MINISTRY OF EDUCATION AND TRAINING NONG LAM UNIVERSITY DEPARTMENT OF BIOTECHNOLOGY BACHELOR THESIS SEROLOGICAL STATUS OF NEWCASTLE DISEASE IN COMMERCIAL CHICKENS IN PROVINCES OF SOUTHERN VIETNAM Major : Biotechnology Student : Nguyen Thi Thoi Course : 2007 - 2011 July, 2011 MINISTRY OF EDUCATION AND TRAINING NONG LAM UNIVERSITY DEPARTMENT OF BIOTECHNOLOGY BACHELOR THESIS SEROLOGICAL STATUS OF NEWCASTLE DISEASE IN COMMERCIAL CHICKENS IN PROVINCES OF SOUTHERN VIETNAM Supervisor Student Dr Karnam Shiv Shankar Nguyễn Thị Thời July, 2011 ACKNOWLEDGEMENTS This chapter of acknowledgements has given me a golden occasion to convey my hearty thankfulness to all of them who have directly or indirectly contributed to this thesis and stretched their helping hands for successful execution of my research project It gives me immense pleasure to express my sincere and heartiest sense of gratitude, abstruse indebtedness and best regards to Respected Managing Director, Uttara Feeds & Foods Pvt Ltd for giving me permission to my research and thesis under his lab I take the privilege in expressing my deep sense of gratitude and indebtedness to Dr Karnam Shiv Shankar, Manager of Venky’s Poultry Disease Diagnostic and Nutrition Laboratory for noble guidance, close supervision, constructive criticism and constant encouragement with relevant suggestions during entire period of my study and research work I take the opportunity of recording my heartfelt thanks to Dr Le Dinh Don, Head of Biotechnology Department of Nong Lam University for his awe - inspiring support, constant encouragement, cooperation, precious help and charming loom during entire study period It is my amusing duty to pronounce my heartily thanks to All staff of Venky’s Vietnam Co., Ltd for their support, cooperation and timely help during research work I would like to extend my sincere thanks to all my teachers and friends of Biotechnology Department of Nong Lam University for their untiring help, never ending support and active cooperation whenever required during the entire study I found my vocabulary to be exhausted to render my most sincere, respected and deepest sense of reverence to my beloved parents for their eternal blessing and affections and innumerable sacrifice, which has brought me up to this stage Nguyễn Thị Thời July, 2011 i SUMMARY The thesis title “Serological Status of Newcastle Disease in commercial chickens in some provinces of southern Vietnam” was conducted in Venky’s Poultry Disease Diagnostic and Nutrition Laboratory at Research Institute for Biotechnology and Environment, Nong Lam University from February to June, 2011 A survey on serological status of Newcastle Disease was carried out in broiler, layers and colour chickens in Vung Tau, Binh Duong, Dong Nai province of southern Vietnam A total of 701 samples were collected from broiler, layers and colour chickens farms were screened for presence of antibody titers against NDV by using haemaglutination inhibition test Of these, 596 samples (85.0%) were positive for NDV antibody titer (HI titers ≥ log23) while 105 samples (15.0%) were negative Positive percentage of ND - HI titers of broiler, colour chickens and layers were 77.2, 80.0 and 95.5 respectively Statistical analysis by Chi - square test revealed that there was significant difference (p < 0.05) in the prevalence of positive NDV antisera from layers in - 18 weeks group (91.7%) and 19 - 40 weeks group (100%) There was significant difference (p < 0.05) in rate of sero - positive colour chickens of different age groups 71.8%, 76.4% and 86.6% colour chickens had protective HI titers in group of - 13 days, 14 - 27 days and 28 - 63 days respectively Seroprevalence rate of NDV antibodies of broilers were 69.6% in group of - 13 days, 71.9% in group of 14 - 27 days and 84.8% in group of 28 - 42 days but the difference was not