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Advances in PoultryScience for One Health
th28 Feb 2020
ISO 9001:2015 Certified
ICAR-National Institute of Animal Nutrition and Physiology
Proceedings of the World VeterinaryPoultry Association (India) Conference 2020
EditorsV Sejian
B BarmanA V Elangovan
J VermaR Bhatta
ICAR-NIANP, Bengaluru
WVPA (INDIA) CONFERENCE 2020
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Proceedings of the World VeterinaryPoultry Association (India) Conference 2020
EditorsV Sejian
B BarmanA V Elangovan
J VermaR Bhatta
Advances in PoultryScience for One Health
th28 Feb 2020ICAR-NIANP, Bengaluru
CitationSejian, V., Barman, B., Elangovan, A.V., Verma, J and Bhatta, R (2020). Proceedings of the World Veterinary Association (India) Conference on “Advances in poultry science for one health”. Published by Director, ICAR-National Institute of Animal Nutrition and Physiology, Bengaluru, India, pp 1- 88.
Compiled and Edited byDr V SejianDr B BarmanDr A V ElangovanDr J VermaDr R Bhatta
Published byRaghavendra BhattaDirector, ICAR-NIANP
February, 2020
©ICAR-NIANP
The authors are solely responsible for the contents of the papers compiled in this volume. The publisher/editors are not responsible for views, opinions and materials expressed by the authors. No part of this compilation may be reproduced, stored in a retrieval system, or transmitted in any form or by any means, electronic, mechanical, photocopying, recording, or otherwise without written permission from the publisher.
PREFACEPoultry is one of the fastest growing segments of the agricultural sector in India
today. While the production of agricultural crops has been rising at a rate of 1.5 to 2 percent per annum, that of eggs and broilers has been rising at a rate of 8 to 10 percent per annum. The total poultry has increased by 16.8% and the total poultry is 851.8 million during 2019. Over 45.8% increase in backyard poultry has been estimated with total backyard poultry of 317.07 Million in 2019. The commercial poultry has increased by 4.5% and the total commercial poultry is 534.74 million.
Poultry production is an important and diverse component since egg and meat are significant components of health and diet for larger population all over the world. The largest proportion of poultry meat and eggs were found to be produced by intensive poultry farming. The increasing demand for poultry products has transformed poultry production activity into a full fledged industry from a mere household/backyard activity. Technological advances have revolutionized the role and the structure of poultry industry in India. The distribution of the poultry population suggests that it is concentrated in some limited pockets. Forty two percent of the total population of poultry is confined to the Southern region, with 22% in the Eastern Region and 20% in Western Region, with only about 16% in the Northern region. Improved poultry breeds account for 59% of the total bird population, contributing about 89% of the total egg production in the country.
Poultry farming is now recognized as one of the most suitable agro-industries in the world. It is an industry with lot of potential for intercropping from trees to other animal husbandry activities. Poultry in India is a powerful tool to fight poverty by creating job opportunities for rural farmers/ land less farmers or small to marginal landholders or weaker section of people in rural areas. It is one of the cheapest and easily accessible nutritious protein. Poultry farming is an ancestral small-scale agricultural operation in our country and from backyard practice; it has now developed as industry. Therefore, the present World Veterinary Poultry Association India chapter conference entitled “Advances in Poultry Science for One Health” assumes special significance, as it would provide a platform to the researchers to deliberate and develop future action plan to meet the challenges of poultry industry to sustain the poultry production in the country.
The editorial board is thankful to all the contributors who have come forward to share their scientific findings, knowledge and opinion through this publication. This publication is the outcome of dedicated efforts by all member of the editorial committee. We are sure this conference publication would serve as an excellent reference material for the students and researchers involved in poultry science research.
Editors
Dr.JeetendraVermaPresident
World Veterinary Poultry Association (India)
Feb 22, 2020MESSAGE
It gives me an immense pleasure in extending cordial welcome to all delegates, dignitaries and guests participating in the one day Annual meeting of World Veterinary Poultry Association (India) and the National conference on “Advances in Poultry Science for One Health” on 28th February 2020 at ICAR- National Institute of Animal Nutrition and Physiology, Bengaluru.
It is a matter of great satisfaction that our scientific association is organizing its first annual meeting. The association aims to strengthen research and education in poultry production and harness the knowledge for the welfare of farming communities of the country, especially the poultry farmers. It is imperative to note that the theme of this national conference “Advances in Poultry Science for One Health” is very pertinent in today’s context. The poultry industry is a high profit sector which however is highly susceptible to losses especially due to disease outbreaks. Such outbreaks pose a threat for other livestock species and humans too as some of the poultry diseases are of zoonotic importance. Thus there are increasing programs focusing towards one health initiative which aims to create common ground for several disciplines in order to establish more holistic approaches to diseases shared by more than one species. With this objective the World Veterinary Poultry Association (India) chapter paved the way to organize its first conference to focus on recent advancement and innovations in the field of poultry science which would also be discussed by global experts. I am confident that the deliberations from experts from different fields would certainly be helpful in designing new strategies to tackle these critical issues paving way for sustainable poultry production in the country.
On behalf of the WVPA (India) family, I take this opportunity to profusely thank all thelocal ICAR-NIANP organizers for accepting the task of organizing this mega event and providing very impressive scientific program addressing priority issues of poultry production research. I also take this opportunity to thank all the office bearers of WVPA (India) for their valuable and support in organizing this mega event. It is also the matter of great contentment that we are going to release the conference compendium containing the lead lectures and scientific abstracts could serve as good reference materials for different stakeholders involved in poultry research.
I wish the WVPA (India) Conference 2020 a grand success.
(Jeetendra Verma)
1] Immune Evasion Strategies of Poultry VirusesAuthors: Abdul Rahman Omar
Lead Paper
3
2] Current and Future Challenges of the Poultry Vaccine IndustryAuthors: Marcelo Paniago
Lead Paper
6
3] Newcastle Disease: Current Situation and Strategies For VaccinationAuthors: Jose Luis Losada Torres
Lead Paper
8
4] Development Of Indianized Immune Complex Vaccine -VIPx : Effective Approach For Intervention of Novel vvIBDV InfectionAuthors: Sanjay Gavkare
Lead Paper
11
5] Recombinant Technology for ND and MD Vaccine strainsAuthors: Vijayakumar E
Lead Paper
13
6] Immune modulation and enhancement of immunity by the use of an adjuvanted solvent (Hipramune®T) in a live coccidiosis vaccine for breeders and layers (Evalon®)Authors: Pagès M, Bech G, March R, Sitjà M, Lillehoj H, Dardi M, Rubio J, del Cacho E
Lead Paper
15
7] Black Soldier Fly Larvae: A Novel Tool to Convert Poultry Waste to WealthAuthors: Nadeem Mohamed, Niranjan B H
Lead Paper
16
8] Avian Disease Diagnosis – Dilemma Between Investigator and InvestorAuthors: Dhinakar Raj D
Lead Paper
18
9] Genetic Analysis of Humoral and Cell Mediated Immune Response in Japanese QuailsAuthors: Monika M, Rokade J J, Narayan R, Snehasmita Panda, Gopi M, Jyotirmoy Saharia
Poultry Breeding and Genetics
23
10] Tanuvas Namakkal Gold Quail- A Potent Egg Laying VarietyAuthors: Moorthy M, Amutha R, Kannan D, Rajendran K, Kannan V
Poultry Breeding and Genetics
24
11] Comparative Assessment of Fertility and Hatchability of Six Breeds of Chicken Reared in University Poultry and Duck FarmAuthors: Nimmi P, Supriya R, Binoj Chacko, Beena C Joseph, Sankaralingam S, Anitha P
Poultry Breeding and Genetics
25
12] Effect of Probiotic and Essential Oil on Broiler Performance - An Alternative to Antibiotic Growth PromoterAuthors: Karthigan M, Tarjan K
Biotic and Abiotic Stress and Welfare
29
13] Efficacy of Bacillus subtilis PB6 on Intestinal Health and Broiler Economic IndicesAuthors: Sai Ram Kumar., Venket M Shelke, Partha Pratim Das, Swati Karki
Biotic and Abiotic Stress and Welfare
30
14] Bacillus subtilis PB6 - A Complete Solution for Dysbacteriosis in BroilersAuthors: Venket M Shelke, Saravanan Sankaran, Chandrasekar S
Biotic and Abiotic Stress and Welfare
31
15] Chicken-Chimera: Primordial Germ Cells Mediated Poultry Germplasm ConservationAuthors: Adnan Naim, Sushobhan Dash, Mishra S K, Debasis Nayak, Chittaranjan Jena
Biotic and Abiotic Stress and Welfare
32
16] Evaluation of Embryonic and Early Post-Hatch Thermal Conditioning on the Thermotolerance Ability of Broiler ChickensAuthors: Aswathi P B, Bhanja S K, Darshana B Bhaisare, Manish Mehra, Akshat Goel, Junaid N
Biotic and Abiotic Stress and Welfare
33
17] Effect of Premature Moulting On Certain Biochemical Parameters In Commercial Layer HensAuthors: David I C G, Ravi Kiran Gorti, Reddy I J, Manpal Sridhar
Biotic and Abiotic Stress and Welfare
34
18] Gut Health and Serum Characteristics of Commercial Broilers Fed with Unconventional Feed (Soya Oil Cake)Authors: Sonale N S, Rajendran K, Moorthy M, Kathirvalen C
Gut Health, Feed Additives and Management
37
19] Nutritional evaluation of Single Cell Protein as a Protein Source in Broiler ChickenAuthors: Hombegowda G P, Suresh B N, Shivakumar M C, Ravikumar P, Ezhil Subbain
Gut Health, Feed Additives and Management
38
20] Study on Various Categories of Feed Acidifiers in Broiler ChickensAuthors: Emili Vinolya R, Rajalekshmi M
Gut Health, Feed Additives and Management
39
21] Effect of Acidifiers on Enhancing the Water Quality and Performance of Broiler ChickensAuthors: Nabila Fathima S, Rajendra Moorthy R, Ravichandran M
Gut Health, Feed Additives and Management
40
CONTENTS
22] Cost Effective Pathogen Control Program Ensuring Safe FeedAuthors: Arun Chandra Mazumder, Venkat M Shelke, Sarwar Ali
Gut Health, Feed Additives and Management
41
31] Polyphenols as an Additive Agent to Improve Oxidative Stability of Chicken Semen During Low Temperature StorageAuthors: Gopi M, Beulah P V, Prabakar G, Jaydip R J, Gautham K, Tyagi J S, Mohan J
Gut Health, Feed Additives and Management
50
32] Nano Nutraceuticals Preparation, Characterization and its Effect on Antioxidant Enzymes and Growth Performance in Broiler ChickensAuthors: Shwetha H S, Narayana Swamy M, Rajendran D, Preedia E, Babu, Prakash N, Jayanaik, Srikrishna Isloor, Kalmath, G P, Veena M P
Gut Health, Feed Additives and Management
51
33] Use of Medium Chain Fatty Acids as Replacement to Antibiotics in Broilers DietAuthors: Pratik Jadhav, Manwar S, Gole M, Langote G, Rathod A, Paradhi K
Gut Health, Feed Additives and Management
52
34] Effect of Dietary Supplementation of a Novel Phytogenic Blend on Organ Weights in Broiler ChickensAuthors: Oso O A, Umaya R S, Malik P K, Thirumalaisamy G, Awachat V B
Gut Health, Feed Additives and Management
53
35] Effect on Supplementation of Moringa oleifera Leaf Powder on Gut Health and Litter Quality in BroilersAuthors: Jayanti L Agashe, Manwar S J, Khose K K,Wade M R
Gut Health, Feed Additives and Management
54
36] Effect of pre-starter amino acid and trace mineral supplementation on growth performance and immune response in broiler chickenAuthors: Awachat V B, Elangovan A V, David C G, Ghosh J, Gowda N K S, Mishra A, Bhanja S K, Majumdar S
Gut Health, Feed Additives and Management
55
37] Dietary supplementation of Dicosan® vis-a-vis AGP improves gut health of commercial male broiler chickensAuthors: Rishikesh Jagzap
Gut Health, Feed Additives and Management
56
38] In ovo establishment of gut microbiome in chicksAuthors: Kolte A P, Saravanakumar M, Dhali A, Malik P K, Awachat V B, Jayaram C, Trivedi S, Elangovan A V, Bhatta R
Gut Health, Feed Additives and Management
57
39] Prevalence of Poultry Diseases in Wayanad District, KeralaAuthors: Dhanya T, Prasanna K S, Pradeep M, Alin Moideen T K, Elizabeth V, Shruthi A, Sachin, Anagha A M, Ansar F, Anoopraj R, Hamza P, Ajith J G
Disease Surveillance and Biosecurity
61
40] Study on Occurrence of ESBL Producing E.coli and Salmonella spp. in Poultry and its Farm Environment from Wayanad District, KeralaAuthors: Hema Persis Andrews, Asha K, Prejit, Jess Vergis
Disease Surveillance and Biosecurity
62
41] Studies on Pancreatic Pathology in PoultryAuthors: Brajesh Kumar, Gupta M K
Emerging Diseases of Poultry And Flock Health
65
23] Multienzyme Composite - A Comprehensive Approach for Enhanced Nutrient UtilizationAuthors: Krithika V, Karthigan M, Tarjan K
Gut Health, Feed Additives and Management
42
24] Effect of Dietary Supplementation of Antibiotic Growth Promoters vs Synbiotics on the Performance and Gut Integrity of Broilers and Detection of their Residues in Meat by HPLC,Authors: Rayala Reddy V, Ravinder Reddy V, Panda A K, Chinni Preetam V, Baswa Reddy P
Gut Health, Feed Additives and Management
43
25] Evaluation of Rice Polish on the Production Performance of Commercial Broiler ChickenAuthors: Shubhankar K S, Moorthy M, Shamsudeen P, Sakthivel P C
Gut Health, Feed Additives and Management
44
26] Histopathological Studies on Pancreatic Pathology in PoultryAuthors: Brajesh Kumar, Gupta M K
Gut Health, Feed Additives and Management
45
27] Evaluation of Pro-nutrient Effect of Combination of Lauric Acid and Glutamate in Broiler ChickensAuthors: Prabakar G, Gopi M, Nirmal Kumar C, Mahadevan K, Shankar N, Rajesh Kanna M
Gut Health, Feed Additives and Management
46
28] Effect of Different Water Sanitizers on Physico-Chemical and Microbiological Characteristics of Drinking Water in Commercial LayersAuthors: Sangeetha P V, Kannan D, Amutha R, Arthanari Eswaran M
Gut Health, Feed Additives and Management
47
29] Effects of Dietary Supplementation of Essential Oils as an Alternative to Antibiotic Growth Promoter on Livability and Economics of Commercial BroilersAuthors: Rajalekshmi C, Mishra R K, Savaliya F P, Bhagora N G
Gut Health, Feed Additives and Management
48
30] Effects of Dietary Supplementation of Essential Oils as an Alternative to Antibiotic Growth Promoter on Performance Parameters and Carcass Traits of Commercial BroilersAuthors: Rajalekshmi C, Mishra R K, Lunagariya P M, Patel A B
Gut Health, Feed Additives and Management
49
CONTENTS
CONTENTSCONTENTS42] Fowl Adenovirus Induced Gizzard Erosion and Incidence of Feed Passage Syndrome in Commercial Broiler Chicken
Authors: Amit Kumar Das, Venket M ShelkeEmerging Diseases of Poultry And Flock Health
66
43] Effect of Tinospora cordifolia on Infectious Bursal Disease Virus Induced Pathology in CockerelAuthors: A A Chavan, P M Sonkusale, S S Shinde, M P Kaore, M M Kadam and N V Kurkure
Emerging Diseases of Poultry And Flock Health
67
44] Comparative Assessment of Route of Induced Infection with Avibacterium paragallinarum in Chicken Employing Conventional Intra-Nasal Inoculation vis-a-vis Intra-Ocular and Nebulizer TechniqueAuthors: Sidhartha Deshmukh, Rajanbir Singh, Harmanjit Singh Banga Kathirvalen
Emerging Diseases of Poultry And Flock Health
68
45] Molecular Epidemiology of Newcastle Disease in India During Year 2019Authors: Bulbule N R, Pardeshi R B, Deshmukh V V, Kaje V S, Gupte A P, Chawak M M
Emerging Diseases of Poultry And Flock Health
69
46] A Case of Osteoma in an African Lovebird (Agapornis)Authors: Ansar Fasaludeen, Pradeep M, Anagha A M, Hamza Palekkoden, Anoopraj R, Prasanna K S, Ajith Jacob George
Emerging Diseases of Poultry And Flock Health
70
47] Ovarian Adenocarcinoma with Cystic Ovaries in a Duck (Anas platyrhynchos)Authors: Anagha A M, Pradeep M, Ansar Fasaludeen, Ajith Jacob George, Hamza Palekkodan, Anoopraj R, Nikhil S Rao, Prasanna K S
Emerging Diseases of Poultry And Flock Health
71
48] An Outbreak Of Degenerative Myopathy And Sarcocystis Infection in Layer ChickensAuthors: T K Alin Moideen, M. Pradeep, Dhanya Thomas, Elizabeth Varghese, A. Shruthi, Sachin, R. Anoopraj, Hamza Palekkodan, K.S Prasanna, Ajith Jacob George
Emerging Diseases of Poultry And Flock Health
72
49] An Outbreak of Favus in A Breeder Poultry Flock in GoaAuthors: Susitha Rajkumar, Nibedita Nayak, Chethan Kumar H B, Chakurkar E B
Emerging Diseases of Poultry And Flock Health
73
50] Molecular characterization of Newcastle disease virus in Central IndiaAuthors: Megha Kaore, Seema Shinde, Kurkure N V
Emerging Diseases of Poultry And Flock Health
74
51] Molecular Characterisation of Ornithobacterium rhinotrachealeAuthors: Megha Kaore, Seema Shinde, Kurkure N V
Emerging Diseases of Poultry And Flock Health
75
52] A Rare Case of Feather Folliculoma in a Japanese QuailAuthors: Azhaguraja M, Simmi Tomar N, Babu Prasath, Beulah Pearlin V
Emerging Diseases of Poultry And Flock Health
76
53] An Out Break of Infectious Bursal Disease in Commercial ChickenAuthors: Chandravathi T, Rama Devi V, Ramesh Babu T, Charitha P
Emerging Diseases of Poultry And Flock Health
77
54] Role of Micro RNAs in Diagnosis of Avian DiseasesAuthors: Rajeshwar Khandare, Sonal Saxena, Sameer Shrivastava, Noopur Dutta, Basant, Purnima Gogoi, Dipankar Bhist, Uzma Jabeen
Emerging Diseases of Poultry And Flock Health
78
55] Fatty Liver Hemorrhagic Syndrome in Desi BirdsAuthors: Charitha R, Chandravathi T, Ramadevi V, Sudha Rani Chowdary Ch , Samatha V
Emerging Diseases of Poultry And Flock Health
79
56] Recycling of Poultry Waste Through Bioconversion Using BSF Larvae for Sustainable Livestock ProductionAuthors: Niranjan B H, Md Nadeem Fairoze, Mahesh, Yandageri S, Elangovan A V
Developments in Poultry Products Technology
83
57] Optimization of Meat Particle Size in Development of Chicken Seekh Kebab Formulated with Spent Hen MeatAuthors: Mukesh Tukaram Nampalle, Ashim Kumar Biswas, Jaydip Jaywant Rokade, Jyotirmoy Saharia, Suchismita Pradhan, Monica Madeswaran
Developments in Poultry Products Technology
84
58] Food Safety Concern on Determination of Antibiotic Residues in Fresh and Cooked Broiler Meat by High Performance Liquid Chromatography (UV-Detector)Authors: Rayala Reddy V, Ravinder Reddy V, Baswa Reddy P ,Panda A K , Chinni Preetam V
Developments in Poultry Products Technology
85
59] Does Poultry Corona virus infection spreads to human beings?Authors: Sreenivas Gowda R N
Lead Paper
86
Lead Papers
Immune Evasion Strategies of Poultry Viruses
Abdul Rahman Omar
Institute of Bioscience, Faculty of Veterinary Medicine
Universiti Putra Malaysia, 43400 UPM Serdang, Selangor, Malaysia
aro@upm.edu.m
Introduction
Currently, the poultry industry is threatened by either more virulent viruses of endemic
diseases or by exotic and emerging diseases that can cause major economic losses to this
sector. Catastrophic diseases caused by velogenic Newcastle disease virus (NDV) and highly
pathogenic avian influenza (HPAI) H5N1 are not easy to overlook. The real challenge is to
confront immunosuppressive diseases, which act in concert with other factors such as
management, environment, and nutrition, forming a continuous threat to the poultry industry.
Besides, poultry with immunosuppression are highly susceptible to secondary infections and
unable to induce an effective response to vaccinations. The viruses that cause
immunosuppressive effects in chickens are oncogenic viruses such as Marek’s disease virus
(MDV), reticuloendotheliosis virus (REV), and avian leukosis virus (ALV). However,
nononcogenic viruses such as infectious bursal disease virus (IBDV) and chicken anemia virus
(CAV) are also able to invade the immune system by impairing the specific functions of immune
responses. Other viruses especially velogenic NDV, HPAI, pathogenic adenoviruses, and
reoviruses have also been reported able to modulate the immune responses, however, their
mechanisms of actions are not well-defined. In general, viruses that cause immunosuppression
can disrupt the immune system by directly destroying specific population of immune cells
and/or by releasing various mediators that impaired the immune functions of other cells. This
paper reviews the current updates on selected non-oncogenic immunosuppressive viral
diseases and their effects on the immune system of the poultry.
