Italian Ryegrass Silage Research Articles

Fermentation Profile, Bacterial Community Structure, Co-Occurrence Networks, and Their Predicted Functionality and Pathogenic Risk in High-Moisture Italian Ryegrass Silage

This study aimed to assess the fermentation characteristics, bacterial community structure, co-occurrence networks, and their predicted functionality and pathogenic risk in high-moisture Italian ryegrass (IR; Lolium multiflorum Lam.) silage. The IR harvested at heading stage (208 g dry matter (DM)/kg fresh weight) was spontaneously ensiled in plastic silos (10 L scale). Triplicated silos were opened after 1, 3, 7, 15, 30, and 60 days of fermentation, respectively. The bacterial community structure on days 3 and 60 were investigated using high-throughput sequencing technology, and 16S rRNA-gene predicted functionality and phenotypes were determined by PICRUSt2 and BugBase tools, respectively. After 60 days, the IR silage exhibited good ensiling characteristics indicated by large amounts of acetic acid (~58.7 g/kg DM) and lactic acid (~91.5 g/kg DM), relatively low pH (~4.20), acceptable levels of ammonia nitrogen (~87.0 g/kg total nitrogen), and trace amounts of butyric acid (~1.59 g/kg DM). Psychrobacter was prevalent in fresh IR, and Lactobacillus became the most predominant genus after 3 and 60 days. The ensilage process reduced the complexity of the bacterial community networks in IR silage. The bacterial functional pathways in fresh and ensilaged IR are primarily characterized by the metabolism of carbohydrate and amino acid. The pyruvate kinase and 1-phosphofructokinase were critical in promoting lactic acid fermentation. A greater (p < 0.01) abundance of the “potentially pathogenic” label was noticed in the bacterial communities of ensiled IR than fresh IR. Altogether, the findings indicated that the high-moisture IR silage exhibited good ensiling characteristics, but the potential for microbial contamination and pathogens still remained after ensiling.

Effects of TMR Feeding Using Italian Ryegrass Silage and Hay of Those Dried by Hot-Air Dry System on Rumination Activity, Milk Productivity and Economic Feasibility in Holstein Dairy Cows

Effects of TMR Feeding Using Italian Ryegrass Silage and Hay of Those Dried by Hot-Air Dry System on Rumination Activity, Milk Productivity and Economic Feasibility in Holstein Dairy Cows

Forage quality and fermentation dynamics of silages of Italian ryegrass (Lolium multiflorum Lam.) wilted for varying periods.

This trial was conducted to explore the impact of different wilting time of Italian ryegrass (IRG) in the field on the nutritional quality and fermentation characteristics of its silage. The harvested IRG was directly wilted in the field for 0 day (W0), 1 day (W1), 2 days (W2), and 3 days (W3), respectively, and tedded every 6 hours. And the preserved IRG was sampled at 1, 2, 3, 5, 10, 20, 30, and 45 days after ensiling and three replicates per treatment. With the extension of wilting, the dry matter (DM) content and pH value of wilted IRG gradually increased (p<0.05). There was a downward trend in; neutral detergent fiber (NDF), acid detergent fiber (ADF), and hemicellulose with the increase of wilting time, but only W2 and W3 were significantly different from W0 (p<0.05). Crude protein (CP), in vitro DM digestibility), total digestible nutrients (TDN), and relative feed value decreased significantly with the increase of wilting time (p<0.05), except for W1. After 45 days of ensiling, W1 had the highest CP, TDN, and the lowest ADF and NDF. During ensiling, the increase of acetic acid and the decrease of WSC in W0 and W1 were similar, but the accumulation rate of lactic acid in W0 was faster than that in W1, resulting in the lowest pH value in W0. After 5 days of ensiling, the ratio of lactic acid to acetic acid in W1 stabilized at around 3:1, while W0 kept changing. Italian ryegrass that wilted in the field for 1 day effectively improved the dynamic changes in CP, TDN, ADF, and NDF and fermentation quality of silage. Therefore, in practice, W1 was more recommended in production of IRG silage.

Effects of Italian ryegrass silage-based total mixed ration on rumen fermentation, growth performance, blood metabolites, and bacterial communities of growing Hanwoo heifers.

