PSIX-10 Effects of Bacillus subtilis ATCC PTA-122264 on macronutrient digestibility and fecal characteristics, metabolites, and microbiota of healthy adult dogs subjected to an abrupt diet change
Authors: Oba Patrícia M , Swanson Olivia , Kang Yifei , Mioto Julio , Menton John , Vinay Elena , Millette Mathieu , Kelly Melissa , Swanson Kelly S 
Published: September 2024

Abstract
Probiotic use in dogs is increasing, with spore-forming Bacillus species gaining interest due to their viability during processing, storage, and in the gastrointestinal tract. B. subtilis is known for its adaptable metabolism that may be useful in pets subjected to abrupt dietary changes. The objective of this study was to determine dietary apparent total tract macronutrient digestibility (ATTD) and fecal characteristics, metabolites, and microbiota of healthy adult dogs supplemented with B. subtilis ATCC PTA-122264. All procedures were approved by the University of Illinois IACUC prior to experimentation. Dogs [n = 12; 6 ± 1 yr; 8.71 ± 0.91 kg body weight (BW)] were used in a replicated 3x3 Latin square design (n = 12/group) and fed to maintain BW. In each 42-d experimental period, dogs were allotted to one of three treatments: Control (kibble diet only), Low [kibble diet + 1×109 colony-forming units (CFU) of B. subtilis], and High (kibble diet + 5×109 CFU of B. subtilis). After a 22-d adaptation phase, a 5-d fecal collection phase was followed by a blood collection phase. On d 28, dogs were abruptly changed to a canned diet and fed for 14 d, with fecal samples collected on d 30 (2 d post change), 34 (6 d post change), 38 (10 d post change), and 42 (14 d post change). Fecal microbiota data were evaluated using QIIME2. All other data were analyzed using the Mixed Models procedure of SAS, with P &lt; 0.05 being considered significant. B. subtilis supplementation of the kibble diet did not influence food or energy intake, fecal output, ATTD, most serum metabolites, hematology, fecal characteristics, fecal bacterial alpha and beta diversity indices, and dysbiosis index, demonstrating the safety of the strain in dogs. However, fecal Streptococcus and Blautia abundances were higher in High dogs than Low or Control dogs, while E. coli abundance was less in Low dogs than High and Control dogs. The abrupt change to a canned diet led to significant alterations in fecal characteristics, dysbiosis index, and bacterial abundances, which were not influenced by B. subtilis supplementation. Bacterial alpha diversity measures were affected, with observed ASV (amplicon sequence variants) and the Fisher Index reducing, and the Shannon Index increasing by dietary change. The abrupt dietary change caused a distinct shift in microbial communities, as evidenced by principal coordinates analysis plots. The relative abundances of various fecal bacterial taxa, including Bifidobacterium, Faecalibaculum, Turicibacter, and Lactobacillus were decreased, while Collinsella, Ruminococcus gnavus group, and Escherichia-Shigella were increased with the dietary change. Our data support the notion that daily supplementation of 1 – 5×109 CFU of B. subtilis ATCC PTA-122264 in healthy dogs is safe and does not affect markers of general health and fecal characteristics, warranting further exploration.