THAI NGUYEN UNIVERSITY UNIVERSITY OF AGRICULTURE AND FORESTRY NGUYEN THI VAN HISTOPATHOLOGICAL ALTERATIONS ON RAT AFTER EXPOSED TO DELTAMETHRIN BY INHALATION TREATEMENTS BACHELOR THESIS Study Mode: Full-time Major : Environmental Science and Management Faculty : International Training and Development Center Batch : 2012-2016 Thai Nguyen, 05/12/2016 n DOCUMENTATION PAGE WITH ABSTRACT Thai Nguyen University of Agriculture and Forestry Degree Program : Bachelor of Environmental Science and Management Student name : Nguyen Thi Van Student ID : DTN1253150042 Thesis Title : HISTOPATHOLOGICAL ALTERATIONS ON RAT AFTER EXPOSED TO DELTAMETHRIN BY INHALATION TREATEMENTS Supervisor (s): Dr. Nguyen The Hung Dr.Mulawarman Abstract: Deltamethrin is a synthetic pyrethroid widely used as the insecticide of choice especially for local vector mosquitoes in most countries. This experimental study aimed to evaluate the morphologic changes in the lungs caused by the inhalation of this insecticide. The study was performed on nine Sprague-Dawley rats.
There were three rats which were used as control group (without Deltamethrin inhalation), the other six rats were used as treated group which was exposed to 1:10 dilution of deltamethrin aerosol spray for 1 hour each day. After 8, 16, 24 days, a group of three rats (including 1 control and two-treated rats) was sacrificed and tissue samples taken from the lungs were processed for both light microscopy and transmission electron ii n microscopy. This experiment was finished after 21 days. Light microscopic examination revealed heavy congestion, marked perivascular edema, and lymphoplasmocytic infiltration with focal nonspecific interstitial pneumonia, foamy macrophage accumulation, emphysema, peribronchial lymphoid tissue hyperplasia, and focal hemorrhage.
Ultrastructurally, the ciliated cells of the airways appeared swollen with a few structurally abnormal cilia. Alveolar lining cells revealed mild degeneration and a slight hyperplasia in type II cells. Increases in the number of collagen bundles and edema in the alveolar septa were also noted. Keywords: Deltamethrin, histopathology, Biochemical parameters, lung Number of Pages: 38 page Date of Submission: December 05th, 2016 Supervisor’s signature iii n ACKNOWLEDGEMENT After an intensive period of three months; writing this note of thanks is the final statement of my thesis.
It has been a period of intense learning for me, not only in the scientific arena, but also on a personal level. Writing this thesis has had a big impact on me. I would like to reflect on the people who have supported and helped me so much throughout this period. First of all, I would like to express my deepest gratitude to my supervisor, Dr.
Arinafril of Sriwijaya University, Indralaya, Indonesia. I have been amazingly fortunate to have an advisor who gave me the freedom to explore on my own, and at the same time the guidance to recover when my steps faltered. He taught me how to formulate question and express ideas. His valuable support helped me overcome many critical situations to finish this research work.
I hope that one day I would become as good an advisor to my students as Dr. Arinaril has been to me. I am grateful to my other supervisor Dr. Nguyen The Hung for his encouragement and practical advice.
I am also thankful to him for reading my reports, giving important comments on my views and helping me understand and enrich my ideas. In addition, I am also thankful to him for encouraging me the use of correct grammar and consistent notation in my writings and for carefully reading and commenting on countless revisions of this manuscript. Krisna Murti in Department of Anatomical Pathology insightful comments and constructive criticisms at different stages of my research were thought- provoking and she helped me focus on my ideas. I am grateful to her for holding me to iv n a high research standard and enforcing strict validations for each research result, and thus teaching me how to do research.
Mulawarman from Sriwijaya University, Indralaya, Indonesia, my advisor has been always there to listen and give substantial advice. I am deeply grateful to him for the long discussions and lectures on related topics that helped me improve my knowledge in the area and helped me sort out the technical details of my thesis. In addition, special thanks should be offered to the Rector of Sriwijaya University, Prof. Badia Perizade, for granting my internship acceptance.
