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CAN THO UNIVERSITY COLLEGE OF AQUACULTURE AND FISHERIES NGUYEN QUANG TRUNG EFFECTS OF QUINALPHOS INSECTICIDE ON PHYSIOLOGY, BIOCHEMICAL CHANGES AND GROWTH OF COMMMON CARP (Cyprinus carpio Linnaeus, 1758) Major: AQUACULTURE Code: 62.62.03.01 ABSTRACT OF THESIS OF DOCTOR DEGREE OF AQUACULTURE Can Tho, November 2013 1 The study was conducted at Department of Nutrition and Fisheries Processing, College of Aquaculture and Fisheries, Can Tho University Supervisor: Asso.Dr. Đo Thi Thanh Huong – Can Tho University Commentator 1: Commentator 2: Commentator 3: Thesis will be defensed at final thesis commentary council at: ……………………………………………………………………… At: o’clock date i TABLE OF CONTENTS AT: O’CLOCK DATE I TABLE OF CONTENTS II LIST OF FIGURE VI VI CHAPTER 1: INTRODUCTION 1 1.1 Introduction 1 1.2 Objective 1 1.3 Contents 2 1.4 Signification of thesis 2 1.5 New results of thesis 2 CHAPTER 2: LITERATURE 4 2.1 Chemical 4 2.2 Active of mechanism of AChE 4 Figure 2.3: Active mechanism of AChE (A) and mechanism 4 of inhibition of organophosphorus pesticide (B) 4 (Richard and David, 2008) 4 CHAPTER 3: METHODOLOGY 5 3.1 Experimental site 5 ii 3.2 Materials 5 3.3 Experimental fish 5 3.4 Experimental chemical 5 3.6 Feed 5 3.7 Methods 5 3.7.1 Survey on pesticide use in Can Tho city 5 3.7.2 Determination of LC50- 96 hrs of quinalphos on common carp 5 Experiment was conducted in glass tank of 50 L, 10 fish/tank, weight from 8-10 g. During the experiment, no exchange of water, no aeration and no feeding was applied 5 3.7.3 Effects of quinalphos on respiratory physiology of common carp 6 3.7.3.1 Oxygen comsumption 6 Experiment included in five treatments: control 10%, 20%, 50% and 75% value of LC50-96 hrs., six replicates, 2 fish/2 L. Oxygen comsumption was determined by method of closed-vessel. There were two cases: contamination design before 24 hours and direct design 6 Experiment included in five treatments: control 10%, 20%, 50% and 75% value of LC50-96 hrs., six replicates, 4 fish/2 L. Oxygen comsumption was determined by method of closed-vessel. There were two cases: contaminating design before 24 hours and direct design. When observed mortality of 50% in vessel, sampling oxygen to determine oxygen threshold 6 3.7.4 Effects of quinalphos on ChE sensitivity and inhibition threshold of common carp 6 3.7.5 Effects of quinalphos on haematological parameters, ChE activity, digestive enzymes and growth of common carp 7 3.7.5.1 Effects of quinalphos on haematological parameters, and enzyme activities 7 3.7.6 Effects of quinalphos on cholinesterase activity and growth performances of common carp cultured in rice-field 8 3.7.6.1 Implementary method 8 3.8 Methods of analysis 8 3.8.1 Measurable method of haematological parameters 8 3.8.2 Measurable method of oxygen consumption 8 3.8.3 Analytic method of biochemical parameters 8 3.8.4 Measurable methods of growth parameters 9 iii 3.8.5 Measurable method of environmental parameters 9 3.8.6 Measurable method of quinalphos residue 9 3.9 Method of data treatment 9 VALUE OF LC50 WAS DETERMINED BY METHOD OF PROBIT. ONE-WAY ANOVA AND TWO-WAY ANOVA WERE USED TO COMPARE THE DIFFERENCES BETWEEN TREATMENTS AT A 5% SIGNIFICANT LEVEL, USING SOFTWARE SPSS 11.5 9 CHAPTER 4: RESULTS AND DISCUSSIONS 10 4.1 Survey on pesticide use in Can Tho city 10 4.1.1 General information 10 4.1.2 Information on Autumn-Summer crop 10 OM 2514 was the most common variety (26,7%). Most of farmers began Autumn-Summer crop from Lunar March to June (88,9%). Rice growth duration was 95 days 10 4.1.3 Technical parameters on rice-fish system 10 4.1.4 