This study utilized Italian ryegrass silage (IRGS) - based total mixed ration (TMR) as feedstuff and evaluated its effects on rumen fermentation, growth performance, blood parameters, and bacterial community in growing Hanwoo heifers. Twenty-seven Hanwoo heifers (body weight [BW], 225.11 ± 10.57 kg) were randomly allocated to three experimental diets. Heifers were fed 1 of 3 treatments as follows: TMR with oat, timothy, and alfalfa hay (CON), TMR with 19% of IRGS (L-IRGS), and TMR with 36% of IRGS (H-IRGS). Feeding high levels of IRGS (H-IRGS) and CON TMR to heifers resulted in a greater molar proportion of propionate in the rumen. The impact of different TMR diets on the BW, average daily gain, dry matter intake, and feed conversion ratio of Hanwoo heifers during the growing period did not differ (p > 0.05). Furthermore, the blood metabolites, total protein, albumin, aspartate aminotransferase, glucose, and total cholesterol of the heifers were not affected by the different TMR diets (p > 0.05). In terms of rumen bacterial community composition, 264 operational taxonomic units (OTUs) were observed across the three TMR diets with 240, 239, and 220 OTUs in CON, L-IRGS, and H-IRGS, respectively. IRGS-based diets increased the relative abundances of genera belonging to phylum Bacteroidetes but decreased the abundances of genus belonging to phylum Firmicutes compared with the control. Data showed that Bacteroidetes was the most dominant phylum, while Prevotella ruminicola was the dominant species across the three TMR groups. The relative abundance of Ruminococcus bromii in the rumen increased in heifers fed with high inclusion of IRGS in the TMR (H-IRGS TMR). The relative abundance of R. bromii in the rumen significantly increased when heifers were fed H-IRGS TMR while P. ruminicola increased in both L-IRGS and H-IRGS TMR groups. Results from the current study demonstrate that the inclusion of IRGS in the TMR is comparable with the TMR containing high-quality forage (CON). Thus, a high level of IRGS can be used as a replacement forage ingredient in TMR feeding and had a beneficial effect of possibly modulating the rumen bacterial community toward mainly propionate-producing microorganisms.

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

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. 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). 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.

The metabolome and bacterial composition of high-moisture Italian ryegrass silage inoculated with lactic acid bacteria during ensiling

BackgroundWith its high nutritional value and productivity, Italian ryegrass as a biomass feedstock constantly supplies rumen degradable nitrogen and digestible fiber to ruminants. However, biofuel production is easily reduced during ensiling due to the high-moisture content of Italian ryegrass, leading to economic losses. Lactic acid bacteria inoculants could improve lignocellulosic degradation and fermentation quality and decrease dry matter loss during the bioprocessing of silage. Therefore, this study analyzed the effects of Lactobacillus buchneri TSy1-3 (HE), Lactobacillus rhamnosus BDy3-10 (HO), and the combination of HE and HO (M) on fermentation quality, bacterial community and metabolome in high-moisture Italian ryegrass silage during ensiling.ResultsThe results showed that the pH value was significantly lower in the HO groups than in the other treatments at the end of ensiling, and the dry matter and acetic acid contents were significantly higher in the HO group than in the other inoculated groups. All inoculants decreased the diversity of the bacterial community and significantly increased the relative abundance of Lactobacillus. Inoculation with HO significantly improved the concentrations of organic acids, dipeptides, ferulic acid, apigenin, and laricitrin. Compared with Lactobacillus buchneri TSy1-3 (HE), HO significantly upregulated the flavonoid compounds in the flavone and flavonol biosynthesis pathway.ConclusionsOverall, these findings suggest that inoculation with HO was beneficial for the development of Italian ryegrass as a biomass feedstock, improving fermentation quality, accelerating changes in bacterial community composition and increasing biofunctional metabolites in high-moisture Italian ryegrass silage.

Effects of different additions to Italian ryegrass (Lolium multiflorum Lam.) on silage quality

This research was carried out to determine the effects of different additions (urea and molasses) used with Italian ryegrass (Lolium multiflorum L.) silage on fermentation, in vitro gas production, microbiological properties, in vitro digestibility parameters, and relative fodder quality (RFQ) in silages made under laboratory conditions. The Italian grass (Lolium multiflorum L.) used in the study was chopped to an approximate size of 2–3.0 cm. Amounts of 0, 2, and 4% molasses and 0, 0.5, and 1% urea were added to the fresh material as a percentage of dry matter. Because of the urea, crude protein (CP) of Italian ryegrass silage increased, but the content of neutral detergent fibre (NDF), acid detergent fibre (ADF), and acid detergent lignin (ADL) decreased. While the addition of urea decreased the acetic acid and butyric acid concentrations of the silage, it increased the pH, lactic acid, and ammonia (NH3) content. Molasses addition increased in vitro gas production and organic matter digestibility (OMD); urea increased metabolic energy (ME) and the net energy lactation (NEL) values of silages. Urea and molasses both increased in vitro digestibility parameters, microbial protein production and synthesis, and relative fodder quality of the silage. As a result of the research, it was determined that urea and molasses could be used at contents of 1.5% and 4%, respectively, in Italian ryegrass silage.

