Evaluating bacterial colonization of a developing broiler embryo after in ovo injection with a bioluminescent bacteria.

In ovo injection of probiotics has been of interest for achieving early health benefits. However, there is limited research demonstrating where bacteria could migrate within the embryo after injection. The objective of this study was to evaluate bacterial colonization or migration after in ovo injection of broiler embryo with bioluminescent Escherichia coli. Injection using 106 CFU/mL nonpathogenic E. coli was applied to amniotic and air cell regions on day 18 of incubation. On days 18, 19, 20, and 21 the amnion, skin, lung, gastrointestinal tract (GIT), bursa, and spleen were collected. On day 21, the GIT was separated into crop, duodenum, jejunum, ileum, and ceca sections. All tissues were visualized using anin vivo imaging system to confirm the presence of bioluminescent E. coli. Samples were homogenized, 10-fold serially diluted, and spread onto appropriate agar to determine bacterial loads in all tissues. Results indicated that eggs injected into the amnion had significantly high numbers of E. coli cells in all tissues compared to air cell injected and control treatments 2 h post-injection (P < 0.0001). E. coli was also found on the lungs, spleen, and bursa of eggs injected either in the amnion or air cell (P < 0.05). Results indicated that in ovo injection into the amnion was more efficient than air cell injection, yielding a higher bacterial concentration in the evaluated tissues, specifically the ileum and ceca. Future research using bioluminescent probiotic bacteria may establish sites of preference for different probiotics leading to site-specific application that can maximize their overall impact when in ovo injected.