Economic efficiency of rice and rice-fish 10 4.1.5 Present status of pesticide use 10 4.1.6 Information on agents of pesticide trade 10 4.2 Acute toxicity of quinalphos insecticide on common carp 11 4.2.1 Environment parameters during the experiment 11 4.2.2 Acute toxicity of quinalphos insecticide on common carp 11 4.3 Effects of quinalphos on respiratory physiology 11 4.3.1 Oxygen consumption 11 4.3.2 Oxygen threshold 11 Increased concentration of decamethrin increased oxygen consumption of common carp (James et al., 1994). Murty (1988) and Đo Thi Thanh Huong (1997) stated that oxygen consumption increased at the beginning of exposing to insecticide because respiratory activity increased but decreasing after 12 4.4 Effects of quinalphos on ChE sensitivity and inhibition threshold 12 4.4.1 Environment parameters during the experiment 12 iv 4.4.2 ChE sensitivity 12 4.4.3 ChE inhibition threshold 12 4.5 Effects of quinalphos on haematological parameters, ChE activity, digestive enzymes and growth of common carp 12 4.5.1 Effects of quinalphos on haematological parameters and enzyme activities of common carp 12 4.5.2 Effects of quinalphos on haematological parameters, cholinesterase activity, digestive enzymes and growth performances of common carp cultured in tank 16 4.5.2.1 Environmental parameters and quinalphos residue 16 4.5.2.2 Effects of quinalphos on haematological parameters 16 4.5.2.3 Effects of quinalphos on ChE activity and digestive enzymes activities of common carp 17 4.6 Effects of quinalphos on ChE activity and growth of common carp cultured in rice-field 19 4.6.1 Environmental paramters during the experiment 19 4.6.2 Effects of quinalphos on ChE activity 19 4.6.3 Effects of quinalphos on growth, survival and yield of common carp cultured in rice-field 19 4.7 General discussion 20 CHAPTER 4: CONCLUSIONS AND RECOMMENDATIONS 22 4.1 Conclusions 22 4.2 Recommendations 22 REFERENCES 23 LIST OF RESEARCH WORK 27 LIST OF TABLE Table 4.9: Survival of common carp exposing to quinalphos 10 Table 4.25: Growth of common carp …………………………. 17 v LIST OF FIGURE Figure 2.3: Active mechanism of AChE 3 Figure 4.1: Changes of brain ChE activity after 28 days ………. 12 Figure 4.2: Brain ChE inhibition after 28 days………………… 12 Figure 4.3: Changes of muscle ChE activity after 28 days ……. 12 Figure 4.4: Muscle ChE inhibition after 28 days ………………. 12 Figure 4.5: Changes of gill ChE activity after 28 days………… 13 Figure 4.6: Gill ChE inhibition after 28 days …….……………. 13 vi CHAPTER 1: INTRODUCTION 1.1 Introduction Quinalphos is among the most widely used organophosphorus insecticides (OPs) in agriculture in India and some other countries among them Viet Nam (Chebbi et al., 2009). Haematological parameters such as hemoglobin, hematocrit, erythrocyte,…can be used to find physiological reaction in fish when environment was pollution (Dethloff et al, 2001). AChE inhibition affected to respiration, swimming activity, feeding and aquatic animal behaviours leading to imbalance, convulsion and even death (Peakall, 1992). Digestive enzymepatterns can reflect the feeding rate and digestive capacity of fish; hence, digestive enzyme activity can be used as bio-indicators of growth and health status of fish (Debnath et al., 2007; Suarez et al., 1995; cited by Li et al., 2010). Common carp Cyprinus carpio was the most common species in rice-field in Mekong Delta and Can Tho (Nguyen Van Hảo et al., 2001; Vromant et al., 2002). Quinalphos was widely used in agriculture and residue of quinalphos can affect to fish (Das et al., 2000). In order to study effects of insecticide on physiological parameters, biochemical changes in tank and rice-field, this study was conducted at College of Aquaculture and Fisheries – Can Tho University. 