PSX-14 Effect of starch supplementation on rumen bacterial abundance in <i>Bos taurus taurus</i> steers consuming low-quality forage

Abstract Low-quality forage provides a limited supply of ruminally available nitrogen and is generally resistant to digestion, inhibiting microbial growth in the rumen and ultimately decreasing forage utilization. Supplementation with protein is well documented to increase forage utilization by promoting microbial growth. Even when protein requirements are met, additional energy, often provided as starch, may be required to meet performance expectations. The objective of this study was to evaluate the effect of increasing starch supplementation on rumen bacterial populations in Bos taurus taurus steers supplemented protein and consuming low-quality forage. Ruminal samples were collected from ruminally-cannulated Angus steers [n = 5; body weight (BW) = 375 ± 45 kg] used in a 4 × 4 Latin square fed a basal diet of King Ranch Bluestem Hay (3.5% CP, 73% NDF) ad libitum, and supplemented with one of four isonitrogenous (130 mg N/kg BW) supplements with increasing starch (2% starch = 100% soybean meal; 20% starch = 73% soybean meal, 26.2% corn, 0.8% urea; 38% starch = 47% soybean meal, 51.6% corn, 1.4% urea; 56% starch = 19% soybean meal, 78.6% corn, 2.4% urea). Rumen content samples were collected via rumen cannula 4 h after supplementation and immediately frozen. Microbial DNA was extracted; full length 16S rRNA amplicon libraries were prepared using Kinnex kits and sequenced on the PacBio Revio platform. Reads were analyzed using DADA2 and QIIME2. Statistical analysis was performed with the GLIMMIX procedure of SAS 9.4 (SAS Inst. Inc., Cary, NC). Firmicutes and Bacteriodata were the most abundant phyla in all treatments (82% and 11%, respectively), and were not affected by starch supplementation (P ≥ 0.45). Actinobacteriota and Euryarchaeota abundance responded quadratically (P &amp;lt; 0.01) to increasing starch provision. Christensenellaceae R-7 group, Rikenellaceae RC9 gut group, Lachnospiraceae XPB1014 group, Butyrvibrio, Lachnospiraceae NK3A20 group, and Prevotella represented the most abundant genera for all treatments. Increasing supplemental starch resulted in a quadratic response (P &amp;lt; 0.01) for abundance of Christensenellaceae R-7 and Lachnospiraceae XPB1014 groups. Relative abundance of Butyrvibrio was 4.05% for steers fed 2% starch supplement and increased to 18.24 and 11.85% for 20 and 38% starch, respectively and then decreased to 4.34% for steers fed 56% starch supplement (quadratic, P &amp;lt; 0.01) Abundance of Pseudobutyrvibrio followed a similar but less dramatic pattern (quadratic, P &amp;lt; 0.01). In contrast, abundance of Streptococcus and Methanobrevibacter decreased with the first two levels of starch and then increased with 56% (quadratic; P &amp;lt; 0.01). In conclusion, responses to the first two increments of starch supplementation suggest increased fibrolytic and decreased methanogenic activity. These results are supported by the previously reported increase in TDOMI for 20 and 38% starch 49.0 and 48.2 g/kg MBW, respectively versus 45.7 and 44.3 g/kg MBW for 2 and 56%, respectively.