorbents and the absorption capacity occurred The observation of histopathological changes was also conducted on gill tissue using Hematoxylin and Eosin Staining Methods The observed changes in gills were congestion, curling bend of secondary lamellae, epithelial proliferation of secondary lamellae, epithelial rupture of secondary lamellae, fusion of secondary lamellae, and telangiectasia Thus, this study provided that peanut shell as natural adsorbents was effective to absorb water that had high content of copper Water pollution, peanut shell, Copper, Cu, Keywords: Copper-(II)-Sulfate, CuSO4, natural adsorbent, histopathology, Oreochromis niloticus, Nile Tilapia, toxicity, gills Number of pages: 64 Date of Submission: September 23rd, 2019 ii ACKNOWLEDGEMENT Alhamdulillah, I praise and thank Allah SWT for giving me the strength, knowledge, ability and opportunity to complete this research study and to persevere and accomplish it satisfactorily From bottom of my heart, I would like to express my deepest appreciation to all those who provided me the opportunity to complete this research First and foremost, I would like to express my sincere gratitude and deep regards to my supervisor: Assoc Prof Dr Nguyen The Hung of Thai Nguyen University of Agriculture and Forestry who guided me wholeheartedly when I implemented this Also, I want to express my thanks to Dr Ho Ngoc Son, the second supervisor, for his supervision, encouragement, advice, and guidance in writing this thesis Besides my supervisors, I would like to thank Dr.-phil Arinafril of Sriwijaya University, Indralaya, Indonesia and Ph.D Dr Krisna MURTI in Department of Anatomical Pathology, who kindly assisted me with the histopathological detection in this dissertation and was very patient with my knowledge gaps In addition, formal thanks should be offered to the Rector of Sriwijaya University, Prof Dr Ir H Anis Saggaf, MSCE, for granting my internship acceptance I would also like to acknowledge with much appreciation to the Dean of Faculty of Agriculture in Sriwijaya University, Prof Dr Ir Andy Mulyana, M Sc., who gave the permission to conduct my research in Faculty of Agriculture, Sriwijaya University iii Special thanks to Mochamad Syaifudin, S Pi., M Si., Dade Jubaedah, S Pi., M Si., and Dr Mohammad Amin, S Pi., M Si., from Budidaya Perairan, Faculty of Aquaculture, Sriwijaya University My sincere thanks also go to Mrs Nurhayani, Mrs Ana, Ms Naomi, other staffs and friends in Laboratory of Budidaya Perairan and Laboratory of Teknologi Hasil Perikanan, Sriwijaya University for helping and providing me necessary equipment as well as knowledge for fish anatomy My parents, Mohammad Burhanuddin and Yuliani, deserve special mention for their inseparable support and prayers Finally, special thanks to my friends for their love and moral support throughout my study and I would like to thank everybody who was important to the successful realization of this thesis, as well as expressing my apology that I could not mention personally one by one Thai Nguyen, September 23rd, 2019 Future Environmentalist, Rizky Gustiani Vidya iv TABLE OF CONTENT Acknowledgement iii TABLE OF CONTENT v List of Figures List of Tables PART I INTRODUCTION 1.1 Background and Rationale 1.2 Research’s Objectives 1.3 Research Questions and Hypotheses 1.4 Limitations PART II LITERATURE REVIEW 2.1 Copper 2.1.1 Copper-(II)-Sulfate 10 2.1.2 Toxic Effects of Copper-(II)-sulfate 13 2.2 Low-cost Adsorbent .13 2.2.1 Peanut Shell .15 2.3 Histopathological Effects .17 2.4 Test Species 19 PART III METHODS .21 3.1 Place and Time 21 3.2 Materials & Equipment 21 3.2.1 Toxicity Testing 21 3.2.2 Adsorbent Testing .21 3.2.3 Histopathology Examination .22 v 3.3 Preparation 23 3.3.1 Fish Preparation .23 3.3.2 Preparation of Cu2+ Solution 23 3.3.3 Adsorbent Preparation .24 3.4 Methods 24 3.4.1 Toxicity Testing 24 3.4.2 Adsorbent Experiment Using Fish as Bioindicator 24 3.4.3 Adsorbent Adsorption Examination 25 3.5 Histopathological Examination 26 3.5.1 Fixation .26 3.5.2 Tissue Processing 