Use of rice husk as natural adsorbent to treat wastewater containing iron II sulfate and its toxicity test on nile tilapia

THAI NGUYEN UNIVERSITY UNIVERSITY OF AGRICULTURE AND FORESTRY DEASY AMALIA SARI BACHELOR THESIS USE OF RICE HUSK AS NATURAL ADSORBENT TO TREAT WASTEWATER CONTAINING IRON II SULFATE AND ITS TOXICITY TEST ON NILE TILAPIA Study Mode : Full-Time Major : Environmental Science and Management Faculty : Advanced Education Program Batch : 2014-2018 Thai Nguyen, September 2018 DOCUMENTATION PAGE WITH ABSTRACT Thai Nguyen University of Agriculture and Forestry Degree Program Bachelor of Environmental Science and Management Student Name Deasy Amalia Sari Student ID DTN1454290076 Thesis Title Use of Rice Husk as Natural Adsorbent to Treat Wastewater Containing Iron II Sulfate and Its Toxicity Test on Nile Tilapia Supervisor (s) Dr.-phil Dipl.-Ing.agr Arinafril Prof Tran Van Dien Abstract : The problem of clean water now becoming a global concern Water is a source of life and is used in various ways such as for cooking, bathing, washing, farming, and other activities The needs of clean water become difficult to be fulfilled due to the presence of water problems One of the water problems occur is water pollution where the water contains harmful substances such as heavy metals and the number is exceeds the maximum contaminant level of iron that allowed in the water One of heavy metals that often found in the water is iron (Fe) Maximum allowed iron content in the water is by not exceeding 0.1mg/L High iron content in the water makes turbid water has smells, causes rust on household appliances, and it affects to the health problems Histopathology examination on fish gill indicates that excessive levels of Fe can damage the tissues of the fish gills The alterations detected were as Telangiectasia, Fusion of Secondary Lamellae, Epithelial Proliferation of Secondary Lamellae and Congestion Treatment to the water is must be carried out in order to decrease the excess iron content However, the processing is quite difficult to and requires a large cost The use of rice husk is a good alternative way to i treat the water containing high iron content Rice husk as a natural adsorbent has the ability to absorb heavy metals Adsorbent test treatment resulted in the decrease of iron levels that contain in the water It showed that rice husk as a natural adsorbent has the ability to absorb Fe ions and effective in handling water problems including water that contains high of iron content Rice husk, bioadsorbent, water pollution, histopathology, Keywords Oreochromis niloticus Number of Pages 46 Date of Submission Supervisor’s Signature ii ACKNOWLEDGEMENT From bottom of my heart, I would like to express my deepest and solid gratitude to ALLAH SWT, Almighty God for giving me the strength and courage as the time gone by, for fading away doubts and leading me to finish this bachelor thesis I would like to express my sincere gratitude and deep regards to my supervisor: Dr.-phil Arinafril of Sriwijaya University, Indralaya, Indonesia, who guided me wholeheartedly when I conducted this research, gave additional knowledge, worthy indications, comments and guidance from the start to the end of this study I also want to thank my second supervisor, Prof Tran Van Dien, for his supervision, encouragement, advice, and guidance in writing this thesis Likewise, I would like to thank Ph.D Dr Krisna Murti in Department of Anatomical Pathology, who was very patiently assisted me with the histopathology examination for this study Furthermore, an acknowledgement also goes to the Rector of Sriwijaya University, Prof Dr Ir H Anis Saggaf, MSCE., for acknowledging the internship acceptance I would also like to say thank to the Dean of Faculty of Agriculture in Sriwijaya University, Prof Dr Ir Andy Mulyana, M Sc., who gave me the permission to use all the necessary facilities for the experiment that has been conducted at Aquaculture Laboratory and Fish Product Technology Laboratory, Faculty of Agriculture, Sriwijaya University, Inderalaya Campus In addition, to Prof Dr Mohammad Amin, S.Pi., M.Si., Prof Mochamad Syaifudin, S.Pi., M.Si., PhD, and Prof Dade Jubaedah, S.Pi., M.Si., from Budidaya Perairan, Faculty of Aquaculture, Sriwijaya University, Inderalaya Campus Thank you for the additional knowledge and guidance during the experimental time iii I also place my gratitude to Mrs Nurhayani, Mrs Ana, Ms Naomi, other staffs and