Lactic Acid Bacteria Inoculants Research Articles

A meta-analysis to observe silage microbiome differentiated by the use of inoculant and type of raw material.

Silage fermentation is naturally carried out by lactic acid bacteria (LAB) to mainly produce lactic acid (LA) and other organic acids as preservatives. Along with fermentation time, the growth of LAB will replace and suppress undesirable microorganisms. This meta-analysis study aimed to explore silage microbiome differentiated by LAB inoculants and type of raw materials. A total of 37 articles with 185 studies and 475 datasets were used for building up the meta-database. Data were subjected to the mixed model methodology. The parameters observed were silage quality and silage microbiome post-ensiling process. Results revealed that four bacterial genera along with Weissella dominated the post-ensiling process. The addition of lactic acid inoculants in the silage has increased the abundance of Lactobacillus spp. and decreased the Shannon index significantly. Moreover, the abundance of both L. plantarum and L. buchneri increased, and subsequently, Weissella, Pseudomonas, Proteobacteria, pH value, ammoniacal nitrogen (NH3-N), coliforms, and the yeasts were decreased significantly due to the addition of LAB inoculants in silage (p < 0.05). Environmental factors such as temperature affected the existence of Pseudomonas, Exiguobacterium, and Acinetobacter. However, the dry matter, LA, acetic acid (AA), the ratio of LA to AA, and the LAB population were enhanced significantly (p < 0.05). Among the LAB types, the lowest abundance of Pseudomonas was due to the LAB group, while the lowest abundance of Weissella and Proteobacteria was due to the addition of the combined LAB group. In conclusion, the addition of LAB is effectively enhancing the silage microbiome and silage quality by altering bacterial diversity and the metabolic products of the silage materials for safe preservation.

Effects of Different Types of LAB on Dynamic Fermentation Quality and Microbial Community of Native Grass Silage during Anaerobic Fermentation and Aerobic Exposure

Silage of native grasses can alleviate seasonal forage supply imbalance in pastures and provide additional sources to meet forage demand. The study aimed to investigate the effects of Lactobacillus plantarum (LP), Lactobacillus buchneri (LB), and Lactobacillus plantarum in combination with Lactobacillus buchneri (PB) on the nutritional quality, fermentation quality, and microbial community of native grass silage at 2, 7, 15, and 60 days after ensiling and at 4 and 8 days after aerobic exposure. The results showed that dry matter content, crude protein content, the number of lactic acid bacteria, and lactic acid and acetic acid content increased and pH and ammonia nitrogen content decreased after lactic acid bacteria (LAB) inoculation compared with the control group (CK). LP had the lowest pH and highest lactic acid content but did not have greater aerobic stability. LB maintained a lower pH level and acetic acid remained at a higher level after aerobic exposure; aerobic bacteria, coliform bacteria, yeast, and molds all decreased in number, which effectively improved aerobic stability. The effect of the compound addition of LAB was in between the two other treatments, having higher crude protein content, lactic acid and acetic acid content, lower pH, and ammonia nitrogen content. At the phylum level, the dominant phylum changed from Proteobacteria to Firmicutes after ensiling, and at the genus level, Lactiplantibacillus and Lentilactobacillus were the dominant genera in both LAB added groups, while Limosilactobacillus was the dominant genus in the CK treatment. In conclusion, the addition of LAB can improve native grass silage quality by changing bacterial community structure. LP is beneficial to improve the fermentation quality in the ensiling stage, LB is beneficial to inhibit silage deterioration in the aerobic exposure stage, and compound LAB addition is more beneficial to be applied in native grass silage.

Effects of small-scale silo types and additives on silage fermentation and bacterial community of high moisture alfalfa on the Qinghai-Tibetan Plateau

Alfalfa is cultivated throughout the world, but its preservation remains challenging to small farms in cold climates. This study aimed to investigate the effects of small-scale silo types and additives on the silage quality of high moisture alfalfa on the Qinghai-Tibetan Plateau. Alfalfa with moisture of about 80% was ensiled in small farm bucket silos (FBS) or wrapped bale silos (WBS) at ambient temperature (5–15 °C) for 180 days. At ensiling, forage was treated without any additive (CK) or with a chemical additive (SB; a mixture of sodium benzoate, potassium sorbate, and sodium nitrite) or a lactic acid bacteria (LAB) inoculant (PBB; a mixture of Lactiplantibacillus plantarum, Bifidobacterium brevis and Pediococcus pentosaceus). Among the CK silages, higher (P < 0.05) acetic acid levels, bacterial diversity indices and relative abundance of L. plantarum were found in WBS vs FBS. Overall, silage treated with SB had the highest crude protein (CP) content, the lowest amylase-treated neutral detergent fiber (aNDF) and acid detergent fiber (ADF) contents and dry matter (DM) loss. Compared with CK, treatments with SB and PBB increased (P < 0.05) lactic acid content and decreased (P < 0.05) final pH value and butyric acid content of silage. In addition, treatment with PBB reduced (P < 0.05) the relative abundance of Lactilactobacillus sakei and increased (P < 0.05) the similarity of silage samples from different silos. There was negligible difference in fermentation quality of silage occurred between the types of silos. However, small WBS tended to increase the relative abundance of L. plantarum for more desirable silage fermentation. Application of additives at ensiling enhanced the preservation of silage nutrients with low DM loss. Based on the above results, small WBS were recommended for producing high-quality alfalfa silage on the Qinghai-Tibetan Plateau.

