Selective Thermotolerant Lactic Acid Bacteria Isolated From Fermented Juice of Epiphytic Lactic Acid Bacteria and Their Effects on Fermentation Quality of Stylo Silages.

The aim of the present study was to isolate and identify lactic acid bacteria (LAB) from fermented juice of tropical crops such as Napier grass, Ruzi grass, Purple guinea grass, Stylo legume, and Leucaena and their application to improve the quality of tropical crop silage. Fifteen strains of LAB were isolated. The LAB strains were Gram-positive and catalase-negative bacteria and could be divided into three groups, i.e., Pediococcus pentosaceus, Lactiplantibacillus (para)plantarum, and Limosilactobacillus fermentum according to the biochemical API 50CH test. Based on the analysis of 16S rRNA sequence, the strains isolated in the group L. (para)plantarum were distinguished. Two isolates (N3 and G4) were identified as Lactiplantibacillus plantarum. Three isolates (St1, St2, and St3) were identified as L. paraplantarum. In addition, the identification of other isolates was confirmed in the group P. pentosaceus (R1, R4, R5, R8, R11, and L1) and the group L. fermentum (N4, G6, G7, and N4). All selected strains were able to grow at 50°C. All LAB strains showed antimicrobial activity against Escherichia coli ATCC 25922, Shigella sonnei ATCC 25931, Pseudomonas aeruginosa ATCC 27853, and Bacillus cereus ATCC 11778. Four selected LAB strains (St1, St3, N4, and R4) were tested for their capacity to successfully ensile Stylo legume (Stylosanthes guianensis CIAT184). Stylo silages treated with LAB were well preserved, the NH3-N and butyric acid contents were lower, and the lactic acid content was higher than those in the control (p < 0.05). The acetic acid content was the highest in R4-treated silage among the treatments (p < 0.05). The crude protein (CP) content of St1-silage was significantly (p < 0.05) higher than the others. The inoculation of thermotolerant LAB selected from fermented juice of epiphytic lactic acid bacteria (FJLB) was found to be highly instrumental to obtain well-preserved silage from the Stylo legume.

Effects of Lactic Acid Bacteria Isolated From Rumen Fluid and Feces of Dairy Cows on Fermentation Quality, Microbial Community, and in vitro Digestibility of Alfalfa Silage.

The objective of this study was to select lactic acid bacteria (LAB) isolated from the rumen fluid and feces of dairy cows, and evaluate their effects on silage quality of alfalfa after 30 or 60 days of ensiling. One hundred and four LAB strains were isolated from rumen fluid and feces of six dairy cows, of which four strains (Lactobacillus plantarum F1, L. plantarum F50, Lactobacillus salivarius L100, and Lactobacillus fermentum L120) and one commercial inoculant (GFG) isolated from forage were employed for further study. The silages treated with F1 had the lowest (P < 0.05) pH value and the highest (P < 0.05) lactic acid (LA) content in all treatments. Besides, higher (P < 0.05) in vitro digestibility was also observed in F1-treated silage after 60 days of ensiling. The microbial analysis showed that the Lactobacillus abundance in the F1-treated silages increased to 60.32%, higher than other treatments (5.12-47.64%). Our research indicated that strain F1 could be an alternative silage inoculant, and dairy cows could be a source for obtaining excellent LAB for ensiling.

Effects of different ratios and storage periods of liquid brewer's yeast mixed with cassava pulp on chemical composition, fermentation quality and in vitro ruminal fermentation.

