Infectious bronchitis

Medicina pediátrica en pequeños animales

PRESENTATION BROCHURE

MAIN CHALLENGES IN P

ULTRY FARMING

Infecti us br nchitis 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

MAIN CHALLENGES IN P

Infectious bronchitis

ULTRY FARMING

Infecti us br nchitis 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

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 €

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.

Tsekhmister/shutterstock.com

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 controlled totally. Knowing its most hazardous features is required to minimise the great impact which has in poultry farming.

hkeita/shutterstock.com

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.

The authors Cintia Hiromi Okino Graduated 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 Diagnostics 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 Montassier Graduated 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 articles about Avian Infectious Bronchitis virus in national and international journals.

Monique Silva de França Graduated in Veterinary Medicine from São Paulo State University (UNESP - Brazil) and concluded 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 University 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. Shivaprasad Graduated in Veterinary Sciences from University of Agricultural Sciences (India), he concluded 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 countries on invitation primarily for teaching.

Hélio José Montassier Graduated in Veterinary Medicine at São Paulo State University (UNESP-Brazil), he concluded 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 develops research on avian respiratory viruses, including evaluation of immune responses, development 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 Santos Graduated in Veterinary Medicine and concluded MS in Veterinary Pathology at São Paulo State University (UNESP - Brazil).

He is technical manager of Animal Health innovation area, looking for development and application 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 Rech Graduated in Veterinary Medicine from Universidade do Estado de Santa Catarina (UDESCBrazil), 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.

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www.grupoasis.com/promo/infectious_bronchitis

MAIN CHALLENGES IN P

ULTRY FARMING

Infecti us br nchitis 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

Table of contents 1. Introduction 2. IBV variability

5. Specimen sampling and diagnostic methods Bird selection

IBV variants

Blood sampling

Classification of IBV strains

Swab sampling

IBV genotypes IBV serotypes

Necropsy and tissue sampling

IBV pathotypes

Filter card paper

IBV protectotypes

3. Immune responses Innate immune responses Adaptive immune responses Cell-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 pathogenesis Macroscopic lesions Microscopic lesions IBV cell tropism

Diagnostic methods Viral isolation Molecular methods Immunohistochemistry Serological methods

6. Prevention and vaccine failure IBV vaccines Oculo-nasal vaccination route (live attenuated vaccine) Spray vaccination (live attenuated vaccine) Drinking water vaccination (live attenuated vaccine) Injectable vaccination (inactivated vaccine)

Vaccine failure IBV vaccines worldwide

7. References

Introduction Âť 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

5

Âť IB is a worldwide infectious and highly contagious disease which causes significant 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.

1

Introduction

Infectious Bronchitis Virus

100 95 92 100 100

99 100

100

100

54 100

100 61

92

100 78

100 100

0,2

Genus

Miniopterus bat coronavirus 1 Miniopterus bat coronavirus HKU8 Porcine epidemic diarrhea virus Scotophilus bat coronavirus 512 Human coronavirus 229E Human coronavirus NL63 Rhinolophus bat coronavirus HKU2 Alphacoronavirus 1

Alphacoronavirus

Bovine coronavirus Mebus Mouse hepatitis virus A59 Human coronavirus HKU1-A SARS-related coronavirus Tor2 Rousettus bat coronavirus HKU9-1 BF-0051 Tylonycteris bat coronavirus HKU4-1 B04f Pipistrellus bat coronavirus HKU5 LMH03f MERS coronavirus Hu/Jorda-N3/2012

Betacoronavirus 1 Murine coronavirus Human coronavirus HKU1 SARS-related coronavirus Rousettus bat coronavirus HKU9 Tylonycteris bat coronavirus HKU4 Pipistrellus bat coronavirus HKU5 To be established

Betacoronavirus

Infectious bronchitis virus Beaudette Beluga whale coronavirus SW1

100

Species

Miniopterus bat coronavirus 1A AFCD62 Miniopterus bat coronavirus HKU8 AFCD77 Porcine epidemic diarrhea virus CV777 Scotophilus bat coronavirus 512/2005 Human coronavirus 229E Human coronavirus NL63 Amsterdam 1 Rhinolophus bat coronavirus HKU2-GD/430/2006 Transmissible gastroenteritis virus PUR46-MAD

Munia coronavirus HKU13-3514 Bulbul coronavirus HKU11-934 Thrush coronavirus HKU12-600

Avian coronavirus Beluga whale coronavirus SW1 Munia coronavirus Bulbul coronavirus Thrush coronavirus

A B D

6

C

Gammacoronavirus Deltacoronavirus

Figure 1. Taxonomy of IBV. 23

INTRODUCTION

Chickens of all ages are naturally affected by IBV, but the disease is more severe in young chicks, causing some mortality. IBV infection also affects other gallinaceous and non-gallinaceous species. 8,18 Although the consequences of viral replication in these hosts remain unknown, apparently, most of these infections are subclinical.

