Campylobacter jejuni Response When Inoculated in Bovine In Vitro Fecal Microbial Consortia Incubations in the Presence of Metabolic Inhibitors

Dana K Dittoe; Robin C Anderson; Nathan A Krueger; Roger B Harvey; Toni L Poole; Tawni L Crippen; Todd R Callaway; Steven C Ricke

doi:10.3390/pathogens12121391

Campylobacter jejuni Response When Inoculated in Bovine In Vitro Fecal Microbial Consortia Incubations in the Presence of Metabolic Inhibitors

Pathogens. 2023 Nov 26;12(12):1391. doi: 10.3390/pathogens12121391.

Authors

Dana K Dittoe¹, Robin C Anderson², Nathan A Krueger³, Roger B Harvey², Toni L Poole², Tawni L Crippen², Todd R Callaway⁴, Steven C Ricke⁵

Affiliations

¹ Department of Animal Science, University of Wyoming, Laramie, WY 82071, USA.
² United States Department of Agriculture, Agricultural Research Service, Southern Plains Agricultural Research Center, College Station, TX 77845, USA.
³ Agricultural Sciences, Blinn College, Bryan, TX 77833, USA.
⁴ Ruminant Nutrition, Ruminant Microbiology, and Preharvest Food Safety, Department of Animal and Dairy Science, University of Georgia, Athens, GA 30602, USA.
⁵ Meat Science and Animal Biologics Discovery Program, Department of Animal and Dairy Sciences, University of Wisconsin, Madison, WI 53706, USA.

Abstract

Infection with the foodborne pathogen Campylobacter is the leading bacterial cause of human foodborne illness in the United States. The objectives of this experiment were to test the hypothesis that mixed microbial populations from the bovine rumen may be better at excluding Campylobacter than populations from freshly voided feces and to explore potential reasons as to why the rumen may be a less favorable environment for Campylobacter than feces. In an initial experiment, C. jejuni cultures inoculated without or with freshly collected bovine rumen fluid, bovine feces or their combination were cultured micro-aerobically for 48 h. Results revealed that C. jejuni grew at similar growth rates during the first 6 h of incubation regardless of whether inoculated with the rumen or fecal contents, with rates ranging from 0.178 to 0.222 h^-1. However, C. jejuni counts (log₁₀ colony-forming units/mL) at the end of the 48 h incubation were lowest in cultures inoculated with rumen fluid (5.73 log₁₀ CFUs/mL), intermediate in cultures inoculated with feces or both feces and rumen fluid (7.16 and 6.36 log₁₀ CFUs/mL) and highest in pure culture controls that had not been inoculated with the rumen or fecal contents (8.32 log₁₀ CFUs/mL). In follow-up experiments intended to examine the potential effects of hydrogen and hydrogen-consuming methanogens on C. jejuni, freshly collected bovine feces, suspended in anaerobic buffer, were incubated anaerobically under either a 100% carbon dioxide or 50:50 carbon dioxide/hydrogen gas mix. While C. jejuni viability decreased <1 log₁₀ CFUs/mL during incubation of the fecal suspensions, this did not differ whether under low or high hydrogen accumulations or whether the suspensions were treated without or with the mechanistically distinct methanogen inhibitors, 5 mM nitrate, 0.05 mM 2-bromosulfonate or 0.001 mM monensin. These results suggest that little if any competition between C. jejuni and hydrogen-consuming methanogens exists in the bovine intestine based on fecal incubations.

Keywords: Campylobacter jejuni; anti-methane compounds; fecal suspensions; rumen incubations.

Grants and funding

This research received no external funding.