Dynamics associated with fermentation and aerobic deterioration of high-moisture Italian ryegrass silage made using Lactobacillus plantarum and caproic acid.

AIMS: To determine the fermentation quality, aerobic stability, and chemical composition of Italian ryegrass silage prepared with Lactobacillus plantarum (LP), caproic acid (CA), and their combination during ensiling and feed-out phase. METHODS AND RESULTS: Six treatments: control (CON), LP, 0.15% caproic acid (LCA), 0.2% caproic acid (HCA), LCA + LP, and HCA + LP were employed for 30 days ensiling and an 8-days aerobic stability test. LP had similar pH value and lactic acid content with LCA + LP, while the contents of NH3-N and total VFAs in LCA + LP were significantly lower than those in LP and CON, and the fermentation quality of LCA + LP performed best among all silages. As air-exposure extended, contents of water-soluble carbohydrates (WSC), lactic, and acetic acids decreased, while pH, and NH3-N content increased significantly. The population of lactic acid bacteria gradually decreased in contrast to increased counts of aerobic bacteria and yeasts. Compared with LCA, 0.2% CA delayed the aerobic deterioration as judged by a slower increase in pH and high residual of WSC and lactic acid, and negligible ethanol content and anaerobe spores counts remained in HCA at the end of air exposure. Compared with CON (73 h), LP showed less aerobic stability (38 h), whereas HCA and HCA + LP prolonged aerobic stability for 210 and 152 h, better than LCA (109 h) and LCA + LP (146 h). CONCLUSIONS: Lactobacillus plantarum apparently improved the fermentation quality, and combined with CA exhibited greater efficiency in inhibiting undesirable microorganism during ensiling. CA at 0.2% optimally extended the aerobic stability.

SLI-GNN: A Self-Learning-Input Graph Neural Network for Predicting Crystal and Molecular Properties.

Since the structures of crystals/molecules are often non-Euclidean data in real space, graph neural networks (GNNs) are regarded as the most prospective approach for their capacity to represent materials by graph-based inputs and have emerged as an efficient and powerful tool in accelerating the discovery of new materials. Here, we propose a self-learning-input GNN framework, named self-learning-input GNN (SLI-GNN), to uniformly predict the properties for both crystals and molecules, in which we design a dynamic embedding layer to self-update the input features along with the iteration of the neural network and introduce the Infomax mechanism to maximize the average mutual information between the local features and the global features. Our SLI-GNN can reach ideal prediction accuracy with fewer inputs and more message passing neural network (MPNN) layers. The model evaluations on the Materials Project dataset and QM9 dataset verify that the overall performance of our SLI-GNN is comparable to that of other previously reported GNNs. Thus, our SLI-GNN framework presents excellent performance in material property prediction, which is thereby promising for accelerating the discovery of new materials.

Dynamics in fermentation quality, bacterial community, and metabolic profile during silage fermentation of late-harvested elephant grass.

The present study aimed to evaluate the effects of delayed harvest and storage length on fermentation products, bacterial community, and metabolic shifts of elephant grass silage. The late-harvested elephant grass (LG) was naturally fermented (NLG) for 1, 3, 7, 15, 30, and 60 days, respectively. After 60-day ensiling, NLG displayed homolactic fermentation with low pH value, butyric acid, and ammonia nitrogen concentrations, and high lactic acid concentration, and ratio of lactic acid to acetic acid. Pseudomonas, Sphingomonas, and Pantoea dominated the bacterial community in LG, but Lactobacillus, Lactococcus, and Pediococcus were the advantageous genera in a 3-day and 60-day NLG. The correlation heatmap revealed that Acetobacter was positively related to acetic acid, ethanol, ammonia nitrogen, and butyric acid concentrations. There were distinct differences in the KEGG (Kyoto Encyclopedia of Genes and Genomes) metabolic profiles of fresh and ensiled LG. Ensiling suppressed the metabolism of amino acid, vitamins, and energy, while promoted the metabolism of carbohydrate. The LG can be well-fermented without additives, but its low crude protein content should not be ignored when applied in agricultural practice. The ensiling process remarkably affected the fermentation quality, bacterial community, and metabolic profiles of NLG.

Exploiting the anaerobic fermentation of alfalfa as a renewable source of squalene.

