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Microbial Community and Fermentation Characteristics of Native Grass Prepared Without or With Isolated Lactic Acid Bacteria on the Mongolian Plateau

ABSTRACT: This study aimed to isolate and identify lactic acid bacteria (LAB) from the native grass and naturally fermented silage from the Mongolian Plateau. The effect of selected strains on bacterial community and quality of native grass silage was also studied. Strains XM2, 265, and 842 could grow normally at 15°C–30°C, pH 4.0–8.0, and NaCl 3 and 6.5%; they were identified as Lactiplantibacillus plantarum subsp. plantarum, Pediococcus acidilactici, and Latilactobacillus graminis, by sequencing 16S rRNA, respectively. The three strains (XM2, 265, and 842) and one commercial additive (L) were used as inoculants and singularly added to the native grass. Compared to the control, the dry matter content was significantly (p < 0.05) lower in L and XM2 groups. The water-soluble carbohydrate content was significantly (p < 0.05) higher in control than in other groups. Compared with the control, the crude protein and ammonia nitrogen contents were significantly (p < 0.05) higher and lower in the LAB-treated groups, and the acid and detergent fiber contents were significantly (p < 0.05) reduced in the L and XM2 groups than those in other groups. There was a significant (p < 0.05) difference in the pH value, lactic acid content, and lactic acid-to-acetic acid ratio in L and XM2 groups than in other groups. Compared with the control, the number of LAB was significantly (p < 0.05) higher in LAB-treated silages, whereas no significant (p > 0.05) differences were observed in yeast and aerobic bacteria in all groups. Compared to the control, the Shannon index was significantly (p < 0.05) reduced. Simpson and Chao1 were significantly (p < 0.05) increased. Principal coordinate analysis based on the unweighted UniFrac distance showed clear separation of the bacterial community in fresh materials and LAB-treated silages. Besides, compared to the control, the principal coordinate analysis of LAB-treated silages was also separate. After 30 days of fermentation, the relative abundance of Firmicutes increased and was the primary phylum in all silages. Compared with the control, the abundance of Firmicutes and Proteobacteriawas significantly (p < 0.05) higher and lower in L and XM2 groups. In contrast, no significant differences were observed among control, 265, and 842 groups. At the genus level, the relative abundance of Lactobacillus, Enterobacter, Pediococcus, and Weissella was increased and dominated the native grass fermentation. Compared with the control, the abundance of Lactobacillus was significantly (p < 0.05) higher in L, XM2, and 842 groups, while no significant (p > 0.05) differences were observed between the control and 265 groups. The abundance of Pediococcus was higher than that in other groups. Consequently, the results demonstrated that LAB significantly influenced silage fermentation by reconstructing microbiota, and Lactobacillus was the dominant genus in the native grass silages. Furthermore, the results showed that strain XM2 could effectively improve the silage quality, and it is considered a potential starter for the native grass silage.

SUBMITTER: You S

PROVIDER: S-EPMC8517267 | biostudies-literature |

REPOSITORIES: biostudies-literature

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Project description:IntroductionTo prepare high-quality silage, we studied the chemical composition, silage fermentation, characterization, and identification of lactic acid bacteria (LAB) associated with the silage fermentation of native grass on the Inner Mongolian Plateau.MethodsLAB were isolated from fresh native grass and their silage, and silages were prepared using a small-scale fermentation system with 2-3 cm length in plastic bags.ResultsThe dominant species of native grasses used were Stipa baicalensis, Leymus chinensis, Cleistogenes squarrosa, Melissilus ruthenicus and Pulsatilla turczaninovii, which contained 47.83-59.43 % moisture, 55.12-67.74 % neutral detergent fiber (NDF), and 8.72-14.55 crude protein (CP), and these nutrients did not change greatly during ensiling. Good preservation with a relatively low pH (below 4.44) and high (p < 0.05) lactic acid content (>0.58) was obtained after ensiling. Based on the morphological and biochemical characteristics, these isolates were divided into 12 groups (A-L). All isolate strains were gram-positive and catalase-negative bacteria that produce lactic acid from glucose. Group A-K were cocci, while group L was rod-shaped. Group A-E formed D-lactic acid, but group H-K formed L-lactic acid, and other groups formed DL-lactic acid. Group A-E were heterofermentative, and Group F-L were homofermentative types of LAB. According to the 16S rRNA gene sequences analysis, strains were identified as genus Leuconostoc (A, B, and C), Weissellla (D, E), Pediococcus (F, G), Enterococcus (H, I, J and K), and Lactiplantibacillus (L). Enterococcus (E.) faecium (29.17%, percentage of total isolates) and Pediococcus (P.) acidilactici (18.75%) were the most frequently occurring dominant species.DiscussionThis study suggests that the native grasses contained abundant LAB species, and they can be used as good-quality silages in animal husbandry. In addition, the strains P. acidilactici and E. faecium were the most frequently isolated from native grass silages as dominant species which can be a potentially excellent inoculant for native grass silage.