OBJECTIVE: This study aims to evaluate the chemical composition, fermentation quality and in vitro ruminal fermentation of various ratios and storage periods of liquid brewer's yeast (LBY) mixed with cassava pulp (CVP). METHODS: Four mixtures of fresh LBY and CVP were made (LBY0, LBY10, LBY20, and LBY30 for LBY:CVP at 0:100, 10:90, 20:80, and 30:70, respectively) on a fresh matter basis, in 500 g in plastic bags and stored at 30 to 32°C. After storage, the bags were opened weekly from weeks 0 to 4. Fermentation quality and in vitro gas production (IVGP) were determined, as well as the dry matter (DM), organic matter (OM), crude protein (CP), ether extract (EE), neutral detergent fiber, acid detergent fiber and acid detergent lignin contents. RESULTS: The contents of CP and EE increased, whereas all other components decreased, in proportion to LBY inclusion (p<0.01). The DM and OM contents gradually decreased in weeks 3 and 4 (p<0.05), while EE contents were lowest in week 0. The pH, ammonia nitrogen per total nitrogen (NH3-N/TN) and V-score in each mixture and storage period demonstrated superior fermentation quality (pH≤4.2, NH3-N/TN≤12.5%, and V-score>90%). The pH increased and NH3-N/TN decreased, with proportionate increases of LBY, whereas the pH decreased and NH3-N/TN increased, as the storage periods were extended (p<0.01). Although IVGP decreased in proportion to the amount of LBY inclusion (p<0.01), in vitro organic matter digestibility (IVOMD) was unaffected by the mixture ratios. The highest IVGP and IVOMD were observed in week 0 (p<0.01). CONCLUSION: The inclusion of LBY (as high as 30%) into CVP improves the chemical composition of the mixture, thereby increasing the CP content, while decreasing IVGP, without decreasing fermentation quality and IVOMD. In addition, a preservation period of up to four weeks can guarantee superior fermentation quality in all types of mixtures. Therefore, we recommend limiting the use of CVP as a feed ingredient, given its low nutritional value and improving feed quality with the inclusion of LBY.

Effects of graded levels of liquid brewer's yeast on chemical composition and fermentation quality in cassava pulp and rice straw-based total mixed ration silage.

This study aimed to evaluate the effect of liquid brewer's yeast (LBY) addition on chemical composition and fermentation quality of mixture of LBY and cassava pulp (CVP) with rice straw (RS) in different ratios during preservation periods. Four mixtures of LBY, CVP and RS were made, that is mixture ratio of LBY : CVP : RS of 0% LBY, 20% LBY, 35% LBY and 50% LBY were 0:70:30, 20:50:30, 35:35:30 and 50:20:30 as fresh matter, respectively. The bags were opened at weeks 0, 1, 2, 4 and 8 after storage. The contents of dry matter, organic matter, crude protein (CP), ether extract (EE), neutral detergent fiber and acid detergent fiber ranged 36.4-40.0, 88.9-90.8, 4.0-12.0, 1.1-1.3, 58.8-61.6 and 37.6-40.0, respectively, and the contents of CP and EE increased and the other components decreased in proportion to LBY inclusion (P < 0.01). 50% LBY had the highest (P < 0.05) pH (4.81) and ammonia nitrogen per total nitrogen (NH3 -N/TN) (7.40%) and the lowest V-score (90.3). Propionic and butyric acid contents were 0.01% or lower in each mixture and storage period. There were rapid pH decrease and NH3 -N/TN increase during the first week of the storage period. The increases of NH3 -N/TN and acetic acid content and decreases of pH, lactic acid content and V-score during the preservation were more drastic as LBY inclusion increased. Although higher proportion of LBY produced higher CP and lower fiber contents in the mixture, attention should be paid for the reduction of fermentation quality during longer storage periods.

The effects of additives in napier grass silages on chemical composition, feed intake, nutrient digestibility and rumen fermentation.

The effect of silage additives on ensiling characteristics and nutritive value of Napier grass (Pennisetum purpureum) silages was studied. Napier grass silages were made with no additive, fermented juice of epiphytic lactic acid bacteria (FJLB), molasses or cassava meal. The ensiling characteristics were determined by ensiling Napier grass silages in airtight plastic pouches for 2, 4, 7, 14, 21 and 45 d. The effect of Napier grass silages treated with these additives on voluntary feed intake, digestibility, rumen fermentation and microbial rumen fermentation was determined in 4 fistulated cows using 4×4 Latin square design. The pH value of the treated silages rapidly decreased, and reached to the lowest value within 7 d of the start of fermentation, as compared to the control. Lactic acid content of silages treated with FJLB was stable at 14 d of fermentation and constant until 45 d of ensiling. At 45 d of ensiling, neutral detergent fiber (NDF) and acid detergent fiber (ADF) of silage treated with cassava meal were significantly lower (p<0.05) than the others. In the feeding trial, the intake of silage increased (p<0.05) in the cow fed with the treated silage. Among the treatments, dry matter intake was the lowest in the silage treated with cassava meal. The organic matter, crude protein and NDF digestibility of the silage treated with molasses was higher than the silage without additive and the silage treated with FJLB. The rumen parameters: ruminal pH, ammonia-nitrogen (NH3-N), volatile fatty acid (VFA), blood urea nitrogen (BUN) and bacterial populations were not significantly different among the treatments. In conclusion, these studies confirmed that the applying of molasses improved fermentative quality, feed intake and digestibility of Napier grass.