Metagenomics insights into the effects of lactic acid bacteria inoculation on the biological reduction of antibiotic resistance genes in alfalfa silage

Antibiotic resistance genes (ARGs) are a new type of pollutant and pose major threats to public health. However, the distribution and transmission risk of ARGs in alfalfa silage as the main forage for ruminants have not been studied. This study first deciphered the effects of Lactobacillus plantarum MTD/1 or Lactobacillus buchneri 40788 inoculations on distribution and transmission mechanism of ARGs in alfalfa silage by metagenomics. Results showed that multidrug and bacitracin resistance genes were the dominant ARGs in ensiled alfalfa. The natural ensiling process increased the abundances of bacitracin, beta_lactam, and aminoglycoside in alfalfa silage with 30% DM, and vancomycin in alfalfa silage with 40% DM. Meanwhile, prolonged wilting increased ARG enrichment in fresh alfalfa. Interestingly, alfalfa silage inoculated with L. plantarum MTD/1 or L. buchneri 40788 reduced the abundances of total ARG, and multidrug, MLS, vancomycin, aminoglycoside, tetracycline, and fosmidomycin resistance genes by reductions of the host bacteria and the enrichment of ARGs located in the plasmid. The hosts of ARG in alfalfa silage were mainly derived from harmful bacteria or pathogens, and some of the clinical ARGs were observed in alfalfa silage. Basically, the combined effect of microbes, MGEs, and fermentation quality was the major driver of ARG transfer and dissemination in microecosystem of ensiling, where the microbes appeared to be the crucial factor. In summary, inoculation with the present lactic acid bacteria could reduce ARG abundance in ensiled alfalfa, and a better effect was observed in L. plantarum-treated silage than in L. buchneri treated silage.

Effect of different dry matter content on fermentation characteristics and nutritional quality of Napier grass silage with novel lactic acid bacteria strains.

To investigate the characteristics of different LAB strains isolated from subtropics and their effects on Napier grass (Pennisetum purpureum Schum.) silage with two dry matter (DM) levels, sugar fermentation pattern, and growth profiles of three screened lactic acid bacteria (LAB) strains [Pediococcus pentosaceus (PP04), Weissella cibaria (WC10), and Lactobacillus plantarum (LP694)] were characterized, and then used either individually or in combination at 1.0×106 cfu g-1 fresh weight to inoculate grass having 15% or 25% DM. Treatments were applied: (1) no inoculant (control); (2) PP04; (3) WC10; (4) LP694; (5) M-1 (PP04: WC10=2:1); (6) M-2 (PP04: LP694=1:2); (7) M-3 (WC10: LP694=2:1); (8) M-4 (PP04: WC10: LP694=2:1:1). The results showed that all inoculations increased LAB, DM recovery, and lactic acid (LA) concentration, while decreasing pH, the ammonia nitrogen/total nitrogen (NH3-N/TN), and butyric acid (BA) concentration compared to control group in both DM. However, the effect of inoculations was very limited at 15% DM. Silages with inoculants achieved higher silage quality at 25% DM than 15% DM. The different LAB inoculants result in significant differences in silage quality, while W. cibaria decreased the pH and inhibited the growth of undesirable bacteria and those characteristics were not affected by the DM content.

Response of fermentation quality and microbial community of oat silage to homofermentative lactic acid bacteria inoculation.