Feeding Mixed Silages of Winter Cereals and Italian Ryegrass Can Modify the Fatty Acid and Odor Profile of Bovine Milk

The utilization of corn silage in animal diets is becoming a challenge, due to the crop’s reduced yield as a result of climate change. Alternative silage types, such as mixtures of Italian ryegrass and winter cereals, may be a good complement to corn silage in diet formulation. Therefore, it is important to investigate how these alternative sources influence milk fatty acid and odor profile, as well as how these quality parameters could be efficiently evaluated. In this study, a corn silage-based control (CTR) and four experimental (EXP) diets—which contained winter cereals (WC), as well as WC with Italian ryegrass (IRG) silages in different proportions—were fed to Holstein-Friesian cows (n = 32) in a single-blinded efficacy study during a series of 4-week periods, with 2 weeks of adaption to each feed before the main trial. Milk from each trial was subjected to fatty acid (FA) analysis and odor profiling through the utilization of gas chromatography and an electronic nose, respectively. The results show that milk FAs in the EXP-3 and EXP-4 groups (which contained mixed silages using WC) changed the most when compared with other groups. Moreover, with a 7 kg/day inclusion rate of WC + IRG and of the WC silages in the diets of the EXP-2 and EXP-3 groups, respectively, the milk from these groups had their n6:n3 ratio reduced, thus indicating possible health benefits to consumers. The odor variation between the milk of the WC + IRG and WC groups was greater than the variation between the milk of the CTR and EXP groups. The main volatile compound responsible for the odor of the CTR milk was ethyl-butyrate, whereas 2-propanol and butan-2-one dominated the WC milk; the milk samples of the WC + IRG groups were influenced largely by ethanol. The study proved that with a 7 kg/day inclusion of mixed silages including winter cereals plus Italian ryegrass, the FA and odor profile of bovine milk could be modified.

Effect of different regions on fermentation profiles, microbial communities, and their metabolomic pathways and properties in Italian ryegrass silage.

Italian ryegrass is less studied in northern China due to high-quality forage grass has not been fully utilized. Full utilization of high-quality forage grass helps to alleviate the shortage of forage grass in winter and spring season and guarantee stable development of livestock production. Consequently, this study was aimed to evaluate the effects of different regions in northern China on the fermentative products, bacterial community compositions, and metabolic pathways and metabolites of Italian ryegrass silage. The Italian ryegrass was harvested from three regions (Ordos-WK; Hohhot-AK; Ulanqab-SYK) and ensiled for 60 days. Single molecule real-time (SMRT) sequencing and ultra-high performance liquid chromatography-mass spectrometry (UHPLC-MS/MS) were used to analyze bacterial communities and metabolites, respectively. After 60 d of fermentation, the SYK group had the lowest pH (4.67), the highest lactic acid contents (95.02 g/kg DM) and largest lactic acid bacteria populations (6.66 log10 cfu/g FM) among the treatment groups. In addition, the SYK group had the highest abundance of Lactiplantibacillus plantarum (63.98%). In SYK group, isoquinoline alkaloid biosynthesis was the significantly enriched (p < 0.05) and high-impact value (0.0225) metabolic pathway. In AK group, tryptophan metabolism the was the significantly enriched (p < 0.001) and high-impact value (0.1387) metabolic pathway. In WK group, citrate cycle (TCA cycle) was the significantly enriched (p < 0.001) and high-impact value (0.1174) metabolic pathway. Further, Lactiplantibacillus plantarum was positively correlated with cinnamic acid, tetranor 12-HETE, D-Mannitol, (2S)-2-amino-4-methylpentanoic acid L-Leucine, guanine, isoleucyl-aspartate and 3,4-Dihydroxyphenyl propanoate, but negatively correlated with isocitrate and D-mannose. In conclusion, this study can improve our understanding of the ensiling microbiology and metabolomics in different regions to further regulate the fermentation products and promote livestock production.