Immunosuppression
The damage to the immune system can be due to many intrinsic and extrinsic factors
such as infectious agents, mycotoxin, nutritional deficiencies, and management failures,
which can lead to stress, or a combination of them resulting in reduced effectiveness of the
immune system. Infectious disease driven modulation of the immune system and function
often associated with virus-induced apoptosis and/or necrosis of lymphoid cells, and virus-
induced modulation in the regulation of immune responses through the production of
interferons and cytokines which can lead to immunosuppression.
Successful poultry production depends on a significant degree of the prevention of
infectious diseases, the most important being immunosuppressive viral diseases. Even
subclinical infection with some of these pathogens can cause immunosuppression.
Immunosuppression can be either antigen-specific or generalized unresponsiveness towards
infection. The impact of the latter on poultry production is far greater than the former. However,
many viruses can cause both types of immunosuppression. Immunosuppressed chickens
usually are more prone to respiratory disease outbreaks, resulting in higher mortality and
morbidity, respond poorly to vaccination and medication, and exhibit increased post-
vaccination reactions. The term immunosuppression is often used as an excuse for poor
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Proceedings of the World Veterinary Poultry Association (India) Conference 2020 3
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performance in a flock when the actual cause is not known.
Non-oncogenic virus immune evasion strategies
Chicken infectious anemia virus (CAV) and infectious bursal disease virus (IBDV) are
lymphotropic viruses that can cause generalized impaired immune responsiveness, leading to
increased susceptibility to other diseases, thus regarded as the most important viral diseases of
the immune system. These small, non-enveloped viruses are very stable and can resist
relatively high temperature. CAV induced apoptosis of the lymphoid and erythroblastoid
progenitor cells, causing immunosuppression, hemorrhages, and anemia; thus resulting in an
increased prevalence of secondary infections. Meanwhile, IBDV is a lymphotropic virus that
targets B cells, macrophages and dendritic cells causing primarily depletion of IgM+ B cells.
Chickens infected with the virus especially very virulent strains showed a severe lesion in the
bursa of Fabricius and increased production of pro-inflammatory cytokines and IFN.
Other viruses such as reovirus, adenovirus, NDV and AIV have also been reported to
evade the immune functions via different mechanisms of actions. Most viral-induced systemic
infections modulate the expressions of interferons (IFN), a potent antiviral innate immunity
mediator. Besides, cytokines and IFN produced during virus infection modulate the key
component of the adaptive immunity, the B and T cell responses. However, infectious with
virulent viruses such as velogenic NDV, HPAI and vvIBDV can result in the excessive
production of proinflammatory cytokines and IFN and may contribute to morbidity and mortality
during the associated infection. Recently, we have shown viruses such as NDV and IBDV
(virulent versus vaccine strains) able to differential modulate another key component of the
innate immunity, NK cells found in the intestinal mucosa (IEL-NK) by modulating the expression
of activating receptors and inhibitory receptors.
Control of immunosuppressive viral diseases – A continuing challenge
In the field, vaccination failure is a complex event involving various factors associated
with vaccine strains and vaccination program, the virulence of field pathogens, and the host
immune competence. In many situations, immunosuppressive agents, primarily IBDV, and
CAV play an important role in increasing the susceptibility of chickens to opportunistic infections
and/or suppressing effective vaccine-induced responses.
The cornerstones of vaccination are of course B cells, which synthesize and secrete
antibodies that could neutralize invading agents or serve as anchor molecules for cellular
cytotoxicity. Even though B cells can respond to antigens, T cells are necessary for an optimal B
cell response, as an optimal response will only be achieved if at the same time an efficient T
helper response is induced. Therefore, every vaccination program should also aim at T cell
activation to ensure its success. However, almost all viral-induced immunosuppression
impaired T cell responses.
Effective control of immunosuppressive poultry diseases involves complex interactions
of management, nutrition, and flock health program. Biosecurity is an important component of
health management, and a vaccination program is the most important component of flock
health program. Since important immunosuppressive diseases of poultry are due to viral
infections, vaccination would be the focus of prevention. Undoubtedly, diagnostic tools,
Proceedings of the World Veterinary Poultry Association (India) Conference 20204
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vaccines, and vaccination equipment have improved over the years through the use of
innovative technology. However, vaccine is not evolution-proof, and it may enhance virus
evolution especially in the absence of a robust immunity; allowing wild-type viruses to be
transmitted through vaccinated chickens. The development of vaccine-induced immunity
depends on a complex set of interactions between macrophages, antigen-presenting cells,
lymphocytes, and cytokines. Immunosuppression can interfere with these interactions resulting
in a negative impact on the success of vaccination programs.
Proceedings of the World Veterinary Poultry Association (India) Conference 2020 5
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Current and Future Challenges of the Poultry Vaccine Industry
Marcelo Paniago
Ceva Animal Health Asia
marcelo.paniago@ceva.com
According to Food and Agriculture Organization of the United Nations (FAO), the world
population will be over 9 billion people by 2050 and, at the same time, the average per capita
income per country is projected to be a multiple of today’s level. This trend means that demand
for food will grow remarkably in the next 3 decades. Without a doubt, a big part of this gigantic
challenge falls on the shoulders of Asian producers as nearly 60% of the global population will
be in this continent.
In the poultry industry, some additional challenges lie ahead of producers such as labor
scarcity, consumers’ pressure on food safety and reduction of the use of antimicrobial drugs,
increasing concerns with animal welfare and environmental impact of the activity, water
scarcity, high disease pressure and many others.
In order to overcome these challenges and become more efficient, companies have
invested heavily on modernization of their production facilities (farms, hatcheries and
processing plants) with a crescent adoption of new technologies. Today, very large farms with
environmentally controlled houses and high levels of automation are seen everywhere.
However, this modern poultry industry has brought new challenges to producers. For
example, most of the farms are located in the same producing areas around big consumers’
centers and these new housing systems are able to hold much higher stocking densities. Not
surprisingly, it resulted in a very high poultry population density in most producing areas and the
risk of diseases challenges increases considerably in such circumstances.
In fact, an estimated one-fifth of livestock around the world are lost to diseases (GHI,
2016) and it is not different in the poultry industry. Therefore, preventing these losses through
biosecurity and vaccination became one of the pillars of animal production. Likewise the poultry
industry proper, vaccine manufacturers have invested massively to develop products that
address the major concerns of the producers.
Since Edward Jenner inoculated James Phipps with cow pox in 1796 and this procedure
was called “vaccination” (vacca = cow, in Latin), the evolution of this method of disease
prevention has developed remarkably. In 1870, Pasteur, working with a chicken disease - Fowl
Cholera, developed the concept of artificial attenuation of a microorganism giving origin to the
first vaccine using a live attenuated bacterial agent. Later on, in 1920’s, Gaston Ramon
developed a method to inactivate antitoxins with formalin, creating toxoids that were used for
immunization. Additionally, his work was pivotal for the development of the vaccine adjuvants.
The first generation of vaccines, thus, includes mostly killed or attenuated live microorganisms.
In 1953, Watson & Crick revealed to the world “the secret of life” and therefore new
horizons to the biological researches were opened. More recently, with the outstanding
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Proceedings of the World Veterinary Poultry Association (India) Conference 20206
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development of the knowledge on molecular technology, the possibility of developing better
vaccines became a reality.
Today, vaccines are developed based on two main drivers: the evolution of the poultry
production and the progress of molecular biology. There are several vaccines commercially
available that were developed using cutting-edge technologies such as immune complex,
vector, virus-like particles, sub-unit, gene-deletion, reverse genetic and others. Molecular
biology has dramatically enlarged the range of possibilities offered to creative researchers.
Many tracks can be followed and many concepts are waiting to be proven, developed and
turned into products.
In spite of all the available technologies to researchers, the so-called ideal vaccine
doesn’t exist. In fact, the mere concept of “ideal vaccine” would differ slightly according to the
disease that it is targeted to, the type of birds, the size of the farms and other factors. However,
in general, the requirements for such vaccine are similar: to be safe (not induce post-
vaccination reactions), to be applied in the hatcheries through in-ovo or subcutaneous route, to
be able overcome passive immunity, to induce long-lasting immunity with broad spectrum of
protection, to be compatible with other vaccines, to be differentiated from field infection (DIVA-
able), to be sold in large presentations, to be cost-effective, to mention a few.
Finally, there have been much more changes in vaccinology during the past 10 years
than during the previous 30 years. The limits for the creativity of researches today lies on
outdated regulations that will have to be, sooner or later, modernized to take into consideration
such innovations that will help producers to meet the colossal challenge of feeding more than 9
billion mouths in few decades.
References
Food and Agriculture Organization of the United Nations (FAO) website. www.who.int
Gardin, Y., Palya, V. and Elattrache, J. Innovations in poultry vaccinology. Ceva Animal Health, 2014
Jackwood, M.W. History of recombinant vaccines – Advances in Veterinary Medicine, v41 p517-522,
1999
Jorge, S. and Dellagostin, O.A. The development of veterinary vaccines: a review of traditional methods
and modern biotechnology approaches. Biotechnology Research and Innovation Volume 1, Issue 1,
January–December 2017, Pages 6-13.
OIE. The application of biotechnology to the development of veterinary vaccines. Guideline 3.3. OIE
Trerestrial Manual, 1338-1347, 2012
Truong, L., Morash, D., Liu, Y. et al. Food waste in animal feed with a focus on use for broilers. Int J Recycl
Org Waste Agricult 8, 417–429 (2019). https://doi.org/10.1007/s40093-019-0276-4
Zeigler, M and Steensland, A. Sustainability in an uncertain season. - Global Agricultural Productivity
R e p o r t ® ( G A P R e p o r t ® ) , G l o b a l H a r v e s t I n i t i a t i v e ( G H I ) , 2 0 1 6 . A c c e s s e d a t
https://www.globalharvestinitiative.org/GAP/2016_GAP_Report.pdf
Proceedings of the World Veterinary Poultry Association (India) Conference 2020 7
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Proceedings of the World Veterinary Poultry Association (India) Conference 20208
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Newcastle Disease: Current Situation and Strategies For Vaccination
Jose Luis Losada Torres
Hipra
jose.losada@hipra.com
INTRODUCTION
The Newcastle disease is a challenge worldwide causing economic losses because of
high mortalities, drop of production and economic restrictions. It is endemic in Latin America,
Asia, Africa and parts of Europe. It is considered exotic in few countries such as United States,
Brazil and parts of Western Europe.
It is a caused by a Paramixovirus serotype 1 (APMV-1), the APMV-1 is classified in two
groups, Class I and Class II, class I has just one genotype, was isolated in wild birds and it is
asymptomatic. Class II contains at least 18 genotypes (I – XVIII), those genotypes in some
cases can be divided in subgroups, the genotype XII is divided in in subgroups XIIa and XIIb).
According OIE Newcastle disease is defined as an infection of birds caused by a virus of
avian paramyxovirus serotype 1 (APMV-1) that meets one of the following criteria for virulence:
a) The virus has an intracerebral pathogenicity index (ICPI) in day-old chicks (Gallus
Gallus) of 0.7 or greater
or
b) Multiple basic amino acids have been demonstrated in the virus (either directly or by
deduction) at the C-terminus of the F2 protein and phenylalanine at residue 117, which is the N-
terminus of the F1 protein. The term 'multiple basic amino acids' refers to at least three arginine
or lysine residues between residues 113 to 116. Failure to demonstrate the characteristic
pattern of amino acid residues as described above would require characterisation of the
isolated virus by an ICPI test.
We have many options regarding live vaccines in order to control NDV field pressure,
and for clinical signs decreasing, but many times vets are not sure about what options should
they use and what pros and cons could they expect.
EPIDEMIOLOGY AND PATHOGENESIS
As in all pathologies affecting poultry the first movement should be decided based on
the disease itself, by knowing pathology and epidemiology of NDV in this case. The two main
reservoirs of the disease are backyard birds and wild birds (especially aquatic). The migration
routes of wild birds have the potential to spread the virus. The transmission is horizontal and
depends on the organs in which the virus multiplies. Birds with respiratory disease spread the
virus mainly through the air by aerosols from contaminated mucus, which may be inhaled by
others susceptible birds, and in lower quantity by faeces.
Some NDV viruses show tropism to intestinal replication, although they are able to
replicate as well in respiratory and reproductive tissues. These viruses are transmitted mainly
by ingestion of contaminated faeces, contaminated feed or water, or by inhalation of small
infective particles produced from dried faeces. The disease spreads by fomites, people and
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poultry products from the contaminated poultry farms included dead birds and natural fertilizer
from the poultry litter farms. The NDV is able to survive in the carcases or excretions on the litter
from several weeks at cool ambient or several years if held frozen temperatures. Virus is shed
from the very early infection, during clinical stages and for a limited period during
convalescence. The period between infection and clinical signs appearance vary from 2 to 15
days, for the purposes of the OIE Terrestrial Animal Health code is 21 days.
CONTROL BASICS
The vaccination program is one of the tools to control the clinical signs of the disease
and reduce virus shedding. The most usual vaccination programs against NDV are based in live
and killed vaccines (mainly in log life birds) and with a combination of just live vaccines, live +
killed vaccines or live + vectored vaccines. Live vaccines will provide local immunity (cell
mediated immunity and local humoral immunity) few days after vaccination, necessary to
protect the mucosa, and humoral systemic immunity after 2 weeks. Inactivated vaccines will
provide higher and more stable humoral immunity but they do not develop a strong cell
mediated response. The best route of vaccination with live vaccines against Newcastle disease
is by using the main route of infection (respiratory), because of limitations on the drinking water
vaccination method (temperature of water, chemicals and time of vaccination), the vaccines
with respiratory tropism are the LaSota strain based vaccines or B1 strain vaccines. Depending
on field pressure challenge, we should adapt our vaccination program to the different kind of
products that we can find in the market. As an example, we could say B1 type vaccines are
mainly used as priming at hatchery, or Lasota and Cloned vaccines are mainly used for
revaccination in farm.
Why are we using different strains with different purposes?
Mainly because, vaccines with better efficacy performance (LaSota and cloned) may
show some post vaccine reaction when they are not used properly, while B1 will show a better
performance in terms of safety, but lower efficacy when compared with LaSota and cloned
vaccines. Between LaSota and cloned vaccines, we should bear in mind that finally, a product
as Hipraviar® CLON is a LaSota strain, but cloned, this means, by cloning procedure, we look
for the best option from a parent LaSota virus, we select a virus population with increased
efficacy and reduced post vaccine reactions, we could call it an “improved” LaSota strain.
However, what really makes the differences is how the lab did select the correct cloned strain, in
order to keep LaSota efficacy.
CONCLUSIONS
We can conclude that each cloned vaccine should be considered individually, as the
selection process seems to be very important in order to keep LaSota´s efficacy and decreasing
post vaccine reactions. Other key point to assets vaccines against NDV is virus titer, as has
been highlighted several times (Cornax, I., Miller, P.J. and Afonso, C.L. 2012). All Hipra´s live
vaccines against ND are formulated always under a minimum titer of 10�,� EID50, providing an
optimal balance between efficacy and safety, and with a wide range of products ready to be
used under all circumstances.
It is clear that the most logical choice is the HIPRAVIAR® CLON and HIPRAVIAR®
Proceedings of the World Veterinary Poultry Association (India) Conference 2020 9
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CLON/H120 (strain CL79). CL79 strain is a Cloned LaSota strain, designed to give a lower
reaction compared to the conventional LaSota, with an IPIC of 0.15, and the same efficacy, as
per the trial above. The strategies for vaccination should take into account the local reality of the
pressure of infection of the region, biosecurity of the farm, maternal antibodies of the birds and
the route of application for live vaccines. The normal strategy is the use of inactivated and live
vaccines.
REFERENCES
Newcastle Disease Virus Vaccine Strains: Immunogenicity is not Influenced by ICPI. In: Orsi
MA, Doretto Júnior L, Reischak D, da Silva LHA, Spilki FR, Buzinaro MG, Arns CW Brazilian
Journal of Poultry Science
Cornax, I., Miller, P.J. and Afonso, C.L. (2012) Characterization of Live LaSota Vaccine
Strain–Induced Protection in Chickens upon Early Challenge with a Virulent Newcastle
Disease Virus of Heterologous Genotype, genotypes III, VIg, VIb, VIId, and IX. AVIAN
DISEASES 56:464–470.
Alexander, D. 2003. Newcastle Disease and Other Avian Paramixoviridae Infections. In:
Diseases of Poultry, Cap 2. 11th Edition. Saif, Barnes, Glisson, Fadly, Mc Dougald, Swayne
(eds.). AAAP Iowa State University - USA. 63-87.
Miller P.J and Koch G., 2013. Newcastle Disease, Other Avian Paramyxoviruses, and
Metapneumovirus Infections. Newcastle disease. In: Diseases of Poultry, 13th Edition. Ed. By
Swayne, D.E., Glisson, J.R., Mc Dougald, L.R., Nolan, L.K., and D.L. Suarez. Edit. Willey-
Blackwell Publishing. Iowa, USA p. 89-107.
Umali, D.V., Ito, H., Shirota, K., Ito, T., & Katoh, H. (2015). Atypical velogenic Newcastle
disease in a commercial layer flock in Japan. Poultry Science, 94 (5), 890-897.doi: 10.3382 / ps
/ pev011
Khorrajiya JH, Pandey S, Ghodasara PD, Joshi BP, Prajapati KS, Hodasara DJ, Mathakiya RA
(2015) Patho-epidemiological study on genotype-XIII Newcastle disease virus infection in
commercial vaccinated layer farms. Veterinary World 8 (3): 372-381.
Proceedings of the World Veterinary Poultry Association (India) Conference 202010
Lead Papers
Development Of Indianized Immune Complex Vaccine - VIPx : Effective Approach For Intervention of Novel vvIBDV Infection
Sanjay Gavkare
Ventri Biologicals, Venkateshwara Hatcheries Pvt. Ltd.,Pune
sanjay.gavkare@venkys.com
IBD is one of the economically most important disease, which is highly contagious and
acute viral disease that is characterized by destruction of lymphoid cells in the bursa of fabricius
causing sever immunosuppression. IBDV was first recognized as a disease entity in “Gumboro”
district of Delaware State in USA in 1957 by A.S. Cosgrove. The intrinsic property of the RNA
virus with low proofreading activity of their viral replicases, leads to quick evolution. Probably
due to this evolutionary selection process in the field, antigenic variants of IBDV were
discovered in the early 1980s and very virulent IBDV (vvIBDV) strains emerged in the late
1980s. Since 1992, IBDV is evolving in India resulting in severe economic losses which were
reported to break through high levels of maternal antibodies in commercial flocks having 60-
100% mortality in young chicks.
As the virus is constantly changing, there is a need to molecularly identify which critical
amino acids have changed. The region in VP2 that is typical for pathogenic strains (S-W-S-A-S-
G-S) occurs in all the Indian vvIBDV strains. All four differences in amino acids 222 A, 256 I, 294
I, and 299 S are characteristics of the vvIBDV strains, which are not present in other less virulent
strains. Substitutions at residues 73 (T to I), 160 (V to I), 212 (D to N) and 384 (V to I) were found
to be novel in the recent Indian VVIBDV isolates, who’s relevance to antigenicity/virulence
needs to be established. The phylogenetic tree prepared based on the amino acid sequence
shows clustering of all these isolates slightly deviating from the other vvIBDV sequences
reported previously from India and other countries.
Successful and effective vaccination depends upon understanding what IBD challenge
exists on the farm, knowing the bird’s maternal antibody status and selecting the appropriate
vaccine. Most commercially available conventional live IBDV vaccines are based on classical
virulent strains. Those classified as ‘‘mild’’ vaccines exhibit only poor efficacy in the presence of
certain levels of maternally derived antibodies and against vvIBDV. ‘‘Intermediate’’ and
‘‘intermediate plus’’ or ‘‘hot’’ vaccines have a much better efficacy and may break through
higher levels of maternally derived antibodies, but they can induce moderate to severe bursal
lesions and, thus, cause corresponding levels of immunosuppression. They may not fully
protect chickens against infection by the vvIBDV strains or by antigenic variants. Safety and
efficacy of this type of vaccine still remain a major concern. As field challenge increases, the
strength of vaccines should also increase. The Ventri’s IBDI+ attenuated vaccine clustered with
the vvIBDV strains from India, have proven to be more effective in protecting the flocks against
vvIBD outbreaks, since two decades compared to other conventional IBD vaccines (Sridevi and
Daral Jackwood., 2007).
Though effective vaccines are available, the practical on-farm administration of the
conventional live vaccines, to a large number of animals is also a technically demanding
process. The difficulties include farm-to-farm variability of chicks, farming conditions, skills in
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vaccination crews, etc. which should not be underestimated when assessing the results of
vaccination programmes.
