PRESENTATION
BROCHURE
MAIN CHALLENGES IN P
ULTRY FARMING
Sampling Cristina Gómez Martínez Sonia Téllez Peña
MAIN CHALLENGES IN POULTRY FARMING
Sampling
MAIN CHALLENGES IN P
ULTRY FARMING
Sampling Cristina Gómez Martínez Sonia Téllez Peña
This book describes the ideal protocols for the collection of samples from live animals and of environmental samples from the facilities, and addresses important aspects of sample collection. It also provides information about the samples of choice for the diagnosis of the most common diseases in commercial poultry flocks.
TARGET AUDIENCE:
✱✱ Production animal vets. Poultry ✱✱ Animal production technicians ✱✱ Veterinary students FORMAT: 17 x 11 cm NUMBER OF PAGES: 80 NUMBER OF IMAGES: 50 BINDING: paperback, wire-o
RETAIL PRICE
€35
Authors CRISTINA GÓMEZ MARTÍNEZ Degree in veterinary science from the University of Extremadura (Spain) and master’s degree in food safety from the Madrid College of Veterinary Surgeons and Complutense University of Madrid. Head of the Project Management Unit at the VISAVET-UCM Centre. SONIA TÉLLEZ PEÑA PhD in veterinary medicine from the Complutense University of Madrid (UCM), Spain. She was a researcher at the VISAVET Health Surveillance Centre, UCM (VISAVET-UCM), and now works as a researcher in a private company.
KEY FEATURES:
➜➜ Includes a large number of images and diagrams. ➜➜ Small and easy-to-handle format. ➜➜ Can be used for quick reference in daily practice.
MAIN CHALLENGES IN POULTRY FARMING | Sampling
Presentation of the book Correct sample collection is key to determine the serological status of a flock and be able to assess or check the efficacy of an established vaccination programme, make a timely diagnosis, conduct epidemiological surveillance or ensure cleaning and disinfection measures are correctly applied on the farm. For this reason, this book describes the ideal protocols for the collection of blood, faeces or respiratory tract secretion samples from live animals and of environmental samples from the facilities. This book addresses the most important aspects of sample collection and will allow farm technicians to carry out this task in an appropriate manner, thereby ensuring the viability of the specimens and, therefore, that these can be used for the intended purpose. It includes, among other aspects, the key elements for the correct collection of specimens, the minimum data that should accompany them and the appropriate conditions of preservation and submission to the laboratory. Finally, it provides information about the samples of choice for the diagnosis of the main diseases in poultry, which will undoubtedly be of great help in the event of a suspected outbreak. Moreover, the large number of images and diagrams and the book’s small format will allow readers to use it for quick reference and help them to easily assimilate the information it contains, thus turning it into an essential tool on poultry farms.
Cristina Gómez Martínez Head of Project Management VISAVET Health Surveillance Centre, Complutense University of Madrid
The authors Cristina Gómez Martínez Cristina Gómez Martínez holds a degree in veterinary science (2007) from the University of Extremadura (Spain) and a master’s degree in food safety (2010) from the Madrid College of Veterinary Surgeons and the Complutense University of Madrid. Her master’s dissertation was titled “Implementation, evaluation and improvement of control plans as the foundation for a system based on HACCP principles to reduce the prevalence and the risk of introduction and dissemination of food-borne zoonoses in poultry meat”. Her professional career has involved work in both the public and private sectors. She began her collaboration with the Health Surveillance Centre of the Complutense University of Madrid (VISAVET-UCM) in 2010, joining the Foodborne Zoonoses and Antimicrobial Resistance Unit to manage research projects in the poultry sector. Currently, she is the head of the Project Management Unit of the VISAVET-UCM Centre. In the past five years, Cristina Gómez has coauthored five articles in specialist magazines, over ten book chapters and a book. In addition, she has given nine presentations at national congresses in the field of animal health and food safety. She has participated in more than 15 research projects supported by public funding and collaborations with government and private companies.
MAIN CHALLENGES IN POULTRY FARMING | Sampling
Sonia Téllez Peña She graduated (1998) and obtained her PhD (2003) in veterinary medicine from the Complutense University of Madrid (UCM), Spain. She joined the Department of Animal Health of the UCM Faculty of Veterinary Medicine in 1999 to complete her doctoral thesis “Detection and characterisation of Salmonella in reptiles and amphibians”. After completing her PhD, she worked as a researcher at the VISAVET Health Surveillance Centre, UCM (VISAVET-UCM), where she focused on the surveillance of microorganisms responsible for food-borne zoonotic diseases and antimicrobial resistance.
hkeita/shutterstock.com hkeita/shutterstock.com
She has worked as a researcher in a private company since 2014.