BACKGROUND: The use of alfalfa is a promising response to the increasing demand for squalene. Ensiling could enhance the squalene content of fresh alfalfa and silage. To investigate and exploit the anaerobic fermentation of forage as a new squalene source, alfalfa was ensiled without (CON) or with molasses (ML) and sunflower seed oil (SSL) for 10, 40, and 70 days. RESULTS: Naturally ensiled alfalfa was of poor quality but had up to 1.93 times higher squalene content (P < 0.001) than fresh alfalfa. The squalene-producing bacteria were found to be cocci lactic acid bacteria (LAB). Adding ML and SSL decreased squalene content (P = 0.002 and P < 0.001) by 6.89% and 11.6%, respectively. Multiple linear regression models and correlation analysis indicated that squalene synthase was the key enzyme for squalene synthesis. The addition of ML and SSL altered the structure of LAB communities, mainly decreasing the relative abundance of cocci LAB, which was responsible for squalene synthesis, and changing the fermentation products (lactic acid, propionic acid, and ammonia-N) influencing the squalene-related enzymes, thereby decreasing squalene production. Compared with squalene production from the reference bacteria (Pediococcus acidilactici Ch-2, Rhodopseudomonas palustris, Bacillus subtilis, engineered Escherichia coli), alfalfa silage had the potential to be a new squalene source. CONCLUSION: Natural ensiled alfalfa was a promising source for squalene, and ensiling was a potential pathway to obtain novel high-yield squalene bacteria. © 2022 Society of Chemical Industry.

The effects of epiphytic microbiota and chemical composition of Italian ryegrass harvested at different growth stages on silage fermentation.

BACKGROUND: The influence of epiphytic microbiota and chemical composition on fermentation quality and microbial community of Italian ryegrass silage was evaluated. Italian ryegrass harvested at the filling stage (FS) and the dough stage (DS) was sterilized by gamma-ray irradiation and inoculated as follows: (I) FS epiphytic microbiota + irradiated FS (FF); (II) FS epiphytic microbiota + irradiated DS (FD); (III) DS epiphytic microbiota + irradiated DS (DD); (IV) DS epiphytic microbiota + irradiated FS (DF). RESULTS: After 60 days of ensiling, silage made from irradiated FS had a lower pH and ammonia nitrogen (NH3 -N) content and a higher lactic acid (LA) content than that made from irradiated DS. Similarly, silage inoculated with the epiphytic microbiota of DS had a lower pH and NH3 -N content and a higher LA content than that inoculated with the epiphytic microbiota of FS. However, LA-type fermentation (lactic acid:acetic acid > 2:1) was presented at DF and DD. The principal coordinates analysis showed that the distance between FF and DF and FD and DD was closer than other treatments, suggesting that the microbial community of silages made from irradiated FS (or DS) was more similar. CONCLUSION: The epiphytic microbiota played a more important role in the fermentation type, whereas the chemical composition had a great influence on the contents of fermentation end-products. However, chemical composition had a stronger effect on the microbial community of silage than the epiphytic microbiota. © 2022 Society of Chemical Industry.

Assessing the impact of phyllosphere microbiota on dynamics of in-silo fermentation of Italian ryegrass harvested at heading and blooming stages.

BACKGROUND: The present study aimed to investigate the relationship between the phyllosphere microbiota of Italian ryegrass (Lolium multiflorum Lam.) harvested at heading (H) [> 50% earing rate or 216 g kg-1 fresh weight (FW)] and blooming (B) (> 50% bloom or 254 g kg-1 FW) stages and in-silo fermentation products, and the composition, abundance, diversity and activity of bacterial community. In total, 72 (4 treatments × 6 ensiling durations × 3 replicates) laboratory scale (400 g) silages of Italian ryegrass were prepared: (i) irradiated heading stage silages (IRH) (n = 36) were inoculated with phyllosphere microbiota inoculum (2 mL) eluted from fresh Italian ryegrass at either heading (IH) (n = 18) or blooming (IB) (n = 18) stages; (ii) irradiated blooming stage silages (IRB) (n = 36) were inoculated with either IH (n = 18) or IB (n = 18). Triplicate silos of each treatment were analyzed after 1, 3, 7, 15, 30 and 60 days of ensiling. RESULTS: In fresh forage, Enterobacter, Exiguobacterium and Pantoea were the three major genera at heading stage, and Rhizobium, Weissella and Lactococcus were the most abundant genera at blooming stage. Higher metabolic activity was found in IB. After 3 days of ensiling, the large amounts of lactic acid in IRH-IB and IRB-IB can be attributed to the higher abundances of Pediococcus and Lactobacillus, 1-phosphofructokinase, fructokinase, l-lactate dehydrogenase and glycolysis I, II and III. CONCLUSION: The composition, abundance, diversity and functionality of the phyllosphere microbiota of Italian ryegrass at different growth stages could remarkably affect silage fermentation characteristics. © 2023 Society of Chemical Industry.