friends in Aquaculture Laboratory and Fish Product Technology Laboratory, Sriwijaya University Thank you for helping, providing me necessary equipment as well as teaching additional knowledge during my experiment at the laboratories The unconditional love from my family, to my parents, Hidayat and Maydaria, my elder sister, Dewi Permatasari, my elder brother Muhammad Alfarizi, and my younger sister, Metha Maya Sari, who gave me strength, support and positive thoughts to finish what I have started and lead me to the successful completion of this study Nevertheless, I would like to thank my best friends – Nur, Givanni, Rani, Annisa, Ara, Dhiya and Ica; my bachelor thesis mates – Amana and Vidya; and all Indonesian friends in the Vietnam Thank you for the unending support and giving the positive thoughts until I finish this study Thai Nguyen, 24th September, 2018 Student Deasy Amalia Sari iv TABLE OF CONTENT ACKNOWLEDGEMENT iii LIST OF FIGURES LIST OF TABLES PART I INTRODUCTION 1.1 Background and Rationale 1.2 Objectives 1.3 Research Questions and Hypotheses 1.3.1 Research Questions 1.3.2 Hypotheses 1.4 Limitations PART II LITERATURE REVIEW 2.1 Iron 2.1.1 Iron (II) Sulfate 2.2 Rice Husk 2.3 Natural Adsorbent 2.3.1 Rice Husk as Adsorbent 10 2.4 Test Species – Oreochromis niloticus 10 2.5 Histopathological Effects 11 PART III METHODOLOGY 13 3.1 Place and Time 13 3.2 Equipment and Materials 13 3.3 Fish preparation, Preparation of Fe2+ Solution and Adsorbent Preparation 14 3.3.1 Fish Preparation 14 3.3.2 Preparation of Fe2+ Solution 14 3.4 Methods 15 3.4.1 Toxicity Experiment 15 3.4.2 Heavy Metal Adsorption Test using Fish as Indicator 15 3.4.3 Adsorbent Adsorption Examination 15 3.4.4 Histopathology Examination 16 PART IV RESULTS AND DISCUSSIONS 18 4.1 Preliminary Toxicity Test 18 4.2 Heavy Metal Adsorption Test using Fish as Indicator 19 4.3 Adsorbent Adsorption Examination 22 4.4 Histopathological Observation of Fish Gills 24 v PART V CONCLUSION 36 REFERENCES 37 APPENDICES 42 vi LIST OF FIGURES Figure Adsorbent Adsorption Result 23 Figure Normal structure of gills 24 Figure Fish gill exposed to Iron (II)- sulfate (FeSO4) with concentration of 5.821mg/L without adsorbent treatment (x100) 25 Figure Fish gill exposed to 5.821mg/L iron (II)- sulfate (FeSO4) with adsorbent treatment (x100) .26 Figure Fish gill exposed to 5.2389 mg/L iron (II)- sulfate (FeSO4) with adsorbent treatment (x100) .28 Figure Fish gill exposed to 4.6568mg/L iron (II)- sulfate (FeSO4) with adsorbent treatment (x100) .29 Figure Fish gill exposed to 6.4031mg/L iron (II)- sulfate (FeSO4) with adsorbent treatment (x100) .30 Figure Fish gill exposed to to 6.9852mg/L iron (II)- sulfate (FeSO4) with adsorbent treatment (x100) .32 LIST OF TABLES Table Iron (II) Sulfate Property Table Iron (II) Sulfate Potential Health Effects Table Rice Husk Property Table List of Equipment Used 13 Table List of Material Used 14 Table Preliminary Toxicity Test Result 18 Table Fish Mortality 20 Table Effect of Iron (II) Sulfate towards Nile Tilapia in 24 hours 21 Table Adsorbent Adsorption Examination Result 23 Table 10 Gills Alterations 35 PART I INTRODUCTION 1.1 Background and Rationale Heavy metals that lead to the water pollution is currently becoming a global concern Lack of clean water is often found in some places in the world Needs of clean water must be fulfilled and one of clean water sources is by using ground water that obtained from well and borehole water However, deficiency of using groundwater commonly can cause several ailments to human health immediately while the contaminants of gas as well as mineral exceeds the maximum allowed level (Suharno, 2017) The presence of dissolved iron in the water can affect the stinking metal flavor, cause the growth of iron bacteria, create reddish-brown staining on clothing, bathtub and other equipment, or even blockage on pipelines In addition, iron that has a concentration greater than 25 mg/L will give a sense of metallic, astrinogent or medicine taste (Dharma, 2002) Maximum contaminant level of Iron that allowed in the water is 0.3mg/L (U.S Environmental Protection Agency, 2013) Some heavy metals removal technologies such as chemical precipitation, reverse osmosis, ion exchange, ultrafiltration, electrodialysis, as well as phytoremediation are generally used for industrial wastewater management Nevertheless, these heavy metals removal methods are usually inexpensive and inadvisable (Ahalya, Ramachandra and Kanamadi, 2003; Yus and Mashitah, 2014) Cost effectiveness is the main attraction of metal biosorption, and it should be kept that way The use of bioadsorben as natural adsorbent material is one of the effective ways to treat wastewater containing hazardous compound (Viera and Volesky, 2000) One of natural bioadsorbents is Rice husk Rice husk is a product in the rice milling industry It is also one of the most important agricultural residues in a big quantity amount On weight basis of the whole rice, it represents around 20% of the whole rice produced (Daifullah, Girgis and Gad, 2003) In the study by Kumar and Bandyopandhyay (2005), rice husk has been reported for their ability to bind metal ions The use of rice husk which is not utilized after the harvest time as a natural adsorbent is a good alternative way to treat wastewater problems Iron (II)- Sulfate (FeSO4) 6.9852mg/L with Adsorbent Treatment Figure Fish gill exposed to to 6.9852mg/L iron (II)- sulfate (FeSO4) with adsorbent treatment; Congestion (Cs); Epithelial proliferation of secondary lamellae (EP); Fusion of Secondary lamellae (F) magnification x100 a a Congestion (Cs) Magnification x400 b b Epithelial proliferation of secondary lamellae (EP) Magnification x400 32 c c Fusion of secondary lamellae (F) Magnification x400 In figure 8, with 6.9852mg/L of iron (II)- sulfate (FeSO4) and adsorbent treatment, the histopathological changes observed in gill of treated fish shows adverse disorders such as congestion, epithelial proliferation of secondary lamellae and fusion of secondary lamellae The histopathological section in O.niloticus considered to understand and recognize direct evidence referring to any adverse effect in the structural organs on fish due to environmental pollution Meanwhile, fish gill is a good indicator of aquatic environmental pollution, where it is as the only respiratory system of the fish The gill epithelium is the controlling site of gas ex-change, ionic-regulation, balance of acidbase, and nitro-genous waste excretion for fishes (Houlihan, Rankin and Shuttleworth, 1982) Gills having important functions to the fish where it plays role in the process of respiration, osmoregulation and excretion, it remains as well as the indicator to external environment that has high sensitivity to changes in the quality of the surrounding area, thus are deliberated as the main target of chemical and other contaminants (Fernandez & Mazon, 2003) Through histopathological examination, gills of fish that exposed to iron (II)sulfate (FeSO4) at different concentration showed the morphological changes and structural damage Most of the histopathological alteration displayed on this research was telangiectasia ( Figure 3, 4, 7), telangiectasia is a change in gills which arising due to physical and chemical trauma and a condition which rupture of the cell at the end of 33 the lamellae and the secondary lamellae (Robert, 2001) The breakage of lacuna due to rupture of pillar cells that causes the buildup of red blood cells in secondary lamella is affected by the water that contains high heavy metal content whose the content is exceeds the allowed standard of iron in the water The iron discharge will build extensive damage comprise of clogging action and block the respiration system of fishes (EPA, 1993) Fish gills as a respirator organ is high risk to the danger of water containing heavy metals considering it is the organ that contacts most quickly to the water Several damages are shown from the examination and one of it was congestion (Figure 3, 5, 8) Congestion is a condition in which the erythrocytes (red blood cells) are stuck in the blood vessels and causing a clumps or dots at the end of the secondary lamellae (Figure 3.1, 5.1, 8.1) Another changes detected was epithelial proliferation of secondary lamellae (Figure 3, 4, 5, 7, 8) and fusion of secondary lamellae (Figure 3, 4, 5, 6, 7, 8) The epithelium on each branchial arch separated within dorsoventral columns of filaments where right angles to the branchial arches are flattened, with dorsal as well as ventral rows of secondary lamellae have more development on each filament, stretching out at right angles to the filamental side (Figure 3.4, 4.1, 5.2, 7.2, 8.2) Secondary lamellae is the place of gas exchange, upon blood to water diffusion distances lower than one micrometer in active species and 1-10 µm in other stagnant fish species (Hughes, 1984) While