MINISTRY OF EDUCATION AND TRAINING MINISTRY OF AGRICULTURE AND RURAL DEVELOPMENT
VIETNAM UNIVERSITY OF AGRICULTURE
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TRAN ANH TUAN
Pathogenicity Study
ACUTE HEPANCREATIC NECROSIS OF SOME SPECIES OF VIBRIO BACTERIA IN SHRIMP FARMING IN BRAID WATER AT
SOME NORTHERN PROVINCES
SPECIALTY: AQUACULTURE CODE: 60.62.03.01
SCIENTIFIC INSTRUCTOR:
Dr. PHAN THI VAN
HANOI - 2014
COMMITMENT
I hereby certify that the data and research results in this thesis are honest and have never been published in any other work.
I certify that all information cited in the thesis has been properly cited.
Thesis author
Tran Anh Tuan
ACKNOWLEDGEMENTS
First of all, I would like to sincerely thank the Board of Directors, Department of International Cooperation and Training - Aquaculture Research Institute 1, the Board of Directors of Hanoi University of Agriculture and the Veterinary Agency of Region III for supporting, helping and creating all conditions for me to successfully complete this course.
I would like to express my deepest gratitude to my supervisor, Dr. Phan Thi Van, who has wholeheartedly guided, instructed and helped me throughout the process of writing my thesis.
Hereby, I would like to thank the Board of Directors of the Center for Environmental Monitoring and Warning and Disease Prevention in the Northern Region - Aquaculture Research Institute 1 and the staff of the project "Emergency Shrimp in 2013" for creating the best conditions for me to carry out the project.
My sincere thanks go to my family, friends and colleagues who have helped and encouraged me in my studies as well as in my life.
Hanoi November 20, 2014 Thesis author
Tran Anh Tuan
INDEX
COMMITMENT i
ACKNOWLEDGEMENTS ii
TABLE OF CONTENTS iii
LIST OF TABLES
LIST OF FIGURES
LIST OF ABBREVIATIONS vii
INTRODUCTION 1
1 Urgency of the topic 1
2 Research objectives 2
3 Research content 2
Chapter 1: DOCUMENT OVERVIEW 3
1.1 Some characteristics of the research subject 3
1.1.1 Classification and morphological characteristics of Vibrio bacteria 3
1.1.2 Distribution and culture characteristics 4
1.1.3 Biochemical properties 5
1.2 Research status on diseases caused by Vibrio spp bacteria in shrimp 8
1.2.1 In the world 8
1.2.2 In Vietnam 15
1.3 Situation of acute hepatopancreatic syndrome in brackish water shrimp 18
1.3.1 The situation of acute hepatopancreatic syndrome in the world 18
1.3.2 Situation of acute hepatopancreatic necrosis syndrome in Vietnam 18
1.4 Studies related to hepatopancreatic necrosis in cultured shrimp 20
1.4.1 Shrimp hepatopancreatic necrosis caused by MBV and HPV 20
1.4.2 Necrotizing Hepatopancreatitic Necrosis of Shrimp (NHP) 20
1.4.3 Hepatopancreatic necrosis caused by microsporidia 21
1.4.4 Hepatopancreatic necrosis due to toxin 21
1.4.5 Acute hepatopancreatic necrosis 23
Chapter 2: OBJECTS, CONTENT AND METHODS OF RESEARCH. 27
2.1 Research objects and contents 27
2.1.1 Research subjects 27
2.1.2 Research content 27
2.2 Time and place of research 27
2.2.1 Research time 27
2.2.2 Research location 27
2.3. Materials and research methods 27
2.3.1 Determining AHPND 27
2.3.2 Isolation and identification of bacteria (Frerichs and Millar (1983, 1993) 30
2.3.3 Determining the virulence of bacteria 33
2.3.4 Antibiogram (Kibry-Bauer) 36
2.4 Data processing method 36
Chapter 3: RESEARCH RESULTS 37
3.1 Determination of the presence of bacteria in shrimp infected with AHPND 37
3.2 Identification of virulent Vibrio strains 40
3.3 Antibiogram test for virulent Vibrio strains identified 46
CONCLUSION AND RECOMMENDATIONS 51
1. Conclusion 51
2. Proposal 51
REFERENCES 52
LIST OF TABLES
No. TABLE NAME PAGE
Table 1.1 Biochemical characteristics of some Vibrio spp bacteria that cause diseases in aquatic animals (Buller, 2004) 7
Table 2.1 How to perform and read biochemical reactions 32
Table 3.1 Bacterial isolation results 37
Table 3.2 Results of sample analysis by tissue method 39
Table 3.3 Results of bacterial re-isolation and histological analysis of experiment 1 41
Table 3.4 Results of bacterial re-isolation and histological analysis of experiment 2 42
Table 3.5 Results of re-isolation of experimental bacteria 3 44
Table 3.6 Results of histopathological analysis of experiment 3 45
Table 3.7 Average sensitivity of some bacterial species to some antibiotics 47
LIST OF IMAGES
NO. NAME OF IMAGE PAGE Image 1.1: Bacteria V. parahaemolyticus 3
Figure 1.2: V. vulnificus 3 bacteria
Figure 1.3: V. harveyi 4 bacteria
Figure 1.4: V. alginolyticus 4 bacteria
Figure 1.5: V. harveyi bacterial colony 5
Figure 1.6: V. Parahaemolyticus 5 bacterial colony
Figure 1.7: Gram stain of V.Vulnificus 5 bacteria
Figure 1.8: Indole reaction on V. parahaemolyticus 5
Figure 1.9: Signs of AHPND identification by histopathological method 24
Figure 2.1: Experimental setup 36
Figure 3.1: Isolation rate of Vibrio species in Nghe An 38
Figure 3.2: Isolation rate of Vibrio species in Nam Dinh
Figure 3.3: Microscopic image of bacteria 39
Figure 3.4: Biochemical test of V. paraheamolyticus re-differentiated 45
Figure 3.5: Tissue structure of shrimp infected with AHPND 46
Figure 3.6: Antibiogram of V.paraheamolyticus (12,020) 49
Figure 3.7: Antibiogram of V. vulnificus and V.paraheamolyticus (13.014) 49
LIST OF ABBREVIATIONS
AHPND: Acute Hepatopancreatic Necrosis Dsease CTVK: Bacterial Formula
Mekong Delta: Mekong Delta EMS: Early Mortality Syndrome
HPV: Hepatopancreatic Pavovirus
IHHNV: Infectious Hypodermal and Hematopoietic Necrosis Virus IMNV: Infectious Myonecrosis Virus
MBV: Monodon Baculovirus
NACA: Network of Aquaculture Centers in Asia-Pacific NHP: Necrotizing Hepatopancreatitis
NTTS: Aquaculture
PCR: Polymerase Chain Reaction Pesticides: Pesticides TSV: Taura Syndrome Virus
YHV: Yellow Head Virus