Project description:To investigate community synergy of lactic acid bacteria (LAB) and cleaner fermentation of oat silage, oat silages were prepared with or without (control) commercial LAB inoculants LI1 (containing Lactiplantibacillus plantarum, Lentilactobacillus buchneri, Lacticaseibacillus paracasei, and Pediococcus acidilactici) and LI2 (containing Lactiplantibacillus plantarum and Lentilactobacillus buchneri). The microbial community, LAB synergy, and cleaner fermentation were analyzed at 1, 3, 6, 15, 35, and 90 days of ensiling. The LAB inoculant improved fermentation quality, with significantly (P < 0.05) lower pH, ammonia nitrogen content, and gas production and higher lactic acid and acetic acid contents than those of the control. Enterobacteriaceae was the main bacterial community in early stage of fermentation, which utilizes sugar to produce CO2 gas, causing dry matter (DM) and energy loss. As fermentation progressed, the microbial diversity decreased, and the microbial community shifted from Gram-negative to Gram-positive bacteria. The inoculation of multispecies LAB displayed community synergy; Pediococcus acidilactici formed a dominant community in the early stage of fermentation, which produced an acid and anaerobic environment for the subsequent growth of Lentilactobacillus and Lacticaseibacillus species, thus forming a LAB-dominated microbial community. The predicted functional profile indicated that the silage inoculated with LI1 enhanced the carbohydrate metabolism pathway but inhibited the amino acid metabolism pathway, which played a role in promoting faster lactic acid production, reducing the decomposition of protein to ammonia nitrogen, and improving the fermentation quality of silage. Therefore, oat silage can be processed to high-quality and cleaner fermented feed by using an LAB inoculant, and LI1 showed better efficiency than LI2. IMPORTANCE Oat natural silage is rich in Enterobacteriaceae, increasing gas production and fermentation loss. Lactic acid bacteria interact synergistically to form a dominant community during ensiling. Pediococci grow vigorously in the early stage of fermentation and create an anaerobic environment. Lactobacilli inhibit the harmful microorganisms and result in cleaner fermentation of oat silage.

Project description:Feed deficiency during the long cold period of a year is one of the major problems that the traditional year-round animal grazing system has faced on the Qinghai-Tibetan plateau (QTP) since ancient time. Therefore, ensiling the grasses from grassland could be a desirable feeding regime to preserve high quality forage and to alleviate the seasonal unbalanced feed supply problem on this plateau. The present study was designed to investigate dynamics of bacterial community and natural fermentation quality of ensiled Elymus nutans collected from grasslands in four different areas with different elevations [Tianzhu County (TZ), 2965 m; Golog Prefecture (GL), 3763 m; Damxung County (DX), 4228 m, and Nagqu Prefecture (NQ), 4752 m] on the QTP. The bacterial community was characterized by using the PacBio single molecule with real-time sequencing technology (SMRT). The harvested fresh E. nutans grasses were ensiled in vacuum-sealed polyethylene bags for 14, 30, 60, and 90 days. Obvious differences in the epiphytic bacterial community of the fresh E. nutans samples from the four areas were observed, which resulted in various bacterial community dynamics and fermentation qualities of ensiled E. nutans. Higher fermentation quality was observed in silage samples from Nagqu than in those from the other areas (P < 0.05). Lactic acid bacteria (LAB) involved in fermentation of E. nutans from low altitude areas consisted of Pediococcus pentosaceus, Lactobacillus sp., Leuconostoc mesenteroides, and Lactobacillus coryniformis, whereas major LAB species involved in the fermentation of E. nutans silage from high altitudes included L. mesenteroides, Lactobacillus brevis, and Lactobacillus sp. Correlation analysis between bacterial composition and fermentation quality of E. nutans silages made from the four different areas in the QTP indicated that the LAB species responsible for silage fermentation in different areas were totally different, which was mainly due to the different epiphytic bacterial compositions in fresh E. nutans before ensiling. The present results provide important information on revealing the bacterial communities and fermentation quality of ensiled E. nutans, and on future screening of LAB isolates for making high quality silage in order to alleviate feed shortage of the traditional year-round grazing system on the QTP.