The immune complex vaccines (Icx) concept has emerged to be a promising solution to
the current challenges. These vaccines consist of a mixture of a certain amount of IBDV-
specific antibodies obtained from the sera of hyperimmunized chickens and infectious IBD
vaccine virus. The major advantage is that they are suitable for in-ovo vaccination at day 18 of
incubation and can also be delivered by subcutaneous injection at 1 day old in the hatchery. In
addition, it has been shown that these vaccines were effective in the presence of maternally
derived antibodies. Considering the overall dynamics of the vvIBDV prevalent in India, the
challenge would be to identify the best strain and antibodies to be used in the immune complex
vaccine so as to provide the most promising solution to such challenges. So, Ventri Biologicals
explored the development of Immune complex vaccine (VIPx) using well established, proven
efficacious Ventri’s IBDI+ strain along with bursal disease antibodies. The trial studies with
VIPx have proved to be a promising alternative and comparatively the best efficacious vaccine
that controls the prevailing vvIBDV challenges in India.
Keywords:
VIPx; Immune-complex; Intermediate Plus
Proceedings of the World Veterinary Poultry Association (India) Conference 202012
Lead Papers
Recombinant Technology for ND and MD Vaccine strains
Vijayakumar E
MSD Animal Health, Pune
vijayakumar.ettigounder@merck.com
Early Protection... Always Producing More:
§ Recombinant vaccines are based on new technologies of molecular engineering
§ It is all based on the technology of the recombination of the DNA (genetic engineering)
§ Consists of a vector (virus or bacteria) expressing foreign antigens
§ Genes that codify for 1 or more antigens (proteins) are introduced in the vector’s genome
§ The immune response (cellular & humoral) is against the vector, including the inserted
antigen(s), without the need of using the causative agent
§ Innovax ND SB1 -First Generation Single Construct Recombinant vaccine
§ Innovax ND SB1 – HVT is a vector and in which part of ND Virus F gene of ND Clone is
inserted.
Natural ND Infection:
§ Expression of F gene in NDV responsible for F-protein on virus surface
§ F protein is attached to the host cell
§ F protein fuses with receptor cell membrane (Host cell)
§ Initiates the infection of Host cell
How Recombinant (Innovax ND SB1) works:
§ Insertion of the F gene of NDV in HVT genome
§ F protein is attached to the host cell
§ F protein fuses with receptor cell membrane (Host cell)
§ The HVT part will still induce Marek’s protection up to the level of related to HVT
§ The F-protein will induce F-antibodies and The F antibodies will neutralize the F protein of
ND virus “blocks” (prevents) the infection of the chicken after a NDV field exposure.
Advantages of a recombinant vaccine:
§ Absence of post vaccination reaction
§ No interference with ND MDA or other vaccines
§ No dissemination of vaccine virus
§ Convenient and homogenous administration (in-ovo or day-old hatchery)
§ Act as a marker vaccine (DIVA: Differentiate Infected from Vaccinated Animals)
§ Ease vaccination program 2 in 1
§ Development of immunity depends of the replication of the vector
§ Need of priming with conventional ND vaccines in high challenge areas
§ Serological monitoring is different
Efficacy of ND Sb1:
ND Challenge study shows that Protection from 2 weeks of age increasing to full protection* by
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4 weeks of age.
Challenge studies with vvNDV strains from different genotypes and geographic areas.
USA: Texas GB – Genotype II
Mexico: Chimalhuacan – Genotype V
Thailand: CU2 – Genotype VII
UK: Herts – Genotype IV
As reference the Hitchner B1 and LaSota strains belong to genotype II
Innovax ND -IBD – Second Generation Double construct:
§ 3 in 1 Protection (MD, ND, IBD)
§ HVT is a vector in which immunogenic part of ND Virus -F protein and immunogenic part of
IBD Virus -VP2 Gene inserted.
How Recombinant ( Innovax ND -IBD) works :
§ Vaccination s/c route ---- Infection of T and B cells ---- Expression of HVT, F, VP2 proteins ---
Induction of immune response.
§ ND, IBD, MD- Safety and Efficacy study done
Storage, Preparation and Application:
§ Recombinant vaccine storage - Cell associated vaccines (CA)
§ Stored in LN2 ( -196 C)
§ Thawing Time – 90 to 120 C
§ Thawing Temp – 27 C
§ Administrative Time- 45 Min
Summary:
Recombinant vaccines perfect fit for Innovatory product, Technology and Automation
Proceedings of the World Veterinary Poultry Association (India) Conference 202014
Lead Papers
Immune modulation and enhancement of immunity by the use of an adjuvanted solvent (Hipramune®T) in a live coccidiosis vaccine for breeders and layers (Evalon®)
1 1 1 1 2 3 3 4Pagès M , Bech G , March R , Sitjà M , Lillehoj H , Dardi M , Rubio J , del Cacho E1Hipra Scientific S.L.U., Avda. La Selva 135, 17170, Amer (Girona), Spain
2Animal Biosciences and Biotechnology Laboratory, USDA, Beltsville
3Hipra, Avda. La Selva 135, 17170, Amer (Girona), Spain
4Department of Parasitology, University of Zaragoza, Spain
shyongwey.ong@hipra.com
Objectives
In order to study the mechanisms by which an adjuvant could modulate the immune
mechanisms that are triggered in the host after coccidiosis vaccination, a study was performed
in which birds were vaccinated with a live attenuated coccidiosis vaccine (EVALON®) at one
day of age, with or without a selected adjuvant, to study important markers of Th1 and Th2
responses.
Materials and Methods
EVALON® is a live coccidiosis vaccine against avian coccidiosis. It is composed of five
attenuated strains: Eimeria acervulina, E. brunetti, E. maxima, E. necatrix and E. tenella.
Eimeria parasites have a complex life cycle that trigger a response by the host’s immune
system. However, they also elude the immune system, producing chemokines that can slow or
inhibit the host’s immune response. In this study, four treatment subgroups received
EVALON®, EVALON® in combination with HIPRAMUNE®T or phosphate buffered solution
(control group). Five birds from each subgroup were used to obtain intestinal lymphocytes from
the mucosa and Peyer’s patches at different time points post-vaccination (7, 23 and 43 days).
The lymphocytes were then incubated in appropriate medium and stimulated overnight with
Eimeria whole antigen. Later, the lymphocytes were fixed and stained using monoclonal
antibodies marked with fluorescein and studied with flow cytometry to detect lymphocytes
producing IL-2, IFN-�, IL-4 and IL-10.
Results and Discussion
The results obtained in the first experiment indicated that HIPRAMUNE®T is able to
increase the level of Th1 cytokines, as indicated by the results obtained for IL-2 (significantly
higher levels were detected at days 23 and 43 for the mucosa and at day 43 for the Peyer’s
patches) and IFN-� (significantly higher levels were detected at days 7 and 23 for the mucosa
and at days 23 and 43 for the Peyer’s patches). By contrast, the levels of IL-4 and IL-10 at days
23 and 43 were equal or lower in the EVALON® with HIPRAMUNE®T group compared to the
EVALON® alone. These results, combined with the results recorded for IL-2 and IFN-�, confirm
HIPRAMUNE®T’s ability to stimulate a cellular immune response. Therefore, it is hypothesised
that EVALON® administered together with HIPRAMUNE®T is able to polarise the immune
response towards a Th1 response with more intensity, as the study indicates, than the live
vaccine without the adjuvant. In vaccines designed for layers and breeders, it is of paramount
importance to provide extended protection throughout the life cycle.
Keywords
Coccidiosis; Immune modulation; Adjuvant; Vaccine; Attenuation
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Proceedings of the World Veterinary Poultry Association (India) Conference 2020 15
Lead Papers
Black Soldier Fly Larvae: A Novel Tool to Convert Poultry Waste to Wealth
Nadeem Mohamed, Niranjan B H
Department of Livestock Products Technology, Veterinary College, Karnataka Veterinary,
Animal & Fisheries Sciences University, Bangalore - 560 094
prof.nadeem@gmail.com
The Indian poultry sector has achieved great strides in terms of production and
productivity but is far behind with respect to waste management. This is an important cause of
concern from the point view of pollution potential, socio-economic issues encountered, house
fly menace and stray dog menace. A number of exercises tried in particular on one side to
combat the fly menace which threatened the very existence of the farms and on the other side to
handle the onslaught of the local public and the authorities due to ever increasing population
and the menace of the stray dogs. Hence, it is the need of the hour to evolve novel strategies to
convert this waste into a byproduct of commercial value. The harmless larvae of the Black
Soldier Fly(BSF) have a great potential to solve two of the modern agricultures’ growing
problems namely, serve as an alternative feed ingredient for animal feeds and disposal of
organic wastes, byproducts and side streams. The BSF larvae could very well be employed as a
novel tool as they have been recognized for their waste salvaging ability across different
organic waste streams following successful trials of artificial farming of BSF larvae in
confinement. These include livestock manures from confined large animal operations; palm
kernel waste, pig liver, kitchen waste, rendered fish and human night soil. This study was
designed to plan a strategy towards pollution control in the context of poultry operations in India
in converting the waste into commercially viable output. Hence, the waste generated from
different streams of poultry operations were considered for efficient and effective salvaging in
the concept of generating wealth from waste.
The experiment was designed to evaluate the waste salvaging ability of BSF larvae
extensively for the material reduction, bio conversion and FCR across test substrates
comprising of hatchery waste, slaughter waste and litter waste in different combinations (T1 to
T12). The performance of the test substrates were compared with that of the domestic kitchen
substrate (T0) in terms of larval performance and the profile of the resultant residue of the test
substrates following the introduction of 7 day old BSF larvae. All the 13 test substrates were
heterogeneous with respect to their attributes such as moisture, nutrient content and pH.
Overall, the test substrate T12 comprising of litter and poultry slaughter waste (litter included
both cage and deep litter at 1:1 ratio and poultry slaughter waste included both intestine and
head & legs at 1:1 ratio - on w/w basis) proved to be the best substrate for larval performance
from among the test substrates. Following the introduction of the 7 day old larvae in to the
various test substrates, the larvae performed well in T0, T7, T8, T9, T10, T11 and T12.
However, all the larvae released into the substrates T1, T2, T3, T4, T5 and T6 perished (100 per
cent) in the initial days of the study itself. The possible reason for the total mortality was
attributed to the swift drying of the substrate despite the mild sprinkling of the water. Time
residence is the term designated to describe the duration taken by 50 per cent the larvae to
emerge as pre-pupae, which indirectly reflected the nutritive value of the test substrate for the
effective larval growth. The larvae reared on T12 got metamorphosed by 20.33±0.33 days and
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Proceedings of the World Veterinary Poultry Association (India) Conference 202016
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became prepupae. In contrast, the time residence of the larvae reared on T9 and T7 stretched
up to 68.67±1.76 and 78.00±1.73 days.
The larvae reared on T9 and T7 recorded the longest period, 90.67±2.90 and
99.00±1.70 days, respectively to become pre-pupae was suggestive of the lowest nutritive
value of the respective test substrates. Even the larvae reared on T0 took less time than the
combination of different litters (T7, T8 and T9) despite lower nutrient profile, which was
attributed to the high caloric value owing to the starch content of domestic kitchen food waste.
The bioconversion was maximum in the test substrate T12 (31.83±1.49 per cent) than other
waste and it was minimum in T9 (12.69±0.32 per cent), possibly due to higher nutritive value of
T12. Despite the superior nutritive value of T9 over T0, the bioconversion was at the minimum
level which was attributed to the higher calorie content of T0. Material reduction was recorded
the highest in T0 and lowest in T8 which was due to the availability of more soluble
carbohydrates in kitchen food waste than the litter waste, which predominantly consisted of
lignin and cellulose, which the larvae found it difficult to utilize. The FCR was highest in T0,
which implied that the larvae had to consume more quantum of test substrate T0 as it had
comparatively less nutrients which might be the probable reason for its high material reduction
capability; whereas, the FCR was the lowest in T12.
The residue analysis was carried to know the effects of the larvae on the test substrates
and vice versa. Further, the residues were analyzed for their manure value in terms of Nitrogen,
Phosphorous, Potassium, total carbon content and C/N ratio. It was demonstrated that there
were high levels of N, P and K in the substrates with low carbon percentage along with
comparatively low C: N ratio. If these substrates were acted upon by the larvae, the levels of
these inorganic nutrient elements in the residue could have been further reduced. The test
substrates T0, T7, T8, T9, T10, T11 and T12 contained unutilized substrate biomass and larval
excreta. The analyses revealed that there were lower levels N, P and K in the residues
compared to the first block (T1 to T6). The Carbon and C: N ratio too was comparatively higher
that had more value for further value addition of organic material in the form of composting. The
observations with respect to proximate profile of the larvae revealed no significant difference
(P≤0.05) in protein content of the pre-pupae obtained from across the test substrates. But, the
correlation analysis between the protein content of the pre-pupae and that of the test substrates
revealed a correlation coefficient value (R2=0.364) which was statistically significant (P≤0.05).
In addition, the difference with respect to the EE content of the pre-pupae harvested from
among the test substrates was statistically significant (P≤0.05).
The livability of the larvae in test substrate was recorded in the study revealed that the
survivability was the highest at 100 per cent in T12 whereas; in T8 it was 87.5 per cent. The
difference in survivability across test substrates was attributed to the balanced nutritive value
and the substrate factors like type and combinations of the test substrates. It could be
concluded that the BSF larvae performed good on wastes harvested from different poultry
operations in combination (T0, T10, T11 and T12) in terms of material reduction, bioconversion
and feedstock conversion efficiency. The larvae reared on the substrates performed well in
terms of their early self harvest, quantitative and qualitative traits of the larvae.
Proceedings of the World Veterinary Poultry Association (India) Conference 2020 17
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Avian Disease Diagnosis – Dilemma Between Investigator and Investor
Dhinakar Raj G
Centre for Animal Health Studies, TANUVAS, Chennai - 600 051
dirtrpvb@gmail.com
With diagnostic science giant-leaping tremendously, the diagnostic platforms available
for disease diagnosis is ever-increasing. Ideally a diagnostic test should classify all subjects
who have the disease as “diseased” and vice versa. In practice, this is never the case!
The nucleic acid based detection methods such as the polymerase chain reactions
(PCR), its many variants and isothermal amplification methods have encompassed the earlier
technologies due to its speed, sensitivity and discriminating power. Among the protein based
diagnostic platforms, enzyme linked immunosorbent assays (ELISA) and Lateral flow assays
(LFA) have versatile applications. Various types of ELISA such as Indirect ELISA, Competitive
ELISA, Immuno Capture ELISA are commonly formulated as kits. With such a large choice
available, choosing the right test “fit-for-that purpose” is becoming challenging. 'Fit for purpose'
means that the test has to be validated to such a level to show that the results can be interpreted
to have a defined meaning in terms of diagnosis or any other biological property being
examined.
The most common ‘fit-for purpose’ applications defined by OIE for diagnostic assays are:
Ÿ Use in demonstrating freedom from infection in a defined population
Ÿ Use to certify freedom from infection or presence of the agent in individual animals or
products for trade purposes / prior to movement.
Ÿ Contribute to the eradication of disease or elimination of infection from defined populations.
Ÿ Confirm diagnosis of suspect or clinical cases
Ÿ Estimate prevalence of infection - Surveillance
Ÿ Determine immune status of individual animals or populations post-vaccination.
Use of diagnostic tests is context-based. A diagnostic test is used to determine or
confirm the presence or absence of a disease when a subject shows signs or symptoms of the
disease (Ruling in). This requires use of a test with reliable positive results for which a test with
high specificity (i.e. few false positives) is used. A screening test identifies asymptomatic
individuals who may have the disease (Ruling out). This requires use of a test with reliable
negative results for which a test with high sensitivity (i.e. few false negatives) is used.
A useful mnemonic to remember is "SPIN and SNOUT." SPIN stands for "SPecific tests
rule IN the condition when they're positive." SNOUT stands for "SeNsitive tests rule OUT the
condition when they're negative."
In addition to the intrinsic complications offered by the available assays, their individual
pros and cons, the contexts in which they need to be applied, the other additional challenge is
the difference in priorities of the investigator (scientists) and the investor (the commercial
companies). For example, when the investor would be interested mainly in ensuring that their
vaccine is able to protect against field isolates (protectotypic classification), the investigator
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Proceedings of the World Veterinary Poultry Association (India) Conference 202018
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would be more interested in their genomic and serologic patterns.
Due to the ease of performance of molecular techniques such as PCR and the
outsourcing facilities available for sequencing, most investigators are resorting to ‘PCRing’ their
field samples obtained, sequencing them, deriving phylogenetic classifications in comparison
with other related sequences available and coming to the conclusion on the circulation of novel
viruses, variants, new genotypes (serotypes) etc. This is essentially true for viruses such as
Infectious bronchitis, Newcastle disease virus, Fowl adeno viruses, Infectious laryngotracheitis
etc. Further, such works are published in scientific journals with low impact factors and leads the
industry to ponder on importation of new vaccines. It is emphasized that more valid and robust
scientific analysis spread over a wide geographical area are all critical before making decisions
of this kind.
Proceedings of the World Veterinary Poultry Association (India) Conference 2020 19
Lead Papers
Poultry Breeding and Genetics
ABSTRACTS
Genetic Analysis of Humoral and Cell Mediated Immune Response in
Japanese Quails
Monika M*, Rokade J J, Narayan R, Snehasmita Panda, Gopi M, Jyotirmoy Saharia
ICAR-CARI, Izzatnagar, Bareilly, Uttar Pradesh-243 122
monikakaniozhi@gmail.com
Background
Generally, broiler quails are slaughtered at about 5 to 6 weeks of age for economic
reasons. Selection for high body weights under modern intensive stressful conditions is often
accompanied by the increased incidence of diseases, which in turn impairs genetic
improvement in production traits. Information on the family on total immune-competence status
will increase the overall efficiency of production. The present investigation is designed to study
the effect of selection on immune-competence status in Japanese quails.
Materials and methods
A total of 450 Japanese quails, belonging to 40 sires of developed varieties of Japanese
quails (CARI-Ujjawal, CARI- Suneheri, CARI-Brown) were selected. The immunity traits
evaluation was done at the age of 6 weeks (HA Titre) & 7 weeks (PHAP). Humoral immunity will
be evaluated as antibody titre (HA) against 1% SRBC suspension (Siegal et al., 1980). The in-
Vivo Cell-mediated immune response (CMI) response to PHA-P mitogen evaluated by the
method of Cheng and Lamont (1988). The data obtained in different studies were subjected to
least squares analysis (JMP) and for heritability estimation within varieties by the full-sib
method as per Wombat mixed model analysis by REML.
Results
Varieties shows a highly significant difference (P<0.001) for HA titre. CARI-Ujjawal
shows the highest titre values as 4.01 when compared to CARI-Brown (3.82) and CARI-
Suneheri (3.70). No significant difference was observed due to the effect of sex on HA titer. For
cell-mediated immune response also CARI-Ujjawal shows the highest footpad index values as
0.40 when compared to CARI-Brown (0.38) and CARI-Suneheri (0.37). Sex has not shown any
significant impact on the footpad index values. The heritability estimates of immune-
competence traits showed that they were moderate to low in the estimation. HA titre, the range
of h² estimates was 0.258 to 0.443 for all the varieties and for PHAP, the observations were
between 0.011 to 0.171.
Conclusion
The heritability estimates observed in the present investigation revealed that the antibody titre
in CARI- Brown, and CARI-Suneheri was influenced by genetic variation and existence of a
distinct additive component which again can be improved through selection of sires for disease
resistance. The low heritability of the footpad index revealed that immune cell (T-cells)
proliferative activity has low genetic variation among the sires and offered less scope for its
improvement through the selection of sires.
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Proceedings of the World Veterinary Poultry Association (India) Conference 2020 23
Poultry Breeding and Genetics
Tanuvas Namakkal Gold Quail- A Potent Egg Laying Variety
M Moorthy*, Amutha R, Kannan D, Rajendran K, Kannan V
Veterinary College and Research Institute, Namakkal, Tamil Nadu - 637 002
moorthy.m@tanuvas.ac.in
The TANUVAS Namakkal gold quail was released by Tamil Nadu Veterinary and
Animal Sciences University, Chennai in the year 2011. Presently the bird stock is maintained at
the Department of Poultry Science, Veterinary College and Research Institute, Namakkal and
the germplasm is supplied to the needy farmers. It is a potent egg laying variety produced by
crossing male line (produced from four different lines) and Manchurian gold variety as female
line. The quails are maintained in cage system from day-old age onwards. The birds are fed with
Japanese quail chick mash (0 to 2 weeks) and grower mash (3 to 6 weeks) which was
standardized for egg type quails. It attains sexual maturity at the age of 40 days with a feed
consumption of 450 to 500g/bird with a body weight of 180 to 200g. The artificial lighting is given
two hours in the early morning and two hours in the late evening to have a total photo-period of
16 hours light during laying period. This quail variety can be used for egg production up to 54
weeks with a production of 257 eggs and with an average egg weight of 13.2g. No deworming
and vaccination is practiced during the entire period from 0 to 54 weeks of age. The peak egg
production is attained at the age of 10 to 12 weeks. The daily feed consumption during the egg
laying period is 30 to 35g/bird with a feed conversion ratio of 456g per dozen eggs. g the sires
and offered less scope for its improvement through the selection of sires.
