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Medicina pediátrica en pequeños animales
PRESENTATION BROCHURE
MAIN CHALLENGES IN P ULTRY FARMING
Infecti us br nchitis
Cintia Hiromi OkinoMaria de Fátima Silva Montassier
Monique Silva de FrançaH. L. Shivaprasad
Hélio José MontassierIgor Leonardo dos Santos
Raquel Rubia Rech
Infectious bronchitis is an acute and highly contagious respiratory
disease of chickens characterised by respiratory signs comprising
gasping, coughing, sneezing, tracheal rales and nasal discharge.
Besides, severe respiratory distress may occur in young chickens.
Respiratory distress, decrease in egg production, and loss of
internal egg quality and egg shell quality have been reported in
layers. Some strains of the virus produce severe kidney damage
and may be associated with high mortality. According to the current
situation (worldwide distribution), it is essential to analyse and
update this severe condition. The authors (highly experienced in this
topic) have developed a complete review including images, tables,
graphs, etc. The precise information and the atlas format make the
contents more understandable and affordable for the readers.
Infectious bronchitis
MAIN CHALLENGES IN P ULTRY FARMING
Infecti us br nchitis
Cintia Hiromi OkinoMaria de Fátima Silva Montassier
Monique Silva de FrançaH. L. Shivaprasad
Hélio José MontassierIgor Leonardo dos Santos
Raquel Rubia Rech
AUTHORS: Cintia Hiromi Okino, Maria de Fátima Silva Montassier, Monique Silva de França, H. L. Shivaprasad, Hélio José Montassier, Igor Leonardo dos Santos, Raquel Rubia Rech.
FORMAT: 17 x 11 cm.
NUMBER OF PAGES: 88.
BINDING: Paperback, wire-o. RETAIL PRICE
30 €
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Infectious bronchitis
Presentation of the book
Infectious bronchitis is an acute and highly contagious respiratory disease of chickens characterised by respiratory signs comprising gasping, coughing, sneezing, tracheal rales and nasal discharge.
In addition, severe respiratory distress may occur in young chickens. Respiratory distress, decrease in egg production, and loss of internal egg quality and egg shell quality have been reported in layers. Some strains of the virus produce severe kidney damage and may be associated with high mortality.
Moreover, this pathology is considered to have worldwide distribution, so it is not contro-lled totally. Knowing its most hazardous features is required to minimise the great impact which has in poultry farming.
According to the current situation, it is essential to review and update this severe problem. The authors have developed a complete review in a didactic and graphic way, including images, tables, graphs, etc., accompanied by a short text, to make the handbook more understandable and affordable. This precise information will help the veterinarians to know everything about this topic and tackle it properly.
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The authors
Cintia Hiromi OkinoGraduated in Veterinary Medicine and concluded MS and PhD in Veterinary Pathology at São Paulo State University (UNESP- Brazil).
Dr. Okino works at the Avian Virology Laboratory, Embrapa Swine and Poultry, a Brazilian Agricultural Research Corporation, where she develops research on avian respiratory viruses (Avian Infectious Bronchitis virus, Avian Metapneumovirus, Newcastle disease virus and Avian Influenza virus). The research includes evaluation of Immune Responses and Molecular Diag-nostics for these diseases. She has published articles about Avian Infectious Bronchitis virus in national and international journals and she won the “José Maria de Lammas Filho award” in 2013 during the Brazilian Poultry Sciences meeting (FACTA).
Maria de Fátima Silva MontassierGraduated in Pharmacy Sciences from Universidade Federal de Pernambuco (UFPE Brazil), she concluded MS in Microbiology at Sao Paulo State University (UNESP- Brazil) and PhD in Microbiology at Institute of Biomedical Sciences - University of São Paulo (USP-Brazil).
Dr. Montassier has developed post-doctoral research at the Viral Immunology Laboratory, Sao Paulo State University (UNESP- Brazil), where she is involved on studies of viral isolation and genome sequencing of Avian Infectious Bronchitis virus. She has published several arti-cles about Avian Infectious Bronchitis virus in national and international journals.
Monique Silva de FrançaGraduated in Veterinary Medicine from São Paulo State University (UNESP - Brazil) and con-cluded PhD in Veterinary Pathology at University of Georgia (USA). She is board certified by the American College of Poultry Veterinarians and American College of Veterinary Pathologists.
Dr. França is an Assistant Professor at Poultry Diagnostic and Research Center at Univer-sity of Georgia (USA), where she is involved with diagnostic service for the poultry industry, research and teaching. She has published several articles about various avian diseases in international journals.