1.2 Objective - General objectives: Search scientific data of toxicity of insecticide on aquatic animals in order to recommend farmers suitable pecticide use in rice-field basically. - Specific objectives: Search effects of quinalphos insecticide to change physiological parameters (haematology and respiratory physiology), biochemical changes (enzyme activities) and growth performances of common carp. As a result, physiological parameters or biochemical changes as a biomarker for organophosphorus insecticides like quinalphos in rice-filed. 1 1.3 Contents a) Survey on pesticide use in Can tho city. b) Determination of LC 50 -96 hrs. of quinalphos on common carp. c) Effects of quinalphos on respiratory physiology of common carp Cyprinus carpio. d) Effects of quinalphos on ChE sensitivity and inhibition threshold of common carp. e) Effects of quinalphos on haematological parameters, cholinesterase activity, digestive enzymes and growth performances of common carp cultured in tank. f) ) Effects of quinalphos on cholinesterase activity, digestive and growth performances of common carp cultured in rice-field. 1.4 Signification of thesis The results of thesis are scientific basic to recommend farmers suitable pecticide use in rice-field. 1.5 New results of thesis Thesis determined quinalphos changed haematological parameters and respiratory physiology (oxygen consumption and threshold). Quinalphos considerably inhibited the brain, muscle, gill and liver ChE activities of common carp exposed to sublethal concentrations in 96 hours of acute test and 28 days and 60 days of long test in tank and 44 days in rice-filed. ChE activity was completly recovered at least 21 days in tank and 14 days in rice-field. Thesis also determined GST activity of common carp didn’t play an important role for decontamination. Quinalphos considerably inhibited digestive enzymes. Thesis determined brain was the most sensitive organ of common carp. ChE inhibition affected to swimming behaviour of fish. ChE inhibition threshold causing died fish was 95%. Thesis determined quinalphos affected to growth performmances, feed conversion rate and survival rate of common carp cultured in tank. Recommended dose affected to growth performmances, survival rate and productivity of common carp as compared to the control in rice-field. Concentrations of quinalphos which gradually decreased during the experiment were under the limit of determination (<LOD) at least 14 days. 2 3 [...]... of Aquaculture, Can Tho University 5 Nguyen Van Hao, Nguyen Trong Hien, Pham Đinh Khoi, Nguyen Tho Đan and Don Griffiths, 2001 The primery results on rice-fish farming system development in Tien Giang International Workshop Yearbook on rice-fish farming Agricultural Publish 159 p 6 Tran Thien Anh, Nguyen Thi Kim Ha, Nguyen Quang Trung, Đo Thi Thanh Huong, Nguyen Thanh Phuong, 2012 Effects of quinalphos... FL, p143 25 Nguyen Quoc Thinh, Tran Minh Phu, Do Thi Thanh Huong, Patrick Kestemont, Nguyen Thanh Phuong, Marie-Louise Scippo, 2012 kinetics of quinalphos residues elimination in water and fish tissues from tank cultured silver barb (Barbonymus gonionotus) AQUA 2012, Prague, Czech Republic Sep 1-5, 2012 26 Peakall, D., 1992 Animals biomakers as pollution indicators, Chapman and Hall, London, UK 27 Richard... Products Division, Worthington Diagnostic System, Freehold, NJ, USA 27 LIST OF RESEARCH WORK Published articles from research work 1) Nguyen Quang Trung and Đo Thi Thanh Huong, 2012 Effects of quinalphos on cholinesterase enzyme