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Proceedings of the World Veterinary Poultry Association (India) Conference 202024
Poultry Breeding and Genetics
Comparative Assessment of Fertility and Hatchability of
Six Breeds of Chicken Reared in University Poultry and Duck Farm
Nimmi P*, Supriya R, Binoj Chacko, Beena C Joseph, Sankaralingam S, Anitha P
Department of Poultry Science, College of Veterinary and Animal Sciences, Mannuthy,
Thrissur, Kerala - 680 651
drnimmijaleel@gmail.com
A study was conducted to compare the fertility and hatchability of six breeds of
chicken reared in deep litter system in University Poultry and Duck farm, KVASU,
Mannuthy. A total of 18422 eggs of Australorp (ALP), 37677 eggs of Kadaknath (DCK),
11022 eggs of Tellichery (DCT), 205720 eggs of Gramasree (GS), 17571 eggs of
Rhode Island Red (RIR) and 17426 eggs of White Leghorn (WLH) were assessed in
around 25 hatches. Infertility was lower in ALP (5.29%) and DCT (6.04%), intermediate
in GS (7.11%), RIR (7.82%) and WLH (7.87%) and higher in DCK (14.23%). There was
no significant difference in percentage of dead germs among the breeds. Dead in shell
was lower in DCT (8.63%), WLH (9.56%) and GS (10.36%), intermediate in ALP
(14.40%) and higher in DCK (17.30%) and RIR (17.58%). The hatchability on total egg
set was higher in DCT (83.13%) followed by GS (80.86%), WLH (80.04%), ALP
(78.16%), RIR (71.90%) and least in DCK (66.16%). Hatchability on fertile egg set was
higher in DCT (88.58%) followed by GS (87.05%), WLH (86.83%), ALP (82.50%) and
lower in RIR (78.01%) and DCK (77.09%). It was concluded that Tellichery breed
showed better performance on hatchability even though Australorp had higher
percentage of fertility. Rhode Island Red had highest percentage of dead in shell.
Kadaknath had lowest percentage of hatchability and fertility. The poor hatch
performance of Kadaknath might be due to inbreeding.
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Proceedings of the World Veterinary Poultry Association (India) Conference 2020 25
Poultry Breeding and Genetics
Biotic, Abiotic Stress and Welfare
Effect of Probiotic and Essential Oil on Broiler Performance
An Alternative to Antibiotic Growth Promoter
Karthigan M*, Tarjan K
Kemin Industries South Asia Pvt. Ltd., C3, 1st street, Ambattur Industrial Estate, Chennai,
Tamil Nadu-600 058
drscsekar@gmail.com
Antibiotic growth promoters (AGPs) is being used in animal production for decades, to
improve growth performance and controlling disease. There is a growing concern on the use of
AGPs because of the development of antibiotic resistances in pathogenic bacteria. Therefore,
there is a need for suitable AGP alternatives. Kemin Industries is currently having a probiotic
product (Bacillus subtilis – PB6) "CLOSTAT™" and an essential oil-based product (Blend of
carvacrol and thymol) "Orsential™ " as an AGP alternatives. An in vivo trial was conducted to
evaluate the growth performance of broiler birds for supplementation of CLOSTAT™ Dry and
Orsential™ Dry in comparison with AGP (bacitracin methylene disalicylate (BMD)). Cobb 430
male chicks were used for a period of 0 to 42 days in deep litter system using complete
randomized design. A total of 600 birds were randomly divided into 4 groups with 10 replicates
each and 15 birds per replicate. The groups consisted of, 1) control (no product), 2) AGP (BMD
15%) at 500 g/ton, 3) CLOSTAT™ at 500 g/ton and 4) Orsential™ at 150 g/ton. The parameters
such as body weight, feed consumption ratio (FCR), mortality, and European efficiency factor
(EEF) were analyzed. Both CLOSTAT™ and Orsential™ treated groups showed significant (P
< 0.05) improvement in body weight and FCR as compared to control. For the treatment groups,
a body weight gain of around 40 grams and improvement of 5 to 6 points FCR was observed
over the control and a body weight gain of 20 g and FCR of 2 to 3 points against BMD was
observed. There was no mortality observed in the trial. The European efficiency factor (EEF) for
BMD, CLOSTAT™ and Orsential™ treated groups were 11, 23 and 20 points higher than the
control group respectively. This study indicates that supplementation of CLOSTAT™ and
Orsential™ improves overall performance of broilers and can be used as potential AGP
replacement in poultry farming.
Keywords
AGP alternative; Probiotics; Essential oil; Animal trial
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Proceedings of the World Veterinary Poultry Association (India) Conference 2020 29
Biotic and Abiotic Stress and Welfare
Efficacy of Bacillus subtilis PB6 on Intestinal Health and Broiler
Economic Indices
Sai Ram Kumar*, Venket M Shelke, Partha Pratim Das, Swati Karki
Kemin Industries South Asia Pvt. Ltd., C3, 1st street, Ambattur Industrial Estate, Chennai,
Tamil Nadu - 600058
drscsekar@gmail.com
Poultry industry is looking towards modern trends of rearing and management practices
to get maximum productivity at a sustainable cost. A major challenge affecting broiler
production performance is poor intestinal health and an impaired gut microbiota. Intestinal
health management ensures optimum microbiota and intestinal integrity for nutrient absorption.
Several nutritional and pathological factors lead to imbalance in microbiota and disruption of the
intestinal health, which results in poor performance. Clostridium perfringens is one of the main
disruptive agent causing necrotic enteritis (NE) which affects the intestinal health. Also, C.
perfringens shows major impact on heavy losses in terms of economics, mortality and uneven
body weight in birds. Bacillus subtilis PB6, an active ingredient in CLOSTAT™ SP has direct
killing effect on Clostridium perfringens. The main objective of trial was to check the efficacy of
CLOSTAT™ SP on the intestinal health condition and economic indices in commercial broilers.
The trial consisted of 2 groups, viz. control group treated without any antibiotics or probiotics in
water and treatment group with CLOSTAT™ SP at 1gm/ 2 liter of drinking water. The birds were
divided and segregated as 2,800 birds in control group and 4,700 birds in treatment group,
which were reared for 39 days. Productivity parameters, viz., body weight, feed conversion ratio
(FCR) and mortality were recorded on weekly basis followed by final report after complete
culling of both the control and treated flocks. Also, the litter condition and lesion scoring were
observed during the trial after 4th and 5th weeks of age. Finally, FCR was calculated for both the
groups. Results indicated that the productivity parameters of the treated flock were
considerably better than control. The trial results strongly supported the fact that usage of
CLOSTAT™ SP in broilers improved the overall performance with better economics.
CLOSTAT™ SP usage had shown a reduced 8 points FCR and 0.9% mortality in treated group
over control at the completion of trial. The litter condition was poor in control group when
compared to treatment. The lesion scoring studies revealed that the level of dysbacteriosis on
28th day was 1.67 and 0.33 in control and CLOSTAT™ SP groups respectively. Whereas, the
dysbacteriosis level on 35th day was observed to be 1.33 and 0.33 in control and CLOSTAT™
SP groups respectively. Thus, the present study showed that using CLOSTAT™ SP offers
better performance over the control group in all the performance parameters.
Keywords
CLOSTAT™ SP; Dysbacteriosis; Necrotic Enteritis; Lesion scoring; Broiler
Performance; Economic Benefits
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Proceedings of the World Veterinary Poultry Association (India) Conference 202030
Biotic and Abiotic Stress and Welfare
Bacillus subtilis PB6 - A Complete Solution for Dysbacteriosis in
Broilers
Venket M Shelke*, Saravanan Sankaran, Chandrasekar S
Kemin Industries South Asia Pvt. Ltd., C3, 1st street, Ambattur Industrial Estate, Chennai,
Tamil Nadu - 600 058
drscsekar@gmail.com
Intestinal health is very much essential for economically viable broiler production. Due
to rapid growth, unconventional feed raw materials and excessive usage of antibiotics, the
intestinal health of broiler chicken is compromised in South Asian countries. Coccidiosis and
Necrotic enteritis are the two major and primary diseases for intestinal health upset. Apart from
these two, one of the widespread syndromes causing poor intestinal health in broilers without
causing mortality is referred as Dysbacteriosis or Nonspecific bacterial enteritis. This condition
is also called as small intestinal bacterial over growth (SIBO) resulting in undigested feed
particles in the droppings, diarrhea and wet litter which in turn cause poor performance. The
study assessed the occurrence of Dysbacteriosis and Coccidiosis in South Asian broiler farms
for the years 2017 and 2018. Also compared the effect of CLOSTAT™ containing Bacillus
subtilis PB6 on controlling Dysbacteriosis and in relation with coccidiosis challenge. For
Coccidiosis, the lesion scoring method developed by Johnson and Reid and for Dysbacteriosis,
the macroscopic scoring method developed by Teirlynck, et al were used for this study. Totally
208 and 224 broiler houses were analyzed from 47 broiler producers across South Asia for
2017 and 2018 respectively. The broiler age range selected for this study was 15 to 45 days with
average age of 26 days based on the occurrence of Coccidiosis and Dysbacteriosis. The
average Dysbacteriosis score in CLOSTAT™ users was 0.63 which was significantly lower
(P≤0.05) than non CLOSTAT™ users which was 1.00 in 2017. In 2018, the Dysbacteriosis
score in CLOSTAT™ users was 0.66 which was also significantly lower (P≤0.05) than the non
CLOSTAT™ users which was 1.10. But the overall average coccidiosis score was more than
one in both the years and there was no significant difference (P≥0.05) between CLOSTAT™
users and non-users. This study revealed that CLOSTAT™ containing Bacillus subtilis PB6 is a
complete solution in controlling Dysbacteriosis despite adverse conditions including
coccidiosis challenge.
Keywords
CLOSTAT™; Dysbacteriosis; Coccidiosis; Lesion scoring
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Proceedings of the World Veterinary Poultry Association (India) Conference 2020 31
Biotic and Abiotic Stress and Welfare
Chicken-Chimera: Primordial Germ Cells Mediated Poultry Germplasm
Conservation
Adnan Naim , Sushobhan Dash , Mishra S K , Debasis Nayak , Chittaranjan Jena1* 1 2 3 1
1KIIT- Technology Business Incubator, KIIT University, Bhubaneswar - ODISHA
2Central Avian Research Institute, Regional Centre, Bhubaneswar - ODISHA
3Discipline of BioSciences and Biomedical Engineering, Indian Institute of Technology
Indore, M.P.
adnan.kiittbi@gmail.com
Avian Influenza outbreaks, global climate change including biotic and abiotic stress has
badly affected the productivity of genetically rich and economically valuable chicken lines.
Currently, such important chicken breeds are maintained as live birds and therefore, remain at
the risk of influenza virus attacks and other diseases, which lead to complete eradication of the
whole population of a particular chicken breed. An alternative to current chicken breed
conservation strategy is to isolate the early embryonic germ cells like Primordial Germ Cells,
from a target chicken breed and cryo-preserve them in liquid nitrogen. The cryo-preserved
PGCs can be retrieved back in the future and can be transplanted into a recipient chicken
embryo host, which will give rise to a Chimera. A germline chimera chicken will have a genotype
of the donor chicken breed and a phenotype of the host. Later, by back-crossing the germline
chimera, one can successfully reconstitute the target chicken breed. Our research work
objective was to explore the potential of Primordial Germ Cells (PGCs) from some indigenous
poultry breeds as a source of poultry germplasm conservation. We successfully isolated the
PGCs from circulating blood and gonads of native chicken breeds. We also successfully
cultured the PGCs under in-vitro conditions and cryo-preserved them. We use Periodic Acid
Schiff' (PAS) stain method and Stage Specific Embryonic Antigen -1 (SSEA-1) to characterize
the isolated and cultured PGCs. We also successfully established the technology of getting a
germline chimera as we found the exogenous PGCs (donor PGCs labeled with tracking dye)
colonizing in the recipient's gonads. In conclusion, we reported the first time in India,
establishing a technology of avian germplasm bio-banking by harvesting, in-vitro culturing and
cryo-preserving the Primordial Germ Cells. Our future aim is to reconstitute a target chicken
population by back-crossing the PGC mediated germline chimera.
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Proceedings of the World Veterinary Poultry Association (India) Conference 202032
Biotic and Abiotic Stress and Welfare
Evaluation of Embryonic and Early Post-Hatch Thermal Conditioning on
the Thermotolerance Ability of Broiler Chickens
Aswathi P B*, Bhanja S K, Darshana B Bhaisare, Manish Mehra, Akshat Goel, Junaid N
Department of Poultry Science, College of Veterinary and Animal Sciences,
Kerala Veterinary & Animal Sciences University, Pookode, Wayanad, Kerala.
aswathi@kvasu.ac.in
Broilers are highly susceptible to heat stress owing to heavy body weight and limited
heat dissipation mechanisms. Exposure to various thermal-hygrometric conditions in neonatal
may result in epigenetic adaption in birds leading to better thermotolerance. Experiment to
improve thermotolerance of broilers at finisher age was attempted by thermal conditioning at 2
stages (1) embryonic thermal manipulation (ETM) with 39°C + RH 60-70% for 6hrs/per day from
ED 11 to 15 (2) Post-hatch thermal manipulation (PHTM at 37°C + RH 60% 6hrs/per day for first
2 weeks. After 35th day post-hatch, birds were exposed to thermal stress for a period of 7 days
(THI >90; i.e. 37±1°C & RH 60-70% for 6 hrs per day). A control was maintained at standard
environmental condition. Body weight was significantly higher (P<0.001) for ETM and PHTM
group on 35th (1342.2g and 1348.3g) and 42nd day (1759.72g and 1748.75g) against control
(1317.5g and 1622.40g on respective days). Better feed conversion ratio was recorded at 35th
day of ETM and PHHS (1.69 and 1.80) and 42nd day post-hatch (2.05 and 2.11) than the control
group (1.8 and 2.25). Serum cholesterol level was significantly lower (P<0.05) on 35th and 42nd
day in ETM (131.0 mg/dl and 133.7 mg/dl) and PHTM (130.9 mg/dl and 136.7 mg/dl) group
chicks than the control group (133.0 mg/dl and 142.4 mg/dl). On 42nd day, H:L ratio of ETM
(0.55) and PHTM (0.52) were significantly lower than control (0.60). Down regulation of HSP70
gene expression in brain tissue was observed after heat stress challenge in ETM and PHTM
with reference to control group indicating epigenetic adaptation and improved stress tolerance
in birds subjected to early age temperature manipulation. In conclusion, early stage thermal
manipulation (ETM and PHTM) is a cost effective mechanism to improve the thermotolerance of
broilers resulting in better growth rate and survivability.
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Proceedings of the World Veterinary Poultry Association (India) Conference 2020 33
Biotic and Abiotic Stress and Welfare
Effect of Premature Moulting On Certain Biochemical Parameters In
Commercial Layer Hens
David I C G*, Ravi Kiran Gorti, Reddy I J, Manpal Sridhar
ICAR-National Institute of Animal Nutrition and Physiology, Bengaluru
corbon.david@gmail.com
Commercial layer hens lay around 330 eggs over a productive period of 55 weeks by
reaching peak production when they are 33-35 weeks old. Often the laying hens are moulted to
improve production and quality of egg. At times when layer hens are exposed to stressful
conditions arising from shortage of feed, water, disease etc., premature moulting occurs during
the production cycle. Moulting causes oxidative stress at cellular level and the exact
physiological responses in different organs is not fully understood. This study compared the
effect of premature moulting on egg production, FCR, and changes in the circulating levels of
uric acid, fatty-acid binding protein 6 (FABP-6) and membrane potential of mitochondria (MMP)
isolated from the duodenum and uterine gland.
Bv300 strain hens were reared in individual cages and fed on standard layer ration (CP
16% and ME 11.28 MJ/Kg) ad libitum with free access to water from 18 to 50 weeks of age.
Several of the birds began to moult between 30th and 40th weeks of age. Fifteen moulted birds
were selected and hen-day egg production (20-40 weeks) was compared with non-moulted
birds (n=15). Blood samples were collected on 20 and 40 weeks of age to study the uric acid,
ileal fatty-acid binding protein 6 (IFBP-6). Six birds from each group were sacrificed on 40th
week and membrane potential of mitochondria isolated from the duodenum and uterine gland
were estimated. The hen day egg production between 20-40 weeks of age was significantly
(P>0.05) lower in the prematurely moulted group compared to non moulted group. The
circulating levels or uric acid and FABP-6 were significantly (P>0.05) higher in moulted birds.
These changes were associated with a concomitant decrease in the membrane potential of the
mitochondria isolated from the duodenum and uterine gland.
We conclude that increased uric acid levels beyond physiological limits by acting as a
pro-oxidant could affect the intestinal integrity (FABP-6) coupled with loss of mitochondrial
membrane integrity (low MMP thus ATP synthesis). These changes might affect the nutrient
absorption and secretion at intestinal and egg shell gland collectively to alter egg laying
potential of layer hens moulted prematurely.
Keywords
Layer hens; Pre-mature moult; FABP-6; Membrane potential; Uric acid
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Proceedings of the World Veterinary Poultry Association (India) Conference 202034
Biotic and Abiotic Stress and Welfare
Gut Health, Feed Additives and
Management
Gut Health and Serum Characteristics of Commercial Broilers Fed with
Unconventional Feed (Soya Oil Cake)
Sonale N S *, Rajendran K, Moorthy M, Kathirvalen C
Suguna Foods Pvt. Ltd., Nashik, Maharashtra
dr.nssonale@gmail.com
Incorporation of conventional feed ingredients like maize, soybean meal, fish meat, etc.
in broiler feed has increased the cost of feed enormously. With the current shortage and rising
costs of conventional feed ingredients, scientists have advocated the use of agro-industrial by-
products as unconventional feedstuff eg. soya oil cake. A study was undertaken to assess the
effect of dietary supplementation of soya oil cake on gut health and serum biochemistry of
commercial broilers fed with different levels of soya oil cake in the EC housing system. Soya oil
cake has a long shelf life, a high level of gross energy (9400 Kcal/kg), less than 0.5% moisture
and less than 2% free fatty acid (as oleic acid) compared to other vegetable oils.250
commercial, one-day-old, broiler chicks were randomly grouped into five treatments with five
replicates containing ten chicks per replicate. The chicks were reared from day-old to thirty-five
days of age EC house. The treatment groups were fed with basal diet with supplementation of
100 percent vegetable oil (rice bran oil-T1) and rice bran oil is replaced with 25 percent soya oil
cake (T2), 50 percent soya oil cake (T3), 75 percent soya oil cake (T4) and 100 percent soya oil
cake (T5). The data collected were subjected to statistical analyses suggested by Snedecor
and Cochran (1989). Supplementation of soya oil cake at the level of 75 percent on broiler diet
(T4) showed significantly (P<0.05) increased level of serum protein. The 50 percent of the soya
oil cake supplemented group had shown significantly (P<0.05) higher level of serum albumin
and serum albumin: globulin ratio. The serum lipid profile viz. total cholesterol, high-density
lipoprotein, low-density lipoprotein, and triglycerides did not show a significant difference
among the treatment. The mean (± S.E.) intestinal histo-morphology of broilers as influenced
by feeding different levels of soya oil cake shows that the intestinal villi height and villi height:
crypt depth showed significant difference (P<0.01) among the treatment groups, but the crypt
depth showed no significant difference among the treatment groups.T5 (1334.55µm) recorded
significantly (P<0.05) higher intestinal villi height and also significantly (P<0.05) higher
intestinal villi height: crypt depth ratio than with other groups. Based on the results of the
experiment, soya oil cake could be replaced with any other vegetable oil up to 75% for better
serum characteristics and intestinal morphology.
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Proceedings of the World Veterinary Poultry Association (India) Conference 2020 37
Gut Health, Feed Additives and Management
Nutritional evaluation of Single Cell Protein as a Protein Source in
Broiler Chicken
Hombegowda G P*, Suresh B N, Shivakumar M C, Ravikumar P, Ezhil Subbain
Department of Animal Nutrition, Veterinary College, Hassan, Karnataka - 573 202
hombesiri6195@gmail.com
Single cell proteins (SCP) are produced from microorganisms utilizing agro-industrial
waste and have the potential to serve as alternative source to meet the demand of protein rich
feed ingredients required for poultry industry. In this direction, the effect of SCP on the growth
performance, carcass characteristics, blood biochemical and gut health status of broiler
chickens was evaluated. Day-old straight-run broiler chicks (Cobb, n=400) were assigned to
ten dietary treatments. Ten iso-nitrogenous and iso-caloric experimental diets containing 0, 2.5,
5, 7.5 and 10 % SCP replacing either soybean meal (T1 to T5) or fish meal (T6 to T10) were
formulated. Each diet, prepared for starter (0-14 days), grower (15-21 days) and finisher (22-42
days) phases, was offered to four replicate groups of 10 chicks each. The results revealed
higher (P<0.01) body weight gain in groups fed 2.5 and 5 % SCP diets and lower (P<0.01) body
weight gain and feed consumption in the groups fed 7.5 and 10% SCP diets compared to those
fed the control during different phases and over the entire experiment. The feed conversion
ratio was better in 2.5 and 5 % SCP incorporated diets and poorer in 7.5 and 10 % SCP
incorporated diets during different phases and cumulatively. On 42nd day, the carcass
evaluation revealed that the inclusion of SCP up to 10% by replacing either soybean meal or fish
meal had no significant (p>0.05) effect on dressing percentage, drumstick percentage,
abdominal fat percentage and meat to bone ratio. However, the breast meat yield was higher
(p<0.01) in broilers fed 2.5 and 5% SCP diets while the broilers fed on 7.5 and 10% 0SCP dies
showed higher (p<0.01) thigh percentage. The duodenum and jejunum villi length, crypt depth
and villi length to crypt depth ratio were similar between control and 2.5 or 5 % incorporated
diets fed groups however significantly (p<0.01) lower at 7.5 or 10% SCP incorporation in diets.