H. L. ShivaprasadGraduated in Veterinary Sciences from University of Agricultural Sciences (India), he conclud-ed MS and PhD in Physiology and Genetics at the Poultry Science Department, The Ohio State University (USA) and MS in Pathology at Purdue University (USA). He is board certified by the American College of Poultry Veterinarians.
Infectious bronchitis
Dr. Shivaprasad is a Professor of Avian Pathology at the California Animal Health and Food Safety Laboratory System, University of California, Davis (USA), where he is involved with diagnostic service, teaching and research. He was the senior author of the best manuscript published on Veterinary Pathology Journal in 2011. He has travelled to more than 30 coun-tries on invitation primarily for teaching.
Hélio José MontassierGraduated in Veterinary Medicine at São Paulo State University (UNESP-Brazil), he conclud-ed MS and PhD in Microbiology at Institute of Biomedical Sciences - University of São Paulo (USP-Brazil) and post-doctoral research at Institute for Animal Health (UK).
Dr. Montassier is an Associate Professor of Veterinary Immunology at UNESP, where he deve-lops research on avian respiratory viruses, including evaluation of immune responses, deve-lopment of vaccines, molecular diagnosis and protein expression in different vectors. He has published several articles about viral immunology in international journals and book chapters.
Igor Leonardo dos SantosGraduated in Veterinary Medicine and concluded MS in Veterinary Pathology at São Paulo Sta-te University (UNESP - Brazil).
He is technical manager of Animal Health innovation area, looking for development and appli-cation of new products and services on the Brazilian Poultry industry. He has large experience of technical services, monitoring avian vaccine protection and identification of challenge by avian pathogens on the field, including avian necropsies and field biosecurity.
Raquel Rubia RechGraduated in Veterinary Medicine from Universidade do Estado de Santa Catarina (UDESC-Brazil), she concluded MS and PhD in Veterinary Pathology at Universidade Federal de Santa Maria (UFSM-Brazil).
She is a Clinical Assistant Professor at the Department of Veterinary Pathobiology - Texas A&M University (USA), where she teaches general and systemic pathology to veterinary and graduate students. Her primary focus in research is to explore pathogenesis of infectious diseases, using various pathologic techniques, especially immunohistochemistry. She has published several articles about pathobiology in national and international journals.
www.grupoasis.com/promo/infectious_bronchitis
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Communication services
MAIN CHALLENGES IN P ULTRY FARMING
Infecti us br nchitis
Cintia Hiromi OkinoMaria de Fátima Silva Montassier
Monique Silva de FrançaH. L. Shivaprasad
Hélio José MontassierIgor Leonardo dos Santos
Raquel Rubia Rech
Table of contents
1. Introduction
2. IBV variability
IBV variants
Classification of IBV strainsIBV genotypes
IBV serotypes
IBV pathotypes
IBV protectotypes
3. Immune responses
Innate immune responses
Adaptive immune responsesCell-mediated (CMI) and humoral immune responses
Decay of maternal antibodies
Humoral immune responses
Importance of memory CMI and humoral immune responses
4. Clinical signs, pathology and pathogenesis
Clinical signs
Pathology and pathogenesisMacroscopic lesions
Microscopic lesions
IBV cell tropism
5. Specimen sampling and diagnostic methods
Bird selection
Blood sampling
Swab sampling
Necropsy and tissue sampling
Filter card paper
Diagnostic methodsViral isolation
Molecular methods
Immunohistochemistry
Serological methods
6. Prevention and vaccine failure
IBV vaccinesOculo-nasal vaccination route (live attenuated vaccine)
Spray vaccination (live attenuated vaccine)
Drinking water vaccination (live attenuated vaccine)
Injectable vaccination (inactivated vaccine)
Vaccine failureIBV vaccines worldwide
7. References
5
» The first observation of infectious bronchitis (IB) was in North Dakota (USA) in 1930, being reported in 1931 as a highly contagious disease of young chicks with respiratory signs. 5 years later, a virus was demonstrated as the causative agent of this disease, being named infectious bronchitis virus (IBV). Thereafter, many cases were reported in several countries around the world. 10
» IB is a worldwide infectious and highly contagious disease which causes signifi-cant economic losses for poultry industry. IBV primarily infects and damages the respiratory tract, but also affects the genito-urinary tract, leading to reduced feed consumption and weight gain, and/or drop in egg production.
