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 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

ObjectiveThis study investigated the association of enzyme-producing microbes and their enzymes with starch and hemicellulose degradation during fermentation of total mixed ration (TMR) silage.MethodsThe TMRs were prepared with soybean curd residue, alfalfa hay (ATMR) or Leymus chinensis hay (LTMR), corn meal, soybean meal, vitamin-mineral supplements, and salt at a ratio of 25:40:30:4:0.5:0.5 on a dry matter basis. Laboratory-scale bag silos were randomly opened after 1, 3, 7, 14, 28, and 56 days of ensiling and subjected to analyses of fermentation quality, carbohydrates loss, microbial amylase and hemicellulase activities, succession of dominant amylolytic or hemicellulolytic microbes, and their microbial and enzymatic properties.ResultsBoth ATMR and LTMR silages were well preserved, with low pH and high lactic acid concentrations. In addition to the substantial loss of water soluble carbohydrates, loss of starch and hemicellulose was also observed in both TMR silages with prolonged ensiling. The microbial amylase activity remained detectable throughout the ensiling in both TMR silages, whereas the microbial hemicellulase activity progressively decreased until it was inactive at day 14 post-ensiling in both TMR silages. During the early stage of fermentation, the main amylase-producing microbes were Bacillus amyloliquefaciens (B. amyloliquefaciens), B. cereus, B. licheniformis, and B. subtilis in ATMR silage and B. flexus, B. licheniformis, and Paenibacillus xylanexedens (P. xylanexedens) in LTMR silage, whereas Enterococcus faecium was closely associated with starch hydrolysis at the later stage of fermentation in both TMR silages. B. amyloliquefaciens, B. licheniformis, and B. subtilis and B. licheniformis, B. pumilus, and P. xylanexedens were the main source of microbial hemicellulase during the early stage of fermentation in ATMR and LTMR silages, respectively.ConclusionThe microbial amylase contributes to starch hydrolysis during the ensiling process in both TMR silages, whereas the microbial hemicellulase participates in the hemicellulose degradation only at the early stage of ensiling.

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Alfalfa (Medicago sativa L.) is viewed as a difficult crop to ensile, primarily because of its high buffering capacity, low concentration of water soluble carbohydrates (WSC), high concentration of moisture and vulnerability to the undesirable secondary clostridial fermentation (Zheng, Niu, Jiang, Zuo, & Xu, 2017). Thus, inoculating lactic acid bacteria (LAB), wilting and ensiling with other materials are often used to enhance the fermentation quality of alfalfa silage

Based on a field investigation and laboratory trials, this study compared the invasive ecology of aquatic and terrestrial alligator weed (Alternanthera philoxeroides G.) ecotypes. The aquatic ecotype showed a stronger eco-adaptation than the terrestrial ecotype. Alligator weed performed well in wetland and riparian habitats, where it was so competitive that only one-tothree other plant types grew successfully in its company. In contrast, it exhibited poor growth in upland fields and parks, where the terrestrial ecotype was dominated by 6-11 other alien plant species. The aquatic ecotype exhibited significantly greater growth inhibition of Microcystis aeruginosa. The ecotypes differed in their pattern of allelopathic inhibition of water blooms.The allelopathic effect of the aquatic ecotype persisted for a period exceeding 20 days, whereas the terrestrial ecotype's allelopathic effect subsided in <5 days. The aquatic ecotype had higher levels of antioxidant compounds, including soluble protein and flavones, and the activity of protective enzymes was also higher, which included superoxidase dismutase, peroxidase and catalase. Alligator weed's unique characteristics derive mainly from its strong allelopathic potential and tolerance of eutrophication in aquatic habitats.