Developmental stages in microbiota, bile acids, and clostridial species in healthy puppies

Background: The fecal microbiota, fecal bile acid concentrations, and abundance of Clostridium perfringens and Clostridium difficile are altered in acute and chronic gastrointestinal disease in adult dogs. However, less is known in young puppies.

Hypothesis/objectives: To determine composition of the fecal microbiota, assess development of fecal bile acid profiles, and determine the abundance of Clostridial species in puppies, young adult dogs, and adult dogs.

Animals: Healthy puppies from a whelping kennel (n = 53) and healthy client-owned dogs <1 year old (n = 20) were separated into 6 age groups, then compared to client-owned dogs over 1 year of age (n = 13).

Methods: Prospective observational study. Naturally voided fecal samples were analyzed by quantitative polymerase chain reaction to measure bacterial abundances. Fecal bile acids were quantified using gas chromatography-mass spectrometry.

Results: Puppies up to 5 to 6 weeks of age had increased Dysbiosis Index (median [min-max]: 5.39 [1.32-8.6], P < .001), increased abundance of C. difficile (4.1 [0.01-4.85] log DNA, P < .001), decreased secondary bile acid concentrations (0.61 [0.28-5.06] μg/mg, P = .006), and decreased abundance of C. hiranonis (0.84 [0.01-6.71], P = .005) compared to adult dogs (-4.62 [-8.36 to -0.61], 0.01 [0.01-0.01], 4.12 [0.32-8.94], and 6.02 [5.06-7.00], respectively). Secondary bile acid concentration positively correlated with C. hiranonis abundance (ρ = 0.77; P < .001).

Conclusions and clinical importance: The increase in secondary bile acids and simultaneous decrease of C. difficile and C. perfringens after 5 to 6 weeks of age warrants further investigation into regulatory impacts that secondary bile acids could have on clostridial species in dogs.