significant (p > 0.05) There was no significant difference (p > 0.05) in positive for specific immunity of serum samples between different regions The result showed that the level of protection in layers was more satisfactory than that of broiler and colour chickens Keywords: Newcastle disease virus, Haemagglutination, Haemagglutination inhibition ii TABLE OF CONTENTS Page ACKNOWLEDGEMENTS SUMMARY ii TABLE OF CONTENTS iii LIST OF ABBREVIATIONS v LIST OF FIGURES, TABLES vi Chapter INTRODUCTION 1.1 Introduction 1.2 Objectives .2 1.3 Performances Chapter LITERATURE REVIEWS 2.1 Newcastle disease 2.1.1 History .3 2.1.2 Etiology .3 2.1.2.1 Genome of NDV 2.1.2.2 Important viral proteins and biologic properties .3 2.1.3 Epidemiology 2.1.3.1 Hosts .6 2.1.3.2 Pathogenesis 2.1.3.3 Sources of NDV 2.1.3.4 Spread of NDV .7 2.1.4 Clinical signs .7 2.1.5 Lesions 2.1.6 Immunity 2.1.6.1 Cell - mediated immunity 2.1.6.2 Humoral immunity 2.1.6.3 Local immunity .9 2.1.6.4 Passive immunity 2.1.6.5 Immunosuppression 2.1.7 Diagnostics 2.1.7.1 Serology 2.1.7.2 Virus isolation 10 iii 2.1.7.3 Pathogenicity tests 10 2.1.7.4 Molecular techniques 11 2.1.8 Prevention 11 2.1.8.1 Vaccination 11 2.1.8.2 Biosecurity and hygiene 12 2.1.9 Zoonosis 12 2.2 Haemagglutination and hsaemagglutination inhibition test 13 2.2.1 Haemagglutination 13 2.2.2 The haemagglutination inhibition test 14 2.3 Studies about prevalence of antibody against NDV in vaccinated chickens .15 Chapter MATERIALS AND METHODS 17 3.1 Time and place to thesis 17 3.2 Materials 17 3.2.1 Subject 17 3.2.2 Equipment 17 3.2.3 Reagents 17 3.3 Method of study 18 3.4 Technical 18 3.4.1 Preparation of sera samples 18 3.4.2 Preparation of 1% CRBC 18 3.4.3 HA and HI test 18 3.5 Statistical analysis 20 Chapter RESULTS AND DISCUSSIONS 21 4.1 Prevalence of antibodies against NDV in commercial chickens 22 4.2 Prevalence of antibodies against NDV in broiler chickens 22 4.3 Prevalence of antibodies against NDV in colour chickens 25 4.4 Prevalence of antibodies against NDV in layers 26 4.5 Prevalence of antibodies against NDV in commercial chickens in relation 29 Chapter CONCLUSIONS AND FURTHER STUDIES .30 5.1 Conclusions 30 5.2 Further studies 30 REFERENCES 31 APPENDIX iv LIST OF ABBREVIATIONS AIV : Avian Influenza virus APMV : Avian paramyxovirus C : Control CRBC : Chicken red blood cells ELISA : Enzyme - linked immunosorbent assay F : Fusion HA : Haemagglutination HAU : Haemagglutinin units HI : Haemaglutination Inhibition HN : Haemagglutinin - neuraminidase IB : Infectious Bronchitis IBD : Infectious Bursal Disease ICPI : Intracerebral pathogenicity index IVPI : Intravenous pathogenicity index MDA : Maternally derived antibody MDL : Maternal antibody level MDT : Mean death time NC : Negative control ND : Newcastle Disease NDV : Newcastle Disease Virus PBS : Phosphate buffered saline PC : Positive control RBC : Red blood cells RT-PCR : Reverse - transcription polymerase chain reaction SD : Standard Deviation v LIST OF FIGURES, TABLES Page Figure 2.1 Diagrammatic representation of Newcastle disease virus Figure 3.1 Diagrammatic presentation of procedure of method of study 17 Figure 3.2 Photograph showing haemagglutination test 21 Figure 3.3 Photograph showing haemagglutination inhibition test 21 Figure 4.1 Percentage of HI titers in different broiler 24 Figure 4.2 Percentage of HI titers in different colour 26 Figure 4.3 Percentage of HI titers in different layers 28 Table 3.1 Protocol of the haemagglutination test 19 Table 3.2 Protocol of the haemagglutination inhibition test 20 Table 4.1 Distribution of NDV antibodies in commercial chickens 22 Table 4.2 Comparison of prevalence of antibody to NDV .23 Table 4.3 Distribution of HI titers against NDV in serum samples of broiler 23 Table 4.4 Comparison of prevalence of antibody to NDV .25 Table 4.5 Distribution of HI titers against NDV in