Tài liệu 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 do 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 1. Supervisor’s Signature 2. 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 ............................................................................................................................ 1 LIST OF TABLES .............................................................................................................................. 2 PART I. INTRODUCTION ................................................................................................................ 3 1.1. Background and Rationale ................................................................................................... 3 1.2. Objectives ........................................................................................................................... 4 1.3. Research Questions and Hypotheses .................................................................................... 4 1.3.1. Research Questions ........................................................................................................... 4 1.3.2. Hypotheses ........................................................................................................................ 5 1.4. Limitations .......................................................................................................................... 5 PART II. LITERATURE REVIEW..................................................................................................... 6 2.1. Iron ..................................................................................................................................... 6 2.1.1. Iron (II) Sulfate ................................................................................................................ 7 2.2. Rice Husk ............................................................................................................................ 8 2.3. Natural Adsorbent ............................................................................................................... 9 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 1. Adsorbent Adsorption Result .................................................................23 Figure 2. Normal structure of gills ........................................................................24 Figure 3. Fish gill exposed to Iron (II)- sulfate (FeSO4) with concentration of 5.821mg/L without adsorbent treatment (x100) ......................................25 Figure 4. Fish gill exposed to 5.821mg/L iron (II)- sulfate (FeSO4) with adsorbent treatment (x100) .....................................................................26 Figure 5. Fish gill exposed to 5.2389 mg/L iron (II)- sulfate (FeSO4) with adsorbent treatment (x100) .....................................................................28 Figure 6. Fish gill exposed to 4.6568mg/L iron (II)- sulfate (FeSO4) with adsorbent treatment (x100) .....................................................................29 Figure 7. Fish gill exposed to 6.4031mg/L iron (II)- sulfate (FeSO4) with adsorbent treatment (x100) .....................................................................30 Figure 8. Fish gill exposed to to 6.9852mg/L iron (II)- sulfate (FeSO4) with adsorbent treatment (x100) .....................................................................32 LIST OF TABLES Table 1. Iron (II) Sulfate Property ................................................................................7 Table 2. Iron (II) Sulfate Potential Health Effects ........................................................8 Table 3. Rice Husk Property ........................................................................................9 Table 4. List of Equipment Used ................................................................................ 13 Table 5. List of Material Used .................................................................................... 14 Table 6. Preliminary Toxicity Test Result .................................................................. 18 Table 7. Fish Mortality............................................................................................... 20 Table 8. Effect of Iron (II) Sulfate towards Nile Tilapia in 24 hours ........................... 21 Table 9. Adsorbent Adsorption Examination Result ................................................... 23 Table 10. Gills Alterations ......................................................................................... 35 2 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. 3 Biomass composed of cellulose, hemicellulose, protein, fat and lignin, can be used as bioadsorbents. It composed of active groups in the form of phenolic, hydroxy, carboxyl, amine, phosphate groups, which can be a simple treatmeant or as adsorbents which have the ability to bind heavy metal ions. (Kim et al., 2005; Jhonson, Jain and Prasad, 2008 cited in Lesmana et al., 2009). A huge amount of rice husk usually burnt by in situ processs, produce CO2 and pollution in many forms. Hence, the use of rice husk not individually would provide a low cost sorbent in activated carbon nor synthetic in ion exchange and easily to find, but also maintain the environment from numerous pollution problems (Kumar, 2009). The aim of this study is to investigate the effectiveness of using rice husk as natural adsorbent to treat wastewater containing iron (II) sulfate by conducting the toxicity test on Nile Tilapia fish. To see how much the iron (II) sulfate that can be adsorbed by the natural adsorbent through the adsorbent adsorption examination. To determine the damage that might be occured by the pollutan (Fe), histopathological examination will be conducted on fish gills organ. 