activity and glutathione-S-transferase of common carp (Cyprinus carpio) Journal of Science - Can Tho University 2012: Volume 22a 131-142 2) Nguyen Quang Trung and Đo Thi Thanh... Trung and Đo Thi Thanh Huong, 2012 Effects of quinalphos on sensitivity of cholinesterase enzyme and physiological parameters of common carp (Cyprinus carpio) Journal of Science - Can Tho University 2012: Volume 24a 167-178 Published articles related to research work 1) Tran Thien Anh, Nguyen Thi Kim Ha, Nguyen Quang Trung, Đo Thi Thanh Huong, Nguyen Thanh Phương, 2012 Effects of quinalphos on cholinesterase... gonionotus Journal of Science - Can Tho University 2012: Volume 22a, 269-279 Oral and poster in domestic and International Conferences 1) Nguyen Quang Trung, Đo Thi Thanh Hương, 2011 Effects of quinalphos on cholinesterase activity and glutathione-S-transferase of common carp (Cyprinus carpio) Yearbook of Student and Young Staff Conference on National Scientific Research, 2011 2) Nguyen Quang Trung, S... Master of Aquaculture, Can Tho University 3 Ngo Van Ngoc, Le Thanh Hung and Hakan Berg, 2001 Survey on effects of pesticide use in rice-fish system in Nha Be and Moc Hoa, Viet Nam International Workshop Yearbook on rice-fish farming Agricultural Publish 159 p 4 Nguyen Thi Que Tran, 2010 Effects of Kinalux 25EC insecticide (active ingredient of quinalphos) on enzymes of Pangasianodon hypophthalmus Degree... comprehensively - The study can use ChE activity as biomarker in correlation with concentrations of quinalphos residue in water 23 REFERENCES Vietnamese literature 1 Đo Thi Thanh Huong, 1997 Effects of Basudin 50EC on changes of physiological and haematological parameters of common carp, tilapia and silver barb Degree of Master of Aquaculture, Nha Trang University 2 Đo Van Buoc, 2010 Effects of quinalphos insecticide... site The study was carried out at College of Aquaculture and Fisheries, Cantho University and Thoi Hung commune, Co Do district, Can Tho city from May 2009 to July 2012 3.2 Materials The study used main materials such as: - Composite tank (capacity of 500 L, 2 m3, 4 m3), glass tank 84 L - Spectrophotometer, centrifugal machine and so on 3.3 Experimental fish Common carp (8-12 grams/individual) were... behaviours, inhibited digestive enzymes, considerably inhibited ChE activity of common carp for acute test and long-term test resulting in affecting growth parameters, FCR and survival rate of fish Consequently, ChE acitivity could be used to detect contaminated fish by organophosphorus insecticide like quinalphos The results of the study were suitable for the statement of Fulton and Key (2001) These authors... silver barb Barbodes gonionotus Journal of Science - Can Tho University 2012: Volume 22a, 269-279 7 Trinh Binh Dy, Pham Thi Minh Đuc, Phung Xuan Binh, Le Thu Lien and Hoang The Long, 2006 Physiology Volume 1 Medical Publish English literature 8 Ali, H Abdul JAFFAR and V Jaya Rani, 2009 Effect of phosalone on haematological indices in the tilapia, Oreochromis mossambicus Turk J Vet Anim Sci, 2009; 33(5): . THO UNIVERSITY COLLEGE OF AQUACULTURE AND FISHERIES NGUYEN QUANG TRUNG EFFECTS OF QUINALPHOS INSECTICIDE ON PHYSIOLOGY, BIOCHEMICAL CHANGES AND GROWTH OF COMMMON CARP (Cyprinus carpio Linnaeus,. study was carried out at College of Aquaculture and Fisheries, Cantho University and Thoi Hung commune, Co Do district, Can Tho city from May 2009 to July 2012. 3.2 Materials The study used. Can Tho (Nguyen Van Hảo et al., 2001; Vromant et al., 2002). Quinalphos was widely used in agriculture and residue of quinalphos can affect to fish (Das et al., 2000). In order to study effects