serum samples of colour 25 Table 4.6 Comparison of prevalence of antibody to NDV .27 Table 4.7 Distribution of HI titers against NDV in serum samples of layers 27 Table 4.8 Comparison of prevalence of antibody to NDV in 29 Table 4.9 Comparison of prevalence of antibody to NDV in 29 vi Chapter INTRODUCTION 1.1 Introduction Poultry industry is an emerging agri - business and has established its position as fastest growing segment in the agriculture sector of world Poultry industry is emerging in Vietnam from seventies Many small and large scale chicken farms are rapidly growing in Vietnam Vietnam is an agriculture country with 70% of population living in rural and 90% of household keep poultry The growth of this profitable subsector is interrupted by a number of infection diseases Such diseases include Newcastle disease (ND), Infectious bronchitis (IB) andInfectious bursal disease (IBD) etc Newcastle disease is a highly contagious and commonly fatal viral poultry disease Newcastle Disease was discovered in Newcastle upon Tyne, England in 1926 It was widespread throughout the rest of the world Its spread is normally either via newly introduced birds, selling or giving away sick and carrier birds ND is enzootic in most countries in Africa, Asia and South America, where it is a major constraint against the development of both industrial and village poultry production (Aldous and Alexander, 2001) In Vietnam the ND is the most important cause mortality in chicken (Nguyen, 1992) Every year, ND occurs in many localities and is responsible for economic losses in poultry industry of Vietnam These losses will be due to losses in productivity and death of poultry Vaccination is practiced widely and is the recommended method for prevention Farmers or persons in charge of vaccination are likely to believe that the chicken flocks will be protected after vaccination But apparent ideal ND vaccination programs not always guarantee protection of chickens flocks against ND due to incautious handling of vaccines and so on So, seromonitoring of humoral immune response in vaccinated chicken flocks is necessary for controlling the ND (Rahman et al., 2002) Arising from above, I conducted a research on “Serological Status of Newcastle Disease in commercial chickens in some provinces of southern Vietnam” with the following objective in commercial poultry 1.2 Objectives  To determine the serological status of ND in vaccinated chickens  To investigate the influence of different factors (age, type of chickens, regions) on NDV antibody prevalence in commercial chickens 1.3 Performance Collection of samples from different poultry farms in some regions of southern Vietnam to perform Haemagglutination test (HA)/ Haemagglutination Inhibition test (HI) to determine antibody titers against NDV Table 4.2 Comparison of prevalence of antibody to NDV between different age groups of broiler chickens Age (day) – 13 Number of samples 23 Specific immunity 16 (69.6%) Non – specific immunity (30.4%) 14 – 27 32 23 (71.9%) (28.1%) 28 – 42 46 39 (84.8%) (15.2%) P > 0.05 In - 13 days of age group, the rate of chickens had specific immunity was the lowest (69.6%) The group 14 - 27 days of age, presence of sero – positive chickens was intermediate (71.9%) and last group 28 - 42 days of age, positive percentage of serum samples was highest (84.8%) and NDV antibody titers were very high up to log210 The differences were not significant (p > 0.05) (appendix 2) Table 4.3 Distribution of HI titers against NDV in serum samples of broiler chickens Age (day) Antibody titers obtained by HI test (log2) 10 Mean Standard HI titer Deviation 4 1 - - 3.8 2.0 14 - 27 32 11 - - - - - 3.2 1.0 28 - 42 46 - 10 10 4.5 2.0 - 13 Number of samples 23 In - 13 days of age HI antibody titers varied from log 21 - log28 (their respective percentages were 13, 17.4, 17.4, 13, 17.4, 