26 3.5.3 Sectioning 26 3.5.4 Staining procedure using Hematoxylin and Eosin 27 PART IV RESULTS AND DISCUSSION 28 4.1 Preliminary Toxicity Test 28 4.2 Adsorbent Experiment Using Fish as Bioindicator 30 4.3 Adsorbent Adsorption Examination 34 4.4 Histopathological Observation of Gills 36 PART V CONCLUSIONS .52 REFERENCES 54 APPENDICES 65 vi LIST OF FIGURES Figure Effect of Contact Time on the Absorption of Cu2+ Ion (mg/L) 35 Figure Normal histological structure of gills 37 Figure Copper-(II)-Sulfate (CuSO4) with concentration of 21.91 mg/L without adsorbent treatment Congestion (Cs); Epithelial proliferation of secondary lamellae (EP); Fusion of secondary lamellae (F); Epithelial rupture of secondary lamellae (ER); Telangiectasia (Tel); Curling bend of secondary lamellae (CB) H&E, magnification x100 .38 Figure Tilapia gills exposed to 17.53 mg/L Copper-(II)-Sulfate (CuSO4) with adsorbent treatment (4a); Tilapia gills evidencing fusion of secondary lamella (4b); Tilapia gills presenting curling bend of secondary lamella (4c); Tilapia gills showing epithelial proliferation of secondary lamellae in which secondary lamellae cells grow excessively (4d); A-D are stained with H&E, magnification of A is 100x while B, C, and D is 400x .41 Figure A) Copper-(II)-Sulfate (CuSO4) with concentration of 19.72 mg/L with adsorbent treatment H&E, magnification x100; B) Tilapia gills (O niloticus) showing congestion and fusion of secondary lamellae H&E, magnification x400; C) Tilapia gills evidencing epithelial proliferation of secondary lamellae H&E, magnification x400 .42 Figure A) Copper-(II)-Sulfate (CuSO4) with concentration of 21.91 mg/L with adsorbent treatment H&E, magnification x100; B) Tilapia gills presenting telangiectasia H&E, magnification x400; C) Tilapia gills showing disintegration of secondary lamellae H&E, magnification x400 43 Figure Tilapia gills exposed to 24.10 mg/L Copper-(II)-Sulfate (CuSO4) with adsorbent treatment (7a); Tilapia gills showing epithelial proliferation of secondary lamellae (7b); Tilapia gills evidencing fusion of secondary lamella (7c); Tilapia gills showing curling bend of secondary lamella (7d); A-D are stained with H&E, magnification of A is 100x while B, C, and D is 400x 45 Figure A) Copper-(II)-Sulfate (CuSO4) with concentration of 26.29 mg/L with adsorbent treatment H&E, magnification x100; B) Tilapia gills presenting curling bend of secondary lamellae and congestion H&E, magnification x400; C) Tilapia gills showing fusion of secondary lamellae H&E, magnification x400; D) Tilapia gills presenting epithelial proliferation of secondary lamellae H&E, magnification x400 47 LIST OF TABLES Table Physical and Chemical of Copper-(II)-Sulfate Material 12 Table The Toxic Effects of Copper-(II)-sulfate 13 Table Types of Adsorbents Based on Surface Area 14 Table Adsorption Capacities for Low-Cost Adsorbents 15 Table Chemical Composition of Peanut Shell 16 Table Preliminary Toxicity Test Result 28 Table The Value of Varied Concentrations 30 Table Effect of CuSO4 (Copper-(II)-sulfate) on Nile Tilapia in Five Replications 31 Table Effect of Copper-(II)-sulfate (CuSO4) to The Behaviour of Nile Tilapia in 24 Hours 32 Table 10 Effect of Contact Time with The Adsorbent Solution of Cu2+ Ion 35 Table 11 Gills Alterations 50 Aydın, H., Bulut, Y., & Yerlikaya, Ç (2008) Removal of copper (II) from aqueous solution by 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Natural Adsorbent of Wastewater Treatment Containing Copper- II- Sulfate (CuSO4) and Its Toxicological Effects on Oreochromis niloticus Supervisor (s) Prof Nguyen The Hung Dr Ho Ngoc Son Supervisors’... that peanut shell as natural adsorbents was effective to absorb water that had high content of copper Water pollution, peanut shell, Copper, Cu, Keywords: Copper- (II) -Sulfate, CuSO4, natural adsorbent, ... positive effects of using copper- (II) -sulfate on plants For example, the status of the soil, the frequency of application and the quantity of copper- (II) -sulfate will decide the effects of copper- (II) -sulfate