A survey of fermentation parameters, bacterial community compositions and their metabolic pathways during the ensiling of sorghum.

AIMS: To investigate the fermentative products, bacterial community compositions and their metabolic pathways of sorghum (Sorghum dochna Snowden) during ensiling. METHODS AND RESULTS: Sorghum was harvested at soft dough stage and ensiled in laboratory-scale silos (10 L capacity). Triplicate silos were sampled after 1, 3, 7, 15, 30 and 60 days of ensiling, respectively. The bacterial communities on day 3 and 60 were assessed through high-throughput sequencing technology, and 16S rRNA gene-predicted functional profiles were analysed according to the KEGG (Kyoto Encyclopedia of Genes and Genomes) using Tax4Fun. The sorghum silages had high lactic acid (~85.4 g kg-1 dry matter [DM]) contents, and low pH (~3.90), butyric acid (~1.27 g kg-1 DM) and ammonia nitrogen (~86.3 g kg-1 total nitrogen [TN]) contents. During the ensiling, glucose was the first fermentable substrate, and the contents of xylose and arabinose were increased. The dominant genus Lactococcus on day 3 was replaced by Lactobacillus on day 60. The metabolism of amino acid, energy, cofactors and vitamins was inhibited, whilst the metabolism of nucleotide and carbohydrate was promoted after ensiling. 1-Phosphofructokinase and pyruvate kinase of bacterial community played important roles in stimulating the lactic acid fermentation. CONCLUSIONS: Knowledge about bacterial community dynamics and their metabolic pathways during sorghum ensiling is important for understanding the fermentative profiles and may promote the production of nutritious and stable sorghum silage. SIGNIFICANCE AND IMPACT OF THE STUDY: The high-throughput sequencing technology combined with 16S rRNA gene-predicted functional analyses revealed the differences in silages not only for distinct bacterial community but also for specific functional metabolites. This could provide some new insights into bacterial community and functional profiles to further improve the silage quality.

Separating the chemical and microbial factors of oat harvested at two growth stages to determine the main factor on silage fermentation.

AIMS: This work evaluated the effects of epiphytic microbiota and chemical components on fermentation quality and microbial community of ensiled oat. METHODS AND RESULTS: Oat harvested at the heading stage (HS) and the milk stage (MS) was sterilized by gamma-ray irradiation and inoculated as the following: (1) HS epiphytic microbiota + sterilized HS (H-H); (2) MS epiphytic microbiota + sterilized HS (M-H); (3) MS epiphytic microbiota + sterilized MS (M-M); and (4) HS epiphytic microbiota + sterilized MS (H-M). After 60-day fermentation, silages inoculated with the epiphytic microbiota of HS had higher acetic acid content than those inoculated with MS. Silage made from sterilized MS had lower pH, ammonia nitrogen and butyric acid contents and higher dry matter, water-soluble carbohydrates and lactic acid contents than that made from sterilized HS. The microbial communities of oat silages were similar, and they were mainly Lactobacillus. CONCLUSIONS: The chemical component rather than the epiphytic microbiota at harvest exerted more effects on oat silages. SIGNIFICANCE AND IMPACT OF THE STUDY: This work reveals the different effects of chemical and microbial factors on the fermentation of silage, which is instructive for us to produce quality silage.

Exploring the ensiling characteristics and bacterial community of red clover inoculated with the epiphytic bacteria from temperate gramineous grasses.