13, 4.3 and 4.3) with the mean HI titer was 3.8 but in 14 - 27 days of age HI antibody titers varied from log 21 - log25 (their respective percentages were 3.1, 25.0, 34.4, 28.1, and 9.4) with mean HI titer was 3.2 Whereas in 28 – 42 days of age HI antibody titers varied from log 22 - log210 (their respective percentages were 15.2, 21.7, 15.2, 21.7, 10.9, 6.5, 2.2, 4.3 and 2.2) with mean HI titer was 4.5 There had wide range in HI titers because the vaccine efficacy of each farm was different 23 35 30 percentage 25 20 - 13 days 15 14 - 27 days 10 28 - 42 days 5 10 HI titers Figure 4.1 Percentage of HI titers in different age groups of broilers `Similar report had been described by Numan et al., 2005 who reported that 98.7% of serum samples were positive for specific immunity against NDV for broilers in Pakistan Mozaffor et al., 2010 reported that 78.04 % of serum samples of broiler in Bangladesh have specific immunity The antibody titer in - 13 days of age was high may be due to high levels of MDV which are transmitted from hens to chicks and protect them at early ages However, the wide range of HI titer (SD: 2.0) showed that the MDL is not uniformity between chicks On the other hand, the birds within 14 – 27 days of age showed low level of serum antibody titers This level is unexpected, since we expected that thought vaccination the level of serum antibody titer should increase at this age There are several possible reasons for these low levels of antibody titers in birds, such as poor quality of vaccine, unsuitable vaccination schedule because high level of maternal antibody could be reason of failure vaccine, presence of immunosuppressive substances in the feed or immnunosuppressive diseases (Mozaffor et al., 2010) The chickens within 28 - 42 days of age showed level of serum antibody titers were increasing might because of booster dose However, there is still 15.2% non - specific immunity in Table 4.2 The reason may be some farmers combine ND and IB vaccine unsuitably that don’t be handled according to the recommendations of vaccine manufacturer That may affect the response to each virus of vaccine The route by which the vaccine is administered can affect the outcome of vaccination The oral 24 vaccination of the chickens is the most practical method of vaccination in flocks of large size In present study, all farmers used administration of vaccine by drinking water Quality and quantity of water to be used for the vaccination of chicken orally is very critical factor, as it decides immune response of the chickens to the vaccination 4.3 Prevalence of antibodies against NDV in colour chickens in relation to age A total of 355 serum samples were collected from commercial colour chickens farms were subjected to HI test Out of them, 284 were found positive for specific immunity against NDV with an overall positive percentage of 80.0 in Table 4.1 Different age groups were examined in detail in Table 4.4 and Table 4.5 Table 4.4 Comparison of prevalence of antibody to NDV between different age groups of colour chickens Age (day) - 13 Number of samples 71 Specific immunity 51 (71.8%) Non – specific immunity 20 (28.2%) 14 - 27 127 97 (76.4%) 30 (23.6%) 28 - 63 157 136 (86.6%) 21(13.4%) P < 0.05 In - 13 days of age group, presence of samples had specific immunity was the lowest intermediate (71.8%) The group 14 - 27 days of age, positive percentage of serum samples was intermediate (76.4%) and last group 28 - 63 days of age, serological evidence of specific immunity was the highest (86.6%) The differences were significant (p < 0.05) Table 4.5 Distribution of HI titers against NDV in serum samples of colour chickens Age (day) 12 10 13 12 1 - Mean HI titer 14 - 27 127 24 34 29 15 14 - - 3.7 1.5 28 - 63 157 - 21 4.6 1.8 - 13 Number of samples 71 Antibody titers obtained by HI test (log2) 10 27 35 31 20 13 