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Dynamic Succession of Microbiota during Ensiling of Whole Plant Corn Following Inoculation with Lactobacillus buchneri and Lactobacillus hilgardii Alone or in Combination.

ABSTRACT: Lactic acid bacteria (LAB) used as silage additives have been shown to improve several fermentation parameters, including aerobic stability. Inoculation with a combination of Lactobacillus buchneri NCIMB40788 and Lactobacillus hilgardii CNCM-I-4785, contributes to an increase in aerobic stability, compared to each strain inoculated independently. To understand the mode of action of the combination on the LAB community, a fermentation-kinetic study was performed on corn. Four treatments, Control, Lb. buchneri, Lb. hilgardii, and a combination of the two strains, were fermented 1, 2, 4, 8, 16, 32, and 64 days. Corn silage inoculated by both strains had a lactate:acetate ratio of 0.59 after 64 days and a higher concentration of lactate than Lb. buchneri. Analysis of the microbiota by 16S and ITS amplicon metasequencing demonstrated that inoculation led to lower bacterial diversity after 1 day, from 129.4 down to 40.7 observed operational taxonomic units (OTUs). Leuconostocaceae represented the dominant population by day 1, with 48.1%. Lactobacillaceae dominated the succession by day 4, with 21.9%. After 32 days, inoculation by both strains had the lowest bacterial alpha diversity level, with 29.0 observed OTUs, compared to 61.3 for the Control. These results confirm the increased fermentation efficiency when the two Lactobacillus strains are co-inoculated, which also led to a specific yeast OTUs diversity profile, with Hannaella as the main OTU.

SUBMITTER: Drouin P

PROVIDER: S-EPMC6955939 | biostudies-literature | 2019 Nov

REPOSITORIES: biostudies-literature

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Dynamic Succession of Microbiota during Ensiling of Whole Plant Corn Following Inoculation with Lactobacillus buchneri and Lactobacillus hilgardii Alone or in Combination.

Drouin Pascal P Tremblay Julien J Chaucheyras-Durand Frédérique F

Microorganisms 20191121 12

Lactic acid bacteria (LAB) used as silage additives have been shown to improve several fermentation parameters, including aerobic stability. Inoculation with a combination of Lactobacillus buchneri NCIMB40788 and Lactobacillus hilgardii CNCM-I-4785, contributes to an increase in aerobic stability, compared to each strain inoculated independently. To understand the mode of action of the combination on the LAB community, a fermentation-kinetic study was performed on corn. Four treatm ...[more]

PMID: 31766494

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Project description:The objective of this study was to assess the impact of Saccharomyces cerevisiae in combination with Lactobacillus buchneri on the fermentation characteristics, aerobic stability, nutritive value, and microbial communities of corn silage. Whole crop corn (39% DM) was either uninoculated (Control) or inoculated with S. cerevisiae and L. buchneri at the following concentrations: S. cerevisiae 104 cfu/g fresh forage (S4), S. cerevisiae 105 cfu/g (S5), S. cerevisiae 104 cfu/g + L. buchneri 105 cfu/g (S4L5), and S. cerevisiae 105 cfu/g + L. buchneri 104 cfu/g (S5L4), and ensiled in mini silos for 118 d, followed by 7 d of aerobic exposure. Changes in fermentation characteristics and nutritive value were assessed in terminal silages. Saccharomyces cerevisiae, L. buchneri, and total yeast, fungal, and bacterial communities in silage were estimated using quantitative PCR. Composition of bacterial and fungal communities during ensiling and aerobic exposure was measured using 16S rDNA and ITS sequencing, respectively. In the first 7 d of ensiling, the concentration of lactic acid rapidly increased (P < 0.01) in all silages, with the pH declining to 4.0 (P < 0.001) and thereafter remaining stable (P = 0.23). Although S4L5 contained a higher (P = 0.03) concentration of acetic acid than Control, other fermentation characteristics were did not differ among terminal silages. Inoculation with S. cerevisiae had no detrimental effect on the aerobic stability of silage, whereas L. buchneri did not prevent spoilage as the pH across all silages averaged 8.0 after 7 d of aerobic exposure. Total yeast (P = 0.42), bacterial (P = 0.13), and fungal (P = 0.89) communities were not altered by the inoculants after ensiling or aerobic exposure. Sequencing identified temporal shifts of bacterial and fungal communities during ensiling and aerobic exposure. Concentrations of S. cerevisiae and L. buchneri in all inoculated silages remained greater (P < 0.01) than Control after ensiling, with numbers of S. cerevisiae increasing after 7 d of aerobic exposure. Bacterial communities in silages inoculated with S. cerevisiae and L. buchneri clustered separately from other silages, an observation that was not apparent for fungal communities. Our findings suggest that aerobic exposure could potentially increase the abundance of S. cerevisiae with probiotic properties in corn silage just prior to feeding.

