Toxicological impact and histopathological response of tilapia after lead (II) nitrate (pb (n03)2) contamination

THAI NGUYEN UNIVERSITY UNIVERSITY OF AGRICULTURE AND FORESTRY MENDOZA, JIMLEA NADEZHDA A TOXICOLOGICAL IMPACT AND HISTOPATHOLOGICAL RESPONSE OF TILAPIA AFTER LEAD (II)-NITRATE (Pb (NO3)2) CONTAMINATION BACHELOR THESIS Study Mode: Full-time Major : Environmental Science and Management Faculty : International Training and Development Center Batch : 2012-2016 Thai Nguyen, September 2016 i DOCUMENTATION PAGE WITH ABSTRACT Thai Nguyen University of Agriculture and Forestry Degree Program : Bachelor of Environmental Science and Management Student name : Mendoza, Jimlea Nadezhda A Student ID : DTN1353110555 Thesis Title : TOXICOLOGICAL IMPACT AND HISTOPATHOLOGICAL RESPONSE OF TILAPIA AFTER LEAD (II)-NITRATE (Pb(NO3)2) CONTAMINATION Supervisor (s): Krisna MURTI, MD., M Biotech Stud., Ph.D Duong Van Thao, Ph.D Abstract: To investigate the effects of environmental contaminants, histopathological response of fish exposed to pollutants have been used as sensitive biomarkers The present study was conducted to assess the histopathological alterations in the gills, heart, dorsal muscles and liver of tilapia Oreochromis niloticus which were kept in aqueous solution of lead nitrate of two concentrations of 0.2 mg/l, 1.0 mg/l for days under laboratory conditions The resultant histopathological changes in the gills, heart, dorsal muscles and liver were recorded by light microscope The observed changes in the treated groups were disintegration of secondary lamellae, atrophy, curling and shortening of secondary lamellae, swelling/ inflammation, desquamation, epithelial lifting, curling bend of secondary lamellae and necrosis in gills Atrophy and splitting i of muscle fibers are recognized as common changes recorded in the heart of experimental fish Atrophy in dorsal muscles and splitting of dorsal muscle fibers, necrotic damage and degradation of muscle fibers were an interesting observation in dorsal muscle tissue of experimental fish Examination of liver sections after exposure showed sinusoidal dilatation and leukocyte infiltration in central veins and in peripheral areas occurred after exposure The damages in histology of gills, heart, dorsal muscles and liver depend on the exposure concentrations tolead (II)-nitrate (Pb (NO3)2) As the exposure concentrations increased, the more adverse damage occurred in the organs Therefore, the present investigation gives a brief account of the toxic effects of heavy metals on fish The present review illustrates that these histopathological alterations would contribute an important role in assessing the harmful effects of lead nitrate As such, fish are used as bio-indicators, providing useful purpose in monitoring heavy metals contamination Hence, implementation of regulations regarding the conservation of aquatic environments must be taken into consideration Keywords: LEAD (II)-NITRATE (Pb (NO3)2), histopathology, Oreochromis niloticus Number of Pages: 56 Date of Submission: September, 2016 Supervisor’s signature ii ACKNOWLEDGEMENT 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 supervisors: Dr Phil Arinafril of Sriwijaya University, Indralaya, Indonesia and Krisna MURTI, MD., M Biotech Stud., Ph.D in the Department of Anatomical Pathology, Faculty of Medicine, Sriwijaya University/Dr Mohammad Hoesin Public Hospital who kindly assisted me with the histopathological detection in this dissertation and was very patient with my knowledge gaps and in guiding me wholeheartedly when I implemented this research I also want to express my thanks to Dr Duong Van Thao, the second supervisor, for his supervision, encouragement, advice, and guidance in writing this thesis Besides my supervisors, I would like to thank Ms Fadila Mutmainnah from Pascasarjana Program Sriwijaya University, Ms Mirna Fitrani, Ms Sefti Heza Dwinanti and Ms Ade Dwi Sasanti of Aquaculture Laboratory, Faculty of Agriculture, Sriwijaya University, Dr Imelda Mendoza Moreno, Mrs Aimee Ciriaco from Laguna State Polytechnic University, Siniloan Campus, Siniloan Laguna for providing me an additional knowledge about Lead nitrate, water quality and fish histology In addition, formal thanks