25 2 Standard Deviation 2.0 30 25 percentage 20 - 13 days 15 14 - 27 days 10 28 - 63 days 5 10 HI titers Figure 4.2 Percentage of HI titers in different age groups of colour chickens In - 13 days of age HI antibody titers varied from log 21 - log29 (their respective percentages were 11.3, 16.9, 14.1, 18.3, 12.7, 16.9, 7, 1.4 and 1.4) with the mean HI titer was 4.0 but in 14 - 27 days of age HI antibody titers varied from log21 - log28 (their respective percentages were 4.7, 18.9, 26.8, 22.8, 11.8, 11.0, 3.1 and 0.8) with mean HI titer was 3.7 Whereas in 28 - 63 days of age HI antibody titers varied from log22 - log210 (their respective percentages were 13.4, 17.2, 22.3, 19.7, 12.7, 8.3, 3.8, 1.3 and 1.3) with mean HI titer was 4.6 In the present study, the chickens in - 13 days of age group showed intermediate level of antibody titers because in chicks the antibody response may also be caused by maternal antibodies, whereas 14 – 27 days of age group was showed low level of serum antibody titers This level is unexpected, since we expected that thought vaccination the level of serum antibody titer should increase at this age The chickens within 28 - 63 days of age showed higher antibody levels than the previous two groups This may be because of booster dose, the HI titer was up to log210 Shuaib et al., 2003 also reported that secondary vaccination yielded HI titers than the single vaccination However, there was still 13.4% non - specific immunity in Table 4.4 Due to differences in flock health and management practices, the manner in which flocks respond can vary considerably Proper house management that provides a stress - free environment is essential for the vaccine program to be successful We collected samples from March to May, dry season, so chickens may be 26 in heat stress leading to reduced immune response Moreover, all the farms from where we have collected samples were open farms, temperature is relatively high The farmer use water without ice and skimmed milk to dilute vaccine which reduces vaccine efficacy 4.4 Prevalence of antibodies against NDV in layers in relation to age A total of 245 serum samples were collected from commercial layers farms were subjected to HI test Among them, 234 were found positive for specific immunity against NDV with an overall positive percentage of 95.5 in Table 4.1 We divided layers into two groups according to their age and different age groups were also examined in detail Table 4.6 Comparison of prevalence of antibody to NDV between different age groups of layers Age (week) – 18 Number of samples 133 Specific immunity 122 (91.7%) Non – specific immunity 11 (8.3%) 19 – 40 112 112 (100%) (0%) P < 0.05 In 133 chickens from to 18 weeks of age, the prevalence of NDV specific antibody titers was 91.7% Whereas all samples from layers 19 to 40 weeks of age group were found positive for specific immunity to ND virus The difference was statistically significant (p < 0.05) (appendix 4) In period from 17 – 18 weeks, layers were shifted cage to lay eggs After that, every months ND vaccine was used for prevention chickens from ND That is reason antibody levels of layers from 19 to 40 weeks were high Table 4.7 Distribution of HI titers against NDV in serum samples of layers Age Number (week) of samples – 18 133 Antibody titers obtained by HI test (log2) 10 Mean Standard HI titer Deviation 9 24 34 24 12 6.4 2.2 19 - 40 112 - - - 22 32 31 15 7.3 1.3 27 30 percentage 25 20 15 - 18 weeks 19 - 40 weeks 10 5 10 HI titers Figure 4.3 Percentage of HI titers in different age groups of layers In - 18 weeks of age, HI antibody titers varied from log 21 - log210 (their respective percentages were 3.8, 4.5, 3.0, 6.8, 6.8, 18.0, 25.6, 18, and 4.5) with mean HI titer was 6.4 Whereas in 19 - 40 weeks of age HI antibody titers varied from log 24 to log210 (their respective percentages