AIMS: To explore the microbiological factors that cause the difference in silage fermentation characteristics between temperate gramineous (Italian ryegrass and oat) grass and legume forage. METHODS AND RESULTS: Through γ-ray irradiation sterilization and bacterial transplantation technology, the sterilized red clover was inoculated as follows: (i) aseptic water (STRC); (ii) epiphytic bacteria on red clover (RCRC); (iii) epiphytic bacteria on oat (RCOT); and (iv) epiphytic bacteria on Italian ryegrass (RCIR). Red clover was ensiled in laboratory-scale silos made of vacuum-packed plastic bags for 1, 3, 7, 14, 30 and 60 days. STRC remained unfermented state based on similar chemical components with fresh red clover. Compared with RCRC and RCIR, higher lactic acid content and ratio of lactic acid to acetic acid, and lower pH, acetic acid and ammonia nitrogen contents were observed in RCOT after 60 days of ensiling. Using next-generation sequencing, higher abundances of Methylobacterium and Sphingomonas were observed in the epiphytic bacteria on red clover. Pediococcus was dominant in RCRC after 3 days of ensiling. Lactobacillus was the most predominant in each group after 60 days of fermentation. RCRC and RCIR had a more hetero-fermentative process, while RCOT possessed a more homo-fermentative pathway. According to the 16S rRNA gene-predicted functional profiles, the metabolism of amino acids was accelerated by the epiphytic bacteria from red clover. CONCLUSIONS: Inoculating the epiphytic bacteria from oat could alter the fermentation characteristics and bacterial compositions of red clover silage. The relative abundance and activity of Methylobacterium, Sphingomonas, Enterobacteriaceae and hetero-fermentative Lactobacillus in red clover silage could be changed by the epiphytic bacteria on temperate gramineous grass. SIGNIFICANCE AND IMPACT OF THE STUDY: The exogenous micro-organisms inhibiting the metabolism of amino acids can be a good potential source to improve the silage quality of legume forage.

Characterization and identification of a novel microbial consortium M2 and its effect on fermentation quality and enzymatic hydrolysis of sterile rice straw.

AIMS: To isolate and enrich lignocellulolytic microbial consortia from yak (Bos grunniens) rumen and evaluate their effects on the fermentation characteristics and enzymatic hydrolysis in rice straw silage. METHODS AND RESULTS: A novel microbial consortium M2 with high CMCase and xylanase activities was enriched and observed to be prone to use natural carbon sources. Its predominant genus was Enterococcus, and most carbohydrate-active enzyme (CAZyme) genes belonged to the glycosyl hydrolases class. The consortium M2 was introduced with or without combined lactic acid bacteria (XA) to rice straw silage for 60 days. Inoculating the consortium M2 notably decreased the structural carbohydrate contents and pH of rice straw silages. Treatment that combines consortium M2 and XA resulted in the highest levels of lactic acid and lignocellulose degradation. The consortium M2 alone or combined with XA significantly (p < 0.01) increased water-soluble carbohydrates (WSCs), mono- and disaccharides contents compared with the XA silage. Combined addition obviously improved the enzymatic conversion efficiency of rice straw silage with higher glucose and xylose yields (23.39 and 12.91 w/w% DM, respectively). CONCLUSIONS: Ensiling pretreatment with the microbial consortium M2 in sterile rice straw improved fermentation characteristics. The combined application of consortium M2 with XA had synergistic effects on promoting the degradation of structural carbohydrates and enzymatic hydrolysis. SIGNIFICANCE AND IMPACT OF THE STUDY: Rice straw is difficult to ensile because of its low WSC and high structural carbohydrate contents. The microbial consortium M2 identified herein exhibits great potential for degrading fibrous substrates, and their combination with XA provides a faster and more effective synergistic strategy for biorefinery of lignocellulosic biomass.

Effect of epiphytic microbiota from napiergrass and Sudan grass on fermentation characteristics and bacterial community in oat silage.

