Najafi, Pardis http://orcid.org/0009-0005-9512-9679
Reimer, Christian http://orcid.org/0000-0002-9697-2511
Gilthorpe, Jonathan D. http://orcid.org/0000-0002-6884-4774
Jacobsen, Kirsten R. http://orcid.org/0000-0002-3211-7890
Ramløse, Maja http://orcid.org/0009-0006-2217-7300
Paul, Nora-Fabienne http://orcid.org/0009-0003-2442-9864
Simianer, Henner http://orcid.org/0000-0002-7551-3797
Tetens, Jens http://orcid.org/0000-0001-5352-464X
Falker-Gieske, Clemens http://orcid.org/0000-0001-9160-1909
Funding for this research was provided by:
Georg-August-Universität Göttingen
Article History
Received: 29 November 2023
Accepted: 2 February 2024
First Online: 21 February 2024
Declarations
:
: The housing of the animals was in accordance with EU Directive 2010/63/EU of 22 September 2010 on the protection of animals used for scientific purposes. The barrier facility where the animals were housed is fully AAALAC accredited and in compliance with the Eighth Edition of the Guide for the Care and Use of Laboratory Animals (NRC 2011). The project was reviewed by the ethics committee at Ellegaard Göttingen Minipigs A/S (Dyrevelfærdkomiteen). Procedures performed are all covered by the animal license granted by the Danish Animal Experiments Inspectorate, Ministry of Food, Agriculture and Fisheries in Denmark. For animals processed before February 2022, the license number was: 2017–15-0201–01202, and as of March 2022, the license number is 2022–15-0201–01167. We ensure that our reporting adheres to the ARRIVE guidelines () for the reporting of animal experiments. The general health and well-being of the GMPs, when not experiencing seizures, were determined to be similar to unaffected GMPs as animals were otherwise normal—clinically and behaviourally. The GMPs were inspected daily by caretaking staff and any sign of a decrease in general welfare or clinical disease was notified to, and assessed by, the attending veterinarian. Animals observed to be experiencing frequent or severe seizures were reported to the attending veterinarian, and then humanely euthanised. This was defined as observation by the attending staff of > 4 seizures per day and/or status epilepticus. Animals were euthanised under current guidelines established by the American Veterinary Medical Association (AVMA). First, general anaesthesia was induced with a combination of sedative and anaesthetic agents: Zoletil mixture (Tiletamine 125 mg and Zolazepam 125 mg dissolved in Xylazine 20 mg/ml) 0.1 ml/kg administered intramuscularly. Once the animals were anesthetised, animals were euthanised with an overdose of pentobarbital (400 mg/ml, 0.25 ml/kg) by intravenous or intracardial injection. Skin biopsies were taken immediately post-mortem. Weighing was performed concerning the inclusion of animals in the study, monthly, and if there were signs of weight loss. In case of weight loss > 10% compared to the starting weight, it was attempted to correct with a different feeding strategy at a weight loss of 20% the minipig was euthanised. GMPs were euthanised and sampled when sufficient data had been collected or humane endpoints met, whichever came first. None of the GMPs in this article was euthanised due to meeting project-defined humane endpoints. Blood samples for whole genome sequencing were taken at the Georg-August-University in Göttingen, Germany. Blood sampling for this study was approved by the animal welfare body (Tierschutzbeauftragte/ Tierschutzkommission) of the University of Göttingen and is documented under reference E1–20. No human participants were involved in the study.
: Not applicable.
: Not applicable.
: Maja Ramløse and Kirsten Rosenmay Jacobsen are employed by Ellegaard Göttingen Minipigs A/S.