were 1.8, 5.4, 19.6, 28.6, 27.7, 13.4 and 3.6) with mean HI titer was 7.3 Similar reports had been described by Numan et al., 2005 who reported 100% of layer chickens were positive for specific immunity against NDV in Pakistan This finding also supports the report of Mozaffor et al., 2010, there had 96.67% samples has specific immunity in layer in Bangladesh The result showed that the level of protection in layers was higher than that of broiler and colour chickens The low level of antibody titers (< log 23) in some vaccinated layer chickens in this study may be due to the poor vaccine quality or vaccine failure There are several possible causes for the vaccine failure in the developing countries like poor manufacturing standards, lack of adequate storage facilities, application of expired vaccines, faulty application and vaccine handling during transportation (Vui et al., 2002) Vaccinated chickens had various levels of antibodies in their blood Antibody levels in 19 – 40 weeks group was less various than those of – 18 weeks group In this study, immune response of layer from 19 to 40 weeks was satisfactory 28 4.5 Prevalence of antibodies against NDV in commercial chickens in relation to regions Table 4.8 Comparison of prevalence of antibody to NDV in chickens for meat group of different regions Regions Chickens for meat Number of chickens < log 23 p HI titers ≥ log 23 Binh Duong 175 30 (17.1%) 145 (82.9%) Dong Nai 158 33 (20.9%) 125 (79.1%) Vung Tau 123 31 (25.2%) 92 (74.8 %) > 0.05 The distributions of NDV antibody titers of serum samples in different regions for broiler, colour chickens are shown in Table 4.8 Broiler and colour chickens belong to chickens for meat group The serological evidence specific immunity was the highest in Binh Duong (82.9%), was intermediate in Dong Nai (79.1%) and was the lowest in Vung Tau (74.8%) Lowest antibody titers in Vung Tau may be cause of their housing, management, low biosecurity and lack of knowledge regarding vaccine and vaccination Differences were no statistically significant (p > 0.05) (appendix 5) Table 4.9 Comparison of prevalence of antibody to NDV in chickens for egg group of different regions Regions Chickens for eggs Number of HI titers chickens < log 23 P Vung Tau 136 (2.9 %) 132 (97.1 %) Dong Nai 109 (6.4 %) 102 (93.6 %) ≥ log 23 > 0.05 The distributions of NDV antibody titers of serum samples in different regions for layers are shown in Table 4.9 Prevalence of NDV specific antibody titers was 97.1% in Vung Tau and 93.6% in Dong Nai However, differences have no statistically significant (p > 0.05) (appendix 6) 29 Chapter CONCLUSIONS AND FURTHER STUDIES 5.1 Conclusions Result from 701 samples screened show that 596 samples (85.0%) were positive for NDV antibody titer while 105 samples (15.0%) were negative Positive percentage of ND - HI titer of broiler, layers and colour chickens were 77.2, 95.5 and 80.0 respectively The level of protection in layers was more satisfactory than that of broiler and colour chickens 5.2 Further studies From this study, we recognized that the farmers were used to using vaccine but it was not really effectively Organize training courses for farmers on how to use vaccine effectively is needed Beside, guiding farmer how to apply biosecurity measures is necessary Because of limited time and scale of thesis, the recorded data is relatively less There should be further surveillance about prevalence of antibody to NDV in more regions, from which to access actual status of seroprevalence to take appropriate means of protecting birds from ND before outbreak should be instituted In addition, we also have survey about antibody level of another disease in poultry such as IBD, IB etc 30 REFERENCES English Aldous, E.W and D.J Alexander 2001 Detection and differentiation of Newcastle disease virus (avian paramyxovirus type 1) Avian Pathol, 30: 117 - 128 Alexander, D J 1988 Newcastle Disease Kluwer Academic Publishers, pp: – 10 Alexander, D J 2003 Newcastle disease, other Paramyxoviruses and Pneumovirus Infections Diseases of poultry Y M Saif, Aly M Fadly, John R Glission, Larry R McDougald, L K Nolan, David E Swayne (eds.) 