1.2. Objectives This study was designed to assess the toxicological effects of iron (II)- sulfate (FeSO4) on Oreochromis niloticus before and after the adsorbent was given as well as to reveal the histopathological alterations on its gills. 1.3. Research Questions and Hypotheses 1.3.1. Research Questions The study was conducted to answer specific questions: 1. How does iron (II)- sulfate (FeSO4) affect the gills of Nile Tilapia before the adsorbent is given? 2. How does iron (II)- sulfate (FeSO4) affect the gills of Nile Tilapia after the adsorbent is given? 3. How is the effectiveness of rice husk for the removal of iron (II)- sulfate (FeSO4)? 4 1.3.2. Hypotheses Hypotheses 1: H0 (Null Hypotheses): Exposure to iron (II)- sulfate (FeSO4) will not result in change in gills histology of Nile Tilapia after the adsorbent is given. HA (Alternative Hypotheses) : Exposure to iron (II)- sulfate (FeSO4) will result in change in gills histology of Nile Tilapia after the adsorbent is given. Hypotheses 2: H0 (Null Hypotheses): Exposure to iron (II)- sulfate (FeSO4) will not result in change in gills histology of Nile Tilapia before the adsorbent is given. HA (Alternative Hypotheses) : Exposure to iron (II)- sulfate (FeSO4) will result in change in gills histology of Nile Tilapia before the adsorbent is given. Hypotheses 3: H0 (Null Hypotheses): Rice husk is not effective for the removal of iron (II)sulfate (FeSO4). HA (Alternative Hypotheses): Rice husk is effective for the removal of iron (II)sulfate (FeSO4). 1.4. Limitations There is a lack of experimental methodology of adsorbent testing directly to the O. niloticus as well as lack of experimental results to the histopathological effects on gills tissues of fish of iron (II)- sulfate (FeSO4) in the literature. 5 PART II. LITERATURE REVIEW 2.1. Iron Iron is the second most plentiful metal in the earthh's crust. Iron elements is barely presence in nature, and usually in the form of ions Fe2+ and Fe3+ that immediately combine with oxygen as well as sulfur containing compounds as an accumulator of oxides, sulfides carbonate and hydroxides. Iron is often generally found in nature, as its oxides forming (Elinder, 1986; Knepper, 1981). The presence of iron in the environment is constant and as necessary component to the animal as well as human. Applied for the synthesis of hemoglobin and myoglobin in the muscle. Requirement of iron in the daily basis is provided from food. When inadequacy disorders are happened, it can be treated with the use of iron salts as a remedy. Iron can be found in almost all organs as well as tissues, in addition the largest amount of iron are stocked in the liver and spleen. Iron salts captured verbally do not poison, nevertheless can also deliver side effects such as obstipation and vomiting (Beata, 2014). An analysis of iron toxicity to the aquatic plants specifically on rice, stated that the growth of aquatic reed species was found prevented by concentration of 1mg/L of total iron (Phippen et al., 2008). The function of iron toxicity within rice involve excessive of Fe2+ from roots, acropetal translocation to the leaves, changing color of rice leaves as well as yield loss (Becker and Asch, 2005). An excessively level of iron infilrate into the human body move across the rate-limiting absorption phase and develop into saturated. Those free irons get through physically into cells of the brain, heart and liver. Whereas the disorder of oxidative phosphorylation from free iron, ferrous iron is transformed to ferric irom that unbind hydrogen ions, hence enlarging metabolic acidity. The free iron might also becomes lipid peroxidation that results in particular loss to mitochondria, microsomes and other cellular organelles (Albretsen, 2006). Iron toxicity on cells has persuaded to iron mediated tissue injuries including cellular oxidizing and decreasing mechanisms and its toxicity against intracelular organelles like mitochondria and lyso-somes. An expansive of free radicals which are able to cause capability cellular damage are delivered by plenty intake of iron. 6 Hydrogen free radicals that produced by iron are able to attack DNA, lead to cellular damage, mutation as well as malignant transformations that will cause various disease (Grazuleviciene et al., 2009). Heavy metals are usually found in both environment and diet. While the amounts are in allowed limits, they are necessary for maintaning good health. In addition, if the amounts presence are above its allowed limits, they might become dangerously toxic. Heavy metal toxicity will decrease energy levels as well as damage the function of brain, lungs, liver, kidney, blood structure and other important organs. If the exposure is in a long-term period of time, that might lead to gradually physical progressing, muscular, as well as neurological degenerative practices and affect several diseases such as multiple sclerosis, Parkinson's disease, Alzheimer's disease and muscular dystrophy. Continuosly exposure of some metals and its compounds might even lead to cancer (Jarup, 2003). 