Table of contents 1. Introduction Reasons for sampling. Importance Keys for correct sampling Biosecurity in sampling Microbiology samples Molecular biology samples Pathology samples Other samples Minimum data to accompany the samples
2. Biological samples Blood and serum Venipuncture of the cutaneous ulnar vein (wing vein) Jugular vein puncture Cardiac puncture Handling of blood samples and serum collection Material for blood and serum collection Excrement Faeces Meconium Upper respiratory tract samples Tracheal swabs Oropharyngeal swabs
3. Environmental samples from the poultry houses Floor and surface samples Sampling material Dust Boot swabs Faeces Microbiological control plates Other samples Water Feed
4. Sample storage and submission Storage conditions and submission of samples to the laboratory Temperature Time
5. Samples of choice for diagnosis Samples of choice for the diagnosis of the major diseases in poultry production
MAIN CHALLENGES IN P
ULTRY FARMING
Sampling Cristina Gómez Martínez Sonia Téllez Peña
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Biological samples
Sampling
Blood and serum Venous or cardiac puncture may be performed for blood and serum sampling. Slight negative pressure should always be applied intermittently to determine when the puncture of the vein or heart has occurred. As guidance, it is safe to draw volumes between 0.3 and 0.6 ml of blood per 100 g of live weight of the bird.
Venipuncture of the cutaneous ulnar vein (wing vein) It is the technique of choice in broilers, laying hens and turkeys over 4 weeks of age. The cutaneous ulnar vein, or wing vein, is located in the depression formed by the biceps brachii and triceps brachii muscles in the ventral part of the humeral region. The puncture is performed by extending both wings dorsally and grasping them firmly to expose the wing membrane. To better visualise the vein, some feathers can be removed and the skin moistened with alcohol (70 %) or another disinfectant (thereby also ensuring asepsis). The needle is inserted at an intermediate point of the vein between the elbow and shoulder joints and opposite to the direction of blood flow (Fig. 1). Care should be taken not to damage the tendon of the pronator muscle, which runs parallel to the vein, or the ulnar nerve, which is below it (Fig. 2).
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Biceps
Humerus Ulnar nerve Triceps
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Cutaneous ulnar vein or wing vein Tendon of pronator muscle
Figure 1. Blood sampling from the wing vein. Image courtesy of AVEZOCA.
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Figure 2. Puncture site for drawing blood from the wing vein.
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Biological samples
Sampling
Jugular vein puncture It can be performed in birds of all ages, but is the technique of choice in day-old chicks. In addition, in day-old chicks, when it is not essential to keep them alive, blood samples can be taken by decapitation. Suction should be slow to prevent the vein from collapsing.
The ideal puncture site is the middle part of the neck. The chick should be held in one hand, and with the thumb of that hand the jugular vein should be pressed so that it dilates (Fig. 3). With the free hand, the needle is inserted against the direction of blood flow of the vein and the blood sample is taken. The vein can also be punctured with the needle and the blood collected by the drip method (Fig. 4).
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Subsequent pressure at the puncture site promotes coagulation and prevents bleeding and the formation of haematomas.
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Figure 3. Exposure of the jugular vein prior to the collection. Image courtesy of AVEZOCA.
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Figure 4. Collection of a blood sample using the drip method.
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Sampling
Cardiac puncture It is an effective and rapid technique indicated for the collection of large volumes of blood (4–10 ml) in young (over 4 weeks) and adult birds. It should be performed with great care and only by experienced personnel, since an error can cause the immediate death of the bird; it is advisable to practice beforehand on recently dead animals.
It can be carried out using two approaches: anterior and lateral. The choice of one or the other will depend on whether it is done alone or with help and on the particular skill of each person.
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When alone, the anterior or thoracic approach will be performed. The bird will be placed lying on its back (at the edge of a table or cage, or on the legs of the technician), with the neck and head extended downward, and the legs will be restrained by holding them with one hand. The needle must penetrate the highest point of the inverted V formed by the clavicle, angled towards the tail and in the same plane as the keel on its right side (Fig. 5). In adult birds, the needle must penetrate completely until entering the heart.