Introduction
Infectious BronchitisIntroduction1
6
Virus Species Genus
Figure 1. Taxonomy of IBV. 23
Miniopterus bat coronavirus 1A AFCD62Miniopterus bat coronavirus HKU8 AFCD77Porcine epidemic diarrhea virus CV777Scotophilus bat coronavirus 512/2005Human coronavirus 229EHuman coronavirus NL63 Amsterdam 1Rhinolophus bat coronavirus HKU2-GD/430/2006Transmissible gastroenteritis virus PUR46-MAD
Bovine coronavirus MebusMouse hepatitis virus A59Human coronavirus HKU1-ASARS-related coronavirus Tor2Rousettus bat coronavirus HKU9-1 BF-0051Tylonycteris bat coronavirus HKU4-1 B04fPipistrellus bat coronavirus HKU5 LMH03fMERS coronavirus Hu/Jorda-N3/2012
Betacoronavirus 1Murine coronavirusHuman coronavirus HKU1SARS-related coronavirusRousettus bat coronavirus HKU9Tylonycteris bat coronavirus HKU4Pipistrellus bat coronavirus HKU5To be established
Infectious bronchitis virus BeaudetteBeluga whale coronavirus SW1
Avian coronavirusBeluga whale coronavirus SW1
Munia coronavirus HKU13-3514Bulbul coronavirus HKU11-934Thrush coronavirus HKU12-600
Munia coronavirusBulbul coronavirusThrush coronavirus
Miniopterus bat coronavirus 1Miniopterus bat coronavirus HKU8Porcine epidemic diarrhea virusScotophilus bat coronavirus 512Human coronavirus 229EHuman coronavirus NL63Rhinolophus bat coronavirus HKU2Alphacoronavirus 1
Alphacoronavirus
Betacoronavirus
Gammacoronavirus
Deltacoronavirus
100
100100
100
100
100100
61
100
10078
100100
0,2
100
95
92
99
54
92
A
BD
C
INTRODUCTION
7
Chickens of all ages are naturally affected by IBV, but the disease is more severe in young chicks, causing some mortality. IBV infection also af-fects other gallinaceous and non-gallinaceous species. 8,18 Although the consequences of viral replication in these hosts remain unknown, ap-parently, most of these infections are subclinical.
Figure 2. Major local sites of replication of IBV in domestic fowl. 8
1. Harderian gland2. Trachea3. Oesophagus4. Lung5. Duodenum6. Ileum
7. Kidney8. Caecum9. Bursa of Fabricius10. Oviduct 11. Rectum
1
2
3
45
6
7
8
910
11
Infectious BronchitisIntroduction1
8
Morphology •Round and pleomorphic•Approximately 120 nm diameter
Genome
•Single strand RNA (ssRNA)•Length: 27.6 kb•Positive sense•Non-segmented
Thermostability •Inactivated after 15 minutes at 56 ºC or after 90 minutes at 45 ºC•Outdoors: survival up to 12 days (spring) and 56 days (winter)
pH stability More stable at pH 6.0-6.5 than 7.0-8.0
Chemical agents Inactivated after contact with most of the usual disinfectants
Table 1. Characteristics of IBV.
INTRODUCTION
9
Figure 3. IBV transmission (horizontal, as vertical transmission has not still been reported).
HEALTHY BIRDINFECTED BIRD (ACUTE PHASE)
Wild birds infected???
Clinical signs
24-48 hours
Rapid transmission
Air-borne
Direct-contact
Mechanic
INFECTED BIRD (CHRONIC PHASE)
Persistence
Faeces ≅20 w
eeks
Infectious BronchitisSpecimen sampling and diagnostic methods5
44
Bird selection
Table 2. Birds collected for each diagnostic method.
Sample Number Observation
Serum (breeders/layers)
30-45 –
Serum (broilers) 18-20 –
Swabs 30Swabs by route (tracheal and/or cloacal)
Tissue 53 birds with clinical signs and 2 birds without clinical signs
Figure 29. Methods for selecting birds in the flock for swabs and/or serum sampling.
Recommended method
Non-recommended methods
Imagine two lines crossing aviary diagonally, and then randomly choosing animals
Picking birds only from the middle
Picking birds only from one sideSelected bird
45
SPECIMEN SAMPLING AND DIAGNOSTIC METHODS
Blood sampling
Figure 30. Blood sampling from the brachial vein. Place the collected blood carefully into a tube, keep tube inclined at room temperature during 5 hours to overnight, and transfer serum to a clean microtube. Keep at 4 °C for short storage time or at -20 °C for longer time. Do not freeze and thaw several times.