2.1.1. Iron (II) Sulfate Iron (II) sulfate or Ferrous Sulfate is a greenish or yellow-brown crystalline solid from synthetic origin and belongs to the pharmacological groups known as hematological agents and iron salts. The main hazard is the threat to the environment (“DrugBank,” n.d; “Cameo Chemicals,” n.d). Stabilized of iron (II) sulfate has a natural composition such as ferrous sulfate stabilized with glycine and malic acid (Salgueiro, Torti and Meseri, 2007). Iron (II) sulfate property as well as its potential health effects displayed in Table 1 and Table 2. Table 1. Iron (II) Sulfate Property Chemical Name Common Name CAS NO. Molecular Weight Chemical Formula Appearance Solubility Density Boiling Point Melting Point Hazardous Iron (II) sulfate (1:1), heptahydrate; sulfuric acid, iron (2+) salt (1:1), heptahydrate. Ferrous Sulfate 7720-78-7 278 FeSO4 Blue green crystals 48.6 g/100 g water @ 50C (122F) 1.90 > 300°C (> 572°F) Decomposes 57°C (135°F) Loses water Yes (Source: MSDS, 2000) 7 Table 2. Iron (II) Sulfate Potential Health Effects Type of Health Effect Inhalation Ingestion Skin Contact Eye Contact Description First Aid Measures Irritation in the respiratory tract. Release to fresh air. Whenever Symptoms might include coughing breathing is difficult, give an and shortness of breath. artificial respiration and oxygen. Get medical attention for further aid. Low toxicity in small number but Generate vomiting immediately higher dosages may cause nausea, as directed by medical vomiting, diarrhea, and black stool. personnel. Do not give anything Pink urine discoloration indicates by mouth to an unconscious strong iron poisoning. Lead to liver person. Get medical attention for damage, coma, and death due to iron further medication. poisoning. Children are more vulnerable to the iron toxic at smaller concentration. Skin irritation. Several symptoms such Rinse skin immediately with as redness, itching, and pain. plenty of soap and running water for at least 15 minutes. Change contaminated clothing and shoes. Get medical attention. Wash clothes and shoes thoroughly before reuse. Irritation, redness, and pain. Flush eyes directly with plenty of water for at least 15 minutes, lifting lower and upper eyelids occasionally and get medical attention for further medication. (Source: MSDS, 2000) 2.2. Rice Husk Rice husk is a product in the rice milling industry and it is one of the most largely agricultural wastes in rice producing. It is approximately 600 million tons of rice paddy is produced globally for each year. It represents on average 20% of the rice paddy produced is husk, annualy creating 120 million tones of total production (Giddel and Jivan, 2007). The outer part of rice husk structured by dentate rectangular elements, which are generally composed of silica coated with a thick cuticle and surface hairs. The inside part and inner epidermis are contain of silica. Besides in agricultural side, rice husk can be applied in industrial processes as well. The applications are including steel foundries, for houses manufacture material and refractory bricks (Bronzeoak, 2003). 8 Rice husk typical analysis shown in Table 1. The composition of each property is depends on rice variety, soil content, climate altitude, as well as geographical localized of the cultivation (Agus, 2002). Table 3. Rice Husk Property Property Bulk density (kg/m3) Hardness (Mohr’s scale) Ash (%) Carbon (%) Hydrogen (%) Oxygen (%) Nitrogen (%) Sulphur (%) Moisture Range 96-160 5-6 22-29 ≈ 35 4-5 31-37 0.23-0.32 0.04-0.08 8-9 (Source: Muthadhi and Kothandaraman, 2007) Considering to the rice husk characteristic properties and chemical composition, it has been found about the application of rice husk on the side of construction, energy production, production of various chemical product, and et cetera (Valchev et al., 2009). Burning process towards rice husk at high temperature will convert rice husk to another form which is rice husk ash that contains silica. Rice husk ash does not only provide sufficient benefits, but it also has significant market value. It has been demonstrated that 85% to 95% amorphous silica can be converted from it (Krishnarao and Subramanian, 2001). 2.3. Natural Adsorbent Adsorbent is a solid substance that has an ability to adsorb other subtances from one to another surface without any covalent bonding. Example of natural adsorbents are activated charcoal, rice husk, penaut shell, and et cetera (Miller, 2003). Several studies have been showed the ability of aqueous solutions such as orange and apple residue, rice husk, peanut and coconut shell, as well as tea are effective in removal toxic substances (Alagumuthu and Rajan, 2010). Several low cost adsorbent resulting from natural and and anthropogenic sources have been investigated for treatment of waste water contaminated by heavy metals. Generally, 9 adsorbent used are natural materials from agricultural waste, industrial byroproducts as well as modified biopolymers (Tripathi, 2015). 