Life Technologies

5 Life Technologies | Animal Health Sampling technique TTR 1. Move the pig into a lateral recumbent position and secure this with appropriate material, e.g., towels ventral and dorsal of the body. 2. Palpate the trachea and hold the larynx with one hand, while palpating the first tracheal rings beyond the larynx with the other hand. 3. Puncture the trachea with a permanent needle in the caudodorsal direction (approx. 1–2 cm caudal of the larynx) and control the success of reaching the trachea by aspirating air instead of blood. 4. Insert the catheter through the needle into the trachea and then push the catheter as deep as possible into the lung. 5. Instill sterile sodium chloride solution (or PBS), and aspirate it after 2–3 breaths of the animal. The amount of solution should not exceed 0.5–0.7 mL per kg of pig. Storage The aspirated lavage fluid should be transferred immediately into a sterile sample collection tube with a twist lock. This tube should be labeled with the animal ID using a permanent marker. Write numbers and letters clearly according to good clinical practice (e.g., 5 not 5). The sample should be stored in a refrigerator until shipment to the laboratory, which should be within 24–36 hours. If this is not possible and only PCR is required, freeze the sample at –20 to –80°C. Keep in mind that no further cultural examination is possible after freezing a sample. If cultural examination is required and transport cannot be organized immediately, the BALF should be centrifuged at 2,000 x g for 10 minutes. After discharging the supernatant, take a swab from the sediment and place this in appropriate transport medium (e.g., Amies), and store the swab at 8°C.

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