Good qualityRemove the needleCollect 3-4 ml of blood Poor quality
Infectious BronchitisSpecimen sampling and diagnostic methods5
46
Figure 31. (a) Tracheal and (b) cloacal swab sampling. Place the swab into the tube containing the transport media. Keep at 4 °C for short storage time or at -20 °C for longer time.
Swab sampling a b
47
SPECIMEN SAMPLING AND DIAGNOSTIC METHODS
The necropsy correlates the gross lesions with the clinical signs of the chicken and the flock, and gives the tentative diagnosis based on the interpretation of the gross lesions. The necropsy is an excellent source for collecting tissues for the laboratory diagnosis and establishes the definitive diagnosis. While performing a necropsy, (1) perform a systematic necropsy technique, (2) examine and collect from the cleanest to the dirtiest organs, and (3) describe the clinical history and the gross findings in the necropsy form. Keep fresh tissue samples at 4 °C for short storage time or at -20 °C for longer time.
Necropsy and tissue sampling
Infectious BronchitisSpecimen sampling and diagnostic methods5
48
Figure 32. Respiratory tract sampling.
Proximal Medial Distal
49
SPECIMEN SAMPLING AND DIAGNOSTIC METHODS
Figure 33. Urogenital and gastrointestinal samples.
a b c
Testes
Kidneys
Oviduct
Cloaca
Colon
Caecum
Caecal Tonsils
Infectious BronchitisSpecimen sampling and diagnostic methods5
50
For histopathologic examination, take samples of 0.5-1 cm thickness and place them into 10 % buffered formalin for at least 1 day (not >5 days). Do not freeze sam-ples to be fixed. Use clean scissors and forceps for taking samples for viral isolation or molecular diagnosis. The trachea, lung, kidney, caecal tonsils of intestinal tract and oviduct are the better sources of virus depending of pathogenesis of disease.
51
SPECIMEN SAMPLING AND DIAGNOSTIC METHODS
Spotting swab or fresh tissue samples on active area of filter card papers could be an alternative method, providing transport of non-infectious material for long periods at room temperature. Filter cards are cotton-based cellulose papers impregnated with chemicals that inactivate many types of viruses and bacteria.
Filter card paper
Figure 34. Spotting a tracheal sample on filter card paper. Keep at room temperature (<41 °C) for up to 15 days. Courtesy of Erich Linnemann.
Infectious BronchitisSpecimen sampling and diagnostic methods5
52
Diagnostic methods
Table 3. Main methods for IBV diagnosis.
Differential diagnoses
Avian influenza virus Avian metapneumovirus
Newcastle disease virus Avibacterium paragallinarum
Infectious laringotracheitis virus Mycoplasma gallisepticum
Method What is identified? When is recommended to collect samples? Examples
Direct•Aetiological agent•Nucleic acids (genome)•Antigen
Preferable at the beginning of clinical signs
•Viral isolation (standard)•Molecular diagnosis•Immunohistochemistry
Indirect Antibodies anti-IBV Monitoring during all life cycle ELISA
53
SPECIMEN SAMPLING AND DIAGNOSTIC METHODS
Viral isolation
Table 4. Characteristics of viral isolation method.
Principle Detection of viable (infecting) virus (classic method)
Methods •Inoculation in embryonated eggs (most common)•Inoculation in tracheal organ culture
Biological material Tracheal swabs, cloacal swabs, fresh tissue samples
Cost ↑
Time 1-3 weeks
Sensitivity ++
Advantages Low to moderate cost
Disadvantages
•In delay on delivery of biological material or inappropriate storage, the virus could be easily inactivated providing a false negative result
•This method cannot differentiate attenuated vaccine virus from field strains of IBV•Only strains adapted to embryonated eggs (usually after several passages) will induce
characteristic lesions of IBV on the embryo
Recommended for Titration of vaccine, 52 viral propagation, isolation for characterization 49
Infectious BronchitisSpecimen sampling and diagnostic methods5
54
Figure 35. Workflow for viral isolation in embryonated eggs.
How to interpret the results?The positive result confirms the presence of infecting IBV. However, this virus could be an attenuated vaccine or a field strain, highlighting the importance of proving flock history and characterization of the viral isolate.
Observation of mortality
Mortality at 24 hours post-inoculation considered
non-specific
1st passage
NP
Biological materialEmbryonated
eggs 9-11 days of incubation
Collect allantoic fluid at 3rd day for sequential
passages
Evaluation of all inoculated eggs
at 7th day
Negative embryos (N) = at least two more passages to confirm resultPositive embryo/s (P) = positive for IBV
Candling eggs during 7 days
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