However, ileum crypt depth and villi length to crypt depth ratio were remained similar among
different treatment groups. Further, inclusion of SCP upto 10% replacing either soybean meal
or fish meal in the broiler diets has not affected total microbial count and pH of intestinal content.
It was concluded that the inclusion of SCP upto 5% replacing soybean meal in broiler diets is
beneficial in improving growth rate with better breast meat yield and intestinal morphology.
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Proceedings of the World Veterinary Poultry Association (India) Conference 202038
Gut Health, Feed Additives and Management
Study on Various Categories of Feed Acidifiers in Broiler Chickens
Emili Vinolya R*, Rajalekshmi M
Kemin Industries South Asia Pvt. Ltd., C3, 1st street, Ambattur Industrial Estate, Chennai,
Tamil Nadu - 600 058
drscsekar@gmail.com
Dietary acidifiers are considered to be an alternative to feed antibiotics which improve
bird's performance by lowering gastric pH to enhance proteolytic enzymes action and by
reducing intestinal pathogenic bacteria. Several types of acidifiers are available such as
organic, inorganic and coated acidifiers. This study focused on the effect of dietary
supplementation of different acidifiers and its effect on broiler performance and intestinal health
under high challenge condition (Feed with 5% animal protein). A 42 days trial was conducted in
a total of 800 1-day old Cobb 400 chicks were randomly segregated into 8 test groups (10
replicates/group and 20 birds /replicate) using complete randomized design in deep litter
system. A corn soya mash diet with 5% meat and bone meal was fed ad libitum. Different
categories of acidifier products such as encapsulated butyric acid (T1), glyceride esters of
butyric acid (T2), phosphoric acid (T3), sodium di formate (T4), blend of organic acids (T5),
blend of organic acid and essential oil (T6), blends of organic acid and medium chain fatty acid
(T7) were tested at their recommended dosages (T1,T2 -300g/MT and other products -
1kg/MT). A control group was kept without acidifier. Bird's performance was measured in terms
of average body weight (ABW), feed conversion ratio (FCR) and mortality rate at every week.
Intestinal health was studied through scoring on bacterial enteritis, measuring the bacterial
population in ileum (Clostridium perfringens, lactic acid bacteria) at the end of trial. The
performance study results indicate that the products T2 and T5 showed significantly higher (P <
0.05) ABW and lower (P < 0.05) FCR by 5 points than control. Mortality was relatively
decreased up to 86% by all treatments expect T4 compared to control. The test products such
as T5, T6, T7 showed significant benefits in all intestinal health studies as follows lower (P <
0.05) bacterial enteritis lower (P < 0.05) C. perfringens, higher (P<0.05) lactic acid bacteria than
control. The study results confirm that the blends of organic acids (with or without other
biomolecules) deliver both performance and intestinal benefits than other products.
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Proceedings of the World Veterinary Poultry Association (India) Conference 2020 39
Gut Health, Feed Additives and Management
Effect of Acidifiers on Enhancing the Water Quality and Performance of
Broiler Chickens
Nabila Fathima S*, Rajendra Moorthy R, Ravichandran M
Kemin Industries South Asia Pvt. Ltd., C3, 1st street, Ambattur Industrial Estate, Chennai,
Tamil Nadu - 600058
drscsekar@gmail.com
Water is an essential nutrient in poultry's health and nutrition. The ideal pH for drinking
water is around 6.5 with hardness of 60 to 180 ppm. In India, poultry drinking water of around
500 ppm hardness, pH of 7.8 and microbial load is commonly seen. The objective of this work
was to study the effect of AcidLAC™ azure (a liquid acidifier product) on enhancing water
quality by pH reduction, antimicrobial efficacy, and performance of chickens provided with 900
ppm hard water. In natural water of hardness 750 ppm, 0.1 mL/L of AcidLAC™ azure reduced
the pH from 7.61 to 6.74. In artificial hard water of varying hardness, 0.2 mL/L of AcidLAC™
azure brought down the pH to around 6.5. AcidLAC™ azure showed inhibition against
Staphylococcus aureus and Escherichia coli, when tested by minimal inhibitory concentration
and zone of inhibition studies. An in vivo performance trial was conducted to evaluate the effect
of AcidLAC™ azure in Cobb 430 birds using complete randomized design. A total of 196-day
old chicks was divided into control and treatment groups with 8 replicates per group and 12 birds
per replicate. The trial consists of control group provided with water alone and treatment group
treated with AcidLAC™ azure at 0.15 mL/L dose for a period of 42 days. At the end of the trial, no
significant difference in body weight was observed between control (2317.92 g) and treatment
group (2275.38 g). Significant improvement in the feed conversion ratio (FCR) was seen in the
treatment group (1.69) compared to control group (1.76) at 95% confidence interval, which
resulted in return on investment of 3:1 in AcidLAC™ azure treated group. The above results
suggest that use of acidifier helps in improving the water quality (pH and microbes), thereby
enhancing the broiler birds' performance.
Keywords
Acidifiers; pH; Hardness; Water quality
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Proceedings of the World Veterinary Poultry Association (India) Conference 202040
Gut Health, Feed Additives and Management
Cost Effective Pathogen Control Program Ensuring Safe Feed
Arun Chandra Mazumder*, Venkat M Shelke, Sarwar Ali
Kemin Industries South Asia Pvt. Ltd., C3, 1st street, Ambattur Industrial Estate, Chennai,
Tamil Nadu - 600 058
drscsekar@gmail.com
Poultry feeds can serve as a carrier for a wide range of pathogenic microorganism
including Salmonella, E. coli and mould that have significant negative impact on birds' health
and egg production. Egg production cost can increase more than 20% due to mortality caused
by disease. A liquid balanced blend of organic acids, essential oils and surfactants, which is
effective against Enterobacteriaceae and mould in feed raw materials can be used to maintain
better feed hygiene. A study was conducted to evaluate the efficacy of liquid blend (T2) in
comparison with the combination of two different products in powder form, recommended as
Salmonella killer and mould inhibitor. Liquid blend was applied by spraying on layer mash feed
during mixing at 1 kg per ton of feed. Dry Salmonella killer and mould inhibitor was used at
1.25kg and 0.5kg per ton of feed by pour on in the mixture respectively. Treated feed was kept at
feed mill for 30 days and feed samples were collected at 2nd, 15th and 30th day after treatment
for microbial analysis. Results indicated that both the groups effectively controlled Salmonella
and E. coli. The mould counts were 8000, 6900 and 2000 cfu/gm of feed for T1 group & 2900,
1900 and 1000 cfu/gm of feed for T2 group at three different storage periods respectively. The
mould count was reduced in T2 when compared to T1 at 3 different storage periods on 2nd, 15th
and 30th day of analysis. Thus, the study shows that liquid blend of organic acids, essential oils
and surfactants is a potential and cost-effective tool for feed hygiene program.
Keywords
Organic acids; Essential oils; Salmonella; E. coli; Mould
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Proceedings of the World Veterinary Poultry Association (India) Conference 2020 41
Gut Health, Feed Additives and Management
Multienzyme Composite- A Comprehensive Approach for Enhanced
Nutrient Utilization
Krithika V, Karthigan M*, Tarjan K
Kemin Industries South Asia Pvt. Ltd., C3, 1st street, Ambattur Industrial Estate, Chennai,
Tamil Nadu - 600 058
drscsekar@gmail.com
The increasing cost of conventional poultry feed ingredients is a major concern, since
feed constitutes about 75% of the overall production cost. This has led to the use of cheaper,
unconventional alternative raw materials. However, the digestibility of these raw materials is
affected by the presence of anti-nutritional factors such as non-starch polysaccharides (NSP),
which are known to reduce the nutrient digestibility in birds by binding higher quantities of water
and increasing intestinal viscosity. Supplementation of exogenous NSP degrading enzymes
(NSPase) help in reducing the intestinal viscosity by enhancing the NSP hydrolysis thereby
improving the digestibility and productivity of animals. Nutrikem™ XL PRO is a blend of
NSPase, lysophospholipids and protease, which work synergistically to enhance the overall
metabolism in birds. The objective of this study was to evaluate the efficacy of Nutrikem™ XL
PRO, in improving the performance of broiler birds fed with diet deficient in metabolizable
energy (ME) and amino acids. A total of 600, Cobb 430 male birds were randomly divided in 3
groups with 10 replicates per group and 20 birds per replicate. The groups consisted of a
positive control with normal corn-soya diet, a negative control with reduction of ME by 150 Kcal
and amino acid by 8% and Nutrikem™ XL PRO treated group at 500 g/ton in the negative
control diet. Significant improvement (P<0.05) in body weight gain (87.4 grams) and FCR (6
points) was observed between Nutrikem™ XL PRO and negative control at the end of 35 days.
About 3 points FCR difference was observed between the positive and negative control groups.
The European efficiency factor (EEF) was observed to be 32 points higher in the Nutrikem™ XL
PRO treated group compared to negative control. Based on the results, it can be concluded that
supplementation with combination of NSPase, lysophospholipids and protease to nutrient
deficient diet can effectively improve broiler performance. This will further facilitate poultry
producers to reformulate their feed with alternative feed stuffs, thereby reducing the feed cost.
Keywords
Non starch polysaccharides; Metabolizable energy; European efficiency factor
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Proceedings of the World Veterinary Poultry Association (India) Conference 202042
Gut Health, Feed Additives and Management
Effect of Dietary Supplementation of Antibiotic Growth Promoters vs Synbiotics on the Performance and Gut Integrity of Broilers and Detection of their Residues in Meat by HPLC
Rayala Reddy V*, Ravinder Reddy V, Panda A K, Chinni Preetam V, Baswa Reddy P
College of Veterinary Science, Rajendra Nagar, Hyderabad
rayalreddy046@gmail.com
A study was carried out to investigate the effect of dietary supplementation of antibiotic
growth promoters (AGPs) – (Oxytetracycline OTC and Chlortetracycline CTC) and synbiotics
on the performance and gut integrity of broilers and detection of their residues in meat by
HPLC.A total of 400 day old commercial male broiler chicks were randomly divided into 10
treatment groups with 8 replications of 5 birds each (10×8×5) and reared up to 42 d of age. The
broilers were fed corn soya based broiler pre-starter (0-14 d), starter (15-28 d) and finisher (29-
42 d) diets. The control diet (T1) was prepared with corn-soya diets without antibiotics, for
remaining treatment groups (T2- T10) diets were formulated by corn-soya diets with different
levels of antibiotics, OTC@ (100 mg/kg (T2,T3), OTC@ 200 mg/kg (T4,T5), CTC @ 100 mg/kg
(T6,T7), CTC @ 200 mg/kg (T8,T9) and synbiotics @250 mg/kg (T10) for 42 days. On 36th day,
antibiotics (OTC, CTC) were withdrawn from T3, T5, T7, and T9 dietary treatments while the
other treatment diets were continued with antibiotics till the end of experiment. During the
experiment weekly body weight, weekly feed intake and FCR were recorded regularly. For the
study of histopathology and ultrastructure morphology (SEM) during slaughtering intestine
samples of 2 cm were collected from mid portion of jejuno-ileal junction. The results indicated
that the body weight gain of broilers were increased significantly (P<0.05) in the dietary groups
on supplementation of OTC, CTC or synbiotics. Supplementation of AGPs or synbiotics did not
have any significant (P>0.05) influence on cumulative feed consumption and feed conversion
ratio of broilers during pre-starter and finisher phases and during overall period (0-42 d). There
was a significant (P<0.05) influence observed in the length of intestinal villi on supplementation
with AGPs and synbiotics in broilers up to 42 d. The higher villi length was found in CTC 200
mg/kg followed by OTC 100 mg/kg diet groups. No significant pathological findings observed in
the intestinal villi of various dietary treatments. It can be concluded that supplementation of
AGPs@200 mg/kg diet improved the performance of broilers and can be practiced in broiler
production following withdrawal periods about 5-7 days and synbiotics can also be used as
alternative to the AGPs in the broiler production. The antibiotic residues analyzed through
HPLC in fresh meat and cooked meat are discussed in the following session.
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Proceedings of the World Veterinary Poultry Association (India) Conference 2020 43
Gut Health, Feed Additives and Management
Evaluation of Rice Polish on the Production Performance of Commercial Broiler Chicken
K S Shubhankar, Moorthy M*, Shamsudeen P, Sakthivel P C
Veterinary College and Research Institute, Namakkal, Tamil Nadu - 637 002
moorthy.m@tanuvas.ac.in
A biological experiment was conducted in broiler chicken from 0 to 35 days of age to
assess the effect of dietary inclusion of rice polish at graded levels on production performances
of commercial broiler chicken. Three hundred, one day-old, sexed, commercial broiler chicks
were randomly grouped into six treatments with five replicates for each treatment, containing
ten chicks per replicate. The treatment groups were fed with basal diet without rice polish (T1),
inclusion of rice polish at 2 (T2), 4 (T3), 6 (T4), 8 (T5) and 10 per cent (T6) replacing maize in
basal diet. Dietary inclusion of rice polish by replacing maize upto 8 per cent levels during first
four weeks and upto 4 per cent at final week gave statistically similar body weight and body
weight gain compared to control group. The final cumulative feed intake was low in 10 per cent
rice polish based ration and the feed conversion ratio was similar for all the levels of rice polish.
Dietary inclusion of rice polish did not influence carcass characteristics, serum lipid profile of
broiler chicken. Dietary inclusion of rice polish at 4 per cent significantly increased the villi height
and crypt depth of ileum. The return over feed cost per bird was highest in T3 (Rs. 30.09)
followed by T2 (Rs. 29.98) and T1 (Rs. 28.11). Based on the result of the study, it could be
concluded that rice polish can be used in broiler pre-starter and starter diet up to 8 per cent and
in finisher diet up to 4 per cent where body weight and body weight gain are comparable to
maize based diet with better return over feed cost per bird in commercial broilers.
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Proceedings of the World Veterinary Poultry Association (India) Conference 202044
Gut Health, Feed Additives and Management
Histopathological Studies on Pancreatic Pathology in Poultry
Brajesh Kumar*, Gupta M K
Arawali Veterinary College, Ranchi, Jharkhand - 834 006
brajesh.kumar.7927@gmail.com
Pancreas is highly innervated and vascular organ playing crucial role in maintaining
health and optimal growth of poultry through its digestive enzymes and hormones. The exocrine
portion consists of acini lined by secretory cells containing enzymes filled zymogen granules
and basally oriented nuclei. The enzymes on proper stimulus are emptied into the duct system
of pancreas connected to lumen of the acini. Duct finally opens in the lumen of the duodenum to
carry out its digestive function. Endocrine pancreas consists of clusters of cells called islets of
Langerhans which constitutes only 1-2% of the pancreas and remain distributed throughout the
exocrine parenchyma. In our study 22.23% (844/3796) birds showed definite pancreatic
pathology out of which 244 pancreas were subjected for through histopathological examination.
Microscopic examination of diseased pancreas revealed several important histopathological
alterations. Histopathological alterations observed during entire study period, affected mostly
the exocrine pancreas. Major histopathological alteration in decreasing order of frequency were
interstitial fibrosis (54.41%), congestion (46.51%), individualization of acinar cells (32.55%),
multifocal necrosis in exocrine pancreas (30.85%), vacuolar degeneration of acinar cells
(27.94%), periductular fibrosis (13.24%) increased level of zymogen granules (13.24%) and
capsuler thickening (5.39%). It was concluded that the most frequent histopathological
alteration found in the study was interstitial fibrosis. It constitutes an important contributory
factor in variable growth rate shown by birds in cases of disease outbreak and consequently
economic loss. It would be beneficial to add preparation of pancreatic enzymes in all cases of
disease out-break for early recovery of birds and thus production potential.
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Proceedings of the World Veterinary Poultry Association (India) Conference 2020 45
Gut Health, Feed Additives and Management
Evaluation of Pro-nutrient Effect of Combination of Lauric Acid and Glutamate in Broiler Chickens
Prabakar G*, Gopi M, Nirmal Kumar C, Mahadevan K, Shankar N, Rajesh Kanna M
SRM Institute of Science and Technology, Faculty of Agricultural Sciences, Kattankulathur,
Chengalpattu, Chennai Tamil Nadu - 603 203
prabavet@gmail.com
Ever since the ban on use of antibiotic growth promoters, the search of pro-nutrients is
keeps moving on. On the background of their characteristic individual activities of lauric acid
and glutamate were selected for the study. The biological trial was conducted by using one
hundred thirty commercial (Vencobb 400Y), straight run, and day-old broiler chicks. These
chicks were randomly grouped into four treatments of thirty two chicks, each treatment had four
replicates and fed with basal diet as T1 (Control), T2 (Control + 0.5% lauric acid), T3 (Control +
0.1% of glutamate), T4 (Control + 0.5% lauric acid + 0.1% of glutamate). Both compounds used
in the study were with a purity of about 99% and were obtained from Hi-media laboratories. The
experimental feed was formulated according to the standards prescribed in Bureau of Indian
Standards (2007).The body weight gain, feed intake, feed conversion efficiency and livability
were calculated. At the end of experimental period (42 days) bird were slaughtered and carcass
characteristics and intestinal length were studied.The supplementation of pro-nutrients and
their combination in broilers (T3) had significant difference (P<0.05) in body weight from third to
fifth weeks, body weight gain at sixth week, numerically higher feed consumption and
significantly (P<0.05) improved the feed conversion ratio in T3 treatment group. The intestinal
length was significantly (P<0.05) higher in T3 group. However, there was no significant
difference (P>0.05) in livability and carcass characteristics. Based on the experimental results,
it could be concluded that 0.5% glutamate fed group showed better production performance in
broilers.
Keywords
Broiler; Lauric acid; Glutamate; Performance; Intestinal length
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Proceedings of the World Veterinary Poultry Association (India) Conference 202046
Gut Health, Feed Additives and Management
Effect of Different Water Sanitizers on Physico-Chemical and Microbiological Characteristics of Drinking Water in Commercial Layers
Sangeetha P V *, Kannan D, Amutha R, Arthanari Eswaran M
Department of Poultry Science, Veterinary College and Research Institute,
Tamil Nadu Veterinary and Animal Sciences University, Namakkal, Tamil Nadu - 637 002
brajesh.kumar.7927@gmail.com
The study was undertaken to know the effect of different water sanitizers on physico-
chemical and microbiological characteristics of drinking water in commercial layers. The
biological experiment was carried out with three hundred and thirty six, 43 weeks old BV 300
commercial layer birds from single hatch and reared up to 45 weeks of age by providing
adaptation period for two weeks. At 46th week, the birds were weighed, leg banded and
randomly assigned into seven treatment groups with six replicates of eight birds each and
reared from 46 to 57 weeks (Three laying periods of 28 days) under standard managemental
practices. The treatment group consisted of T1 - Control (untreated water), T2 - Reverse
Osmosis water (plain), T3 - Chlorine water, T4 - Chlorine dioxide water, T5 - Hydrogen peroxide
water, T6 - Ozonized water and T7 - UV treated water. Throughout the experimental period of
twelve weeks, the experimental water samples were collected aseptically in the sterile
containers from nipple drinker before and during biological trial and were analysed for physico-
chemical parameters such as pH, total dissolved solids, dissolved oxygen, calcium,
magnesium and salt (as chloride) and microbiological parameters such as total bacterial and
coliform count. Result showed that physico-chemical parameters of experimental water
samples were within the normal range whereas significantly (P<0.01) higher and lower values
were recorded in control (T1) and RO (T2) group. Microbiological parameters of drinking water
revealed that total bacterial count was significantly (P<0.01) reduced from 3.36 to 4.68 log
reduction in all treatment groups, except control (5.59) group. Total coliform count was absent
in all treatment groups. It can be concluded that ozonization (T6) is considered as safe and
effective water sanitizer for better production performance with reduced microbial load in
drinking water without considering the initial investment.
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Proceedings of the World Veterinary Poultry Association (India) Conference 2020 47
Gut Health, Feed Additives and Management
Effects of Dietary Supplementation of Essential Oils as an Alternative to Antibiotic Growth Promoter on Livability and Economics of Commercial Broilers
Rajalekshmi C*, Mishra R K, Savaliya F P, Bhagora N G
Anand Agricultural University, Anand, Gujarat
rajalekshmiraju14@gmail.com
An experiment was conducted to investigate the effects of thyme essential oil
and peppermint essential oil as an alternative to antibiotic growth promoter on growth
performance and carcass traits of broiler chicken. Day old broiler chicks (Vencobb,
n=256) were randomly distributed into eight experimental groups with four replicates of
eight birds each. The experimental groups were fed with control diet without
supplementation of antibiotic growth promoter/essential oils (T1), or with
supplementation of bacitracin methylene disalicylate antibiotic at 500 mg per kg diet
(T2), or with supplementation of thyme essential oil at 150 mg per kg diet (T3), 200 mg
per kg diet (T4), 250 mg per kg diet (T5), or with supplementation of peppermint
essential oil at 200 mg per kg diet (T6), 250 mg per kg diet (T7) and 300 mg per kg diet
(T8). The trial was carried out in deep litter pen for 42 days. The mortality pattern of the
birds during the whole experiment was within the permissible limits and revealed no
adverse effects on feeding essential oils in diet. The return over feed cost was highest in
broilers assigned diet with antibiotic growth promoter; however it was statistically
similar to basal diet. Considering the net return over fed cost of broilers, use of thyme oil
was uneconomical. But supplementation of peppermint essential oil @ 200 and 300
mg/kg diet increased the net return over feed cost and was economical.
