Production of Germ-Free Fast-Growing Broilers from a Commercial Line for Microbiota Studies

Abstract

Studies of the gut microbiota contribution to the host physiology and immunocompetence are facilitated by the availability of germ-free animal models, which are considered the gold standard. Nesting birds are ideal models for the production of germ-free animals since there is no need to raise their relatives under sterile conditions. Germ-free chickens are mainly generated from specific-pathogenfree (SPF) experimental lines, which are poorly representative of commercial chicken lines. The method proposed here allowed the production of germ-free chickens from the fast growing broiler line Ross PM3, commonly used by the poultry industry. Eggs were quickly collected after laying at a broiler breeder farm. They underwent a strict decontamination process from the collection to the introduction in a sterile egg hatching isolator. The chicks have been hatched and kept in these sterile isolators during the period necessary to control their sterility. Originally developed for an experimental SPF white leghorn line, the present protocol has been adapted not only to the Ross PM3 broiler line but also to quails. It therefore represents a robust and readily adaptable procedure to other poultry species and nesting birds of economic, biological or ecological relevance.

The Absence of Gut Microbiota Alters the Development of the Apicomplexan Parasite Eimeria tenella

“…To study the effect of the microbiota on E. tenella infection, we used an original model of germ-free broilers that we recently developed in our facilities ( Guitton et al., 2020 ). A previous study reported that a high dose of E. tenella inoculum administered per os (>10,000 sporulated oocysts) led to similar oocyst load in caecal contents in germ-free and conventional chickens ( Visco and Burns, 1972a ).…”

“…Conventional and germ-free Ross PM3 broilers were hatched in the Infectiology of Farm, Model and Wildlife Animals Facility (PFIE, Centre INRAE Val De Loire: ; member of the National Infrastructure EMERG’IN) as described by ( Guitton et al., 2020 ). Briefly, Ross PM3 eggs from two French farms were collected, decontaminated with a 1.5% peracetic acid solution (1.5% Divosan Plus VT53, Johnson Diversey, France).…”

“…Eggs were then incubated, decontaminated a second time as described above and placed in a hatching incubator for production of conventional chicks or an isolator for the production of germ-free animals. Bacteriological controls were performed as described by ( Guitton et al., 2020 ); animals were confirmed to be bacteria-free while conventional chickens developed a microbiota.…”

“…In the present study, we investigated the influence of the absence of microbiota on E. tenella infection. For this purpose, we used a recently developed model of germ-free fast-growing broilers ( Guitton et al., 2020 ). Our results highlight a critical role for the microbiota in sustaining the optimal development of the parasite in the chicken intestinal tract.…”

Sterilization of sealed PVDF pouches containing decellularized scaffold by electrical stimulation

“…Meanwhile, the color of the broth media changed to red in the presence of resazurin, a redox indicator of FTM, indicating that the microorganisms were cultured in the aerobic incubation conditions. [ 30,31 ]…”