AIMS: To investigate the effects of epiphytic microbiota from napiergrass and Sudan grass on ensiling characteristics and microbial community of oat silage. METHODS AND RESULTS: By γ-ray irradiation sterilization and microbiota transplantation technology, the sterilized oat was inoculated as follows: (a) aseptic water (STOT), (b) epiphytic bacteria on oat (OTOT), (c) epiphytic bacteria on napiergrass (OTNP) and (d) epiphytic bacteria on Sudan grass (OTSD). STOT remained in the unfermented state based on similar chemical components with fresh oat. Compared with OTOT and OTSD, higher lactic acid content and ratio of lactic acid to acetic acid, and lower pH, acetic acid and ammonia nitrogen contents were observed in OTNP after 60 days of ensiling. At the late stage, Lactobacillus was the most predominant in each group. Lactococcus was eventually replaced by Lactobacillus in OTSD, whereas Lactococcus was found throughout the whole ensiling process in OTNP. Higher abundance of Weissella was observed in OTSD at the early and late stages. The result of co-occurrence network analysis proved that Lactococcus was pivotal in determining the silage fermentation pattern during ensiling. According to the 16S rRNA gene-predicted functional profiles, the inoculation of epiphytic microbiota from oat enhanced the metabolism of amino acids, whereas the inoculation of epiphytic microbiota from napiergrass and Sudan grass accelerated the carbohydrate metabolism. CONCLUSIONS: The epiphytic microbiota on napiergrass promoted a homo-fermentative process, whereas the epiphytic microbiota on oat and Sudan grass facilitated a hetero-fermentative pattern in oat silage, which was closely related to the abundance and metabolism of Lactococcus, Weissella and Lactobacillus. SIGNIFICANCE AND IMPACT OF THE STUDY: The exogenous microorganisms that promote the carbohydrate metabolism and inhibit the metabolism of amino acids could be a good potential source to improve the silage quality of temperate grass.

Dynamics of the bacterial communities and predicted functional profiles in wilted alfalfa silage.

AIMS: To investigate the fermentation characteristics, bacterial community and predicted functional profiles during the ensiling of wilted alfalfa (Medicago sativa L.). METHODS AND RESULTS: First-cutting alfalfa was harvested at the early bloom stage, wilted for 6 h, and ensiled in laboratory-scale silos (1 L capacity). Triplicate silos were sampled after 1, 3, 7, 15, 30 and 60 days of ensiling, respectively. The bacterial communities of wilted alfalfa and silages on day 3 and 60 were assessed through high throughput sequencing technology, and their functional characteristics were evaluated according to the Kyoto Encyclopedia of Genes and Genomes using Tax4Fun. After 60 days of ensiling, alfalfa silage showed a moderate fermentation quality, indicated by high lactic acid (56.7 g kg-1 dry matter [DM]) and acetic acid (39.4 g kg-1 DM) contents, and low concentrations of butyric acid (2.12 g kg-1 DM) and ammonia nitrogen (128 g kg-1 total nitrogen). Lactobacillus rapidly became predominant on day 3 and increased to 60.4% on day 60. Results of functional prediction analyses showed that the metabolism of amino acid, energy, cofactors and vitamins were reduced, while metabolism of nucleotide and carbohydrate were increased during ensiling. Fructokinase, 1-phosphofructokinase and pyruvate kinase played important roles in producing lactic acid. The production of acetic acid may be correlated with the enhancement of 6-phosphogluconate dehydrogenase and acetyl-CoA synthetase. CONCLUSIONS: Knowledge regarding bacterial dynamics and their metabolic pathways during alfalfa ensiling is important for understanding the fermentation process and may contribute to the production of nutritious and stable alfalfa silage. SIGNIFICANCE AND IMPACT OF THE STUDY: High throughput sequencing technology combined with 16S rRNA gene-predicted functional analyses could provide a new and comprehensive insight into bacterial community dynamics and functional profiles to further improve the silage quality.

Using γ-ray irradiation and epiphytic microbiota inoculation to separate the effects of chemical and microbial factors on fermentation quality and bacterial community of ensiled Pennisetum giganteum.

AIMS: This study aimed to separate the effects of chemical and microbial factors on the fermentation quality and bacterial community of ensiled Pennisetum giganteum. METHODS AND RESULTS: Fresh P. giganteum of two vegetative stages (stage I, GI ; stage II, GII ) was treated as follows: GI epiphytic microbiota + Î³-ray presterilized GI (MI CI ), GII epiphytic microbiota + Î³-ray presterilized GI (MII CI ), GI epiphytic microbiota + Î³-ray presterilized GII (MI CII ), and GII epiphytic microbiota + Î³-ray presterilized GII (MII CII ). Triplicates per treatment were sampled after 30 days of ensiling for chemical and microbial analyses and high-throughput sequencing. Silages made from CII (MI CII and MII CII ) had higher lactic acid concentration and the ratio of lactic to acetic acid, and lower pH and ammonia nitrogen concentration than silages produced by CI (MI CI and MII CI ). Species differential analyses showed that the changes of chemical composition rather than epiphytic microbiota significantly affected the relative abundance of Lactobacillus, Pediococcus and Pantoea in P. giganteum silages. CONCLUSION: These above results manifested that chemical composition was the main factor influencing the fermentation quality and bacterial community of P. giganteum silage in this study. SIGNIFICANCE AND IMPACT OF THE STUDY: The obtained results may, therefore, be the first record to provide an in-depth understanding of the relative contributions of chemical and microbial parameters on fermentation quality and bacterial community, which is of great importance for modulating silage fermentation and improving silage quality.