Keywords
Broiler; Thyme oil; Peppermint oil; Livability; Economics
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Proceedings of the World Veterinary Poultry Association (India) Conference 202048
Gut Health, Feed Additives and Management
Effects of Dietary Supplementation of Essential Oils as an Alternative to Antibiotic Growth Promoter on Performance Parameters and Carcass Traits of Commercial Broilers
Rajalekshmi C*, Mishra R K, Lunagariya P M, Patel A B
Anand Agricultural University, Anand, Gujarat
rajalekshmiraju14@gmail.com
An experiment was conducted to investigate the effects of thyme essential oil and
peppermint essential oil as an alternative to antibiotic growth promoter on growth performance
and carcass traits of broiler chicken. Day old broiler chicks (Vencobb, n=256) were randomly
distributed into eight experimental groups with four replicates of eight birds each. The
experimental groups were fed with control diet without supplementation of antibiotic growth
promoter/essential oils (T1), or with supplementation of bacitracin methylene disalicylate
antibiotic at 500 mg per kg diet (T2), or with supplementation of thyme essential oil at 150 mg
per kg diet (T3), 200 mg per kg diet (T4), 250 mg per kg diet (T5), or with supplementation of
peppermint essential oil at 200 mg per kg diet (T6), 250 mg per kg diet (T7) and 300 mg per kg
diet (T8). The trial was carried out in deep litter pen for 42 days. The results of the study
indicated that the supplementation of thyme oil at 200 mg/kg and peppermint oil at 300 mg/kg
broiler diet significantly (p<0.05) improved body weight and body weight gain. When compared
between thyme oil and peppermint oil supplemented groups, the peppermint oil
supplementation at 300 mg/kg found to be more beneficial. Feed conversion ratio significantly
(p<0.05) improved with peppermint oil supplementation at 300 mg/kg diet. A significant
(p<0.05) reduction in the mean abdominal fat was observed in treatments supplemented with
higher levels of thyme essential oil and peppermint essential oil (T5 and T8).
Keywords
Antibiotic; Broiler; Carcass trait; Essential oil; Performance parameter
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Proceedings of the World Veterinary Poultry Association (India) Conference 2020 49
Gut Health, Feed Additives and Management
Polyphenols as an Additive Agent to Improve Oxidative Stability of Chicken Semen During Low Temperature Storage
Gopi M*, Beulah P V, Prabakar G, Jaydip R J, Gautham K, Tyagi J S, Mohan J
ICAR-Central Avian Research Institute, Izatnagar, Bareilly, Uttar Pradesh - 243 122
gopsgopi72@gmail.com
The ultimate aim of poultry breeder operations is the production of fertile eggs as it has
great economic importance in determining the profitability of production. With the growing
advances in artificial insemination (AI) in the poultry sector, the short term storage of diluted
semen from superior germplasm is getting importance. However, due to higher concentration
and lipid content, the storage of chicken semen is not an easy task. An in vitro study was carried
out to assess the additive effect of polyphenols on sperm quality and antioxidant status in the
seminal plasma at different storage periods under refrigeration condition. The polyphenols
used in the study was extracted from the pomegranate peels using methanol as a solvent. The
extract was further evaporated and dried to obtain polyphenol powder. Forty healthy breeding
roosters of White Leghorn chicken having similar body weight were randomly selected for the
study. The semen samples were collected from each male periodically. The semen samples
were pooled and split equally into three portions for the addition of different concentrations of
polyphenols @ 0, 50 and 100 ppm respectively. Estimation of catalase, inhibition of lipid
peroxidation, reduced glutathione and superoxide dismutase was done at 0, 24 and 48 h
intervals of storage at refrigeration condition. The parameters were repeated for six different
collections. The sperm motility invariably increased in all treatment groups (50, 100 ppm) than
control in both storage conditions. Other physical attributes such as sperm concentration, live
and dead spermatozoa did not differ over control regardless of concentrations and time periods
of storage. Inhibition of lipid peroxide action was found significantly reduced at the end of 48 h of
storage in both the polyphenol treatment groups than control, while there was no change at 0
and 24 h. Polyphenols from pomegranate peels improved spermatozoal motility and
antioxidant activity in seminal plasma and it could be concluded that the polyphenols have the
potentiality to protect the spermatozoa against oxidative damage during storage of chicken
semen.
Proceedings of the World Veterinary Poultry Association (India) Conference 202050
Gut Health, Feed Additives and Management
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Nano Nutraceuticals Preparation, Characterization and its Effect on Antioxidant Enzymes and Growth Performance in Broiler Chickens
Shwetha H S*, Narayana Swamy M, Rajendran D, Preedia E, Babu, Prakash N,
Jayanaik, Srikrishna Isloor, Kalmath G P, Veena M P
Veterinary College, Hebbal, Bengaluru, Karnataka - 560 024
drshwetha.h.s@gmail.com
The present study was undertaken with the objective of preparation of nanocurcumin
and nanoselenium to improve their aqueous-phase solubility and bioavailability to evaluate
antioxdants property and growth performance in broiler chickens. Nanocurcumin was prepared
by solvent-antisolvent precipitation method and nano selenium was prepared by chemical
method. The particle size was characterised by Dynamic Light Scattering (DLS) analysis and
Scanning Electron Microscope (SEM) method. The chicks were reared in deep litter system
under standard managemental conditions. A total of 360 male day old commercial broilers were
divided randomly into four groups with six replicates in each group. Each group was having 90
birds and divided in to 15 chicks per replicate. Control group (GI) was fed on basal broiler diet.
The study groups such as GII, GIII and GIV were fed with nanoselenium, nanocurcumin and
combination of both along with basal diet, respectively. The body weight was recorded regularly
at weekly intervals. Blood samples were collected on 21st, 28th, 35th and 42nd day and serum
was separated for analysis. The DLS results confirmed that the hydrodynamic diameter of
nanocurcumin and nanoselenium is 288.2 nm and 70 nm, respectively. The results confirmed
that the prepared nanoparticles have reduced size and enhanced solubility. Increased activity
of antioxidant enzymes such as SOD, GPx and CAT was consistent with decreased MDA
concentration in supplemented groups compared to control group. Enzyme activity was higher
in all nanoparticles enriched feed supplemented groups. There was significant decrease of
body weight in Group III compared to other treatment groups as well as control group at day 42
and significant increase in body weight of Group II and IV compared to control group. The study
explored the impact of supplementation of nanocurcumin and nanoselenium in antioxidant
capability that imparts natural immunity and supports the growth performance in broiler
chickens.
Keywords
Nanocurcumin; Nanoselenium; Body weight; Antioxidants; Broiler chickens
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Proceedings of the World Veterinary Poultry Association (India) Conference 2020 51
Gut Health, Feed Additives and Management
Use of Medium Chain Fatty Acids as Replacement to Antibiotics in Broilers Diet
Pratik Jadhav*, Manwar S, Gole M, Langote G, Rathod A, Paradhi K
Post Graduate Institute of Veterinary and Animal Sciences, Akola, MAFSU, Nagpur
pratikjadhav2049@gmail.com
This study was conducted to evaluate the influence of dietary supplementation of
medium chain fatty acids (MCFA) in broilers. The day old chicks were equally distributed into
five treatment groups, randomly. There were three replicates of 20 chicks each under each
treatment. The birds under control group (T0) offered basal diet as per BIS (2007). The group
(T1) offered basal diet with antibiotic of BMD group. The group (T2) offered basal diet with
0.25% lauric acid. The group (T3) offered basal diet with 0.25% capric acid. The group (T4)
offered basal diet with 0.25% lauric and 0.25% capric acids. The birds fed diet containing lauric
acid, capric acid or their combination recorded significantly higher (P<0.01) body weight
compared to control. The increase in live weight in birds fed lauric acid over control was 4.37.
Likewise, the broilers fed diets with lauric acid singly or along with capric acid recorded
significantly higher gain in weights compared to other groups. The feed intake was comparable
in all treatment groups though numerically increased feed consumption due to lauric and capric
acid supplementation was evident. The feed conversion ratio was improved in broilers fed with
MCFAs either singly or in dietary combination. The reduction in mortality was recorded in
groups fed combination of MCFAs. The coliform count was numerically reduced, whereas, the
total viable count in group fed MCFAs and their combination were significantly (P<0.05)
reduced. The HI titers did not show significant differences between treatments. The dietary
supplementation of MCFAs was found to improve profit margin in broiler production. In
conclusion, the results of the present experiment suggested that the dietary supplementation of
medium chain fatty acids singly or in combination at 0.25 % level in diets improved growth
performance, gut health and found to be economically beneficial.
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Proceedings of the World Veterinary Poultry Association (India) Conference 202052
Gut Health, Feed Additives and Management
Effect of Dietary Supplementation of a Novel Phytogenic Blend on Organ Weights in Broiler Chickens
Oso O A, Umaya R S*, Malik P K, Thirumalaisamy G, Awachat V B
ICAR-National Institute of Animal Nutrition and Physiology, Bangalore, Karnataka - 560 030
r.umayasuganthi@gmail.com
BackgroundUse of antibiotics as growth promoters (AGPs) in poultry feed is associated with
emergence of antibiotic resistance in pathogens and carryover of residues in poultry products
intended for human consumption. Hence efficacy of a novel phytogenic blend on organ weights
in broiler chickens was explored in an attempt to identify AGP alternatives.
Methodology
One hundred ninety-two numbers of day old, unsexed Cobb chicks were distributed to
four dietary treatments consisting of only basal diet, basal diet supplemented with 355g/ton of
feed grade chlortetracycline and 1% and 2% PB, respectively. Each dietary treatment was fed
to six replicates (8 birds/replicate) and the experiment lasted for 42 days. At the end of trial, two
birds from each replicate were sacrificed and weight of drumstick, thigh, breast and back and
the weights and relative weights of gizzard, lung, liver, heart, pancreas, kidney, spleen, thymus
and bursa were measured.
Results
The weight and relative weight of gizzard, heart, pancreas, spleen, thymus and bursa
were not affected by dietary treatments. Orthogonal effect of PB levels revealed a quadratic
reduction in relative weight of gizzard (P = 0.008) as supplemental levels of PB increased from 0
to 1%, but showed increase in relative weight with 2% PB. Broilers fed diet supplemented with 1
and 2% PB exhibited higher (P < 0.0001) weight and relative weight of lung than other
treatments. Broilers fed diet supplemented with 1 and 2% PB exhibited lower weight (P =
0.0032) and relative weight (P = 0.0002) of liver when compared with the antibiotic group.
Orthogonal effect of PB levels showed a quadratic reduction (P = 0.022) in the relative weight of
liver as supplemental levels of PB increased from 0 to 1%, but recorded higher relative weight
with 2% PB. The relative weight of heart also increased quadratically (P = 0.044) with 1% PB
supplementation, but reduced with 2% PB supplementation. Broilers on negative control diet
recorded the highest weight (P = 0.0002) and relative weight (P < 0.0001) of kidney. Dietary
treatment had no effect on weight of drumstick, thigh and breast meat.
Conclusion
Dietary supplementation with 1% phytogenic blend did not exert any adverse effect on
weight of various organs in broiler chickens.
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Proceedings of the World Veterinary Poultry Association (India) Conference 2020 53
Gut Health, Feed Additives and Management
Effect on Supplementation of Moringa oleifera Leaf Powder on Gut Health and Litter Quality in Broilers
Jayanti L Agashe*, Manwar S J, Khose K K, Wade M R
Indian Veterinary Research Institute, Izatnagar, Bareilly, Uttar Pradesh - 243 122
vetdr.jayanti@gmail.com
The present study was conducted to evaluate effect of supplementation of Moringa
oleifera leaf powder on gut health and litter quality in broilers. The research work was conducted
on day old four hundred straight-run broiler chicks. The day old Vencobb-400 chicks were
randomly distributed into 4 treatment groups and each treatment group had 4 replicates of 25
chicks each. A control group (T0) was fed basal diet adequate in all nutrients as per BIS (2007)
and birds in treatment groups T1, T2 and T3 were offered basal diet containing Moringa oleifera
leaf powder at 0.2, 0.4 and 0.6%, respectively. The digestive tracts were dissected and
described Caecal content samples were taken aseptically and were transported in cold chain to
the laboratory, samples diluted, cultured and checked for the presence of total viable count and
coliform count and compared with the control after incubation. Total count was expressed as
log10 colony-forming units per gram of caecal content (log10 CFU/gram). The litter moisture
percentage was estimated at day 21st and 42nd by obtaining a 100 gram litter subsample which
was placed in tray and oven-dried for 48 hours at 600C. Results of study revealed that, the total
viable and coliform counts in treatment group were slightly decreased than control but having
non-significant difference at 42nd day of experiment. The mean weekly litter moisture
percentages were found significantly reduced at 21st day age. However, the mean weekly litter
moisture percentage was reduced at 42nd day age but differences are statistically non-
significant. Thus, the supplementation of Moringa oleifera leaf powder was found helpful in
boosting up gut health of birds and to maintain optimum litter moisture in broilers.
Keywords
Moringa oleifera leaf powder; broiler; gut health; litter quality
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Proceedings of the World Veterinary Poultry Association (India) Conference 202054
Gut Health, Feed Additives and Management
Effect of pre-starter amino acid and trace mineral supplementation on growth performance and immune response in broiler chicken
Awachat V B*, Elangovan A V, David C G, Ghosh J, Gowda N K S, Mishra A, Bhanja S 1 1
K , Majumdar S
ICAR - National Institute of Animal Nutrition and Physiology, Bengaluru - 560 0301ICAR-Central Avian Research Institute, Izzatnagar, Bareilly - 243 122
vaibhav.cari@gmail.com
The effect of prestarter amino acid and trace mineral supplementation on growth
performance and immune response in commercial broiler chicken has been assessed in this
study. One hundred sixty chicks divided into two groups; one group with post hatch (0-3days of
age) supplementation {25% higher level of amino acid (lysine, methionine, arginine, glutamine
and threonine); 100% higher level of trace mineral (zinc, copper, selenium)] and other group
was control one without post hatch supplementation. At 5 wk of age, body weight gain (1965 vs
1614 g/b) and feed intake (2988 vs 2497 g/b) was higher (P<0.05) due to post hatch
supplementation. Humoral immune response remained non significant (P>0.05). Cell mediated
immune response measured by foot web thickness was significantly (P<0.05) higher in post
hatch supplementation group. Most of the digestive organ weight (% of live weight) and length
(cm/ 100g live weight) did not differ significantly (P>0.05) due to post hatch supplementation at
first to fifth wk of age. It could be concluded that the post hatch supplementation of combination
of selected amino acids (lysine, methionine, arginine, glutamine and threonine) and minerals
(zinc, copper and selenium) for first three days of age was effective in enhancing the growth
performances of broiler chicken.
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Proceedings of the World Veterinary Poultry Association (India) Conference 2020 55
Gut Health, Feed Additives and Management
Dietary Supplementation of Dicosan® vis-a-vis AGP Improves Gut Health of Commercial Male Broiler Chickens
Rishikesh Jagzap
Norel Nbpl India Pvt. Ltd, Pune, India.
rishikesh122@gmail.com
An experimental trial was conducted by supplementation of Dicosan® a commercial
blend of sodium salts of coconut fatty acid distillate by Norel as a replacement in-feed antibiotic
growth promoter (AGP) in the feeding regimen of commercial broiler chickens. The objective
was to study the effect of Dicosan® on histomorphology of small intestine and gut health in
experimental broilers. A flock of 400 male Cobb-430 broiler chickens was divided into four
treatment groups for 35 days comprising of Dicosan® at varied levels in addition to basal diet
(NC). Each dietary group had 10 replicates and each pen housed 10 birds, viz. NC (Negative
control-Basal diet) PC (Positive control= NC + BMD 50ppm), Dicosan 1(1.5 g, 1.0 g and 0.5 g
per kg) and Dicosan 2 (1.0 g, 0.5 g and 0.5 g per kg) of diet during the starter, grower and finisher
phases respectively. All birds were reared according to standard management practices. At 35
days of age 10 birds from each dietary group were sacrificed and the small intestine was taken
for histomorphology and enumeration of bacterial count viz. Salmonella, E coli, Clostridium and
Lactobacillus. The intestinal villus height and crypt depth was measured by histological
techniques and the bacterial numbers were enumerated using method described by APHA
(2001). Statistically significant improvement in villus height and crypt depth was not evident,
however histological examination revealed more compact arrangement of villi in Dicosan 1 and
Dicosan 2 fed birds which should support better digestion and absorption of nutrients,
especially in pathogen challenged birds. No effect was observed on small intestinal lesion
scoring in all treatment groups. Dicosan 1, Dicosan 2 and PC greatly reduced E coli, Clostridium
and Salmonella counts, along with increased lactobacillus count, indicating the competitive
exclusion of harmful pathogens by lactobacilli. Dicosan 1 significantly reduced (P<0.0001) all
types of pathogenic bacteria more effectively than any other group with concomitant rise in
lactobacillus count indicating improved gut micro-environment. Therefore, it may be concluded
that, the dietary supplementation of graded levels of Dicosan, especially Dicosan-1 as against
AGPs improves gut health and overall performance in commercial broiler chickens.
Key wordsBroiler; Gut health; AGP; Dicosan®
References:
AMERICAN PUBLIC HEALTH ASSOCIATION–APHA. 2001. Compendium of Methods
for the Microbiological Examination of foods, 4. ED.
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Proceedings of the World Veterinary Poultry Association (India) Conference 202056
Gut Health, Feed Additives and Management
In ovo Establishment of Gut Microbiome in Chicks
Kolte A P*, Saravanakumar M, Dhali A, Malik P K, Awachat V B, Jayaram C, Trivedi S,
Elangovan A V, Bhatta R
Omics Laboratory, ICAR-NIANP, Bengaluru
atulkolte@gmail.com
The gut microbiome is increasingly recognized for its effect on feed efficiency as well as
immunity in poultry. The new born are known to inherit the gut microbiota from mother as well as
environment. In the present study, an effort was made to identify the vertically inherited gut
microbiome in chicks. The gut microbial diversity was estimated in the intestine of Pre-hatch
(n=3) and post-hatch (n=3) chicks. Briefly, the intestinal portion from the pre-hatch and post-
hatch chick was isolated aseptically, and total DNA was extracted using RBBC method. The V3-
V4 region of the 16s rRNA gene was amplified using forward GCCTACGGGNGGCWGCAG,
and reverse ACTACHVGGGTATCTAATCC primers before sequencing on illumina MiSeq
instrument using 2 x 300 bp chemistry. The high-quality reads were processed using Microbial
Genomics Module of CLC Genomics Workbench 20.0 (Qiagen Inc). The merged reads were
used for reference based OTU picking using Greengenes database (v13_5) at 97% similarity.
Total number of OTUs detected in pre-hatch chicks (2858-3288) were more than the post-hatch
chicks (1652-1978). The higher OTUs in the pre-hatch chicks may indicate diversity acquired
form the hen and may have role in shaping the immune system of the chicks. The pre-hatch
chicks had prevalence of Proteobacteria, Firmicutes and OD1 phylum constituting more than
60% of the microbiome however, in the post-hatch chick, more than 80% of the microbiome was
dominated by Proteobacteria. At family level, the prehatch chicks were dominated by
Lactobacillaceae and post hatch chicks has abundance of Enterobacteriaceae. Post hatch
changes in the microbiome composition could be attributed to the passing of meconium that
facilitate the microbiome members to grow differently in absence of inhibitory substances and
microbial competition. The study indicates the transfer of microflora from hen during egg
formation that establishes in the intestine.
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Proceedings of the World Veterinary Poultry Association (India) Conference 2020 57
Gut Health, Feed Additives and Management
Disease Surveillance andBiosecurity
Prevalence of Poultry Diseases in Wayanad District, Kerala
Dhanya T*, Prasanna, Pradeep K S, Alin Moideen M, Elizabeth T K, Shruthi V, Sachin A, Anagha A M, Ansar F, Anoopraj R, Hamza P, Ajith J G
Department of Veterinary Pathology, College of Veterinary and Animal Sciences, Kerala
Veterinary and Animal Sciences University, Pookode, Wayanad - 673 576
dhanyavetj3ny@gmail.com
According to the 20th livestock census of 2019, poultry population in Kerala was 29.8
million which showed 22.6 % increase from that of 2012 census. This indicates that poultry
farming continues to be one of the most lucrative income generating occupations for farmers in
the state. Out of the total poultry holding, more than 90 % is with marginal farmers and rest
remains with the small scale farmers. Though there are many reasons for mortality in poultry
birds, the proximity to neighboring states of Tamil Nadu and Karnataka and disease
transmission through transportation is a significant problem in Wayanad district. The single
major cause for economic loss in this sector is incidence of diseases due to multiple etiology
and it is necessary to understand the mortality pattern in poultry to reduce huge economic loss
for farmers to take suitable preventive measures time and again. It was found that in spite of
effective deworming drugs available in the market, the most common cause for mortality being
worm infestation and associated anemia and malnutrition. The second most common cause
observed was colibacillosis with main manifestations of fibrino-purulent air sacculitis.
perihepatitis, pericarditis and lymphocytic depletion of bursa and thymus. Other important
infectious causes found in chicken were coccidiosis, infectious bursal disease, mycoplasmosis
and New castle disease. Proliferative diseases such as avian leucosis complex was prevalent
in birds more than 8 months of age. Other conditions observed were gout, fatty liver, hydro
pericardium syndrome, crop impaction and vent pecking. This paper describes the detailed
pattern of mortality in broiler and layer chicken based on retrospective analysis of the post
mortem findings maintained in the Department of Veterinary Pathology, College of Veterinary
and Animal Sciences, Pookode.