Oat (Avena sativa L.) is one of the important forage crops in the world. However, oat grown in Southwest China has higher moisture content and their preservation face significant challenges. In addition, existing commercial lactic acid bacteria (LAB) have poor fermentation effects in hot and humid regions. Consequently, the current study investigated the response of oat fermentation quality and microbial community to self-selected LAB inoculation. The treatments were: CK, sterilized water; LP694, Lactobacillus plantarum 694; LR753, Lactobacillus rhamnosus 753; and LPLR, LP694 combined with LR753, followed by 1, 3, 7, 14, and 60 days (d) of fermentation. The results showed that LAB inoculation significantly raised the lactic acid content, and decreased the level of pH value, acetic acid, and ammonia-N in oat silage. The LR753 group had a significantly higher (p < 0.05) lactic acid content (60.95 g kg-1 DM), and lower pH value (3.95) and ammonia-N content (10.1 g kg-1 DM) followed by the LPLR group. The LR753 showed lower NDF (54.60% DM) and ADF (39.73% DM) contents than other groups. The Lactobacillus was a prevalent genus in LAB-treated groups, and its relative abundance reached maximum in LP694 (69%) on day 3, while in the LR753 group (72%) on 60 days. The Lactobacillus rhamnosus, Lactobacillus plantarum, and Lactobacillus fermentum became the dominant species in LAB-treated groups with fermentation time. The Lactobacillus genus was positively correlated with WSC (R = 0.6, p < 0.05), while negatively correlated with pH (R = -0.5, p < 0.05), and BA (R = -0.5, p < 0.01). Overall, the LR753 group had better fermentation quality and preservation of nutritional components providing theoretical support and guidance for future oat silage production in Southwest China.

Metagenomics analysis reveals the performance of homo- and heterofermentative lactic acid bacteria in alfalfa silage fermentation, bacterial community, and functional profiles.

Alfalfa (Medicago sativa L.) is a kind of roughage frequently utilized as an animal feed but challenging to be ensiled due to its low water-soluble carbohydrate (WSC), high water content, and elevated buffering capacity, thus requiring the application of lactic acid bacteria (LAB) to improve its fermentation. This study employed high-throughput metagenomic sequence technology to reveal the effects of homofermentative LAB, Lactobacillus plantarum (Lp), or Pediococcus pentosaceus (Pp), and heterofermentative LAB, L. buchneri (Lb), or their combinations (LbLp or LbPp) (applied at 1.0 × 109 colony forming units (cfu) per kilogram of alfalfa biomass fresh material) on the fermentation, microbial community, and functional profiles of alfalfa silage after 7, 14, 30, and 60 ensiling days. The results indicated a reduction (P < 0.05) in glucose and pH and higher (P < 0.05) beneficial organic acid contents, xylose, crude protein, ammonia nitrogen, and aerobic stability in Lb-, LbPp-, and LbLp-inoculated alfalfa silages after 30 and 60 d. Also, higher (P < 0.05) WSC contents were recorded in LbLp-inoculated alfalfa silages after 30 d (10.84 g/kg dry matter [DM]) and 60 d (10.92 g/kg DM). Besides, LbLp-inoculated alfalfa silages recorded higher (P < 0.05) LAB count (9.92 log10 cfu/g) after 60 d. Furthermore, a positive correlation was found between the combined LAB inoculants in LbLp-inoculated alfalfa silages and dominant LAB genera, Lactobacillus and Pediococcus, with fermentation properties after 30 and 60 d. In addition, the 16S rRNA gene-predicted functional analyses further showed that the L. buchneri PC-C1 and L. plantarum YC1-1-4B combination improved carbohydrate metabolism and facilitated further degradation of polysaccharides in alfalfa after 60 d of ensiling. These findings reveal the significant performance of L. buchneri and L. plantarum in combination with dominant LAB species in suppressing the growth of Clostridia, molds, and yeasts and improving the fermentation characteristics and functional carbohydrate metabolism of alfalfa after 60 d ensiling, thus suggesting the need for further studies to uncover the diverse performance of the LAB combination and their consortium with other natural and artificial inoculants in various kinds of silages.

Fermentation quality and bacterial ecology of red clover dominated silage modulated by different management factors

Identification of bacterial communities in both the raw material and the subsequent silages provides new insights into understanding the silage fermentation process. The objective was to evaluate how different silage management factors affect silage preservation characteristics, the microbiome, and their correlations. A red clover dominated sward was used as the raw material and ensiled in pilot scale using a 3 × 4 design, with three management conditions including the level of compaction (loose and tight), and further, for the tightly compacted silages contamination with soil and faeces was conducted; and four different additive treatments with different modes of action: Control without additive (CONT), formic and propionic acid-based additive (FPA), homofermentative lactic acid bacteria inoculant (LAB) and salt-based additive (SALT). Samples of the raw material and subsequent silages were taken and routinely analysed, including DNA extraction and PCR amplification using universal primers. Tight compaction reduced slightly the extent of silage fermentation, but contamination with soil and faeces stimulated a non-desired type of fermentation with higher concentrations of ethanol, acetic acid and propionic acid and a higher pH. Use of LAB and SALT had only minimal effects on silage fermentation, but FPA clearly restricted fermentation and resulted in a better fermentation quality (lower pH, ammonia N and acetic acid concentration) of the silages compared to CONT. The FPA silages presented greater diversity of bacterial communities compared to the other silages. Proteobacteria were the most abundant in raw material, followed by Firmicutes, and major shifts happened in these communities during the silage fermentation process. Weissella was found in small amounts in the raw material but it dominated in the silages. The most abundant communities in the silages, such as Weissella, Lactobacillus and Pseudomonas, were correlated with several silage fermentation characteristics. Use of FPA improved fermentation quality of silages, but SALT and LAB differed from CONT to a smaller extent. All additives modified the bacterial profiles of grasses ensiled under different management conditions. The combination of parameters related to silage quality and bacterial communities provided a deeper understanding of the silage fermentation process and how they can be manipulated to obtain better feed quality.