2.3.1. Rice Husk as Adsorbent Rice husk contains large amount of hydrocarbon in the form od cellulose and lignin content. Those can be aplied as a raw material to make activated carbons which have a complex porous structure. There are two methods in term of rice husk activation: physical (thermal) activation and chemical activation. Afterwards, carbonization and activation usually done in a single step by using a chemical agent while physical activation produces activated carbon and it offers low specific area. Activated carbon and or adsorbent from rice husk was investigated effective in term of adsorption process considering its microporous structure (Cristina and Rosa, 2008). Andi (2010) has been reported the optimum time of rice husk in absorbing Pb2+ ion is approximately 85 minutes the optimum contact time for rice husk in absorbing Pb2+ ions is 85 minutes with an absorption capacity of 0.232 mg/g. In coorelated by the longer contact time of adsorption process, the higher Pb2+ ion will be absorbed. Rice husk has an ability of absorption due to its large pores as well as surface area on the surface of rice husk. Rice husk is composed by 80% of organic matter as the main element which are cellulose compounds and lignin as functional group as hydroxyl (OH), carboxyl (COOH), ketones (R-O-R) that will react to bind heavy metals (Dupont, 2005). 2.4. Test Species – Oreochromis niloticus Scientific Classification Kingdom : Animalia Phylum : Chordata Class : Actinopterygil Order : Perciformes Family : Cichlidae Genus : Oreochromis Species : O. Niloticus 10 Nile tilapia (O. niloticus) was one from the early fish species cultured. From illustrations by Egyptian crypts that commend if nile tilapia were cultured over and above 3000 years ago. Tilapia have been knew as "Sain't Peter's fish" in resource to biblical quotation about the fish fed to the large group. In africa, Nile tilapia still be the most largely cultured species. Nile tilapia fish has adaptation ability to the water temperature lower than 10 to 11°C (Thomas and Michael, 1999). 2.5. Histopathological Effects Patnaik et al. (2010), were evaluating histopathological changes on the gills of Cyprinus carpio communis exposed to different concentrations of lead and cadmium. The results of the damage by Lead were fusion and disintegration of primary lamellae as well as extensive vacuolization upon disruption of epithelial lining. On the other side, hyperplasia, congestion, and vaculization were recorded due to the effect of the Cadmium. The effect of heavy metal, was examined on gill morphology of fresh water fish from Garmat Ali River adjacent to Al- Najebyia Power Station Iraq. Morphological gill changes were identified by the adhesion of some secondary lamellae correlated to the imflammatory cells, congestion, and loss of epithelial lining made cappilaries were the only ones left. Fragment of secondary lamellae were detected due to the destruction of structures affected by the heavy metals. In the study by Poleksic et al. (2009), the Sterlet (Acipenser ruthenus L.) were indicated by some heavy metals (Cd, As, Pb, Cr, Hg, Cu, Ni, Fe, Mn, and Zn). The histopathological study was conducted and showed 95% of structural changes comprised by the loss of the secondary lamellae and hyperplasia of the respiratory epithelium. Abalaka (2015) investigated the effect of heavy metals concentration in Tiga dam, Nigeria using Auchenoglanis occidentalis as the indicator. Their bioaccumulation and histopathologic alterations on the fish gills were carried out. The present of some metals were detected as zinc (Zn), cadmium (Cd), lead (Pb) and iron (Fe). Morphological changes showed that the the toxic effect of Pb and Cd affected to the damage of membrane integrity with subsequent loss 11 of membrane-bound enzyme activity arising in cellular death. The alterations changes comprised of hyperplasia and the detachment of the gill epithelial cells followed by the fusion of lamellae caused by the effect toxic contaminations. 12 PART III. METHODOLOGY 3.1. Place and Time This experiment started from May 2017 until September 2017. Toxicity test and heavy metal adsorption test using fish as indicator were conducted at Aquaculture Laboratory, Faculty of Agriculture, Sriwijaya University, Inderalaya Campus. Adsorbent adsorption examination was conducted at Fish Product Technology Laboratory, Faculty of Agriculture, Sriwijaya University, Inderalaya Campus. Histophatological examination was conducted at Laboratory of Anatomical Pathology Laboratorium Barokah Palembang. 3.2. Equipment and Materials Equipment and materials used for this study are listed as in Table 2 and Table 3 below. Table 4. List of Equipment Used Toxicity Testing          Aquarium Aerator Fish strainer Erlenmeyer Beaker Spatula Tweezers Gloves Mask Bioadsorbent Histopathological Testing Testing Apparatus used in  Aquarium laboratory :  Aerator  Graduated cylinder  Erlenmeyer  Glass funnel  Pipettes  Cassettes  Beaker  Dissecting Knife  Shaker  Scissors  Analytical  Scalpel balance  Rubber gloves  Magnetic funnels  Aprons  Gloves  Mask  AAS (Atomic  Microscope slides Absorption Spectroscopy) Instruments used in laboratory:  Histokinet  Shandon histocentre 3  Shandon finesse 325 (microtome)  LAB Line Barnstead 26025 Slide Warmer  Tissue floating bath  OLYMPUS BX51 microscope Additional  Pen  Pencil  Book  Plastic bag 13