should be offered to the Rector of Sriwijaya University, Prof Badia Perizade, for granting my internship acceptance iii I would also like to acknowledge with much appreciation to the Dean of Faculty of Medicine in Sriwijaya University, Dr Mohammad Zulkarnain M Med Sc, PKK., who gave the permission to use all required equipment and the necessary materials to conduct my research in Department of Anatomical Pathology, Faculty of Medicine, Sriwijaya University/Dr Mohammad Hoesin Public Hospital Special thanks to Mrs Ana Nyayu, Mr Mohammad Zainuri, and other staffs and students in Aquaculture Laboratory, Faculty of Agriculture, Sriwijaya University for helping and providing me necessary equipment as well as knowledge for fish anatomy I wish to thank the technicians who work in Department of Anatomical Pathology, Faculty of Medicine, Sriwijaya University/Dr Mohammad Hoesin Public Hospital for their help in tissue preparation namely: Mrs Fitri Faurianty and Madi Santoso, without them, this research could not be accomplished on time My sincere thanks also go to Duong Hong Ngoc, Keraia Vince Geronimo, Phonevilay Soukhy, Tran Cong Phong, Nguyen Thi Van and Do Manh Dung, Jose Alberto Dunca for helping me finish this study Of course, I would like to thank to my Indonesian friends – Rotua Febriani, Hendra Edison and others for their invaluable support and encouragement when I stayed in Palembang Finally, special thanks to my family, my friends for their love and moral support throughout my study Thai Nguyen, 20th September, 2016 Student Mendoza, Jimlea Nadezhda A iv Table of Contents 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 PART II LITERATURE REVIEW 2.1 Lead Compound- Lead (II) Nitrate 2.2 Toxic effects of Lead on organisms 2.2.1 Toxicity 2.2.2 Histopathological Effects 12 2.3 Test species -Oreochromis niloticus 16 PART III MATERIALS AND METHODS 17 3.1 Time and Place 17 3.2 Materials 17 3.3 Equipment 17 3.4 Methods 19 3.4.1 Toxicity testing 19 v 3.4.2 Histopathological Examination 19 PART IV RESULTS 23 4.1 Histopathological observation of gills 23 4.2 Histopathological observations of heart 26 4.3 Histopathological observations of Dorsal muscle 27 4.4 Histopathological observations of liver 29 PART V DISCUSSION AND CONCLUSION 32 5.1 Discussion 32 5.2 Conclusions 41 PART VI REFERENCES 43 vi LIST OF FIGURES Figure Normal histological structure of gills Figure Comparison of the normal and treated gills Figure Histopathological changes observed in heart of experimental fish Figure Comparison of the normal and treated dorsal muscle Figure Comparison the normal and treated liver Figure Comparison of the normal and treated liver LIST OF TABLES Table 1.Physical and Chemical of Lead (II) Nitrate material Table Important routes of exposure to lead PART I INTRODUCTION 1.1 Background and rationale At present, the major challenge is to find ways to sustain activities, working procedures and management of industries towards economic development to promote human health and environmental protection The vulnerability of aquatic ecosystems to heavy metal contamination has been recognized as a serious pollution problem Metals that are deposited in the aquatic environment may accumulate in the food chain and cause ecological damage posing threat to human health and sustainable food supply due to biomagnifications over time The anthropogenic pollutants from industrial, domestic and agricultural waste are ultimately absorbed by aquatic plants and animals Potential damage to ecosystem may originate from exposure to metal pollutants because of extent and location and current ecological services of the lake as well These contaminants come sources such as from human activities that bring economic developments that deliver benefits to society (Molina, 2011) The heavy metal contamination of water bodies as one of the substantial environmental concerns was reported (Agrahari, 2009) and crucial harmful impacts on environmental quality, ecosystem integrity and health of humans have often been linked with improper management of chemical substances and the removal of hazardous materials Although some adverse health impacts of heavy metals have already been well known, heavy metal contamination is increasing in