Two novel screened microbial consortia and their application in combination with Lactobacillus plantarum for improving fermentation quality of high-moisture alfalfa.

AIMS: To enrich lignocellulolytic microbial consortia and evaluate whether a combination of these consortia and Lactobacillus plantarum can facilitate degradation of structural carbohydrates and improve fermentation quality of high-moisture alfalfa silage. METHODS AND RESULTS: Two novel microbial consortia (CL and YL) with high lignocellulolytic potential were enriched, and had higher enzyme activities at slightly acidic conditions (pH 3.5-6.5). Two consortia were inoculated with and without combined L. plantarum (LP) to alfalfa for up to 120 days of ensiling. The two consortia alone or combined with LP significantly (p < 0.05) increased lactic-to-acetic acid ratios and decreased contents of volatile organic acids and NH3 -N as compared to the control. Treatments that combining microbial consortia and LP further resulted in the higher contents of lactic acid (LA), water soluble carbohydrates (WSC) and crude protein, dry matter (DM) recovery, and lower neutral detergent fibre, acid detergent lignin and cellulose contents, with YLP silage showing the lowest pH (4.41) and highest LA content (76.72 g kg-1  DM) and the conversion of WSC into LA (184.03%). CONCLUSIONS: The addition of lignocellulolytic microbial consortia (CL or YL) to alfalfa silages as attractive silage inoculants could improve fermentation quality, and that their combination with L. plantarum appeared more effective on the degradation of structural carbohydrates and conversion of soluble carbohydrates into LA. SIGNIFICANCE AND IMPACT OF THE STUDY: High-moisture alfalfa is difficult to ensile due to its high buffering capacity and low readily fermentable carbohydrate contents. Microbial consortia (CL and YL) can encode a broad selection of multi-functional CAZymes, and their combination with LP could be promising for the degradation of structural carbohydrates simultaneously with improvement fermentation quality, with high performance in LA production.

Evaluating the fermentation quality and bacterial community of high-moisture whole-plant quinoa silage ensiled with different additives.

AIM: To explore the potential of whole-plant quinoa (WPQ) as a high-protein source for livestock feed, this study evaluated the effects of additives on the fermentation quality and bacterial community of high-moisture WPQ silage. METHODS AND RESULTS: High-moisture WPQ was ensiled with one of the following additives: untreated control (C), fibrolytic enzyme (E), molasses (M), LAB inoculant (L), a combination of fibrolytic enzyme and LAB inoculant (EL) and a combination of molasses and LAB inoculant (ML). The fermentation quality and bacterial community after 60 days of ensiling were analysed. Naturally fermented WPQ exhibited acetic acid-type fermentation dominated by enterobacteria, with low lactic acid content (37.0 g/kg DM), and high pH value (5.65), acetic acid (70.8 g/kg DM) and NH3 -N production (229 g/kg TN). Adding molasses alone or combined with LAB inoculant shifted the fermentation pattern towards increased intensity of lactic acid fermentation, lowering the pH value (<4.56), contents of acetic acid (<46.7 g/kg DM) and NH3 -N (<140 g/kg TN) and total abundance of enterobacteria (<16.0%), and increasing the lactic acid content (>60.5 g/kg DM), lactic/acetic acid ratio (>1.40) and the relative abundance of Lactobacillus (>83.0%). CONCLUSIONS: The results suggested that the lack of fermentable sugar could be the main factor of restricting extensive lactic acid fermentation in WPQ silage. Supplementing fermentable sugar or co-ensiling with materials with high WSC content and low moisture content are expected to be beneficial strategies for producing high-quality WPQ silage. SIGNIFICANCE AND IMPACT OF STUDY: High biomass production and high protein content make WPQ to be an ideal forage source for livestock feed. Results of this study revealed the restricting factor for extensive lactic acid fermentation in WPQ silage, which could be helpful in producing high-quality WPQ silage.

