Effect of Bacillus subtilis and Bacillus licheniformis supplementation in diets with low-and high-protein content on ileal crude protein and amino acid digestibility and intestinal microbiota composition of growing pigs

Background： Bacillus spp. seem to be an alternative to antimicrobial growth promoters for improving animals＇ health and performance. However, there is little information on the effect of Bacillus spp. in combination with different dietary crude protein（CP） levels on the ileal digestibility and microbiota composition. Therefore, the objective of this study was to determine the effect of Bacillus spp. supplementation to low-（LP） and high-protein diets（HP） on ileal CP and amino acid（AA） digestibility and intestinal microbiota composition.Methods： Eight ileally cannulated pigs with an initial body weight of 28.5 kg were randomly allocated to a rowcolumn design with 8 pigs and 3 periods of 16 d each. The assay diets were based on wheat-barley-soybean meal with two protein levels： LP（14% CP, as-fed） and HP diet（18% CP, as-fed）. The LP and HP diets were supplemented with or without Bacil us spp. at a level of 0.04%（as-fed）. The apparent ileal digestibility（AID） and standardized ileal digestibility（SID） of CP and AA was determined. Bacterial community composition from ileal digesta was analyzed by Illumina amplicon sequencing and quantitative real-time PCR. Data were analyzed as a 2 × 2 factorial design using the GLIMMIX procedures of SAS.Results： The supplementation with Bacillus spp. did not affect both AID and SID of CP and AA in growing pigs.Moreover, there was no difference in AID of CP and AA between HP and LP diets, but SID of cystine, glutamic acid,glycine, and proline was lower（P 〈 0.05） in pigs fed the HP diets. The HP diets increased abundance of Bifidobacterium spp. and Lactobacillus spp.,（P 〈 0.05） and by amplicon sequencing the latter was identified as predominant genus in microbiota from HP with Bacillus spp., whereas dietary supplementation of Bacillus spp. increased（P 〈 0.05） abundance of Roseburia spp.Conclusions： The HP diet increased abundance of Lactobacillus spp. and Bifidobacterium spp.. The supplementation of Bacil us spp. resulted in a higher abundance of healthy gut associated bacteria without affecting ileal CP and AA digestibility, whereas LP diet may reduce the flow of undigested protein to the large intestine of pigs.

Methodological impact of starch determination on starch content and ileal digestibility of starch in grain legumes for growing pigs

Background： Grain legumes represent a valuable energy source in pig diets due to their high starch content. The present study was conducted to determine the content and apparent ileal digestibility（AID） of starch in different grain legume cultivars for pigs by means of both a polarimetric and enzymatic method for starch determination.Methods： Three experiments were conducted with six barrows each which were fitted with ileal T-cannulas. In total, 18 diets including six different cultivars of faba beans（Vicia faba L.） and peas（Pisum sativum L.）, five different cultivars of lupins（Lupinus luteus L., Lupinus angustifolius L.）, and one diet with a soybean meal（SBM） were fed.Results： The starch content of faba beans and peas was greater（P ＆lt; 0.05） when determined polarimetrically than enzymatically（438 vs. 345 g/kg dry matter（DM） in faba beans and 509 vs. 390 g/kg DM in peas, respectively）.Considerable lower starch contents were obtained in lupins and SBM, with 82 and 48 g/kg DM（analyzed polarimetrically） and ＆lt;1.1 and 3 g/kg DM（analyzed enzymatically）, respectively. Mean values for contents of neutral detergent fiber（NDF） and acid detergent fiber（ADF） in grain legumes ranged from 111 and 79 g/kg DM in peas to248 and 207 g/kg DM in lupins, respectively. Contents of condensed tannins in the colored flowered faba bean cultivars ranged from 2.1 to 7.4 g/kg DM. The AID of starch was greater（P ＆lt; 0.05） in pea than in faba bean cultivars,and using the polarimetric starch determination method resulted in greater（P ＆lt; 0.05） digestibility values than using enzymatic starch analysis（84 vs. 80% in faba beans and 86 vs. 83% in peas）. Moreover, AID of starch differed（P ＆lt; 0.05）within pea cultivars and starch digestibility in faba beans decreased linearly（P ＆lt; 0.05） as the content of condensed tannins increased. However, there was no relationship between contents of NDF and ADF and AID of starch in pea and faba bean cultivars.Conclusion： Both contents and AID of starch in grain legumes can vary as influenced by the analytical method used for starch determination. Generally, starch digestibility is greater when measured by polarimetric rather than enzymatic methods.