Effects of Italian ryegrass silage supplementation on feed intake and behavior of pregnant sows.

This study aimed to assess the behavior and stress status of pregnant sows following supplementation with Italian ryegrass silage (IRS) and the impact of feeding the IRS on feeding costs. Six sows with an initial body weight (BW) of 238.6 ± 5.9 kg were allotted to a 6 × 3 Latin square design with a 5-day acclimatization period followed by a 5-day data collection period. A commercial diet was replaced by IRS on a dry matter (DM) basis up to 0%, 9%, and 13% in the control treatment and the two test treatments, respectively. Apart from collecting data on daily feed intake and BW, urine was collected, and video footage was recorded for the last day of each treatment for analysis of urinary cortisol and behavior. There were no leftovers with all diets and nutrient uptake was unaffected (p > 0.05), while BW gain decreased (p < 0.05) to be a limited range from 1% to 3%, with increased inclusion of IRS. Both the behavior of sows and cortisol concentration were unaffected (p > 0.05). Furthermore, it was estimated that feeding 13% DM of IRS would reduce feed costs by 17%. IRS would be acceptable in replacing up to 13% of the commercial diet and cutting feeding costs.

Effect of Formic Acid and Inoculants on Microbial Community and Fermentation Profile of Wilted or Un-Wilted Italian Ryegrass Silages during Ensiling and Aerobic Exposure

Wilted (around 35% DM) or un-wilted (around 35% DM) Italian ryegrass treated with three additives (formic acid, FA; Lactobacillus plantarum, LP; Lactobacillus buchneri, LB) was utilized to evaluate the effects of the dry matter (DM) contents on the microbial community and fermentation characteristics, which was ensiled for 60 days in a laboratory-scale silo, followed by 3 days of aerobic exposure. Significantly lower pH and higher lactic acid (LA) contents were observed in the LP-treated group ensiled at both DM contents (differences were significant when p &lt; 0.05). The contents of LA, acetic acid (AA), numbers of lactic acid bacteria (LAB) and ammonia nitrogen (NH3-N) in the FA-treated group were significantly lower than those in other treatments (p &lt; 0.05). L. buchneri was the dominant bacteria after 60 days fermentation, while Enterobacteria became prevalent after 3 days of aerobic exposure. L. buchneri was found in the LB-treated group with higher acetic acid. Although the best fermentation quality was observed in the LP-treated silages, the aerobic stability was lowest compared to other groups (p &lt; 0.05). In conclusion, our findings suggest that the DM content of Italian ryegrass affected its epiphytic microbial community and the effectiveness of the different type of additives. Formic acid was more suitable for un-wilted Italian ryegrass silage, L. plantarum had a better effect in wilted Italian ryegrass silage, and L. buchneri prolonged the aerobic stability of Italian ryegrass. DM content and purpose of ensiling should be the key factors for choosing different types of additives for Italian ryegrass silage.

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

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). 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. 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.

Fermentation Profiles, Bacterial Community Compositions, and Their Predicted Functional Characteristics of Grass Silage in Response to Epiphytic Microbiota on Legume Forages.

This study aimed to investigate the effect of epiphytic microbiota from alfalfa and red clover on the fermentative products, bacterial community compositions, and their predicted functional characteristics in Italian ryegrass silage. By microbiota transplantation and γ-ray irradiation sterilization, the irradiated Italian ryegrass was treated as follows: (1) sterile distilled water (STIR); (2) epiphytic microbiota on Italian ryegrass (IRIR); (3) epiphytic microbiota on alfalfa (IRAL); and (4) epiphytic microbiota on red clover (IRRC). The irradiated Italian ryegrass was ensiled for 1, 3, 7, 15, 30, and 60 days. STIR had similar chemical components with fresh Italian ryegrass. IRAL had higher lactic acid concentrations [64.0 g/kg dry matter (DM)] than IRIR (22.3 g/kg DM) and IRRC (49.4 g/kg DM) on day 3. IRRC had the lowest lactic acid concentrations (59.7 g/kg DM) and the highest pH (4.64), acetic acid (60.4 g/kg DM), ethanol (20.4 g/kg DM), and ammonia nitrogen (82.6 g/kg DM) concentrations and Enterobacteriaceae [9.51 log10 cfu/g fresh weight (FW)] populations among treatments on day 60. On days 3 and 60, Lactobacillus was dominant in both IRIR (42.2%; 72.7%) and IRAL (29.7%; 91.6%), while Hafnia-Obesumbacterium was predominant in IRRC (85.2%; 48.9%). IRIR and IRAL had lower abundances of “Membrane transport” than IRRC on day 3. IRIR and IRAL had lower abundances of phosphotransacetylase and putative ATP-binding cassette transporter and higher abundances of arginine deiminase on day 3. IRAL had the highest abundance of fructokinase on day 3. Overall, inoculating epiphytic microbiota from different legume forages changed the fermentative products, bacterial community compositions, and their predicted functional characteristics in Italian ryegrass silage. The microbial factors that result in the differences in fermentative profiles between legume forage and grass were revealed. Knowledge regarding the effect of epiphytic microbiota could provide more insights into the improvement of silage quality.