Keywords
Poultry; Disease; Prevalence; Wayanad
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Proceedings of the World Veterinary Poultry Association (India) Conference 2020 61
Disease Surveillance and Biosecurity
Study on Occurrence of ESBL Producing E.coli and Salmonella spp. in Poultry and its Farm Environment from Wayanad District, Kerala
Hema Persis Andrews*, Asha K, Prejit, Jess Vergis
Department Of Veterinary Public Health, College Of Veterinary And Animal Sciences, Pookode, Wayanad, Kerala
andrewspersis@gmail.com
Intensively reared poultry serve as one of the most important reservoir of drug
resistant microorganisms which are of huge public health significance. Dissemination
of drug resistant genes viz., Extended-Spectrum �-Lactamase (ESBL) harbouring
Gram-negative bacteria among food-producing birds possess health risks to the human
population. A total of 40 samples including cloacal swabs, feed, litter, air and water
samples were collected from layer birds and farm environment of organised farms in
Wayanad. Samples were collected in sterile swabs and transported in Cary Blair media
and air samples were collected in MacConkey agar strip using a microbial air sampler.
Collected samples were brought to laboratory under refrigerated condition. Samples
were pre-enriched in Buffered Peptone Water. From the pre-enriched sample, one mL
of homogenate was streaked on Eosin-Methylene blue agar plates for E. coli and in
Rappaport-Vassiliadis (RV) broth and further streaked to Hektoen-Enteric agar for
Salmonella spp. Confirmation of isolates were carried out using biochemical tests and
by PCR targeting invA gene for Salmonella spp. and uidA gene for E. coli. Isolates were
screened for ESBL production based on standard Double disc diffusion method and
evaluated for the presence of three genes conferring ESBL resistance viz., blaCTX-M,
blaTEM, blaSHV. For detection of biofilm production, resistant isolates were streaked
on Congo red agar. Based on the appearance of colonies on CRA the cultures were
graded for biofilm production as strong, moderate, and weak biofilm producers. A total
of 29 samples were tested positive for E. coli and none of the isolates were positive for
Salmonella spp as per PCR assay. In phenotypic assay for drug resistance, confirmed
E. coli isolates screened were found to be ESBL producers and genotypic assay of
E.coli isolates by employing revealed occurrence of blaCTX-M (12.5%), blaSHV (5%)
and blaTEM (47.5%). High prevalence rate of ESBL resistance conferring genes
among poultry necessitates the adoption of strict public health measures.
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Proceedings of the World Veterinary Poultry Association (India) Conference 202062
Disease Surveillance and Biosecurity
Emerging Diseases of
Poultry and Flock Health
Studies on Pancreatic Pathology in Poultry
Brajesh Kumar*, Gupta M K
Arawali Veterinary College, Ranchi, Jharkhand - 834006
brajesh.kumar.7927@gmail.com
Pancreatic pathology of 3796 birds was studied between December 2017 to November
2018. In our study 22.23% (844/3796) birds showed definite pancreatic pathology. Major gross
pancreatic pathology included bleached appearance, mottled appearance with multifocal white
necrotic lesions, pancreatic deformity, congestion, atrophy or hypertrophy. Bleached pancreas
was major gross pathological alteration (36.97%). Major histopathological alteration in
decreasing order of frequency were interstitial fibrosis (54.41%), congestion (46.51%),
individualization of acinar cells (32.55%), multifocal necrosis in exocrine pancreas (30.85%),
vacuolar degeneration of acinar cells (27.94%) and periductular fibrosis (13.24%). Incidence of
pancreatic pathology was high in IBD (94.74%), IB (93.33%), pasteurellosis (90.48%),
metabolic disorders (82.86%) and Aspergillosis (83.33%). Diseased Broiler birds were found to
be quite prone (75.34%) to develop pancreatic pathology. Maximum pancreatic pathology was
observed in grower (33.69%) followed by chicks (20.05%) and adult (19.54%). Seasonal study
revealed highest pancreatic pathology in monsoon (28.84%) followed by summer (22.57%) and
winter (12.92%). It was concluded that incidence of primary pancreatic pathology is rare in
poultry. However, its involvement as a secondary complication is quite high and constitutes an
important contributory factor in variable growth rate shown by birds in cases of disease
outbreak and consequently economic loss. It would be beneficial to add preparation of
pancreatic enzymes in all cases of disease out-break for early recovery of birds and thus
production potential.
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Proceedings of the World Veterinary Poultry Association (India) Conference 2020 65
Emerging Diseases of Poultry And Flock Health
Fowl Adenovirus Induced Gizzard Erosion and Incidence of Feed Passage Syndrome in Commercial Broiler Chicken
Amit Kumar Das*, Venket M Shelke.
Kemin Industries South Asia Pvt. Ltd., C3, 1st street, Ambattur Industrial Estate, Chennai,
Tamil Nadu-600 058
drscsekar@gmail.com
Several factors related to feed quality as well as infections can predispose broiler birds
to feed passage syndrome. Incidences of feed passage syndrome were observed to be on rise
during the late half of 2019 in Karnataka and Andhra Pradesh, located in Southern part of India.
Affected commercial broiler farms having incidences of feed passage syndrome were visited in
several regions of Karnataka, viz. Bengaluru, Mangalore, Chikmagaluru and Chittoor in Andhra
Pradesh. Low body weight gain and poor feed conversion were observed in flocks having feed
passage syndrome. But there was no increase in mortality. Intestinal health was examined in
these flocks for lesions of coccidiosis and bacterial enteritis, and were scored based on their
severity according to the method of Johnson & Reid. The common observation in all the birds
examined was presence of gizzard erosion. As the gizzard was flaccid, moderate to severe
erosions in the koilin layer of gizzard was observed. In few birds, mild to moderate Eimeria
maxima lesions were observed. Tissue impression on FTA card was collected from gizzards
having erosion on koilin layer. Six such gizzard impression samples were collected from these
regions and submitted for identification of Fowl Adenovirus (FAdV) through polymerase chain
reaction (PCR) method. All the samples were found positive for FAdV. Of these, four samples
were further processed for gene sequencing. Out of these four samples, the fowl adenovirus
serotype 11 was detected in three samples and fowl adenovirus serotype 8a was detected in
one. Thus, it is found that fowl adenovirus induced gizzard erosions in commercial broiler birds
leads to feed passage syndrome. A holistic approach is needed to control the feed passage
syndrome in commercial broiler birds. Identifying predisposing factors is vital for effectively
addressing the feed passge syndrome. In this case, vaccination against fowl adenovirus along
with good anticoccidial program in the feed can help in reducing feed passage syndrome,
leading to improved performance parameters in commercial broiler flocks.
Keywords
Fowl Adenovirus; Gizzard Erosion; Feed passage syndrome; Lesion Scoring;
Predisposing factors
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Proceedings of the World Veterinary Poultry Association (India) Conference 202066
Emerging Diseases of Poultry And Flock Health
Effect of Tinospora cordifolia on Infectious Bursal Disease Virus Induced Pathology in Cockerel
Chavan A A, Sonkusale P M, Shinde S S, Kaore M P, Kadam M M, Kurkure N V*
Nagpur Veterinary College, Seminary Hills, Nagpur, Maharashtra - 440 006
nitinkurkure@rediffmail.com
Study was conducted to investigate the effect of T. cordifolia on pathology of induced
IBDV infection in cockerels. Bursal tissue samples from IBD suspected birds were collected
during natural outbreaks and confirmed by RT-PCR assay by depicting the product size of
643bp of VP-2 gene of IBDV. IBDV positive bursal tissues were homogenised in 10% PBS and
inoculated into 9-11 days old SPF eggs through CAM route. During an experiment, total of 240
cockerels chicks were divided into six equal groups each comprising 40 cockerel chicks viz;
Group A (normal diet), Group B (IBDV infection + normal diet), Group C (IBDV infection + T.
cordifolia stem powder @ 1gm/kg of feed 0-40 day of age), Group D (IBDV infection + T.
cordifolia stem powder@ 2gm/kg of feed 0-40 day of age), Group E (IBDV infection + T.
cordifolia stem powder@ 1gm/kg of feed 28-40 day of age) and Group F (IBDV infection + T.
cordifolia stem powder @ 2gm/kg of feed 28-40 day of age). Cockerels were challenged with
IBDV with dose of 102.0 EID50/0.1 ml per birds by intra-ocular route on 28th day of age.
Results demonstrated that, cockerels supplemented with T. cordifolia and challenged with
IBDV showed significant improvement in growth performance and humoral and cell mediated
immune response and also results in reduction in severity of lesions as compared to control
infection group. Findings of present study support the beneficial effect of T. cordifolia during
IBDV infection in cockerels.
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Proceedings of the World Veterinary Poultry Association (India) Conference 2020 67
Emerging Diseases of Poultry And Flock Health
Comparative Assessment of Route of Induced Infection with Avibacterium paragallinarum in Chicken Employing Conventional Intra-Nasal Inoculation vis-a-vis Intra-Ocular and Nebulizer Technique
Sidhartha Deshmukh*, Rajanbir Singh, Harmanjit Singh Banga
Department of Veterinary Pathology, College of Veterinary Science, GADVASU, Ludhiana,
Punjab-141 004
sid0159@yahoo.com
The known, conventional experimental method of producing clinical infection of
Avibacterium paragallinarum in chicken model is either intra-nasal or intra-sinus inoculation,
which considerably exude clinical symptoms of infectious coryza. Intriguingly, the disease is not
fatal to chicken. There have been several scientific broaching over the usage of conventional
method for not mimicking the natural course of infection. For checking the efficacy of a vaccine
or any effective drug molecule on anvil or pathogenetic mechanism, an authenticate
establishment of clinical form of disease is mandatory, which can reasonably be shown to get
hindered through drug/vaccine mediated interference following in-situ progression of intricate
immune molecules or other any protective mechanism, as commonly enunciated during natural
infection(s), which sans the annals of literature. Further, there have been regional disputes over
undertaking the challenge model and evaluation of clinical signs of this disease of chicken. In
this study, we attempted to simulate the natural way of infection by instilling infection through
ocular drop and or by nebulizer route comparable to that of closely placed in-contact birds as a
challenge method. Ocular way of infection is also very common and incriminates in the
development of disease in the natural cases. The objective of the study was to identify a suitable
method of induced inoculation that can gradually replace both intra-sinuses/ intra nasal
inoculation,with new tested methods without compromising the results per se. The results
indicated, that both ocular and nebulizer route of inoculation method has a potential to
represent like natural course of infection than the intra nasal method, where tissue localization
as well as dispersal of pathogen in the turbinates area, trachea and in lungs, besides cellular
response was fairly appreciated through immunohistochemistry (IHC). This study further
suggested that the effect of such a challenge method can also act as more accurate model for
any pathogenetic and efficacy studies in future where intention to validate different steps of A.
paragallinarum pathogenesis and immune response during infection is desirable.
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Proceedings of the World Veterinary Poultry Association (India) Conference 202068
Emerging Diseases of Poultry And Flock Health
Molecular Epidemiology of Newcastle Disease in India During Year 2019
Bulbule N R*, Pardeshi R B, Deshmukh V V, Kaje V S, Gupte A P, Chawak M M
Poultry Diagnostic Research Center 18 Km, Pune Solapur Road,
Loni Kalbhor, Pune, Maharashtra-412 201
n.bulbule@rediffmail.com
In-spite of continuous vaccination program, Newcastle disease is endemic in India.
Recently several outbreaks in commercial poultry farms were found to be due to genotype XIII
Newcastle disease virus (NDV) isolates. In 2016, it was reported that NDV genotype VII was
isolated from Indian Peafowl from North India. In the present study, disease outbreak
investigation in commercial poultry farms during 2019 from different regions of India was carried
out. More than 2000 samples were collected and tested from different commercial poultry farms
experiencing high mortality (6 to 40%), torticollis, respiratory distress and/or drop in egg
production from all over the India. In some farms prominent gross lesions observed were
tracheal and lung congestion, hemorrhages in both proventricular glands and ceacal tonsils,
raised intestinal ulcers in most of the dead birds. Pooled tissue samples (Trachea, Lung and
spleen) were collected for virus isolation and histopathology. 10% tissue homogenate was
inoculated in 9 day-old embryonated specific pathogen free (SPF) eggs for virus isolation. NDV
isolations were confirmed by heamagglutination activity and RT-PCR and more than 30%
samples were found positive for NDV from processed samples. Representative isolates from
different cases/regions were further characterized for pathogenicity and genotyping studies.
The mean death time of most of the isolates was 48-60 hours. The intracerebral pathogenicity
index values ranged from 1.76 to 1.90, confirming the velogenic nature of the NDV isolates.
Sequencing of the fusion cleavage site identified multiple basic amino acids (112 RRQKRF117
motif), confirming this to be a virulent isolate. A phylogenetic tree was constructed based on the
complete F gene. Most of the isolates from different part of India showed close similarity with
sub-genotype XIIIb and few isolates from Maharashtra and North Indian states showed close
similarity with sub-genotype VIIi. In conclusion this study showed that velogenic NDV strains
belonging to sub-genotype XIIIb has been predominantly responsible for most of the outbreaks.
But isolation of sub-genotype VIIi from few cases is highly related to the NDV circulating in
Pakistan. This is the first report regarding the isolation of the NDV genotype VIIi from
commercial poultry farms of India. These findings will help to make strategies for the prevention
and control of Newcastle disease.
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Proceedings of the World Veterinary Poultry Association (India) Conference 2020 69
Emerging Diseases of Poultry And Flock Health
A Case of Osteoma in an African Lovebird (Agapornis)
Ansar Fasaludeen, Pradeep M, Anagha A M, Hamza Palekkoden, Anoopraj R,Prasanna K S, Ajith Jacob George
Dept. of Veterinary Pathology, College of Veterinary and Animal Sciences, Pookode,
Wayanad, Kerala, Kerala Veterinary and Animal Sciences University
dr.ansarvet@gmail.com
Osteoma is a slow growing benign outgrowth of bone tissue and is found to be
uncommon in avian species. An African love bird of age one and half years was presented to the
Department of Veterinary Pathology, College of Veterinary and Animal Sciences, Pookode with
complaint of lameness, lethargy and emaciation since one month. Upon request of the owner,
the bird was euthanized and examined. The post mortem examination of the carcass revealed
prominent keel bone with multiple hard small masses on its inner aspect and a hard mass on the
right dorso-lateral aspect of pelvic bone. The nodular lesions, spleen and liver were screened
for Avian leucosis complex virus by PCR and was found to be negative. Histopathological
examination of the hard mass revealed densely packed trabeculae consisting of mature woven
bone, cartilage and fibrous tissue with varied degree of mineralisation. The trabeculae
contained proliferating osteoblasts, erythrocytes and myeloid tissue. Based on the
histopathological findings the masses were identified as osteoma. Based on the history of the
flock and laboratory diagnosis it could be concluded that this is a rare case of non-infectious
sporadic case of osteoma.
Keywords
Osteoma; African lovebird; avian tumour
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Proceedings of the World Veterinary Poultry Association (India) Conference 202070
Emerging Diseases of Poultry And Flock Health
Ovarian Adenocarcinoma with Cystic Ovaries in a Duck (Anas platyrhynchos)
Anagha A M, Pradeep M, Ansar Fasaludeen, Ajith Jacob George, Hamza Palekkodan, Anoopraj R, Nikhil S Rao, Prasanna, K S
Dept. of Veterinary Pathology, College of Veterinary and Animal Sciences, Kerala
Veterinary and Animal Sciences University, Pookode, Wayanad, Kerala
anaghaanuam@gmail.com
Ovarian adenocarcinoma is a spontaneously occurring neoplasm in hens and it
serves as a potent animal model for studying human ovarian tumours. But the report of
ovarian adenocarcinoma in ducks is rare. This article reports a case of ovarian
adenocarcinoma with cystic ovaries in a one year old domestic duck. The duck brought
for postmortem examination to Department of Veterinary Pathology, College of
Veterinary and Animal Sciences Pookode Wayanad showed its body cavity, filled with
numerous cystic follicles of varying size containing straw yellow to red coloured fluid.
Wall of a few follicles were highly thickened. Histopathological examination of the ovary
revealed, multiple acinar pattern of epithelial cells. The acini were lined by single layer
of low columnar to cuboidal cells. In a few areas epithelial papillary projections into the
lumen of dilated acini were observed. Immunohistochemical expression of different
receptors were also studied. The other birds in the flock were under constant
monitoring and no further incidence of ovarian adenocarcinoma could be detected
suggesting the spontaneous occurrence of the neoplasm.
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Proceedings of the World Veterinary Poultry Association (India) Conference 2020 71
Emerging Diseases of Poultry And Flock Health
An Outbreak Of Degenerative Myopathy and Sarcocystis Infection in Layer Chickens
Alin Moideen T K*, Pradeep M, Dhanya Thomas, Elizabeth Varghese, Shruthi A, Sachin, Anoopraj R, Hamza Palekkodan, Prasanna K S, Ajith Jacob George
Dept. of Veterinary Pathology, College of Veterinary and Animal Sciences, Kerala Veterinary
and Animal Sciences University, Pookode, Wayanad, Kerala
alinmoideen@gmail.com
Degenerative myopathies such as deep pectoral myopathy and white striping are
usually reported in broiler chicken as a metabolic disease. Even though Sarcocystis affecting
poultry were reported from different countries high mortality were rarely reported. This article
reports a case of high mortality due to severe degenerative myopathy and sarcocystosis in adult
layer birds in a farm from Wayanad district of Kerala. Mortality of 25 adult birds were reported
within a span of 3 days with gross skeletal muscle lesions in a medium sized farm. Detailed post
mortem examination at department of Veterinary Pathology, College of Veterinary and Animal
Sciences, Pookode, revealed skeletal muscle bundles with 1-2 cm long elliptical white stripes,
extending deep in to the muscle mass. On microscopic examination skeletal muscle fibres
showed loss of striations, hypereosinophilia, vacuolar degeneration and fragmentation with
infiltration of macrophages. Multiple basophilic round to oval thick walled cysts containing
bradyzoites were observed in skeletal and cardiac muscle fibres. The characteristic feature of
the cyst and that of bradyzoites was diagnostic for sarcocysts. Deworming and
supplementation of vitamin E in the flock subsided the mortality. A combined effect of
sarcocysts along with vitamin E deficiency might have caused high mortality in the farm. This
incidence warrants the need for early detection and management of Sarcocystis infection in
chickens.
Keywords
Sarcocyst; Myopathy; Vitamin E deficiency; Layer chicken
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Proceedings of the World Veterinary Poultry Association (India) Conference 202072
Emerging Diseases of Poultry And Flock Health
An Outbreak of Favus in A Breeder Poultry Flock in Goa
Susitha Rajkumar*, Nibedita Nayak, Chethan Kumar H B, Chakurkar E B
ICAR-Central Coastal Agricultural Resarch Institute, Old Goa, Goa-403 402
drsusitharajkumar@gmail.com
Favus is a chronic inflammatory skin condition caused dermatophytes group of fungus
mainly Microsporum gallinae. The condition is reported worldwide. Favus caused by M. gallinae
has been reported in the chicken, turkey, duck, quail, and wild birds. The disease is contagious
and zoonotic. In the present case an outbreak of favus was observed in a 40 week old breeder
flock of 700 birds maintained in 2 sheds in a poultry farm in Goa during May 2019. Initially a
whitish colour change was observed on the combs of few birds mainly male birds. Mild
thickening of the affected areas on the comb and presence of white diffuse areas with black
spots were noticed. This was followed by occurrence of thickening, white scaly lesions over
comb and surrounding areas on head and neck. Wrinkled crusty lesions and circular
depressions were seen over comb in some birds in later stages. Mild to severe lesions were
observed on more than 50% of the flock. Feather loss in the head and neck was noticed in
severely affected birds. Skin scrapings from the affected area were examined after treatment
with 10% KOH showed fragmented fungal hyphae. Samples were inoculated in Sabarauds
Dextrose agar, but pathogen could not be isolated. The birds were treated with anti fungal
dusting powder containing Clotrimazole for 2 weeks. In severely affected birds Miconazole
ointment was applied on lesions daily for 2 weeks. The entire flock was given oral multivitamin
supplementation. The condition subsided after 2 weeks and birds were cleared completely of
infection. Poor immunity, overcrowding and damp surroundings are predisposing factors for
fungal infections including dermatophytosis. Goa being a tropical coastal region has a hot
humid weather especially during summer months from March to June which favours the growth
of fungal pathogen in environment. This condition can be prevented by applying adequate
hygienic measures in poultry sheds and premises including maintenance of dry floor and litter,
proper ventilation and exposure to sunlight etc. Immediate treatment and isolation of affected
birds prevent spread of the disease in large flocks.
Keywords
Sarcocyst; Myopathy; Vitamin E deficiency; Layer chicken
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Proceedings of the World Veterinary Poultry Association (India) Conference 2020 73
Emerging Diseases of Poultry And Flock Health
Molecular characterization of Newcastle Disease Virus in Central India
Megha Kaore*, Seema Shinde, Kurkure N V
Department of Pathology, .Nagpur Veterinary College, Seminary Hills, Nagpur,
Maharashtra-440 006
meghakaore7@gmail.com
BackgroundNewcastle disease (ND) is a highly infectious viral disease of domestic and wild avian
species. The virus is rapidly evolving and several new genotypes have been discovered in the
past few years. Thus the characterization of these strains is important to evaluate the
emergence of new genotype, anticipate new outbreaks and develop adequate control
measures.