many parts of the world, mostly in developing countries, even though emissions have reduced in most developed countries over the last thousand decades (Järup, 2003) Bio-accumulation in various tissues of aquatic organisms and rising level of heavy metal toxicity poses serious threats to 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Exposure to lead (II)- nitrate (Pb (NO3)2) contamination will not result in changes in liver histology of Oreochromis niloticus HA (Alternative Hypothesis): Exposure to lead (II)- nitrate (Pb (NO3)2) contamination will result in changes in liver histology of Oreochromis niloticus Hypothesis 3: HO: Exposure to lead (II)- nitrate (Pb (NO3)2) contamination will not result in changes in heart histology of Oreochromis... Exposure to lead (II)- nitrate (Pb (NO3)2) contamination will result in changes in heart histology of Oreochromis niloticus 6 Hypothesis 4: HO: Exposure to lead (II)- nitrate (Pb (NO3)2) contamination will not result in changes in dorsal muscle histology of Oreochromis niloticus HA: Exposure to lead (II)- nitrate (Pb (NO3)2) contamination will result in changes in dorsal muscle histology of Oreochromis... Tilapia? - How does lead (II)- nitrate (Pb (NO3)2) affect the dorsal muscle of Tilapia? 1.3.2 Hypotheses Hypothesis 1: HO (Null Hypothesis): Exposure to lead (II)- nitrate (Pb (NO3)2) contamination will not result in changes in gills histology of Oreochromis niloticus HA (Alternative Hypothesis): Exposure to lead (II)- nitrate (Pb (NO3)2) contamination will result in changes in gills histology of Oreochromis... curling bend of secondary lamellae(c) and necrosis (nec) A-F are stained with H&E, magnification of A, C and E is 100x while B, D, and F is 200x 24 Upon treatment with 0.2 and 1 ppm of lead (II)- nitrate (Pb (NO3)2) some alterations were observed The initial lesions in the gills were manifested in groups exposed to 0.2 and 1 ppm of lead (II)- nitrate (Pb (NO3)2) The anomalies include disintegration of secondary... normal and treated liver A normal architecture of mass of hepatic cells (H) and normal central vein (cv) B moderate atrophy of hepatocytes (H) leading to early necrosis C Foci of necrosis (nec) A – C stained with H&E, magnification of A to C is 40x The histological examination of liver sections after exposure to 0.2 ppm lead (II)- nitrate (Pb (NO3)2) demonstrated moderate atrophy of hepatocytes leading... especially fishes (Markus and McBratney, 2001).Generally, because of its low rate of 4 disposal, lead (Pb) is one of the most crucial toxins in our surroundings which accumulates in the body As indicated by the results of past investigations, the present work was proposed to investigate the impact of lead nitrate on histological changes of the gills, heart, liver and dorsal muscle of the tilapia, Oreochromis... edema and satellitosis These findings showed the detrimental effects of Pb2+ on trace metal metabolism and tissue architecture of Crucain carp (Khan et al., 2014) Sharma and Tamot carried out the study to investigate the histopathological alterations in the liver and kidney of fresh water teleost, Channa striatus after exposure to 28 mg/l (10% of 96 hrs.LC50) of lead nitrate for a period of 30 and 60... Toxic effects of Lead on organisms 2.2.1 Toxicity Table 2 shows the toxicity classification of lead via the designated routes of exposure Table 2 Important routes of exposure to lead Pathway of Exposure Mechanism Ingestion Ingestion of food contaminated by lead, water or alcohol may pose risk for some populations Lead paint is the major source of lead exposure for children (ASTDR, 2005) As lead paint ... Table Physical and Chemical of Lead (II) Nitrate material Chemical Name LEAD NITRATE; Lead dinitrate; Lead( II) nitrate; Plumbous nitrate; Lead nitrate (Pb( NO3)2) Product Names Lead( II) nitrate Molecular... gills of Tilapia? - How does lead (II)- nitrate (Pb (NO3)2) affect the liver of Tilapia? - How does lead (II)- nitrate (Pb (NO3)2) affect the heart of Tilapia? - How does lead (II)- nitrate (Pb (NO3)2)... Student ID : DTN1353110555 Thesis Title : TOXICOLOGICAL IMPACT AND HISTOPATHOLOGICAL RESPONSE OF TILAPIA AFTER LEAD (II)- NITRATE (Pb( NO3)2) CONTAMINATION Supervisor (s): Krisna MURTI, MD., M Biotech