High-throughput sequencing based on a novel Clostridia-specific primer set proved a potentially useful tool to study the clostridial community dynamics, and could aid to elucidate the mechanism by which the clostridial fermentation develops during the ensiling of alfalfa.

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The objective of this study was to assess the use of peach pomace in total mixed ration (TMR) silages and clarify the differences in aerobic stability between TMR and TMR silages caused by lactic acid bacteria (LAB). The TMR were prepared using peach pomace, alfalfa hay or Leymus chinensis hay, maize meal, soybean meal, cotton meal, limestone, a vitamin-mineral supplement, and salt in a ratio of 6.0:34.0:44.4:7.0:5.0:2.5:1.0:0.1 on a dry matter (DM) basis. Fermentation quality, microbial composition, and the predominant LAB were examined during ensiling and aerobic deterioration. The results indicated that the TMR silages with peach pomace were well fermented, with low pH and high lactic acid concentrations. The aerobic stability of TMR silages were significantly higher than that of TMR. Compared with TMR silages with alfalfa hay, TMR silage with Leymus chinensis hay was much more prone to deterioration. Although the dominant LAB were not identical in TMR, the same dominant species, Lactobacillus buchneri and Pediococcus acidilactici, were found in both types of TMR silages after 56 d of ensiling, and they may play an important role in the aerobic stability of TMR silages.

The objective of this study was to gain deeper insights into chemical transformations and the clostridial community dynamics during the ensiling of alfalfa. A factorial experiment was conducted with four alfalfa cultivars [Sanditi (A1), AC Caribou (A2), WL319HQ (A3) and 4030 (A4)] Â three wilting durations [0 h (direct-cut), 2 h and 4 h] Â three storage periods (14, 28 and 56 days). The clostridial community was examined using denaturing gradient gel electrophoresis. High butyric acid and ammonia nitrogen (NH 3 -N) contents and clostridia numbers were observed in direct-cut silage, with higher values observed in A4 than in the other three cultivars. However, butyric acid and NH 3 -N contents and clostridia numbers decreased with wilting regardless of the cultivar. Although Clostridium ghonii and Clostridium sartagoforme were common to all direct-cut and wilted silages, bands for these species were faint. Differences in the appearing time of these species were observed, bands for C. ghonii were found after 14 and 28 days of ensiling, while those for C. sartagoforme were only found after 56 days. In addition, in direct-cut silage, distinct bands for Clostridium perfringens were detected in A1, A2 and A3, while those for Clostridium sporogenes were detected in A4. The inactive spores of clostridia could be observed in 4-h wilted silage as no butyric acid was detected. It was concluded that the enhanced clostridial fermentation of direct-cut silage was attributed mainly to C. perfringens and C. sporogenes, while C. ghonii and C. sartagoforme were involved in the restricted clostridial fermentation of 2-h wilted silage. ARTICLE HISTORY

This study was conducted to examine the effects of Lactobacillus plantarum (LP) and sucrose (S) on clostridial community dynamics and correlation between clostridia and other bacteria in alfalfa silage during ensiling. Fresh alfalfa was directly ensiled without (CK) or with additives (LP, S, LP + S) for 7, 14, 28 and 56 days. Clostridial and bacterial communities were evaluated by next-generation sequencing. Severe clostridial fermentation occurred in CK, as evidenced by the high contents of butyric acid, ammonia nitrogen, and clostridia counts, whereas all additives, particularly LP + S, decreased silage pH and restrained clostridial fermentation. Clostridium perfringens and Clostridium butyricum might act as the main initiators of clostridial fermentation, with Clostridium tyrobutyricum functioning as the promoters of fermentation until the end of ensiling. Clostridium tyrobutyricum (33.5 to 98.0%) dominated the clostridial community in CK from 14 to 56 days, whereas it was below 17.7% in LP + S. Clostridium was negatively correlated with the genus Lactobacillus, but positively correlated with the genera Enterococcus, Lactococcus and Leuconostoc. Insufficient acidification promoted the vigorous growth of C. tyrobutyricum of silage in later stages, which was mainly responsible for the clostridial fermentation of alfalfa silage.

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A 4 x 4 Latin square experiment was conducted to examine abomasal passage of biogenic amines in steers fed silage and their related effects on intake, digestibility, and digestive function. Thirty percent of the dry matter (DM) in the diets consisted of alfalfa forage, which was fed as either hay or silage. The DM from alfalfa silage DM was substituted at 0, 33, 67, and 100% for DM from alfalfa hay and was fed to four ruminally and abomasally cannulated steers. The roughage component of the diet constituted 50% of the DM and consisted of 60% alfalfa silage or hay and 40% tropical corn silage. The concentrate was composed mainly of ground corn. The concentrations of putrescine and cadaverine in abomasal digesta increased as alfalfa silage in the diet increased. Abomasal recovery of biogenic amines, a product of their concentration in abomasal digesta and the passage of DM through the abomasum, was negatively correlated with intake. Abomasal recovery of most amines was 5 to 20% of intake. Abomasal recovery of cadaverine was correlated with depressed intake. Total DM intake was reduced 8.3 to 25.8% as the proportion of alfalfa silage in the diet increased. Frequency of reticular contractions, intake, ruminal DM digestibility, ruminal outflow, volatile fatty acids, and total tract DM digestibility decreased in steers fed diets that contained more alfalfa silage. Ruminal fluid pH and NH3 concentration increased in steers fed more alfalfa silage; however, mass and the DM percentage of ruminal contents decreased linearly. Postprandial insulin concentrations were quadratically related to the proportion of alfalfa hay or silage in the diet. Intraruminal metabolism of biogenic amines is extensive based on the relatively low quantities recovered in abomasal digesta; however, the amounts recovered in abomasal digesta were related to intake depression and associated physiological effects.