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106 11 Nguyen Tien Dung 1992 Poultry production and Newcastle Disease in Viet Nam Newcastle Disease in village chickens, control with thermostable oral vaccines P B Spradbrow (eds), proceeding No 39, pp: 169 - 170, Canberra: Australian Central for international Agricultural Research (ACIAR) 12 Numan, M., Zahoor, M A., Khan, H A., and Siddique, M 2005 Serological status of Newcastle disease in broilers and layers in Faisalabad and surrounding districts Pakistan Veterinary Journal, 25 (2): 55 – 58 13 Rahman M M, Bari A S M, Giasuddin M, Islam M R, Alam J and Sil G C 2002 Evaluation of maternal and humoral Immunity against Newcastle Disease Virus in chicken International Journal of Poultry Science 1, pp: 116 - 163 14 Shuaib, M., Ashfaque, M., Sajjad-ur, R., Mansoor, M.K and Yousaf, I 2003 Comparative immune response of broiler chicks to Newcastle disease vaccine (Lasota strain) Pak Vet J 23: 91 – 93 31 15 Shuaib, M., Khan, H., Sajid-ur-Rehman and Ashfaque, M 2006 Humoral Immune Response to Newcastle Disease Vaccine (Lastoa Strain) in Broilers International Journal of Poultry Science (5): 411 – 414 16 Swayne, D E., Glisson, J R., Jackwood, W M., Pearson, J.E and Road, M W 1987 Isolation and identification of avian pathogens 4th edition, pp: 256 17 Tran Quang Vui, J E Lohr, M N Kyule, K.-H Zessin and M P O Baumann 2002 Antibody Levels against Newcastle Disease Virus, Infectious Bursal Disease Virus and Avian Influenza Virus in Rural Chickens in Viet Nam International Journal of Poultry Science 1: 127 – 132 Internet 18 http://www.fao.org/docrep/005/ac802e/ac802e0o.htm#bm24 19 http://www.oie.int/ fileadmin/Home/eng/ Animal_Health_in_the_World/docs/pdf /NEWCASTLE_DISEASE FINAL.pdf 32 APPENDIX Appendix1Result for HI test of NDV of commercial chickens farms No Number of Address samples 30 Vung Tau Type of Age chicken Colour days chicken Results 6,1,4,3,4,6,4,6,4,6,7,5,2, 6,5,5,7,4,4,4,7,7,9,6,6,5, 3,3,2,4 19 Vung Tau Colour chicken days 1,1,1,1,1,2,2,2,2,2,3,1,1, 4,5,3,2,2,2 20 Vung Tau Colour Chicken 20 days 6,6,6,4,3,1,2,2,5,4,4,2,3, 2,2,3,4,4,6,6 21 Vung Tau Colour Chicken 26 days 5,4,6,6,6,7,5,4,4,2,1,2,4,4, 3,2,5,5,3,3,4 Vung Tau Broiler 28 days 3,2,5,3,3,4,2,3,4 24 Vung Tau Colour Chicken 40 days 2,8,3,6,5,5,4,6,3,4,5,3,4, 4,3,3,6,3,5,5,4,7,5.3 24 Vung Tau Layer 18 weeks 9,8,8,9,10,10,7,10,9,9,8, 8,7,8,8,8,7,7,7,7,9,8,8,7 22 Vung Tau Layer 11 weeks 6,6,6,4,6,6,6,5,7,6,7,7,6, 7,8,4,4,8,7,7,7,7 13 weeks 9,9,9,7,8,6,7,7,9,8,8,9,10 10,8,10,8,7,6,8,6 21 20 Vung Tau Layer weeks 1,2,7,8,5,2,6,6,4,4,5,3,5,5, 6,6,2,4,7,7 10 20 Vung Tau Layer 30 weeks 7,8,6,7,7,6,8,8,8,9,7,7,8,8,6 ,6,7,8,9,5 11 29 Vung Tau Layer 39 weeks 8,4,8,7,7,7,7,7,8,6,9,7,7, 9,8,7,9,7,8,7,8,8,7,7,7,6, 6,9,9 12 25 Dong Nai Colour chicken 23 days 2,3,2,2,4,4,3,4,3,3,2,3,1, 3,2,4,4,4,3,3,4,5,5,2,2 13 22 Dong Nai Colour Chicken 50 days 3,3,3,4,5,2,2,4,2,2,3,5,5, 2,3,3,3,3,6,2,2,2 14 24 Dong Nai Colour chicken 44 days 4,4,3,5,4,5,4,4,4,6,10,4,6 4,4,7,7,6,6,5,5,3,4,4 15 10 Dong Nai Colour chicken 19 days 4,2,3,3,5,4,3,2,2,6 16 10 Dong Nai Colour chicken 23 days 3,3,4,4,4,4,6,7,3,3 17 21 Dong Nai Colour chicken 32 days 4,5,2,7,7,6,4,8,4,6,4,2,2,2, 5,4,3,2,5,5,4 18 25 Dong Nai Colour chicken 38 days 6,3,5,5,2,3,5,5,5,6,2,7,4,8, 3,9,6,6,3,5,3,5,7,6,3 19 21 Dong Nai Colour chicken 17 days 1,2,2,3,3,5,4,6,3,3,4,2,3,1, 2,6,3,5,7,8,5,4 20 23 Dong Nai Layer weeks 8,2,5,5,6,3,7,1,8,7,1,1,3,1, 8,2,6,2,7,7,4,7,3 21 20 Dong Nai Layer 25 weeks 7,8,8,8,9,9,10,6,6,6,6,8,8, 8,9,9,5,6,6,8 22 22 Dong Nai Layer 35 