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Salmonella  colonization of food animals is a concern for animal health and public health as a food safety risk. Various obstacles impede the effort to reduce asymptomatic  Salmonella  carriage in food animals, including the existence of numerous serovars and the ubiquitous nature of  Salmonella . To develop an intervention strategy that is non-specific yet effective against diverse  Salmonella  serovars, we explored the prophylactic use of a cytokine to decrease  Salmonella  in swine by boosting the host’s innate immune system. Granulocyte-colony stimulating factor (G-CSF) is the major cytokine regulating the production, differentiation, function, and survival of neutrophils. Neutrophils play a critical role in the response to  Salmonella;  therefore, we evaluated the vectored-delivery of porcine G-CSF as a prophylactic to reduce  Salmonella  in pigs. Crossbred pigs, 5 weeks of age, were intramuscularly injected with a replication-defective human adenovirus (Ad5) engineered to express porcine G-CSF (Ad5-G-CSF,  n  = 9). Control pigs received the same Ad5 vector lacking the gene encoding G-CSF (Ad5-empty,  n  = 7). Four days later, all pigs ( n  = 16) were intranasally inoculated with 1 × 10 7  colony forming unit (CFU) of  Salmonella enterica  serovar Typhimurium UK1. At 2 and 3 days post-challenge with  Salmonella , Ad5-G-CSF-treated pigs shed significantly less  Salmonella  (~10 3  CFU/g) in their feces than Ad5-empty-treated pigs (~10 4 –10 5  CFU/g;  P  < 0.05). A significant 4-log reduction in tonsil colonization was also observed in the Ad5-G-CSF-treated pigs at 7 days post-challenge ( P  < 0.05). In the gastrointestinal tract, the Peyer’s patch region of the ileum exhibited a significant 0.5-log reduction in colonization in the Ad5-G-CSF-treated pigs ( P  < 0.05). The microbiota of all challenged pigs was assessed by sequencing and analyzing the V1–V3 region of the 16S rRNA gene from fecal DNA samples. The microbial community structure of  Salmonella -challenged pigs was less disturbed post-challenge in the Ad5-G-CSF-treated pigs than the Ad5-empty-treated pigs. This suggests that Ad5-G-CSF administration mitigated changes in the microbial community structure caused by  Salmonella  challenge. Collectively, these data suggest that delivery of a targeted immunostimulant to enhance neutropoiesis may be a strategy to reduce  Salmonella  colonization, potentially during periods of immunological stress.

frontiers in veterinary science

Prophylactic Administration of Vector-Encoded Porcine Granulocyte-Colony Stimulating Factor Reduces Salmonella Shedding, Tonsil Colonization, and Microbiota Alterations of the Gastrointestinal Tract in Salmonella-Challenged Swine