Project description:AimsThis study evaluated changes in epiphytic microbial population of alfalfa (Medicago sativa) during the growing season. First cut forage was harvested to study the effects of an inoculant combining two obligate heterofermentative lactic acid bacteria strains on the bacterial and fungal communities and the fermentation of alfalfa silage.Methods and resultsThe epiphytic microbiome of alfalfa was evaluated 10-times during the growing season. Alfalfa wilted to 395.0 g/kg was treated with water (Control) or with a combination of L. buchneri NCIMB 40788 and L. hilgardii CNCM-I-4785 (LBLH). Mini-silos were opened after 1, 4, 8, 16, 32, and 64 days of ensiling. The relative abundance (RA) of the epiphytic bacterial and fungal families varied during the growing season. After 1 day, Weissella was the most abundant genus and present at similar RA in the two treatments (average 80.4%). Compared with Control, LBLH had a higher RA of Lactobacillus at day 1, 16, 32, and 64, and a lower RA of Weissella from day 8 to 64. Control contained more bacteria belonging to the Enterobacteriales than LBLH up to day 16. Inoculated silage had more acetate than Control at day 32 and 64. The fungal population were similar between treatments. The enhanced development and dominance of Lactobacillus in inoculated silage led to greater accumulation of acetate and propionate, which reduced the numbers of culturable yeasts but did not markedly affect the fungal community structure.ConclusionsThe bacterial community composition of alfalfa stands in the filed changed over time and was affected by cutting. For the ensiling trial, inoculation modified the composition of the bacterial community of alfalfa, increasing the RA of Lactobacillus while reducing the RA of Weissella and of Enterobacteriaceae.Significance and impact of studyInoculation increased the RA of Lactobacillus, hampering the dominance of Weissella in the early stages of ensiling, improving antifungal compounds production and reducing the numbers of culturable yeasts.

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Project description:Production of biofuels, bioproducts, and bioenergy requires a well-characterized, stable, and reasonably uniform biomass supply and well-established supply chains for shipping biomass from farm fields to biorefineries, while achieving year-round production targets. Preserving and stabilizing biomass feedstock during storage is a necessity for cost-effective and sustainable biofuel production. Ensiling is a common storage method used to preserve and even improve forage quality; however, the impact of ensiling on biomass physical and chemical properties that influence bioconversion processes has been variable. Our objective in this work was to determine the effects of ensiling on lignocellulosic feedstock physicochemical properties and how that influences bioconversion requirements. We observed statistically significant decreases (p < 0.05) in the content of two major structural carbohydrates (glucan and xylan) of 5 and 8%, respectively, between the ensiled and non-ensiled materials. We were unable to detect differences in sugar yields from structural carbohydrates after pretreatment and enzymatic hydrolysis of the ensiled materials compared to non-ensiled controls. Based on this work, we conclude that ensiling the corn stover did not change the bioconversion requirements compared to the control samples and incurred losses of structural carbohydrates. At the light microscopy level, ensiled corn stover exhibited little structural change or relocation of cell wall components as detected by immunocytochemistry. However, more subtle structural changes were revealed by electron microscopy, as ensiled cell walls exhibit ultrastructural characteristics such as wall delimitation intermediate between non-ensiled and dilute-acid-pretreated cell walls. These findings suggest that alternative methods of conversion, such as deacetylation and mechanical refining, could take advantage of lamellar defects and may be more effective than dilute acid or hot water pretreatment for biomass conversion of ensiled materials.

Dataset Information

Dynamic Succession of Microbiota during Ensiling of Whole Plant Corn Following Inoculation with Lactobacillus buchneri and Lactobacillus hilgardii Alone or in Combination.

Publications

Dynamic Succession of Microbiota during Ensiling of Whole Plant Corn Following Inoculation with <i>Lactobacillus buchneri</i> and <i>Lactobacillus hilgardii</i> Alone or in Combination.

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