Gamma-ray irradiation and microbiota transplantation to separate the effects of chemical and microbial diurnal variations on the fermentation characteristics and bacterial community of Napier grass silage.

BACKGROUND: To investigate the contributions of chemical and microbial diurnal variations in fermentation characteristics and bacterial community of Napier grass silage, gamma-ray irradiated Napier grass harvested at 07.00 h (AM), 12.00 h (M) and 17.00 h (PM) was inoculated with the microbiota derived from Napier grass harvested at AM, M and PM in a 3 (irradiated forage: AMG , MG and PMG ) × 3 (microbiota: AMM , MM and PMM ) design and then ensiled for 14 and 60 days. RESULTS: Napier grass harvested at various times had different chemical compositions and epiphytic microbiota prior to ensiling. For silages inoculated with the same microbiota, the pH values, residual water soluble carbohydrates and dry matter contents increased, and lactic acid, acetic acid, propionic acid, butyric acid, ethanol and volatile fatty acids contents decreased in PMG and MG silages compared to AMG silages. MM and PMM inoculum promoted lactic acid fermentation as indicated by higher lactic acid contents and lactic/acetic acid ratios in MM and PMM -inculated silages compared to those in AMM -inoculated silages after 60 days of ensiling. During ensiling, epiphytic microbiota affected the Chao1 index, operational taxonomic units (OTUs) number and Shannon index, as well as the abundances, of more than half of the top 10 abundant genera, whereas chemical composition did not affect any of the bacterial diversity and richness indices and only showed significant impacts on the abundances of two genera. CONCLUSION: The results indicated that chemical diurnal variation exerted an influence mainly on the extent of fermentation, whereas microbial diurnal variation affected more the bacterial community and fermentation types during Napier grass ensiling. © 2022 Society of Chemical Industry.

Effects of hexanoic acid on microbial communities, fermentation, and hygienic quality of corn silages infested with toxigenic fungi.

BACKGROUND: This study aims to reveal the effects of hexanoic acid on the microbial communities, fermentation, and the hygienic quality of corn silages with or without fungal infection. Fungal-infested (FI) and non-infested (NFI) whole-crop corn samples were separately ensiled without (control, CON) or with hexanoic acid (Hex, 90% purity) at 0.2 g·kg-1 fresh weight (FW). RESULT: The addition of Hex accelerated the pH decline during the first 5 days of ensiling regardless of fungal infestation. The lactic acid (LA) concentration in Hex silages was lower than that in CON during 45 days of ensiling; however the FI-Hex silage had the highest LA concentration among treatments on day 90. The Hex silage showed lower aflatoxin B1 (AFB1 ), zearalenone (ZEA), and deoxynivalenol (DON) concentrations than CON for FI silages. On day 5, the addition of Hex decreased the relative abundance of Klebsiella, Pantoea, and Enterobacter compared with CON, regardless of fungal infestation. This inhibitory effect lasted until day 90 for NFI silages but disappeared for FI silages on day 90. The fungal infestation resulted in the accumulation of Candida (34.05%) and Wickerhamomyces (19.46%). Hex decreased the relative abundance of Asperigillus, Issatchenkia, and Penicillium for NFI silages on day 5; however, its inhibitory effects were not observed in FI silages on day 5. CONCLUSION: Fungal infestation was associated with poor fermentation and hygienic quality of corn silage. Adding Hex accelerated the pH decline and maintained the antifungal activity until 90 days of ensiling, attenuating adverse effects of fungal infestation on the fermentation and preventing the accumulation of mycotoxins in corn silages. © 2021 Society of Chemical Industry.

Fermentation profile and microbial diversity of temperate grass silage inoculated with epiphytic microbiota from tropical grasses.

