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. Duong Van Thao, Ph.
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 0.0 mg/l for 2 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.
Arinafril of Sriwijaya University, Indralaya, Indonesia and Krisna MURTI, MD. 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., 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. 1 LIST OF TABLES.
Background and rationale. Research questions and hypotheses. Lead Compound- Lead (II) Nitrate. Toxic effects of Lead on organisms.
Test species -Oreochromis niloticus. MATERIALS AND METHODS. Time and Place. Histopathological observation of gills.
Histopathological observations of heart. Histopathological observations of Dorsal muscle. Histopathological observations of liver. DISCUSSION AND CONCLUSION.
43 vi LIST OF FIGURES Figure 1. Normal histological structure of gills. Comparison of the normal and treated gills. Histopathological changes observed in heart of experimental fish.
Comparison of the normal and treated dorsal muscle. Comparison the normal and treated liver. Comparison of the normal and treated liver. 1 LIST OF TABLES Table 1.Physical and Chemical of Lead (II) Nitrate material Table 2.
Important routes of exposure to lead 2 PART I. 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 the 3 biodiversity of ecosystems and human health (Adeniyi et al., 2008; Vinodhini and Narayanan, 2008a; George et al. Fishes have been considered as good bio-accumulators of organic and inorganic toxicants. Fishes are primary sources of protein.
They comprise large components of most aquatic environment functioning as bio-indicator of heavy metal contamination in the aquatic ecosystem.These days, a huge segment of the overall eating routine comprises of nourishments from fresh and salt water bodies. This utilization has had a positive effect on job and livelihood for example, in instances of fish production and every year a huge assortment of goods made out of seafood items are becoming commercially available widely.Heavy metals concentrated in water systems and expand through food chain and aquatic organisms especially fishes and being the primary consumers, they are largely influenced. People on the other hand also are affected by eating fishes from those areas where main food is fish (Afshan et al. Terrestrial and aquatic food chains are suitable for accumulating numerous environmental pollutants up to levels that could be toxic to both anthropogenic and aquatic organisms’ health.
The pollution might cause adverse effects on the ecosystem such as the health of aquatic animals (Ashraj, 2005; Vosyliene and Jankaite, 2006; Farombi et al. Lead, like majority of the heavy metals are being discharged into surface waters affecting the health of aquatic organisms such as fishes.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 niloticus. This study is an important tool that evaluates the impacts of human activities and weighs the adverse effects to health of the people amidst the contributions of developments economically.
This study aims to evaluate the toxic effects of heavy metals such as lead nitrate on the organs of Oreochromis niloticus (Tilapia) to support of predicting potential health consequences to the human and health of fish which could serve as a scientific basis for decisionmaking and policy development and be able to determine the need for urgent measurements which should be done by concerned authorities to protect health of communities consuming fish products. Objectives This study was designed to assess the toxicological effects of lead (II)-nitrate (Pb (NO3)2) on Oreochromis niloticus (Tilapia) as well as to reveal the histopathological alterations on its gills, heart, liver and dorsal muscle under different concentrations of lead (II)-nitrate (Pb (NO3)2). Research questions and hypotheses 1. Research Questions The research aims to answer the follow questions; - How does lead (II)-nitrate (Pb (NO3)2) affect the gills of Tilapia? 5 - 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) affect the dorsal muscle of Tilapia? 1.
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 niloticus. Hypothesis 2: HO (Null Hypothesis): 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 niloticus. HA: 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 niloticus.
Lead Compound- Lead (II) Nitrate Table 1. Physical and Chemical of Lead (II) Nitrate material LEAD NITRATE; Lead dinitrate; Lead(II) nitrate; Plumbous nitrate; Lead nitrate (Pb(NO3)2) Chemical Name Product Names Lead(II) nitrate Molecular Formula HNO3.1 or N2O6Pb Chemical Formula Pb(NO₃)₂ Form White crystal discolored CAS Registry Number 10099-74-8 Molecular Weight 331.