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 60days. 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 60days 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 phytol in forage on phytanic acid content in cow's milk.

ObjectiveBioactive compounds in ruminant products are related to functional compounds in their diets. Therefore, this study aimed to explore the effect of forage sources, Italian ryegrass (IR) silage vs corn silage (CS) in the total mixed ration (TMR), on milk production, milk composition, and phytanic acid content in milk, as well as on the extent of conversion of dietary phytol to milk phytanic acid.MethodsPhytanic acid content in milk was investigated for cows fed a TMR containing either IR silage or CS using 17 cows over three periods of 21 days each. In periods 1 and 3, cows were fed CS-based TMR (30% CS), while in period 2, cows were fed IR silage-based TMR (20% IR silage and10% CS).ResultsThe results showed that there were no differences in fat, protein, lactose, solids-not-fat, somatic cell count, and fatty acid composition of milk among the three experimental periods. There were no differences in the plasma concentration of glucose, triglycerides, total cholesterol, and nonesterified fatty acids among the three experimental periods, while the blood urea nitrogen was higher (p<0.05) in period 2. The milk phytanic acid content was higher (p<0.05) in period 2 (13.9 mg/kg) compared with periods 1 (9.30 mg/kg) and 3 (8.80 mg/kg). Also, the phytanic acid content in the feces was higher (p<0.05) in period 2 (1.65 mg/kg dry matter [DM]) compared with period 1 (1.15 mg/kg DM), and 3 (1.17 mg/kg DM). Although the phytol contents in feces did not differ among the three feeding periods, the conversion ratio from dietary phytol to milk phytanic acid was estimated to be only 2.6%.ConclusionPhytanic acid content in cow’s milk increases with increasing phytol content in diets. However, phytol might not be completely metabolized in the rumen and phytanic acid, in turn, might not be completely recovered into cow’s milk. The change of phytanic acid content in milk may be positively correlated with the change of phytol in the diet within a short time.

Effects of using different roughages in the total mixed ration inoculated with or without coculture of Lactobacillus acidophilus and Bacillus subtilis on in vitro rumen fermentation and microbial population

ObjectiveThis study aimed to determine the effects of different roughages in total mixed ration (TMR) inoculated with or without coculture of Lactobacillus acidophilus (L. acidophilus) and Bacillus subtilis (B. subtilis) on in vitro rumen fermentation and microbial population.MethodsThree TMRs formulations composed of different forages were used and each TMR was grouped into two treatments: non-fermented TMR and fermented TMR (F-TMR) (inoculated with coculture of L. acidophilus and B. subtilis). After fermentation, the fermentation, chemical and microbial profile of the TMRs were determined. The treatments were used for in vitro rumen fermentation to determine total gas production, pH, ammonia-nitrogen (NH3-N), and volatile fatty acids (VFA). Microbial populations were determined by quantitative real-time polymerase chain reaction (PCR). All data were analyzed as a 3×2 factorial arrangement design using the MIXED procedure of Statistical Analysis Systems.ResultsChanges in the fermentation (pH, lactate, acetate, propionate, and NH3-N) and chemical composition (moisture, crude protein, crude fiber, and ash) were observed. For in vitro rumen fermentation, lower rumen pH, higher acetate, propionate, and total VFA content were observed in the F-TMR group after 24 h incubation (p<0.05). F-TMR group had higher acetate concentration compared with the non-fermented group. Total VFA was highest (p<0.05) in F-TMR containing combined forage of domestic and imported source (F-CF) and F-TMR containing Italian ryegrass silage and corn silage (F-IRS-CS) than that of TMR diet containing oat, timothy, and alfalfa hay. The microbial population was not affected by the different TMR diets.ConclusionThe use of Italian ryegrass silage and corn silage, as well as the inoculation of coculture of L. acidophilus and B. subtilis, in the TMR caused changes in the pH, lactate and acetate concentrations, and chemical composition of experimental diets. In addition, F-TMR composed with Italian ryegrass silage and corn silage altered ruminal pH and VFA concentrations during in vitro rumen fermentation experiment.