Fermentation Characteristics, Microbial Compositions, and Predicted Functional Profiles of Forage Oat Ensiled with Lactiplantibacillus plantarum or Lentilactobacillus buchneri

This study aimed to investigate the effects of lactic acid bacteria (LAB) inoculants on the fermentation quality, microbial compositions, and predicted functional profiles of forage oat. The forage oat was inoculated with distilled water, Lentilactobacillus buchneri (LB), and Lactiplantibacillus plantarum (LP) as the control (CON), LB and LP treatments, respectively, and the addition of Lentilactobacillus buchneri (LB) or Lactiplantibacillus plantarum (LP) resulted in 1 × 106 colony-forming units/g of fresh weight. After 30 days of fermentation, the lowest pH (4.23) and the lowest content of ammoniacal nitrogen (NH3-N) in dry matter (DM, 4.39%) were observed in the LP treatment. Interestingly, there was a significant (p &lt; 0.05) difference in lactic acid (LA) concentration among the three treatments. The LP treatment had the highest lactate concentration (7.49% DM). At the same time, a markedly (p &lt; 0.05) elevated acetic acid (AA) concentration (2.48% DM) was detected in the LB treatment. The Shannon and Chao1 indexes of bacterial and fungal communities in all the silage samples decreased compared to those in the fresh materials (FM). Proteobacteria was the dominant phylum in the FM group and shifted from Proteobacteria to Firmicutes after ensiling. Lactobacillus (64.87%) and Weissella (18.93%) were the predominant genera in the CON, whereas Lactobacillus dominated the fermentation process in the LB (94.65%) and LP (99.60%) treatments. For the fungal community structure, the major genus was Apiotrichum (21.65% and 60.66%) in the FM and CON groups after 30 days of fermentation. Apiotrichum was the most predominant in the LB and LP treatments, accounting for 52.54% and 34.47%, respectively. The genera Lactococcus, Pediococcus, and Weissella were negatively associated with the LA content. The genus Ustilago and Bulleromyces were positively associated with the LA content. These results suggest that the addition of LAB regulated the microbial community in oat silage, which influenced the ensiling products, and LP was more beneficial for decreasing the pH and NH3-N and increasing the LA concentration than LB in forage oat silage.

Effects of Lactic Acid Bacteria Inoculants and Stage-Increased Storage Temperature on Silage Fermentation of Oat on the Qinghai–Tibet Plateau

Ensiling is a simple and effective method of alleviating a shortage of forage for ruminants. This study aimed to investigate the effects of lactic acid bacteria (LAB) inoculants and stage-increased temperature on the fermentation characteristics and chemical composition of oat silage on the Qinghai–Tibet Plateau. The silage was treated with local laboratory inoculant (I) and commercial inoculant (S) and stored at ambient temperature (&lt;10 °C) or stage-increased (5, 10 and 15 days) temperatures of 10 °C and 15 °C for 60 days. The results showed that stage-increased storage temperature can improve silage fermentation. Compared with 10 °C, a stage-increased storage temperature of 15 °C effectively (p &lt; 0.05) promoted the fermentation rate of silage by increasing the dominance of Lactiplantibacillus plantarum, with higher lactic, acetic and propionic acid contents and a lower ammonia-N ratio of the total N and final pH value. Compared with S, treatment with I increased the water-soluble carbohydrate and lactic acid contents and decreased the ammonia-N ratio of the total N and final pH value. This work demonstrated that increasing the storage temperature in stages using a warming infrastructure facilitates the preservation of oat silage in cold regions, and the inoculation of lactic acid bacteria could advance silage fermentation on the Qinghai–Tibet Plateau.