1

3 2 4

5

7 10

9

7

11

6 8

1.

2.

3.

4.

5.

6.

Harderian gland Trachea Oesophagus Lung Duodenum Ileum

7.

8.

9.

10.

11.

Kidney Caecum Bursa of Fabricius Oviduct Rectum

Figure 2. Major local sites of replication of IBV in domestic fowl. 8

1

Introduction

Infectious Bronchitis

Table 1. Characteristics of IBV. Morphology

Genome

Thermostability pH stability Chemical agents

• Round and pleomorphic • Approximately 120 nm diameter • Single strand RNA (ssRNA) • Length: 27.6 kb • Positive sense • Non-segmented • 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) More stable at pH 6.0-6.5 than 7.0-8.0 Inactivated after contact with most of the usual disinfectants

8

INTRODUCTION

Clinical signs 24-48 hours Wild birds infected???

Rapid transmission

INFECTED BIRD (ACUTE PHASE)

HEALTHY BIRD

Air-borne Direct-contact

9

Mechanic

Pe rsi ste nc e

s ek we 0 ≅2 es ec a F

INFECTED BIRD (CHRONIC PHASE)

Figure 3. IBV transmission (horizontal, as vertical transmission has not still been reported).

5

Specimen sampling and diagnostic methods

Bird selection

Recommended method

Infectious Bronchitis

Non-recommended methods

Table 2. Birds collected for each diagnostic method.

Number

Observation

Serum (breeders/layers)

Sample

30-45

–

Serum (broilers)

18-20

–

Swabs

30

Swabs by route (tracheal and/or cloacal)

Tissue

5

3 birds with clinical signs and 2 birds without clinical signs

Imagine two lines crossing aviary diagonally, and then randomly choosing animals

Selected bird

Picking birds only from the middle 44

Picking birds only from one side

Figure 29. Methods for selecting birds in the flock for swabs and/or serum sampling.

SPECIMEN SAMPLING AND DIAGNOSTIC METHODS

Blood sampling Collect 3-4 ml of blood

Remove the needle

Good quality

Poor quality 45

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 째C for short storage time or at -20 째C for longer time. Do not freeze and thaw several times.

5

Specimen sampling and diagnostic methods

Infectious Bronchitis

Swab sampling a

b

46

Figure 31. (a) Tracheal and (b) cloacal swab sampling. Place the swab into the tube containing the transport media. Keep at 4 째C for short storage time or at -20 째C for longer time.

SPECIMEN SAMPLING AND DIAGNOSTIC METHODS

Necropsy and tissue sampling 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 째C for short storage time or at -20 째C for longer time.

5

Specimen sampling and diagnostic methods

Proximal

Medial

47

Infectious Bronchitis

Distal

48

Figure 32. Respiratory tract sampling.

SPECIMEN SAMPLING AND DIAGNOSTIC METHODS

a

b

c Caecal Tonsils

Testes

Caecum Oviduct

Kidneys

49

Colon

Cloaca

Figure 33. Urogenital and gastrointestinal samples.

5

Specimen sampling and diagnostic methods

Infectious Bronchitis

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 samples 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.

50

SPECIMEN SAMPLING AND DIAGNOSTIC METHODS

Filter card paper 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.

51

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.

5

Specimen sampling and diagnostic methods

Infectious Bronchitis

Diagnostic methods Differential diagnoses Avian influenza virus

Avian metapneumovirus

Newcastle disease virus

Avibacterium paragallinarum

Infectious laringotracheitis virus

Mycoplasma gallisepticum 52

Table 3. Main methods for IBV diagnosis.

Method Direct Indirect

What is identified?

• Aetiological agent • Nucleic acids (genome) • Antigen Antibodies anti-IBV

When is recommended to collect samples?

Preferable at the beginning of clinical signs Monitoring during all life cycle

Examples • Viral isolation (standard) • Molecular diagnosis • Immunohistochemistry ELISA

SPECIMEN SAMPLING AND DIAGNOSTIC METHODS

Viral isolation Table 4. Characteristics of viral isolation method.

Principle

Detection of viable (infecting) virus (classic method) • Inoculation in embryonated eggs (most common) • Inoculation in tracheal organ culture

Methods Biological material

Tracheal swabs, cloacal swabs, fresh tissue samples

Cost

↑

Time

1-3 weeks

Sensitivity

++

Advantages

Low to moderate cost • 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

Disadvantages

Recommended for

5

53

Titration of vaccine, 52 viral propagation, isolation for characterization 49

Specimen sampling and diagnostic methods 1st passage Biological material

Embryonated eggs 9-11 days of incubation

Observation of mortality

Candling eggs during 7 days

Infectious Bronchitis

Collect allantoic fluid at 3rd day for sequential passages

Evaluation of all inoculated eggs at 7th day

Mortality at 24 hours postinoculation considered non-specific 54

Negative embryos (N) = at least two more passages to confirm result Positive embryo/s (P) = positive for IBV

P N

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.

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