MethodologySamples were collected from a total 25 outbreaks with a history of high mortalities,
respiratory/neurological signs in central India. RNA was isolated from the spleen and trachea.
RT-PCR was carried out by targeting the F gene of NDV. Amplicons were sequenced
commercially and the phylogenetic tree was constructed.
ResultsNucleotide analysis of fusion (F) protein revealed close similarity stains of genotype XII,
VIIg and VIb of NDV.
ConclusionIn spite of regular vaccination vNDV strains of NDV genotype XII, VIIg and VIb are
circulating in field and causing huge economic losses to poultry farmers.
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Proceedings of the World Veterinary Poultry Association (India) Conference 202074
Emerging Diseases of Poultry And Flock Health
Molecular Characterisation of Ornithobacterium rhinotracheale
Megha Kaore*, Seema Shinde, Kurkure N V
Department of Pathology, Nagpur Veterinary College, Seminary Hills, Nagpur,
Maharashtra-440 006
meghakaore7@gmail.com
BackgroundOrnithobacterium rhinotracheale (ORT) is an emerging poultry pathogen which has
been incriminated as a possible additional causative agent in the respiratory disease complex
(RDC) of poultry. Molecular characterization of ORT from flocks suspected to have respiratory
disease complex was carried out.
MethodologyA total of 30 commercial farms monitored for occurrence of RDC. DNA was isolated
from the tracheal sample. Ornithobacterium rhinotracheale was detected by PCR by targeting
the 16srRNA gene. The amplicons were sequenced commercially and the phylogenetic tree
was constructed.
ResultsThe phylogenetic analysis based on the 16S rRNA gene revealed that the ORT in the
present study was closely related to the ESV 55 strain of ORT which belongs to serovar A.
ConclusionORT is considered as an important pathogen in respiratory disease complex.
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Proceedings of the World Veterinary Poultry Association (India) Conference 2020 75
Emerging Diseases of Poultry And Flock Health
A Rare Case of Feather Folliculoma in a Japanese Quail
Azhaguraja M*, Simmi Tomar N, Babu Prasath, Beulah Pearlin V
ICAR- Central Avian Research Institute, Izatnagar, Bareilly, Uttar Pradesh - 243 122
azhagurajamano@gmail.com
Feather folliculoma are the benign cystic growth of the feather follicular epithelium in
birds. A two months old female Japanese quail carcass was presented for necropsy. On
external examination, skin over the back revealed multiple yellowish raised irregularly spherical
firm masses. It contained yellowish white caseous mass with feather remnants.
Microscopically, the mass revealed multiple cystic cavities lined by thick capsule of squamous
epithelium. The lumen contained irregular keratin material and poor downward grown feathers
which were arranged in curls. Infiltrations of heterophils were also observed. Based on the
histopathological appearance, the case was diagnosed as feather folliculoma in a Japanese
quail.
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Proceedings of the World Veterinary Poultry Association (India) Conference 202076
Emerging Diseases of Poultry And Flock Health
An Out Break of Infectious Bursal Disease in Commercial Chicken
Chandravathi T*, Rama Devi V, Ramesh Babu T, Charitha P
NTR College of Veterinary Science, Gannavaram, Andhra Pradesh - 521102
chandrakiran13209@gmail.com
Infectious bursal disease (IBD) is a significant contagious and immunosuppressive
disease and continues to cause economic losses in the poultry industry. IBDV, the cause IBD is
prone to geneticvariation, hence it requires quick diagnosis for planning preventive measures.
The present paper describes an out- break of IBD in 6 week old layer flock. IBD was diagnosed
based on gross and histopathology and molecular techniques. The flock was vaccinated and
the total flock size was about 50000 birds, in that about nearly 4672 birds were dead with a
mortality rate of 9.34% within 2 weeks period. Clinically birds showed dullness, depression,
ruffled feathers, closed eyes, watery to bloody droppings. Grossly, bursa was enlarged,
yellowish creamy in colour with gelatinous covering on bursa in few birds and enlarged,
haemorrhagic in some birds. Other lesions included haemorrhages in thigh and breast muscles
and caecal tonsils. Haemorrhages were also noticed at the junction of proventriculus and
gizzard. Kidneys were enlarged and haemorrhagic and spleen was congested. Histological
sections revealed severe corticomedullary lymphoid depletion with complete loss of
lymphocytes, intrafollicular edema and exudation with heterophilic infiltration. Spleen revealed
mild to moderate lymphoid depletion. On reverse transcription polymerase chain reaction (RT-
PCR) using primers from the hyper variable region of VP2 gene of IBDV yielded an amplicon of
743bp and confirmed the presence of pathogenic IBDV.
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Proceedings of the World Veterinary Poultry Association (India) Conference 2020 77
Emerging Diseases of Poultry And Flock Health
Role of Micro RNAs in Diagnosis of Avian Diseases
Rajeshwar Khandare*, Sonal Saxena, Sameer Shrivastava, Noopur Dutta, Basant, Purnima Gogoi, Dipankar Bhist, Uzma Jabeen
ICAR-IVRI Izatnagar, Bareilly, Uttar Pradesh - 243 112
rajeshwarkhandare@ymail.com
Micro RNA are small non coding endogenous 18-24 nucleotide long sequences that
play vital role in cell development, differentiation, growth, tumorigenesis, immunoregulation,
and viral infection. Recently these miRNAs are being explored as biomarkers in various fields of
disease diagnosis in animals. The available studies on humans, canines and avian shows their
significance in disease diagnosis and prognosis. In Marek's Disease (MD) the expressions of
some cellular miRNAs have been shown to be upregulated (gga-miR-21) or downregulated
(Gga-miR-199-3p, gga-miR-140-3p, and gga-miR-221-5p), and they participate in MDV-1
induced tumors by targeting related genes. A neoplastic disease caused by ALV is Avian
leukemia. ALV-J infects chickens which causes tumor formation in different organs, such as s
bursa, liver, kidney and spleen and shows differentially expression of miRNAs. Infectious bursal
disease (IBD) is a highly contagious and immunosuppressive disease in young chicken.
Inhibition of infectious bursal disease virus infection can be achieved by targeting chicken heat-
shock protein-90 using microRNAs. Reticuloendotheliosis virus (REV) infection of multiple
avian species can lead to a number of diseases such as runting syndrome, immunosuppression
and oncogenesis, causing major economic losses. MicroRNA-155 has been shown to be highly
expressed in REV-infected CEFs in time and dose dependent manner. Infection with influenza
A virus (IAV) leads to higher economic loss and can cross species barriers. MicroRNA-206 has
been shown to be a valuable marker in differentiating IAV expression in broiler and layer. Hence
studies on microRNA are useful in diagnosis and early intervention of various poultry diseases
of economic importance.
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Proceedings of the World Veterinary Poultry Association (India) Conference 202078
Emerging Diseases of Poultry And Flock Health
Fatty Liver Hemorrhagic Syndrome in Desi Birds
Charitha P*, Chandravathi T, Ramadevi V, Sudha Rani Chowdary Ch, Samatha V
Department of Veterinary Pathology, NTR College of Veterinary Science, Gannavaram,
Andhra Pradesh
charithapara49@gmail.com
Fatty liver hemorrhagic syndrome (FLHS) affects chickens including commercial layers
and broiler breeders. It is most common cause of mortality in old laying hens associated with
high energy diets along with restricted exercise and is most common during hot weather. The
present study describes FLHS in 2 desi birds that were brought for necropsy at Department of
pathology, NTR CVSC, Gannavaram .Post-mortem examination revealed abundant fat in the
abdominal cavity. The liver was enlarged, pale yellow and friable. A large blood clot was seen
overlying the liver capsule .Histopathologically section of liver revealed fat vacuoles in
hepatocytes and moderate amount of hemorrhages .Based on gross and histopathological
features FLHS was diagnosed.
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Proceedings of the World Veterinary Poultry Association (India) Conference 2020 79
Emerging Diseases of Poultry And Flock Health
Developments in
Poultry Products Technology
Recycling of Poultry Waste Through Bioconversion Using BSF Larvae for Sustainable Livestock Production
Niranjan B H*, Md Nadeem Fairoze, Mahesh, Yandageri S, Elangovan A V
Veterinary College, Bengaluru, Karnataka-560 024
nirishyla@gmail.com
Poultry sector in India predominantly working on integration model has achieved
production efficiency. However, its progress in the waste management sector is limited. The
search for efficient, economical and user-friendly technology for salvaging waste is on. This
study was one such attempt towards generating valuable byproducts from the various poultry
waste harvested from different poultry operations. A number of treatments (12) were evaluated
against kitchen waste as control, incorporating BSF larvae as a tool for bioconversion. The
findings of the study revealed that the larvae reared on combination of all wastes from different
poultry operations were the best from the point of view of recycling. The protein and fat content
were 43.24±5.13a and 27.55±2.9e percent, respectively with a high bioconversion efficiency of
31.83 percent. The time taken for material recycling and waste reduction was the shortest (24
days). The residue obtained after larval action was also found to be of high manure value with a
C/N ratio of 43.51. The bioconversion of waste was found to be dependent on larval activity in
the substrate, which in turn was dependent on the moisture and nutritional profile of the
substrate. The microbial safety analysis demonstrated that there was complete absence of
Salmonella spp in the residue which ensured public health concern. Thus, it was concluded that
this novel method of salvaging waste through larvae was efficient in converting the waste into
rich feed ingredient of high commercial value.
KeywordsPoultry waste; BSF larva; Bioconversion; Nutritive value; Manure value; Microbial
Safety.
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Proceedings of the World Veterinary Poultry Association (India) Conference 2020 83
Developments in Poultry Products Technology
Optimization of Meat Particle Size in Development of Chicken Seekh Kebab Formulated with Spent Hen Meat
Mukesh Tukaram Nampalle*, Ashim Kumar Biswas, Jaydip Jaywant Rokade, Jyotirmoy Saharia, Suchismita Pradhan, Monica Madeswaran
ICAR-Indian Veterinary Research Institute, Izatnagar, Bareilly U.P. India-243 122
mk.nampalle333@gmail.com
Present study was carried out with the aim of optimizing the meat particle size in
development of chicken seekh kebab (CSK) using spent hen meat. For this, treatments
adopted were control= 8mm+ bowl mixing, T1= 8mm and T2= 4mm. Raw meat batter was
subjected to pH and emulsion stability evaluation. Cooked CSK samples were subjected to
various physicochemical parameters, instrumental colour and sensory attributes. The results
indicated non-significant effect on pH of raw meat batter as well as cooked CSK samples.
However, significant effect on emulsion stability was observed due to lower particle size, and
was highest in T2 samples (P<0.05). Cooking yield of CSK significantly (P<0.05) increased with
lower particle size. Higher cooking yield was observed in T2 (P<0.05). Moisture content of all
CSK samples was non-significant (P>0.05) but numerically higher in T2. Effect of variation in
particle size on instrumental colour values was non-significant (P>0.05). It also revealed
significant effect on sensory attributes (P<0.05). All sensory attributes were significantly higher
for T2 than other CSK samples (P<0.05). A non-significant (P>0.05) difference was observed
amongst control and T1 in all sensory attributes except colour and appearance. On the basis of
these observation it can be concluded that 4mm particle size can be considered as optimum in
development of CSK using spent hen meat.
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Proceedings of the World Veterinary Poultry Association (India) Conference 202084
Developments in Poultry Products Technology
Food Safety Concern on Determination of Antibiotic Residues in Fresh and Cooked Broiler Meat by High Performance Liquid Chromatography (UV-Detector)
Rayala Reddy V*, Ravinder Reddy V, Baswa Reddy P ,Panda A K , Chinni Preetam V
College of Veterinary Science, Rajendra Nagar, Hyderabad
rayalreddy046@gmail.com
The inclusion of Antibiotic growth promoters (AGPs) in the feed is the most common and
cost effective method of enhancing the performance of poultry. The abundant misuse of AGPs
like tetracyclines (TCs) in poultry production results in the presence of their residues in edible
tissues, intended for human consumption, causing a health threat. Hence, the stability of TC
residues in chicken tissues under cooking conditions is an important research area, which
provides valuable information related to health safety aspects. A 42 days study was conducted
to determine the residue levels in fresh and cooked broiler meat by HPLC –UV detector for food
safety concern on public health. In this study 300 day old chicks were divided randomly in to 5
groups of 60 chicks each. All the groups were fed standard basal ration throughout the
experiment. Control group (T0) was without antibiotic supplements, group (T1) was
supplemented with Oxytetracyclines-OTC@ 100ppm, group (T2) with OTC@200ppm,group
(T3) with Chlortetracyclines-CTC@100ppm and group (T4) with CTC@200ppm along with
basal diet. At the end of 42nd day, 100 meat sample of muscle (both thigh and breast) were
collected and processed for HPLC analysis. The principal steps involved extraction in McIIvaine
buffer (pH 3.85) followed by a solid phase clean up step. In HPLC, a reversed phase C18 (Luna-
C18) column was used and compounds were separated at 40°C using a mobile phase of 0.01 M
oxalic acid buffer (pH 3.5)/acetonitrile/methanol (77:15:10, v/v/v) at a flow rate of 1.2 mL/min. A
wavelength of UV detector was set at 360 nm. The detection limit of OTC and CTC in the
method was calculated to be 5.509 ng/mL and 5.309 ng/mL respectively and the minimum
detectable quantity of OTC and CTC was found to be 18.336 ng/mL and 17.696 ng/mL
respectively. The statistical evaluation demonstrated high absolute recoveries of OTC and CTC
from spiked samples at three fortification levels, which were higher than 70% for all drugs. The
mean concentration of fresh muscle of T1 and T2were61.57 µg/kg and 77.65 µg/kg
respectively; T3 and T4were93.58 µg/kg and 95.92 µg/kg respectively. The fresh muscle
samples were cooked in ordinary pressure cooker about 30 min and the mean concentration of
cooked meat of muscle of T1 and T2were 27.06 µg/kg 32.91 µg/kg respectively; T3 and T4 were
20.42 µg/kg and 26.50 µg/kg respectively. The obtained data revealed that there was a
significant reduction (P<0.05) of the residual concentration in muscle samples on ordinary
cooking procedure. Hence sufficient cooking temperature and time can have a significant effect
on the losses of antibiotic residues in meat and provides an additional margin of food safety for
consumers.
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Proceedings of the World Veterinary Poultry Association (India) Conference 2020 85
Developments in Poultry Products Technology
Does Poultry Corona Virus Infection Spreads to Human Beings?
Sreenivas Gowda R N
Karnataka Veterinary, Animal and Fisheries Sciences University, Bidar.
drrns.gowda@gmail.com
There seem to be some confusion and psychic fear among the people that consuming
poultry products will spread corona viral infection to human beings. This is what we are hearing.
Hence this note is to clarify some of their doubts in the poultry business.
It is better to clarify the nature of the corona virus for better understanding of the infection
in various animals and human.
What is Corona virus?
The name of this virus comes from the Crown like spikes it has on its surface —
“corona” is Latin for “crown.(Fig1)” Corona viruses are a large group of viruses that infect
mostly bats, pigs and small mammals. In rare cases, they are zoonotic, meaning they can be
transmitted from animals to humans. Corona viruses are circulating in animals and some of
these corona viruses have the capability of transmitting between animals and humans. We call
this as“spillover . The phenomenon. The present outbreak of corona virus infection in humans
is a classic example.
Which other animals prone get this infection?
Corona virus and corona virus-like infections are described in animals such as swine,
cattle, horses, Camels, cats, dogs, rats, birds, bats, rabbits, ferrets, mink, and various wildlife
species, although many corona virus infections are subclinical or asymptomatic, meaning
without causing any disease. Several important diseases are caused by CoVs in animals and
birds. Infectious bronchitis virus (IBV) was the first CoV identified (in the 1930s) in Chickens.
A pig corona virus caused the deaths of millions of piglets in the United States in 2014. The
feline CoV (FeCoV) causative agent of FIP, a deadly disease of domestic cats. A rodent
Corona virus, mouse hepatitis virus (MHV), has served for many years as a useful model
system for investigating CoV replications and pathogenesis. Canine Respiratory disease
CRCoV is a relatively newly identified cause of contagious respiratory disease in dogs.
Corona virus disease was first described in 1931, with the first corona virus (HCoV-
229E) isolated from humans in 1965. Until the outbreak of severe acute respiratory syndrome in
late 2002, only two human corona viruses (HCoV) were known – HCoV-229E and HCoV-OC43.
Once the SARS corona virus(2002) (SARS-CoV) had been identified, two further human
corona viruses were also identified. Three groups of corona viruses exist: group 1 (HCoV-229E
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Proceedings of the World Veterinary Poultry Association (India) Conference 202086
Lead Papers
Fig1.Electronic microscopic picture of Corona Virus
Proceedings of the World Veterinary Poultry Association (India) Conference 2020 87
Lead Papers
and HCoV-NL63), group 2 (HCoVOC43 and HCoV-HKU1), group 3 (no human CoVsas yet).
SARS-CoV is an outlier to all these three groups, although some place it in group 2.
The evolution of novel corona (nCoV2019) virus causing devastating respiratory
disease in humans in Wuhan city of China, is the reason for mortality of more than 1200 people
and 45,000 patients remaining critical with infection which has created panic world over. The
disease is spreading from person to person through aerosol contamination and hospital
contamination throughout the world in more than 25 Countries.
IsnCovid-19 is similar to SARS and MERS?
In humans, coronaviruses are included in the spectrum of viruses causing the common
cold, and also recently found that human pathogens can cause lethal zoonotic infections like
Severe Acute Respiratory Syndrome(SARS) 2002 and Middle East Respiratory Syndrome
(MERS)2012. Even though these are related the strain of nCoV is different from both SARS
and MERS. The nCov is newly evolved from wild animals and reptiles.” The emergence of new
virus was predicted by scientists (Vincent et al 2007) way back in 2007it self. The presence of
large reservoir of SARS Co-like virus in horseshoe bats, together with culture of eating exotic
mammals in south China is a time bomb. WHO declared it as a new strain and named as
nCovid-19 meaning “novel corona virus disease
Is nCovid-19 present in India?
On 30 January, the Union Ministry of Health and Family Welfare in an official statement
said that students from Kerala, who were studying at Wuhan University and travelled to India,
have been tested positive for the corona virus. Presently they are free form infection, cured and
out of danger.
Is poultry suffer from Corona Virus?
The answer is yes, but these are different strains of corona virus causing three distinct
health problems in chicken. In chicks it causes bronchitis called as Infectious bronchitis (or
familiarly called IB infection). IB caused by Avian RNA Corona virus-Infectious Bronchitis
Virus(IBV).The IBV infected chick gasps for breath and shows respiratory distress. In egg laying
birds (Layer breeder birds) it causes5 to 10 per cent drop in egg production, production of
irregular corrugated shells and thin watery egg albumin. This is because of oviduct infection. It
cause curled embryos leading to poor hatchability. In broiler chicken, the variant IBvv causes
severe kidney damage–“Nephritis nephrosis syndrome”. And cause 15 to 20 percent mortality
among the infected chicks.
Turkey birds also suffer from corona virus infection. Turkey Corona virus were
first recognized in turkeys in the United States in 1951 and were associated with various enteric
disease syndromes, variously termed as “blue comb disease,” “mud fever,” “transmissible
enteritis” and “corona viral enteritis.” This disease is present throughout the world,
essentially wherever turkeys are raised. The virus can also infect turkeys of all ages, but the
most severe enteric disease is evident within the first few weeks of life.
How the corona virus Evolved :
Development of several new IBV strains is due to genetic recombination. Presence of
Proceedings of the World Veterinary Poultry Association (India) Conference 202088
Lead Papers
There are several Corona virus strains originate from different parts of the world.
IB produce Viral quasi species which are the mutant distributions (also termed mutant swarms
or mutant clouds) that are generated upon replication of RNA viruses, thereby producing more
than 50IB Variants.
The Control of Infectious Bronchitis respiratory disease in poultry caused by IB Viral
strains through manufacturing particular vaccine that suits the control a particular strain.(MA5,
H120, AusT)
What types of Vaccines are available to control IB?
The Commercial IB Vaccines are available world over and are manufactured in India
also, therefore IB Vaccine is regularly used in poultry operations in our country. Live and Killed
Vaccines are available. Live vaccines are applied through drinking water, either by spraying or
via eye drops, while killed vaccines are applied intramuscularly.
The frequently asked questions by poultry entrepreneurs and consumers are:
If a person consumes poultry meat of infected birds will it cause respiratory disease in
humans? If the person handle the IB sick birds gets infection? Will IB directly infect farmers?
How safe to eat the meat of IB infected birds?
The answer for all the above is NO. No risks to human health are suspected or
have been demonstrated to arise from poultry corona virus IBV. Neutralising antibodies
have been detected in people working with commercial chicken flocks, but the
significance of this remains unknown. Possibility of transmission of virus from human
to human or human to animal has no evidence. Humans can only transmit IBV to chicks
by mechanical means. Further the Indian cooking habits kill almost all the infectious
agents and hence poultry meat is safe and nutritious.
(All the above facts have been taken from scientific articles published world over
and reports from WHO/CDC documents).
ICAR-National Institute of Animal Nutrition and PhysiologyAdugodi, Bengaluru - 560 030
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