Project description:ObjectiveThe objective of this study was to isolate lactic acid bacteria (LAB) from native grasses and naturally fermented silages, determine their identity, and assess their effects on silage quality and bacterial communities of the native grasses of three steppe types fermented for 60 days.MethodsAmong the 58 isolated LAB strains, Limosilactobacillus fermentum (BL1) and Latilactobacillus graminis (BL5) were identified using 16S rRNA sequences. Both strains showed normal growth at 15- 45°C temperature, 3-6.5% NaCl concentration, and pH 4-9. Two isolated LAB strains (labeled L1 and L5) and two commercial additives (Lactiplantibacillus plantarum and Lentilactobacillus buchneri; designated as LP and LB, respectively) were added individually to native grasses of three steppe types (meadow steppe, MS; typical steppe, TS; desert steppe, DS), and measured after 60 d of fermentation. The fresh material (FM) of different steppe types was treated with LAB (1 × 105 colony forming units/g fresh weight) or distilled water (control treatment [CK]).ResultsCompared with CK, the LAB treatment showed favorable effects on all three steppe types, i.e., reduced pH and increased water-soluble carbohydrate content, by modulating the microbiota. The lowest pH was found in the L5 treatment of three steppe types, at the same time, the markedly (p < 0.05) elevated acetic acid (AA) concentration was detected in the L1 and LB treatment. The composition of bacterial community in native grass silage shifted from Pantoea agglomerans and Rosenbergiella nectarea to Lentilactobacillus buchneri at the species level. The abundance of Lentilactobacillus buchneri and Lactiplantibacillus plantarum increased significantly in L1, L5, LP, and LB treatments, respectively, compared with CK (p < 0.05).ConclusionIn summary, the addition of LAB led to the shifted of microbiota and modified the quality of silage, and L. fermentum and L. graminis improved the performance of native grass silage.

Project description:BackgroundDouzhi, a traditional Chinese fermented beverage, features microbial communities primarily composed of lactic acid bacteria (LAB). As fermented foods continue to gain recognition and popularity, douzhi is attracting growing interest. However, investigation of the critical aspects of douzhi's fermentation processes, including fermentation characteristics and microbial community dynamics, remains vital for enhancing food safety and quality for douzhi, as well as for similar fermented food products.MethodIn this study, we collected douzhi microbial communities from four chain stores, using them as fermentation starter cultures. The microbial dynamics of the fermentation were analyzed, focusing on the inoculation of LAB strains and the transition from a mung bean-based matrix to skimmed milk. The metabolomic profiles of the fermented mung bean matrices were also studied.ResultsDouzhi samples obtained from representative chain stores were found to be overwhelmingly dominated by LAB. When inoculated along with the douzhi community, both LAB strains exhibited notable and substantial reductions in the pH value of the designated mung bean matrices compared to those inoculated indigenous microbiota. Specifically, Lactiplantibacillus plantarum CGMCC 1.1856 retained its population, whereas Pediococcus pentosaceus CGMCC 1.2695 exhibited a decrease in relative abundance. Using skimmed milk as a fermentation substrate instead of the mung bean matrix resulted in significant shifts in microbial communities, particularly leading to an increase in Escherichia sp. The metagenomic analyses and functional predictions illustrated that various metabolic functions were enhanced during the fermentation process due to LAB inoculation. The liquid chromatography-mass spectrometry based metabolomic analysis revealed that the inoculation of Lactiplantibacillus plantarum and Pediococcus pentosaceus in mung bean matrix did not introduce new metabolites but significantly altered the concentration and profile of existing metabolites, especially increased low molecular carbohydrates, which may enhance the nutritional potential of the fermented product.DiscussionThis study examines the microbial dynamics of douzhi microbiota fermentation, emphasizing the role of lactic acid bacteria in enhancing fermentation activity and metabolite profiles. These insights contribute to improving manufacturing processes and ensuring the safety and quality of douzhi and similar fermented foods.

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Microbial Community and Fermentation Characteristics of Native Grass Prepared Without or With Isolated Lactic Acid Bacteria on the Mongolian Plateau

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