weeks 8,8,6,6,7,7,8,6,5,9,10,10,7, 8,10,7,7,7,8,8,6,6 23 22 Dong Nai Layer 35 weeks 8,6,8,9,9,8,7,8,7,7,7,7,9,5, 8,6,4,6,7,5,5,6 24 22 Dong nai Layer 18 weeks 5,8,6,7,7,6,7,7,7,6,6,4,6,7, 9,6,6,4,5,9,8,8 25 23 Binh Duong Broiler days 3,4,2,2,2,2,5,6,5,1,4,1,6,7, 8,5,3,1,3,5,6,3,4 26 32 Binh Duong Broiler 14 days 3,2,3,4,2,3,4,4,5,2,3,4,4,3, 2,5,3,3,4,3,2,5,4,4,3,3,3,2, 4,1,2,2 27 18 Binh Duong Broiler 36 days 4,5,5,4,7,6,5,6,9,5,9,5,5,7, 8,6,7,10 28 19 Binh Duong Broiler 34 days 3,5,2,6,2,2,3,4,4,3,3,4,3,3, 2,6,5,5,2 29 20 Binh Duong Colour chicken 25 days 3,2,3,3,5,7,6,5,5,4,4,2,1,3, 5,6,3,3,3,4 30 22 Binh Duong Colour chicken days 8,3,6,5,4,3,3,3,2,5,5,4,4,6, 6,6,6,4,5,3,3,2 31 17 Binh Duong Colour chicken 42 days 10,5,4,3,1,2,9,8,7,7,7,6,6 7,5,6,4 32 24 Binh Duong Colour chicken 35 days 5,3,7,4,3,3,8,4,2,5,5,2,6,7, 5,4,4,5,4,2,8,4,6,4 ————— 05/07 13:50:22 ———————————————————— Welcome to Minitab, press F1 for help Appendix Comparison of prevalence of antibody to NDV between different age groups of broilers Tabulated statistics: age groups HI titers Using frequencies in ratio Rows: age groups Columns: HI titers - + All a 16 23 5,24 17,76 23,00 b 23 32 7,29 24,71 32,00 c 39 46 10,48 35,52 46,00 All 23 78 101 23,00 78,00 101,00 Cell Contents: Count Expected count Pearson Chi-Square = 2,782 DF = P-Value = 0,249 Likelihood Ratio Chi-Square = 2,849 DF = P-Value = 0,241 Appendix Comparison of prevalence of antibody to NDV between different age groups of colour chickens Tabulated statistics: age groups HI titers Using frequencies in ratio Rows: age groups Columns: HI titers a + All 20 51 71 14,20 56,80 71,00 b 30 97 127 25,40 101,60 127,00 c 21 136 157 31,40 125,60 157,00 All 71 284 355 71,00 284,00 355,00 Cell Contents: Count Expected count Pearson Chi-Square = 8,308 DF = P-Value = 0,016 Likelihood Ratio Chi-Square = 8,454 DF = P-Value = 0,015 Appendix Comparison of prevalence of antibody to NDV between different age groups of layers Tabulated statistics: age groups HI titers Using frequencies in ratio Rows: age groups Columns: HI titers - + All a 11 122 133 5,97 127,03 133,00 b 112 112 5,03 106,97 112,00 All 11 234 245 11,00 234,00 245,00 Cell Contents: Count Expected count Pearson Chi-Square = 9,699 DF = P-Value = 0,002 Likelihood Ratio Chi-Square = 13,875 DF = P-Value = 0,000 Appendix Comparison of prevalence of antibody to NDV in chickens for meat group of different regions Tabulated statistics: regions HI titers Using frequencies in ratio Rows: regions Columns: HI titers - + All b 30 145 175 36,1 138,9 175,0 d 33 125 158 32,6 125,4 158,0 v 31 92 123 25,4 97,6 123,0 All 94 362 456 94,0 362,0 456,0 Cell Contents: Count Expected count Pearson Chi-Square = 2,879 DF = P-Value = 0,237 Likelihood Ratio Chi-Square = 2,860 DF = P-Value = 0,239 Appendix Comparison of prevalence of antibody to NDV in chickens for egg group of different regions Tabulated statistics: regions HI titers Using frequencies in ratio Rows: regions Columns: HI titers - + All v 132 136 6,11 129,89 136,00 d 102 109 4,89 104,11 109,00 All 11 234 245 11,00 234,00 245,00 Cell Contents: Count Expected count Pearson Chi-Square = 1,710 DF = P-Value = 0,191 Likelihood Ratio Chi-Square = 1,704 DF = P-Value = 0,192 * NOTE * cells with expected counts less than ...MINISTRY OF EDUCATION AND TRAINING NONG LAM UNIVERSITY DEPARTMENT OF BIOTECHNOLOGY BACHELOR THESIS SEROLOGICAL STATUS OF NEWCASTLE DISEASE IN COMMERCIAL CHICKENS IN PROVINCES OF SOUTHERN VIETNAM. .. 2002) Arising from above, I conducted a research on Serological Status of Newcastle Disease in commercial chickens in some provinces of southern Vietnam with the following objective in commercial. .. i SUMMARY The thesis title Serological Status of Newcastle Disease in commercial chickens in some provinces of southern Vietnam was conducted in Venky’s Poultry Disease Diagnostic and Nutrition