Parasite isolate, storage and recovery of C. rodentium, animals, culture medium and growth conditions
The T. spiralis isolate ISS534 used in the present study was obtained from domestic pigs in Henan Province, China, and maintained in Kunming mice in the Jilin Provincial Engineering Research Center of Animal Probiotics. C. rodentium ATCC 51459 was provided by Dr Haining Shi (Harvard Medical School, Boston, MA, USA) and cultured in Luria-Bertani medium at 37°C with shaking. Male BALB/c mice (specific pathogen free, SPF) aged 6-8 weeks were purchased from Beijing Hua fu kang Biotechnology. The study protocol was approved by the Jilin Agriculture University Veterinary Research Ethics Committee, and all procedures were performed in strict accordance with the guidelines of the Chinese National Institute of Health Guide for the Care and Use of Laboratory Animals. Care was taken to minimize the number of animals used and to reduce their suffering. Before the experiment, the mice were allowed to eat and drink freely for at least 4 days to adapt to the feeding environment. Throughout the experiment, the mice were allowed to eat and drink freely, and the bedding materials were changed regularly to ensure an excellent feeding environment.