This study was designed to evaluate the effects of epiphytic microbiota from Italian ryegrass (IRIR), Napier grass (IRNP) and Sudan grass (IRSD) on ensiling characteristics and microbial community of Italian ryegrass silage. Each treatment was prepared in triplicate and ensiled in plastic bag silos for 1, 3, 7, 14, 30 and 60 days. The γ-ray irradiation sterilization method, microbiota transplantation and next generation sequencing technology were used. Results indicated that significantly (P < 0.05) higher ratio of lactic acid to acetic acid, and lower acetic acid and ammonia nitrogen contents were observed in IRNP than IRIR and IRSD after 60 days of ensiling. Lactobacillus was the most predominant in each treatment at the late stage of fermentation. Lactococcus was eventually replaced by Lactobacillus in IRSD, whereas higher abundance of Lactococcus was continuously found in IRNP. Co-occurrence network analysis demonstrated Lactococcus was pivotal in determining the silage fermentation pattern of Italian ryegrass. According to the 16S rRNA gene-predicted functional profiles, the metabolism of amino acids was enhanced by the epiphytic microbiota from Italian ryegrass and Sudan grass, while the carbohydrate metabolism was accelerated by the epiphytic microbiota from Napier grass. Overall, IRNP had a homo-fermentative process, whereas IRIR and IRSD possessed a hetero-fermentative pattern. The Lactococcus and heterofermentative Lactobacillus were mainly responsible for this. It also confirmed that the exogenous microorganisms that promote the carbohydrate metabolism and inhibit the metabolism of amino acids could be a good potential source to improve the silage quality of temperate grass.

Effect of storage time on the fermentation quality, bacterial community structure and metabolic profiles of napiergrass (Pennisetum purpureum Schum.) silage.

This study was aimed to investigate the effect of storage time on fermentation characteristics, bacterial community structure and predicted metabolic pathways of napiergrass (Pennisetum purpureum Schum.) silage. First-cutting napiergrass was harvested at the vegetative stage and ensiled in laboratory-scale silos (1 L capacity). Triplicate silos were sampled after 1, 3, 7, 15, 30 and 60 days of ensiling, respectively. The bacterial communities on day 3 and 60 were assessed through the high throughput sequencing technology, and metabolic pathways of bacterial community were predicted according to the Kyoto Encyclopedia of Genes and Genomes (KEGG) via Tax4Fun. Napiergrass silage exhibited an acetic acid-type fermentation, indicated by lower lactic acid contents and ratio of lactic acid to acetic acid, and higher pH, ethanol and acetic acid contents. Before ensiling, the predominant genera in fresh napiergrass mainly included Acinetobacter, Enterobacteriaceae, Enterobacter and Lactococcus. After 60 days of ensiling, high proportions of Enterobacteriaceae, Enterobacter and Lactobacillus were found in napiergrass silages. The metabolism of amino acid, energy, cofactors and vitamins were inhibited, whereas metabolism of nucleotide and carbohydrate were promoted during ensiling. Overall, the combination of high throughput sequencing technology and 16S rRNA gene-predicted functional analyses revealed the differences during the initial and late stages of napiergrass silages not only for distinct bacterial community but also for specific functional metabolites. It could provide a comprehensive insight into bacterial community and functional profiles to further improve the silage quality.

Using a high-throughput sequencing technology to evaluate the various forage source epiphytic microbiota and their effect on fermentation quality and microbial diversity of Napier grass.

Napier grass (Pennisetum purpureum) is well-known due to its high biomass production. The epiphytic microbiota was prepared from Napier, alfalfa, and red clover grass and served as an inoculum. The chopped sterilized Napier grass was inoculated with reconstituted epiphytic microbiota, and treatments were designed as: distilled water (N0); Napier grass epiphytic microbiota (NP); alfalfa epiphytic microbiota (AL); and red clover epiphytic microbiota (RC). The results reveal that the reconstituted epiphytic microbiota bacteria efficiently adapted in Napier grass silage, improved fermentation, and produced lactic acid. The alfalfa-grass inoculum rapidly dropped pH and enhanced the lactic acid (LA) and the ratio of lactic acid-to-acetic acid (LA/AA) during the entire ensiling process. However, red clover attains high lactic acid, while Napier grass produces high acetic acid-type fermentation at terminal silage. After day 60 of ensiling, Lactobacillus proportion was higher in AL (85.45%), and RC (59.44%), inocula as compared with NP (36.41%), inoculum. The NP inoculum terminal silage was diverse than AL and RC inocula and dominated by Enterobacter (16.32%) and Enterobacteriaceae (10.16%) and also significantly (p < 0.05) higher in acetic acid. The present study concluded that AL and RC epiphytic microbiota successfully develop and more efficient than Napier grass microbiota. It is suggested that abundant microbiota isolate from alfala and red clover and develop more economical and efficient inocula for quality fermentation of Napier grass silage in practice.