Effect of Homo-Fermentative Lactic Acid Bacteria Inoculants on Fermentation Characteristics and Bacterial and Fungal Communities in Alfalfa Silage

We evaluated the effects of a homo-fermentative lactic acid bacteria (homo-LAB) inoculant on the fermentation and microbial communities of alfalfa ensiled at two dry matter (DM) contents of 38 and 46% DM. At both DMs, alfalfa was treated or not with an inoculant containing Pediococcus acidilactici, Enterococcus faecium and Lactobacillus plantarum at a targeted application rate of 165,000 cfu/g of fresh weight and stored for 3, 30 and 60 days. Treatment with the inoculant resulted in a lower drop in pH and, in general, higher lactic acid and lower acetic acid when applied to medium DM silage. For the four most abundant microbial genera, increased abundances of Bacteroides and Lactobacillus (p &lt; 0.05), as well as decreased abundances of Muribaculaceae were observed in high DM and inoculated silages. The abundance of Prevotellaceae-UCG-001 was lower in medium DM control silages than in high DM control silages. Inoculation and DM affected abundances of Vishniacozyma (p &lt; 0.05). Increased abundances of Vishniacozyma, as well as decreased abundances of Leucosporidium were observed in medium DM-inoculated silages. Changes in the relative abundance (RA) of the main populations of bacteria and yeasts did explain the fermentation and nutrition differences among treatments.

Recycling of Citric Acid Waste for Potential Use as Animal Feed through Fermentation with Lactic Acid Bacteria and a Mixture of Fibrolytic Enzymes.

Once improperly managed, the citric acid production industry generates waste, which contributes to pollution and other environmental issues. We proposed that, with sufficient quality improvement, citric acid by-product (CAP) might be used for animal feed, thereby reducing the environmental impact. The aim of the present study was to ferment citric acid by-product (CAP) by inoculation with lactic acid bacteria (LAB) and a fibrolytic enzyme mixture for quality improvement and crude fiber reduction in the waste products. LAB inoculants were L. casei TH14, and the additive enzyme used was a fibrolytic enzyme mixture (glucanase, pectinase, and carboxymethylcellulase) of a small-scale fermentation method. The seven treatments employed in this study were as follows: (1) control (untreated), (2) CAP-inoculated L. casei TH14 at 0.01% DM, (3) CAP-inoculated L. casei TH14 at 0.05% DM, (4) CAP-inoculated enzymes at 0.01% DM, (5) CAP-inoculated enzymes at 0.05% DM, (6) CAP-inoculated L. casei TH14 at 0.01% DM with enzymes at 0.01% DM, and (7) CAP-inoculated L. casei TH14 at 0.05% DM with enzymes at 0.05% DM. The samples were taken on days 1, 7, 14, 21, and 28 of ensiling, both before and after. Four replications were used. The results of the chemical composition of the CAP before and after ensilage inoculated with L. casei TH14 did not show any differences in crude protein, ether extract, ash, or gross energy, but the enzymes significantly (p &lt; 0.05) decreased crude fiber and increased nitrogen-free extract. The combination was especially effective at improving the characteristics of CAP, with a reduction in crude fiber from 21.98% to 22.69%, of neutral detergent fiber (NDF) from 16.01% to 17.54%, and of acid detergent fiber (ADF) from 13.75% to 16.19%. Furthermore, the combination of L. casei TH14 and the enzyme increased crude protein from 1.75% to 2.24% at 28 days of ensiling. Therefore, CAP-inoculated L. casei TH14 did not change in chemical composition, while crude fiber, NDF, and ADF decreased when CAP was inoculated with enzyme. The combination of L. casei TH14 and the enzyme is more effective at improving chemical composition and reducing crude fiber and enhancing carbohydrate breakdown in the CAP. Finally, by enhancing the CAP's quality, it may be possible to use it in animal feed and minimize its impact on the environment.

Dry matter recovery, ensiling characteristics and aerobic stability of oat silage treated with microbial inoculants at different temperatures

ABSTRACT To evaluate the effects of temperature and lactic acid bacteria (LAB) inoculants on oat silage in Loess Plateau of China, oat was harvested at dough stage, inoculated without (Control) or with LAB inoculants Synlac I (SLI, Lactobacillus plantarum and Pedioccocus acidilactici) and a selected strain HT1 (L. rhamnosus) and ensiled at 25°C (T25), 35°C (T35) and 45°C (T45). The fermentation quality was measured after 60 d of ensiling and the aerobic exposure was conducted at 30°C for 9 d. The results showed that control silage (stored at 25°C) had better fermentation quality than that ensiled at 35°C or 45°C. High temperature of 45°C resulted in sharp decreases in LAB counts and lactic acid concentration and increases in pH and NH3-N concentration in the control group. Inoculation improved the fermentation quality, and HT1 was more effective than SLI at 35°C and 45°C, while SLI showed better performance at 25°C. All silages displayed mild fluctuation for all treatments at the first 3 d of aerobic exposure, and significant differences were observed among treatments after that. Both control and inoculated silages stored at 25°C showed a sharp pH increase, while HT1 treated silages stored at 35°C and 45°C maintained stable pH and better fermentation quality during the aerobic exposure. In conclusion, SLI was suitable for oat silage fermentation at normal atmospheric temperature (25°C), while HT1 was more effective in improving DM recovery, fermentation quality and aerobic stability of oat silage at high temperature during summer in the Loess Plateau of China.

