Zoletil 100 mg ml

At the end of the observation period (54 days after vaccination), survivor animals were anesthetized by intramuscular injection of an anesthetic combination of tiletamine-zolazepam (Zoletil® 100 mg/ml, Virbac, France, target dose 3 mg/kg) and medetomidine (Medetor®, Virbac, France, target dose 0.05 mg/kg) (28 (link)), then euthanized by intravenous injection of T61. A thorough post-mortem examination was done to detect the presence of macroscopic lesions compatible with ASF. Sixteen different tissues (listed in Table 1), including all major lymph nodes and target visceral organs, were obtained from each necropsied animal and tested by real-time PCR to detect ASFV. Virus isolation was performed from a subset of tissues, using established procedures (29 ). Samples were blind-passaged three times, and plates were examined for hemadsorption during 6 days. If ASF-compatible lesions were found, they were classified based on their distribution and intensity.

Clinical signs, including rectal temperature, were recorded daily and expressed with a quantitative clinical score obtained by adding values for eight clinical signs, as described by Gallardo et al.^{16 (link),17 (link)}. These signs included fever (defined as a rectal temperature higher than 40 °C), anorexia, recumbence, skin hemorrhage or cyanosis, joint swelling, respiratory distress, ocular discharge and digestive findings. The body temperature was measured before each sampling and also, at 3, 6, 10, and 14 days post-inoculation (dpi). At the end of the experimental period (28 dpi), the animals were deeply anesthetized using intramuscular injections with tiletamine-zolazepam (Zoletil® 100 mg/ml, Virbac, France) and medetomidine (Medetor®, Virbac, France), then euthanized by employing intravenous injections of the T61® euthanizing agent (Intervet, Spain). Necropsy was performed on all the animals, and tissue samples (lymph nodes, spleen, liver, lung, heart, kidney, and bone marrow) were simultaneously collected for ASFV detection.

Buccal samples were collected from 30 randomly selected macaques living in KFP and the surrounding urban area during trapping for health screening checkups. The macaques were captured in a soft nylon mesh cage with a wooden frame (150 × 40 × 40 cm) baited with groundnuts and bananas. Tiletamine-zolazepam (Zoletil^® 100 mg/mL, Virbac, Carros, France) was injected intramuscularly to sedate the macaques through 5 mL anesthetic blowpipes after visually assessing a body weight for each macaque in the mesh cage. After anesthesia, the macaques were removed from the cage and vital signs were monitored. Buccal swab samples were then collected aseptically using a sterile buccal swab collection kit with transport media (Deltalab, Barcelona, Spain). All trapping and sampling procedures were conducted by wildlife veterinary specialists from MSU following the protocol approved by the MSU Institutional Animal Care and Use Committee (ethics statement above) and the Thai Department of National Parks. All buccal swab samples were stored at 4°C and taken to the One Health Research Unit, Veterinary Public Health Laboratory, Faculty of Veterinary Sciences at MSU for bacterial testing within 6 h of collection.