I would also like to acknowledge with much appreciation to the Dean of Faculty of Medicine in Sriwijaya University, Dr. Mohammad Zulkarnain, who gave the permission to use all required equipment and the necessary materials to conduct my research in Laboratory of Department of Anatomical Pathology, Dr. Mohammad Hoesin Public Hospital. Particularly, I would like to acknowledge to the members of the Department of Anatomical Pathology, University of Sriwijaya/Dr.
Mohammad Hoesin General Hospital Palembang: Mrs. Fitri Faurianty, Madi Santoso, Eka Susanti, etc… for their guidance and constant supervision as well as for providing necessary information regarding the project and also for their support in completing my research. Special thanks to the technicians who work at the Animal House, Faculty of Medicine, University of Sriwijaya. Especially to Mr.
Parman who provided me rats for free. Without them, my research could not be completed on time. Special thanks to all my friends from Viet Nam and Indonesia also for sharing their experiences, time and commitment especially during finishing this internship program. I am grateful because I have a lot of friends were helped and supported me v n throughout the course of completing the internship program.
My classmate: Jimlea, Ye, Keraia, Ngoc, Dung, Phong and others friends from Indonesia: Umiya, Rotua, Metri, Hendra, Irwan Jani, Dede, Naufal, Arift, Didi, Riko, etc. I greatly value their friendship and I deeply appreciate their belief in me. Most importantly, none of this would have been possible without the love and patience of my family. My immediate familiy, to whom this research work is dedicated to, has been a constant source of love, concern, support and strength all these years.
I would like to express my heart-felt gratitude to my family. Thai Nguyen, December 05th, 2016 Student Nguyen Thi Van vi n TABLE OF CONTENT ACKNOWLEDGEMENT. iv TABLE OF CONTENT. vii LIST OF TABLE.
ix LIST OF FIGURE. Toxic effects of DELTAMETHRIN on organisms. MATERIALS AND METHODS. Time and Place.
For the tissue alteration research. For histopathological examination. DISCUSSION AND CONCLUSION. 31 viii n LIST OF TABLE Table 1.Physical and Chemical Properties of Deltamethrin.
Toxicity classification of Deltamethrin. Equipment used in laboratory. 14 ix n LIST OF FIGURE Figure 1: Bronchiole structure of control and treated rats of week 1. 19 Figure 2: Comparison of bronchioles structure of control and treated rats.
19 Figure 3: Alveoli structures of control and treated rats. 20 Figure 4: The thickness of alveolar wall on the control and treated rats. 21 Figure 5: Development of Bronchiolitis in treated rat. 22 Figure 6: Lymp node of lung of control and treated rats.
23 Figure 7: Inflammatory cells in control and treated lung structure. 24 Figure 8: Swollen alveoli and lymphocyte infiltration in lung of treated rat. 25 Figure 9: Structure of intrapulmonary vascular of control and treated rats. Research rationale Pesticides have become an increasingly serious source of chemical pollution of the environment due to their extensive usage in agriculture, forestry, public health and in veterinary practices.
The most important sources of the animal and human exposure to Deltamethrin are polluted food and water, and it is readily absorbed by oral route. Deltamethrin metabolism and excretion have been extensively studied in rats, mice, and cows, where the pattern does not vary significantly among these species. It is metabolized by liver microsomal esterases and oxidases (Erdoğan et al. Negative effects of Deltamethrin on nervous, hematological, and respiratory systems have been documented (Svobodov et al., 2003) and its biochemical and histopathological effects, at low and high concentrations, have been studied (Velisek et al., 2006 and Yildirim et al.