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BIOLOGICAL SAMPLES
The lateral approach can be performed with the help of another person who immobilises the bird. In this case, the bird will lie on its right side and be immobilised by having its legs held on one side and the wings on the other. The needle must penetrate through the left chest wall in the groove formed by the edge of the pectoral muscle (breast), where the ribs can be felt, about 5 cm above the keel bone and at a 90° angle to the plane of this.
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In both approaches, by applying a slight negative pressure it will be seen that the blood flows easily when the needle enters the heart. Any error can lead to death of the bird due to fatal haemorrhage. Figure 5. Blood sampling by cardiac puncture (anterior thoracic approach with help of another person). Image courtesy of AVEZOCA.
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In all cases in which an incorrect practice of cardiac puncture is detected, the bird should be humanely euthanised.
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Biological samples
Sampling
Handling of blood samples and serum collection In commercial poultry farming, blood sampling is most commonly performed for serological studies. The key steps for obtaining a quality serum sample are detailed below. ◗ Select a representative number of birds (see Introduction). Select healthy birds to determine the serological status of the flock and sick birds for the diagnosis of disease. ◗ Draw a volume of 2 to 3 ml of blood to yield 1.0 to 1.5 ml of serum. ◗ After removing the needle from the syringe, carefully deposit the blood in the coagulation tube without filling it completely, allowing it to run down the side of the tube to prevent haemolysis.
◗ Keep the tubes horizontal and at a temperature between 27 and 38 °C until the clot is formed. This position maximises the surface area of the clot and, therefore, the amount of serum obtained. A higher temperature may damage the sample and a lower temperature may delay clot formation and reduce the serum yield.
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◗ Thereafter, keep samples refrigerated until serum collection or submission to the laboratory. Ideally, serum should be sent but the clotted blood sample may be sent if sudden movements are avoided and submission is made within 8 hours of sampling.
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In turkeys, the normal clotting time is longer than in other birds.
Shaking, exposure to direct sunlight, and freezing of the clotted blood should be avoided since they can cause haemolysis of the clot and, therefore, deterioration of the sample. A good quality serum sample should be clear and slightly yellow (Fig. 6).
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In fattened animals, the serum may appear milky due to the presence of lipids. This is limited by taking the animal’s fasting sample.
Figure 6. Serum. Image courtesy of the VISAVET-UCM centre.
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Sampling
Material for blood and serum collection ◗ Needles of 20–27 gauge and 12.7–25.4 mm (0.5–1.5 inches) length (Fig. 7), depending on the size of the bird and venipuncture site.
Adult turkeys may require larger needles.
◗ Syringes of 3 to 12 ml, depending on the volume of blood to be drawn. ◗ Sterile plastic or glass tubes with hermetic lid (with anticoagulant – sodium citrate, heparin, EDTA – if the sample is whole blood) (Figs. 8 and 9).
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Figure 7. Needles of different calibres. Image courtesy of AVEZOCA. Figure 8. Tube with anticoagulant. Image courtesy of AVEZOCA.
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Figure 9. Tubes without anticoagulant for blood and serum sample collection. Images courtesy of AVEZOCA.
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Biological samples
Sampling
Excrement Faeces Cloacal swabs allow samples of faeces to be taken in live birds under sterile conditions. A sterile swab is gently inserted into the cloaca (its full length in adult birds). The sample is taken using gentle circular movements (Fig. 10).
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Meconium Sampling is performed in day-old birds. The meconium of several birds is collected in a sterile container by gently pressing on the abdomen to excrete it.
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Figure 10. Collection of a faecal sample from the cloaca with a swab. Image courtesy of AVEZOCA.
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Biological samples
Sampling
Upper respiratory tract samples Samples are taken from the upper respiratory tract in live birds using a swab.
Tracheal swabs The bird’s beak is opened, the tongue is depressed and the sterile swab is inserted into the trachea (Fig. 11) avoiding contact with the oral mucosa. Inside the trachea, movements of the swab are made so that it makes contact with the entire tracheal mucosa.
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To avoid confusing the trachea with the oesophagus, the tongue is gently pulled so that the trachea, located ventrally, is exposed in the oral cavity.
Oropharyngeal swabs The swab is introduced in the oral cavity of the bird and the sample is taken by making circular movements over the oral and pharyngeal mucosa.
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Figure 11. Visualisation of the entrance to the trachea. Image courtesy of AVEZOCA.
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