Changes in fermentation pattern and quality of Italian ryegrass (Lolium multiflorum Lam.) silage by wilting and inoculant treatments.

ObjectiveThis study was conducted to investigate the effects of wilting and microbial inoculant treatment on the fermentation pattern and quality of Italian ryegrass silage.MethodsItalian ryegrass was harvested at heading stage and ensiled into vinyl bags (20 cm×30 cm) for 60d. Italian ryegrass was ensiled with 4 treatments (NWNA, no-wilting no-additive; NWA, no-wilting with additive; WNA, wilting no-additive; WA, wilting with additive) in 3 replications, wilting time was 5 hours and additives were treated with 106 cfu/g of Lactobacillus plantarum. The silages samples were collected at 1, 2, 3, 5, 10, 20, 30, 45, and 60 days after ensiling and analyzed for the ensiling quality and characteristics of fermentation patterns.ResultsWilting treatment resulted in lower crude protein and in vitro dry matter digestibility and there were no significant differences in acid detergent fiber (ADF), total digestible nutrient (TDN), water-soluble carbohydrate (WSC), ammonia content, and pH (p>0.05). However, wilting treatment resulted in higher ADF and neutral detergent fiber content of Italian ryegrass silage (p<0.05), and the WNA treatment showed the lowest TDN and in vitro dry matter digestibility. The pH of the silage was higher in the wilting group (WNA and WA) and lower in the additive treatment group. Meanwhile, the decrease in pH occurred sharply between the 3–5th day of storage. The ammonia nitrogen content was significantly lower in the additive treatment (p<0.05), and wilting had no effect. As fermentation progressed, the lactic and acetic acid contents were increased and showed the highest content at 30 days of storage.ConclusionThe wilting treatment did not significantly improve the silage fermentation, but the inoculant treatment improved the fermentation patterns and quality of the silage. So, inoculation before ensiling is recommended when preparing high quality of Italian ryegrass silage, and when wilting, it is recommended to combine inoculation for making high quality silage.

Replacement of Imported Timothy Hay with Domestic Italian Ryegrass Silage in a Horse Feedstuff

Replacement of Imported Timothy Hay with Domestic Italian Ryegrass Silage in a Horse Feedstuff

Effects of Heterofermentative Lactic Acid Bacteria on the Ruminal Fermentation of Italian Ryegrass and Whole Crop Barley Silage

Effects of Heterofermentative Lactic Acid Bacteria on the Ruminal Fermentation of Italian Ryegrass and Whole Crop Barley Silage

Probiotic Characteristics and Antifungal Activity of Lactobacillus plantarum and Its Impact on Fermentation of Italian Ryegrass at Low Moisture

The study aimed to investigate probiotic characteristics, and low moisture silage fermentation capability of selected lactic acid bacteria (LAB) isolated from Alfalfa (Medicago sativa L). Morphological and physiological properties, carbohydrates fermentation, enzymes, and organic acids production, anti-fungal activity, antibiotic sensitivity patterns, and probiotic characteristics (acidic and bile salt tolerances, hydrophobicity and aggregations natures) of LAB were examined. 16SrRNA sequencing was carried out to identify isolated strains. The identified strains Lactobacillus plantarum (KCC-37) and Lactobacillus plantarum (KCC-38) showed intense antifungal activity, survival tolerant in acidic and bile salt environments, cell surface and auto aggregations ability, enzymes and organic acids productions. At ensiled condition, KCC-37 and KCC-38 enhanced acidification of Italian ryegrass silages by producing a higher amount of lactic acid, a key acid for indicating silage quality with less extent to acetic acid and succinic acid at low moisture level than non-inoculated silages. Notably, the addition of mixed strains of KCC-37 and KCC-38 more potentially enhanced acidification of silage and organic acid productions than the single-culture inoculation. The overall data suggested that these strains could be used as an additive for improving the quality of the fermentation process in low moisture silage with significant probiotic characteristics.