Upon arrival, and until the challenge infection, all animals were located in an outdoor animal enclosure. The inside area per cage was 6–8 m2, and the outside area per cage was 17–55 m2. Environmental enrichment items like pull ropes, chewing toys, cloths and dog beds were provided in each cage. If necessary, the inside and outside area of each cage could be separated from each other. The animals were divided by sex in equal groups with a maximum of 5 animals per cage. For the time of vaccine administration, the animals were separated from the others for at least 2–4 h, avoiding possible horizontal transmission. At the time of the challenge, the animals were transferred from the outdoor enclosure to the units of the indoor experimental animal facility at Ceva Innovation Center, Dessau-Rosslau, Germany. The animals were kept in individual cages (ground area: 4 m2). As these units had a total housing capacity for 20 dogs, it was necessary to split the study into 2 parts.
The animals were fed twice a day (Vollmers’ Welpenkost, Gerhard Vollmer GmbH &Co. KG, Spenge, Germany). Water was offered ad libitum. All dogs were observed at least once daily for general health, feed intake and defecation by the staff. In the case of an abnormal clinical finding during the daily animal check, it was determined if the observation was due to a rabies infection and, if not, if the animal needed treatment. In case of severe clinical signs, dogs were humanely killed.
Treatment and sampling in general were done without anesthesia or sedation. Only for the challenge infection, and on single occasions for blood sampling, animals were sedated using Medetomidine (Domitor®, Vetoguinol, Ismaning, Germany) at a dosage of 0.1 mg/kg body weight i.m. during the first part. During the second part of the study, Tiletamin hydrochloride + Zolazepam hydrochloride (Zoletil 100 [50 mg/mL + 50 mg/mL], Virbac Arzneimittel GmbH, Bad Oldesloe, Germany) was used in a dosage of 7.5 mg/kg. Induction of euthanasia was done by Xylazine (2%, 2 mg/kg) and Ketamine (10%, 10 mg/kg) i.m. (Serumwerk Bernburg AG, Bernburg, Germany). For euthanasia, T61® (Intervet GmbH, Unterschleissheim, Germany) was used in deep general anesthesia and was administered at a dosage of 0.3 mL/kg body weight.
At the end of the experiment, all dogs that survived the challenge were humanely killed, as described above, except for the 5 surplus animals that were not challenged. These animals were handed over to new owners through a specialized organization (Labor-Beagle-Hilfe e.V.) after termination of the study.
The animals were fed twice a day (Vollmers’ Welpenkost, Gerhard Vollmer GmbH &Co. KG, Spenge, Germany). Water was offered ad libitum. All dogs were observed at least once daily for general health, feed intake and defecation by the staff. In the case of an abnormal clinical finding during the daily animal check, it was determined if the observation was due to a rabies infection and, if not, if the animal needed treatment. In case of severe clinical signs, dogs were humanely killed.
Treatment and sampling in general were done without anesthesia or sedation. Only for the challenge infection, and on single occasions for blood sampling, animals were sedated using Medetomidine (Domitor®, Vetoguinol, Ismaning, Germany) at a dosage of 0.1 mg/kg body weight i.m. during the first part. During the second part of the study, Tiletamin hydrochloride + Zolazepam hydrochloride (Zoletil 100 [50 mg/mL + 50 mg/mL], Virbac Arzneimittel GmbH, Bad Oldesloe, Germany) was used in a dosage of 7.5 mg/kg. Induction of euthanasia was done by Xylazine (2%, 2 mg/kg) and Ketamine (10%, 10 mg/kg) i.m. (Serumwerk Bernburg AG, Bernburg, Germany). For euthanasia, T61® (Intervet GmbH, Unterschleissheim, Germany) was used in deep general anesthesia and was administered at a dosage of 0.3 mL/kg body weight.
At the end of the experiment, all dogs that survived the challenge were humanely killed, as described above, except for the 5 surplus animals that were not challenged. These animals were handed over to new owners through a specialized organization (Labor-Beagle-Hilfe e.V.) after termination of the study.
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