secondary lamellae fusion is a condition in which lamellae epithelial cells are grow rapidly (hyperplasia) so that the lamellae of gills becomes fused (Figure 3.1, 4.3, 5.2, 6.1, 6.2, 7.3, 8.3) The same case happened on the observation of Hadi and Alwan (2012) in fish gill under the exposure of Alluminium (Al) appeared in several changes of histopathological alterations such as cellular hypertrophy (hyperplasia) in the epithelial row of primary filaments as well as fusion of secondary lamellae Given these structural changes shown that heavy metals produce profound effects on fish gill 34 Table 10 Gills Alterations Treatment Control (without Adsorbent Treatment) Control Control (with Adsorbent Treatment) C – 10% (with Adsorbent Treatment) C – 20% (with Adsorbent Treatment) C + 10% (with Adsorbent Treatment) C + 20% (with Adsorbent Treatment) (•) ■ Telangiectasia •••• Gills Alteration Epithelial Proliferation Congestion of Secondary Lamellae •• • •• • Fusion of Secondary Lamellae •• • • •• • •••• •• ••• • • •• •• : Alterations detected and how many times it was recorded (per slide) : No alterations detected From the table above, it can be seen that the histopathological changes were found in every level of concentration Thus, histopathology examination in all the tissues indicates that iron (II)-sulfate (FeSO4) is toxic to fish and the lower concentration of iron (II)-sulfate (FeSO4) affect to the smaller number of alterations 35 PART V CONCLUSION Histopathology examination to the gills as respiratory system of fish provides evidence that water containing heavy metals has an effect to the change of fish gills Through the examination, changes and damage of the gills can be detected These changes can be mild and even severe damage, depending on how much the exposure to heavy metals is in the environment around the fish The damages found were telangiectasia, congestion, epithelial proliferation of secondary lamellae, and fusion of secondary lamellae Alterations detected are proving if the exposure to iron (II)sulfate (FeSO4) give result in change in gills histology of O niloticus before and after the adsorbent is given In addition, results from atomic absorption spectroscopy (AAS) show that rice husk as natural adsorbents are able to absorb heavy metals for above 30% from the total ion content at its maximum contact time between 60 to 90 minutes of shaken process Decrease in heavy metal content in the water can reduce the risk of death of the fish or may prolong the life of the fish Natural adsorbent can be used as one effective way to treat polluted water considering the materials used are easily to be found It is conclude that rice husk is 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International Microbiology Vol: Yus, A.Y and Mashitah, M.D (2014) Pycnoporus sanguineus as Potential Bioadsorbent for Heavy Metal Removal from Aqueous Solution: A Review School of Chemical 41 APPENDICES 42 1.1 Probit Analysis Confidence Limits 95% Confidence Limits for concentration Probability PROBIT Estimate Lower Bound Upper Bound 0.01 -4.310 0.02 -3.186 0.03 -2.473 0.04 -1.937 0.05 -1.500 0.06 -1.129 0.07 -.803 0.08 -.512 0.09 -.247 0.1 -.003 0.15 1.008 0.2 1.811 0.25 2.500 0.3 3.119 0.35 3.693 0.4 4.237 0.45 4.763 0.5 5.281 0.55 5.799 0.6 6.326 0.65 6.870 0.7 7.443 0.75 8.062 0.8 8.751 0.85 9.555 0.9 10.565 0.91 10.809 0.92 11.074 0.93 11.366 0.94 11.692 0.95 12.063 0.96 12.499 0.97 13.036 0.98 13.749 0.99 14.873 43 Rice Husk before Activated Rice Husk After Activated Weighing of FeSO4 pro analyst FeSO4 Solution 44 Aquarium Condition after Aquarium Condition after 24hour Adsorbent Were added to the Water Separating Process Between Rice Fish Gill Slides after Husk and FeSO4 Solution Histopathological Processes 45 Microscope for Histopathology Histopathology Identification Examination using Microscope 46 ... Thesis Title Use of Rice Husk as Natural Adsorbent to Treat Wastewater Containing Iron II Sulfate and Its Toxicity Test on Nile Tilapia Supervisor (s) Dr.-phil Dipl.-Ing.agr Arinafril Prof Tran Van... alternative way to i treat the water containing high iron content Rice husk as a natural adsorbent has the ability to absorb heavy metals Adsorbent test treatment resulted in the decrease of iron levels... adsorption of heavy metal (Fe) onto rice husk ash grams of rice husk ash were weighed into 250mL erlenmeyer 100mL of * Personal Communication 15 the solution contains 0.87mL of ion FeSO4 were mixed to