Effects of Different Lactic Acid Bacteria Inoculants on Alfalfa Silage Fermentation and Quality

Alfalfa (Medicago sativa L.) is a crucial perennial forage plant with its high protein and mineral content and may be mowed several times through the vegetation period. Along with having a large cultivation area in Türkiye, it constitutes approximately 61% of the total green forage produced. Silage is the best method for keeping grass well and using it as a source of roughage in all seasons. However, ensiling alfalfa, especially with low dry matter content, is very difficult due to its low water-soluble carbohydrate and buffering capacity. This study was carried out to improve the alfalfa plant's silage fermentation process by inoculating new lactic acid bacteria strains. When the alfalfa plant reached 50% flowering, six different lactic acid bacteria strains were inoculated and compared with the uninoculated alfalfa silage. According to the results obtained, it was determined that lactic acid bacteria inoculants improved the fermentation properties of alfalfa silage in general. All inoculated strains caused a significant decrease in pH of the resulting silage. The strain Lactobacillus buchneri (LS-31-1-4) was superior in terms of much dry matter recovery (96.82%) and protein recovery (94.00%). At the same time, Lactobacillus brevis (LS-55-2-2) and Leuconostoc citerum (LS-70-6-1) were the most restrictive strains to yeast and enterobacteria growth in silage, respectively.

Effect of Autochthonous Lactic Acid Bacteria-Enhanced Fermentation on the Quality of Suancai.

The lactic acid bacteria (LABs) used for fermentation have an extremely vital impact on the quality of Suancai, a fermented vegetable. The bacterial diversity and metabolites of inoculated Suancai with LABs, including Lactiplantibacillus plantarum (Lb. plantarum), Levilactabacillus brevis (Lb. brevis), and Leuconostoc mesenteroides (Leu. mesenteroides), were investigated. The inoculation of LABs significantly decreased the pH and the content of nitrite. The Suancai inoculated with LABs had a higher content of the total titratable acidity (TTA) and organic acids than spontaneous fermentation. The LABs inoculation significantly influenced the bacterial community structures, which directly or indirectly caused changes of metabolites. The bacterial community profiles of Suancai inoculated with Lb. plantarum were more similar to spontaneous fermentation. The inoculation of Lb. plantarum, Lb. brevis, and Leu. mesenteroides could increase its abundance in Suancai. Whatever the species inoculated, Lb. plantarum was always the predominant bacterium in Suancai after fermentation. The inoculated LABs were positively correlated with most volatile compounds and amino acids. The inoculated LABs significantly improved the volatile compounds and amino acid content of Suancai. This study could contribute to understanding the function of starters in Suancai fermentation and promote the selection of applicable starters for high-quality Suancai production.

The Use of Removed Mesquite Brush as a Fiber Replacement in Silage Production.

Simple SummaryMesquite is considered an invasive browse species in most of the American Southwest, spreading rapidly into Texas and leading to disruption of native grasslands. Management efforts generate a significant volume of organic debris. Incorporation of this organic material into livestock feeding efforts would represent a sustainable solution to this ecological problem. Our objectives were to (1) determine the nutritive value and fermentation characteristics of silage produced with mesquite biomass, and (2) evaluate potential pre-treatment methods of mesquite biomass prior to ensiling. Inclusion of mesquite biomass in a bermudagrass-based silage was shown to increase fiber concentrations, decrease crude protein and in vitro digestibility, and decrease fermentation quality (pH and volatile fatty acids [VFA]). However, 250 g kg−1 inclusion of mesquite was shown to be similar to grass silage alone. There was no effect of lactic acid bacteria inoculation, though increasing length of incubation did increase VFA production and decrease silage pH. Pre-treatment of mesquite biomass with acid or alkali did not improve ensiling properties. Results are interpreted to mean that mesquite biomass may be effectively incorporated into silage at levels up to 250 g kg−1.Mesquite (Prosopis L.) is considered an invasive browse species in most of the American Southwest. Mechanical intervention removes yields an excess of organic debris. Anecdotal evidence in the past has supported using such browse as feed for livestock. Thus, our objectives were to (1) determine the nutritive value and fermentation characteristics of silage produced with mesquite biomass, and (2) evaluate solvent treatment of mesquite biomass prior to ensiling. In Experiment 1, we evaluated mesquite inclusion rate (0, 250, 500, 750, or 1000 g kg−1 DM), length of fermentation (28, 56, or 84 d), and silage inoculant. In Experiment 2, we evaluated the effects of mesquite pre-treatment with acid (H2SO4) or alkali (NaOH) solutions. Concentrations of NDF, ADF, and ADL, as well as IVTD, decreased (p < 0.05) with increasing mesquite inclusion. However, 250 g mesquite kg−1 DM did not differ from grass silage. There was no effect (p > 0.05) of inoculation, though increasing length of incubation did increase (p < 0.05) VFA production and decrease (p < 0.05) silage pH. Solvent treatment did not improve ensiling properties. Results are interpreted to mean that mesquite biomass may be effectively incorporated into silage at levels up to 250 g kg−1.

