Changes in microbial dynamics and fermentation characteristics of alfalfa silage: A potent approach to mitigate greenhouse gas emission through high-quality forage silage.

Feeding ruminants with high-quality forage can enhance digestibility and reduce methane production. Development of high-quality silage from leguminous plants with lactic acid bacteria can improve digestibility and it mitigate the greenhouse gas emissions. In this study, we developed a high-quality alfalfa silage with improved fermentation index and microbial dynamics using Levilactobacillus brevis-KCC-44 at low or high moisture (LM/HM) conditions and preserved it for 75 or 150 days. Alfalfa fermentation with L. brevis enhances acidification and fermentation characteristics primarily due to the dominance of lactic acid bacteria (LAB) L. brevis (>95%) compared to alfalfa fermented with epiphytic LAB. The inoculant L. brevis improved the anaerobic fermentation indexes resulting in a higher level of lactic acid in both high (10.0 ± 0.12 & 8.90 ± 0.31%DM) and low moisture (0.55 ± 0.08 & 0.39 ± 0.0 %DM) in 75 and 150 days respectively, compared to control silage. In addition, the marginal amount of acetic acid (range from 0.23 ± 0.07 to 2.04 ± 0.27 %DM) and a reduced level of butyric acid (range between 0.03 ± 0.0 to 0.13 ± 02 %DM) was noted in silage treated with LAB than the control. The LAB count and abundance of Levilactobacillus were higher in alfalfa silage fermented with L. brevis. Microbial richness and diversity were reduced in alfalfa silage treated with L. brevis which prompted lactic acid production at a higher level even for a prolonged period of time. Therefore, this L.brevis is an effective inoculant for producing high-quality alfalfa silage since it improves fermentation indexes and provides reproducible ensiling properties.

Effects of Different Lactic Acid Bacteria in Single or Mixed Form on the Fermentative Parameters and Nutrient Contents of Early Heading Triticale Silage for Livestock.

Lactic acid bacteria (LAB) are excellent anaerobic fermenters that produce highly valuable grass-based animal feed containing essential nutrients. In the present study, an ensiling process was used to improve anaerobic fermentation in triticale silage under different moisture conditions with LAB. The triticale was treated with either a single bacterium or combined LAB and then vacuum-sealed. After 180 and 360 days of storage, the silage's fermentation characteristics, microbial changes and nutrient contents were analyzed. The pH of the silage was significantly lower than the control silage. There was a significant difference in the pH values between the silages treated with single or mixed LAB. The LAB treatment led to a substantial increase in lactic acid (LA), a decrease in butyric acid (BA), and marginal levels of acetic acid (AA). The LA content after the mixed LAB treatment was significantly higher than that after the single culture LAB treatment. After single or combined inoculant treatments, the LAB population in the silage increased, while the yeast and mold levels decreased. These findings suggest that the addition of LAB to silage during ensiling could enhance the nutritional quality and reduce unwanted microbial growth. The mixed LAB treatments produced silage with a significantly higher nutritional value than the single LAB treatments.

Quantification of major phenolic and flavonoid markers in forage crop Lolium multiflorum using HPLC-DAD

ABSTRACT The objective of this study was to perform preliminary screening of phytochemical compounds and quantification of major phenolics and flavonoid markers in Italian ryegrass extract using HPLC-DAD. Previously, LC-MS analysis has identified different phenolic acids, including caffeic acid, ferulic acid, p-coumaric acid, chlorogenic acid, dihydroxy benzoic acid, propyl gallate, catechin, and six flavonoids including rutin hydroxide, luteolin, kaemferol, vitexin, narcissoside, and myricetin from Italian ryegrass extract. In the present study, Italian ryegrass silage powder was extracted with ethanol: water for 20 min at 90 °C. The extract targeted optimum yield of phenolic acids and flavonoids. Crude phenolic acid and flavonoids were then purified by solid phase extraction method. Purified fractions were then injected into HPLC with a diode-array detector. Quantified concentrations of isolated phenolic acids and flavonoids ranged from 125 to 220 µg/g dry weight. Limits of detection and limits of quantification for all standards (unknown compounds) ranged from 0.38 to 1.71 and 0.48 to 5.19 µg/g dry weight, respectively. Obtained values were compared with previous literatures, indicating that our HPLC-DAD quantification method showed more sensitivity. This method showed better speed, accuracy, and effectiveness compared to previous reports. Furthermore, this study could be very useful for developing phenolic acids and flavonoids from compositions in Italian ryegrass silage feed for pharmaceutical applications and ruminant animals in livestock industries.

Potential role of marine algae extract on 3T3-L1 cell proliferation and differentiation: an in vitro approach

BACKGROUND: From ancient times, marine algae have emerged as alternative medicine and foods, contains the rich source of natural products like proteins, vitamins, and secondary metabolites, especially Chlorella vulgaris (C. vulgaris) contains numerous anti-inflammatory, antioxidants and wound healing substances. Type 2 diabetes mellitus is closely associated with adipogenesis and their factors. Hence, we aimed to investigate the chemical constituents and adipo-genic modulatory properties of C. vulgaris in 3T3-L1 pre-adipocytes. RESULTS: We analysed chemical constituents in ethanolic extract of C. vulgaris (EECV) by LC-MS. Results revealed that the EECV contains few triterpenoids and saponin compounds. Further, the effect of EECV on lipid accumulation along with genes and proteins expressions which are associated with adipogenesis and lipogenesis were evaluated using oil red O staining, qPCR and western blot techniques. The data indicated that that EECV treatment increased differentiation and lipid accumulation in 3T3-L1 cells, which indicates positive regulation of adipogenic and lipogenic activity. These increases were associated with up-regulation of PPAR-γ2, C/EBP-α, adiponectin, FAS, and leptin mRNA and protein expressions. Also, EECV treatments increased the concentration of glycerol releases as compared with control cells. Troglitazone is a PPAR-γ agonist that stimulates the PPAR-y2, adiponectin, and GLUT-4 expressions. Similarly, EECV treatments significantly upregulated PPAR-γ, adiponectin, GLUT-4 expressions and glucose utilization. Further, EECV treatment decreased AMPK-α expression as compared with control and metformin treated cells. CONCLUSION: The present research findings confirmed that the EECV effectively modulates the lipid accumulation and differentiation in 3T3-L1 cells through AMPK-α mediated signalling pathway.