Moreover, Deltamethrin induced oxidative stress (Yonar and Sakin, 2011). Pesticides of the pyrethroid class, such as deltamethrin, are widely used as insecticides because of their short biodegradation period and their low tendency to accumulate in organisms (Laskowski, 2002). Use of Deltamethrin is extensive in agriculture and forestry because of its high activity against a broad spectrum of insect pests (Glickman and Lech, 1982). There has been increasing concern over the influence of pesticides and man-made chemicals on the normal function of endocrine systems.
Since hormones are involved in fundamental functions of an organism, endocrine disrupting chemicals may have widespread effects, including on development, behavior and reproduction (EDSTAC, 1998). In addition, because so 1 n many individuals are exposed to commonly used chemicals, such as certain pesticides, even seemingly subtle epidemiologic associations may result in large increases in reproductive and other endocrine-related diseases among populations and thus should be of great public health concern (Meeker et al. Data on endocrine function resulting from pyrethroid insecticide exposure are limited, but animal and in studies suggest that some pyrethroid insecticides or their metabolites may possess endocrine disrupting properties. Experimental studies have also implicated pyrethroid insecticides in altered thyroid function (Wang et al., 2002 and Liu et al., 2006), although these associations remain untested in human studies (Meeker et al.
Experimental studies have reported that pyrethroid insecticides affect endocrine function, but the data were limited on histopathology and ultrastructure of thyroid gland toxicity induced by deltamethrin. The aim of this study was to evaluate the toxic effect of deltamethrin on the thyroid gland and the protective effect of lycopene to control the toxicity induced by deltamethrin through histopathological and ultrastructural studies of the thyroid gland, in addition to genotoxic activity by using the comet assay. As a widely used insecticide, we have attempted to better characterize the unpublished adverse effects of deltamethrin threatening the public health and to warn the authorities to control the human exposure of such chemical agents. Reasearch Objectives To assess the historical effects of deltamethrin on lung of rats by inhalation.
Research question and hypotheses 1. Research question How does Deltamethrin affect the lung rat? 1. Hypotheses The Delatametrin caused the degraded tissue of rats by inhalation treatments on the first week, second week and third week. 3 n PART II: LITERATURE REVIEW 2.
Deltamethrin Deltamethrin [(S)-a-cyano-3-phenoxybenzyl(1R,3R)-3-(2,2-dibromvinyl)-2,2- dimethylcyclo propan-carboxylate], one of Type II synthetic pyrethroid insecticides, is similar in structure to natural pyrethrins derived from the chrysanthemum flowers. Deltamethrin was synthesised in 1974 and first marketed in 1977. Deltamethrin is considered the most powerful and therefore the most toxic of the pyrethroids, up to three orders of magnitude more so than some (ETN, 1995). Deltamethrin causes neurotoxicity in insects and mammals by a similar mechanism of action, the modulation of nerve axon sodium channels.
Generally, pyrethroids affect normal production and conduction of nerve signals in the nervous system and act on nerve membranes by delaying the closing of the activation gate for the sodium ion channel. It is commonly used to control caterpillars on apples, pears, and hops, and for the control of aphids, mealy bugs, scale insects, and whiteflies on greenhouse cucumbers, tomatoes, potted plants, and ornamentals (ETN, 1995). Deltamethrin is the active ingredient in Butoflin, Butoss, Butox, Cislin, Crackdown, Cresus, Decis, Decis- Prime, K-Othrin, and K-Otek and it is the first potent and photostable insecticide belonging to the type II pyrethroid group (ATSDR, 2003). Worthing (1983) found that deltamethrin is extremely stable on exposure to air and temperature, and it is more stable in acidic as compared to alkaline media.
All pyrethroids have at least four stereoisomers, with different orientation of the substituents on the cyclopropane ring (or the equivalent part of the phenylacetate) despite the fact that deltamethrin is marketed as a single isomer cis (WHO, 1990). The 4 n isomers have different biological activities. In the summers of 1991 and 1995, the pesticide caused massive eel (Anguilla anguilla) kills in Lake Balaton, Hungary following application for mosquito control. In 1995, the presence of deltamethrin was demonstrated in several other fish species and in sediment samples taken from the lake (Balint et al.