Ensiling pretreatment fortified with laccase and microbial inoculants enhances biomass preservation and bioethanol production of alfalfa stems

This study investigated the potential of ensiling pretreatment fortified with laccase and a lactic acid bacteria (LAB) inoculant on improving the utilization of alfalfa stems for bioethanol production. The alfalfa stems were ensiled with no additives (Con), 0.04 % laccase (LA), a LAB inoculant containing Pediococcus pentosaceus at 1 × 106 fresh weight (FW) and Pediococcus acidilactici at 3 × 105 cfu/g FW (PP), and a combination of LA and PP (LAP) for 120 days. By reshaping the bacterial community structure of alfalfa stem silages toward a higher abundance of Lactobacillus, the addition of laccase and LAB inoculant either alone or in combination facilitated lactic acid fermentation to reduce fermentation losses, as evidenced by low concentrations of ammonia nitrogen (53.7 to 68.9 g/kg total nitrogen) and ethanol (2.63 to 3.55 g/kg dry matter). All additive treatments increased lignocellulose degradation and soluble sugars concentrations of alfalfa stem silages. Due to delignification and polyphenol removal, glucan and xylan conversion (70.3 % vs. 35.7 % and 51.6 % vs. 27.9 %, respectively) and ethanol conversion efficiency (53.9 % vs. 26.4 %) of alfalfa stems were greatly increased by ensiling fortified with LA versus Con, and these variables (79.8 % for glucan, 58.7 % for xylan, and 60.1 % for ethanol conversion efficiency) were further enhanced with a synergistic effect of LA and PP fortification. The spearman correlation analysis revealed that bioethanol fermentation of silage biomass was closely related to ensiling parameters and total phenols. In conclusion, ensiling pretreatment with LA and PP combination offered a feasible way to efficient utilization of alfalfa stems for bioethanol production.

Dynamic Changes in Fermentation Quality and Structure and Function of the Microbiome during Mixed Silage of Sesbania cannabina and Sweet Sorghum Grown on Saline-Alkaline Land.

ABSTRACTProtein-rich Sesbania cannabina and sugar-rich sweet sorghum [Sorghum dochna (Forssk.) Snowden] are characterized by their higher tolerance to saline-alkaline stresses and simultaneous harvests. They could be utilized for coensiling because of their nutritional advantages, which are crucial to compensate protein-rich forage in saline-alkaline regions. The current study investigated the fermentation quality, microbial community succession, and predicted microbial functions of Sesbania cannabina and sweet sorghum in mixed silage during the fermentation process. Before ensiling, the mixtures were treated with compound lactic acid bacteria (LAB) inoculants followed by 3, 7, 14, 30, and 60 days of fermentation. The results revealed that the inoculated homofermentative species Lactobacillus plantarum and Lactobacillus farciminis dominated the early phase of fermentation, and these shifted to the heterofermentative species Lactobacillus buchneri and Lactobacillus hilgardii in the later phase of fermentation. As a result, the pH of the mixed silages decreased significantly, accompanied by the growth of acid-producing microorganisms, especially L. buchneri and L. hilgardii, which actively influenced the bacterial community structure and metabolic pathways. Moreover, the contents of lactic acid, acetic acid, 1,2-propanediol, and water-soluble carbohydrates increased, while the contents of ammonia-N and fiber were decreased, with increasing ratios of sweet sorghum in the mixed silage. Overall, coensiling Sesbania cannabina with >30% sweet sorghum is feasible to attain high-quality silage, and the relay action between homofermentative and heterofermentative LAB species could enhance fermentation quality and conserve the nutrients of the mixed silage.IMPORTANCE The coensiling of Sesbania cannabina and sweet sorghum is of great practical importance in order to alleviate the protein-rich forage deficiency in saline-alkaline regions. Furthermore, understanding the microbial community’s dynamic changes, interactions, and metabolic pathways during ensiling will provide the theoretical basis to effectively regulate silage fermentation. Here, we established that coensiling Sesbania cannabina with >30% sweet sorghum was effective at ensuring better fermentation quality and preservation of nutrients. Moreover, the different fermentation types of LAB strains played a relay role during the fermentation process. The homofermentative species L. plantarum and L. farciminis dominated in the early phase of fermentation, while the heterofermentative species L. buchneri and L. hilgardii dominated in the later phase of fermentation. Their relay action in Sesbania cannabina-sweet sorghum mixed silage may help to improve fermentation quality and nutrient preservation.

Effects of different additives on the bacterial community and fermentation mode of whole-plant paper mulberry silage.

The purpose of this study was to investigate the effects of inoculation with two lactic acid bacteria (LAB) strains (Lacticaseibacillus rhamnosus and Lentilactobacillus buchneri) and the addition of four corn flour proportions (0, 3, 6, and 9%) in different treatments, on the composition and function of the bacterial community in whole-plant paper mulberry silage. The different treatments promoted Lactiplantibacillus, Lentilactobacillus, and Lacticaseibacillus growth, but the microbial species responsible for fermentation differed among the treatments. High species diversity and various Gram-negative bacteria, such as Flavitalea sp., Pantoea agglomerans, Acinetobacter pittii, Turicibacter sanguinis, and Ralstonia pickettii, were found in the uninoculated LAB treatments. A beneficial bacterium, Lactobacillus johnsonii, was discovered for the first time in whole-plant paper mulberry silage. LAB inoculation simplifies the microbial community structure, and beneficial Lactobacillus as a key species aggregates in the inoculated treatment group. However, L. rhamnosus inoculation alone may have limited bacteriostatic activity against in whole-plant paper mulberry silage. Compared with silage lacking corn flour, amino sugar and nucleotide sugar metabolism, galactose metabolism, the phosphotransferase system and the pentose phosphate pathway metabolic activity were increased in corn flour-containing silage. Whole-plant paper mulberry can be used as a high-quality silage to provide high-quality feed resources for sustainable ruminant livestock production. Moreover, additive use is necessary for preparing paper mulberry silage.

Changes in Composition and Diversity of Epiphytic Microorganisms on Field Pea Seeds, Partial Crop Peas, and Whole Crop Peas during Maturation and Ensiling with or without Lactic Acid Bacteria Inoculant.

ABSTRACTThe present study was conducted under the hypothesis that, in field peas, type of plant material, stage of maturity, ensiling, silage additive, and aerobic stress affect the composition and diversity of epiphytic microbial communities. Epiphytic microbial composition and diversity of pea seeds, partial crop peas, and whole crop peas was analyzed at different stages of late maturity, before and after ensiling, and with or without the use of lactic acid bacteria (LAB) as inoculant. Suitable combinations among pea crop variants, maturity stages, and inoculant use for the production of stable silages with sufficient aerobic stability after opening and during feed-out were identified. Genomic DNA was extracted, and 16S and 18S rRNA gene amplicons were sequenced. To assess the quality of the various silages, nutrient concentration, pH value, concentration of lactic acid, short chain fatty acids, and alcohols, and aerobic stability were determined. Pea seeds were barely colonized by epiphytic microorganisms. In partial and whole crop peas, composition and α-diversity (Shannon index) of bacterial communities did not differ between crop variants but differed among maturity stages. Epiphytic eukaryotes were rarely found on partial and whole crop peas. Bacterial composition and α-diversity were affected by ensiling and subsequent aerobic storage. In partial and whole crop peas, plant maturation caused an increase of the relative abundance of naturally occurring LAB (Weissella, Pediococcus, and Lactobacillus spp.). As a possible result, natural LAB support stable ensiling conditions even without the use of inoculants beginning with a maturity of 78 on the BBCH scale. This corresponded with a dry matter (DM) concentration of 341 and 363 g/kg in partial and whole crop peas, respectively. Addition of LAB inoculants, however, reduced ammonia, acetic acid, and butanol concentrations, and supported aerobic stability. Earlier stages of plant maturity (BBCH 76 and 77, 300 g DM/kg or less) were more prone to microbial spoilage. Stable pea seed silages can be produced at a maturity between BBCH 78 (427 g DM/kg) and 79 (549 g DM/kg), but they undoubtedly require LAB inoculation or application of other ensiling agents.IMPORTANCE Field peas are important protein suppliers for human and animal nutrition. They can be grown in many areas of the world, which may reduce imports of protein plants and has beneficial economic and ecological effects. Ensiling is a method of preserving feed that can be implemented easily and cost-effectively at the farm. Peas harvested as seeds, partial crop, or whole crop at different maturities enable a wide range of applications. The study characterized epiphytic microbial communities on peas in terms of composition and diversity depending on the maturity of the plants and feed conservation by ensiling as they play an essential role for the production of silages. Even if this study did not consider year, site, or cultivar effects, the results would show which part of the plant is probably well suited for the production of stable and high-quality silages and at which stage of maturity.