Search

To investigate the effect of 5-aminolevulinic acid (5-ALA) on in vitro rumen gas production, fermentation characteristics, and bacterial community profiles, five levels of 5-ALA (0, 100, 500, 1000, and 5000 mg/kg DM) were supplemented into a total mixed ration (concentrate/forage = 40:60) as substrate in an in vitro experiment. Results showed that as the supplementation level of 5-ALA increased, asymptotic gas production (b) decreased linearly and quadratically (p < 0.01) while the dry matter degradation rate increased quadratically (p < 0.01). Meanwhile, the propionate concentration of 72 h incubation fluid increased linearly (p = 0.03) and pH value increased linearly and quadratically (p < 0.01), while the concentrations of butyrate, isobutyrate, valerate, isovalerate, and NH3-N and the ratio of acetate/propionate (A/P) decreased linearly and quadratically (p < 0.05). There was no significant difference in any alpha diversity indices of bacterial communities among the various 5-ALA levels (p < 0.05). PCoA and PERMANOVA analysis revealed that the bacterial profiles showed a statistical difference between the treatment 5-ALA at 1000 mg/kg DM and the other levels except for 5000 mg/kg DM (p < 0.05). Taxonomic classification revealed a total of 18 and 173 bacterial taxa at the phylum and genus level with relative abundances higher than 0.01% in at least half of the samples, respectively. LEfse analysis revealed that 19 bacterial taxa were affected by 5-ALA levels. Correlation analysis showed that Actinobacteriota was positively correlated with the gas production parameter b, the ratio of A/P, and the concentration of butyrate, isovalerate, and NH3-N (p < 0.05) and negatively correlated with pH (p < 0.05). WPS-2 exhibited a negative correlation with the gas production parameter b, the ratio of A/P, and the concentration of butyrate, valerate, isobutyrate, isovalerate, and NH3-N (p < 0.05), along with a weaker positive correlation with pH (p = 0.04). The Bacteroidales BS11 gut group was negatively correlated with the concentration of propionate but positively correlated with gas production parameter b and the concentration of butyrate and NH3-N (p < 0.05). The Lachnospiraceae NK3A20 group was found to have a positive correlation with gas production parameter b, the ratio of A/P, and the concentration of butyrate, isobutyrate, isovalerate, valerate, total VFA, and NH3-N (p < 0.05), but a highly negative correlation with pH (p < 0.01). Differential metabolic pathways analysis suggested that metabolic pathways related to crude protein utilization, such as L-glutamate degradation VIII (to propanoate), L-tryptophan degradation IX, and urea cycle, increased with 5-ALA levels. In summary, including 5-ALA in the diet might improve energy and protein utilization by reducing the abundance of Actinobacteriota, the Bacteroidales BS11 gut group, the Lachnospiraceae NK3A20 group, and certain pathogenic bacteria and increasing the abundance of WPS-2.

Tea seed oil (TSO) was investigated for its effects on rumen fermentation and in vitro parameters of bacterial communities in water buffalo diets containing Siraitia grosvenorii and soybean residues. TSO was added at rates of 0% (control group (CT)), 0.5% (T1), 1% (T2), and 2% (T3) of the in vitro fermentation substrate weight (dry matter (DM) basis). T2 and T3 had significantly lower acetate and total volatile fatty acid contents but a significantly higher microbial crude protein content than CT. The lowest NH3-N content was observed in T1 and T2. Treatment significantly increased DM digestibility, with the highest percentage observed in T2. T2 showed significantly higher crude protein digestibility than CT. TSO supplementation significantly increased the C18:2n6c, C18:2 trans-10, cis-12, and C20:4n6 concentrations compared to those in CT. The total number of bacteria was significantly lower in T2 than in CT. TSO supplementation decreased the total bacteria, fungi, and methanogen populations but increased rumen microorganism diversity and richness. In conclusion, TSO can regulate the number and flora of rumen microorganisms through antimicrobial activity, thereby affecting rumen fermentation patterns, reducing methane production, and improving nutrient digestibility, and an optimal supplementation rate appears to be achieved with 1% TSO (DM basis).

In this study, we compared the digestibility levels and in vitro fermentation parameters of total mixed rations (TMRs) containing 20% and 40% defatted black soldier fly (Hermetia illucens) larvae (BSF) as a substitute for soybean meal (SBM) in the basal ration (60% roughage/40% concentrated feed) of dairy cows. We evaluated the volatile fatty acid (VFA), total gas production, methane emission, ammonia, pH, carbon dioxide, in vitro dry matter digestibility (IVDMD), and neutral detergent fiber digestibility (IVNDFD) of the TMR0 (basal), TMR20 (20% BSF included), and TMR40 (40% BSF included) diets at the end of 24 and 48 h of incubation. Significantly lower levels of ammonia formation were found in the TMR20 and TMR40 groups at 24 and 48 h (p < 0.001). An increase in total VFA levels was observed in the TMR0 group at 24 h (p < 0.001). The highest IVDMD was determined in TMR20 and TMR40 at 24 h. The highest IVNDFD value was observed in TMR20 at 24 h and in TMR40 at 48 h. The substitution of 20% and 40% of SBM with BSF positively affected IVDMD and IVNDFD (p < 0.001). TMR20 and TMR40 had the highest cumulative gas production at 48 h of incubation (p < 0.05). In conclusion, the use of BSF had a positive impact on digestibility and in vitro rumen fermentation. Therefore, we recommend the use of BSF in formulating dairy cow rations.

This study investigated the effect of co-ensiling increasing levels of artichoke bracts (Cynara cardunculus L.) with berseem (Trifolium alexandrinum L.) (100:0, 75:25, 50:50, 25:75, and 0:100, respectively) on silage quality after 0, 30, 60, and 120 days. Moreover, the in vitro rumen fermentation characteristics and methane (CH4) and ammonia (NH3-N) production were evaluated using a buffalo inoculum source. The results showed that pH of the silage and the concentration of acetic, propionic, butyric acid, and NH3-N significantly decreased (L; p < 0.01) with the increasing amounts of artichoke bracts in the mixture. At 30 and 60 days of ensiling, the highest lactic acid concentration was observed at intermediate proportions of artichoke bracts (p < 0.01). Cumulative gas production was higher in artichoke bracts than in the berseem silage. After 24 h of incubation, the highest value (p < 0.05) of truly dry matter, organic matter, natural detergent fiber degradability, and NH3-N concentration was recorded with 500 g/kg of forage mixtures. As the artichoke bract concentration increased, the partitioning factor and ruminal pH declined linearly (p ≤ 0.05). No significant differences were observed for total volatile fatty acids and volatile fatty acids molar proportions. In summary, co-ensiling artichoke bracts with berseem at a ratio of 1:1 might be a promising and easy method for the production of high-quality silage from legume forage with positively manipulating rumen fermentation.

Gamma-aminobutyric acid (GABA), known as the most abundant inhibitory neurotransmitter in the mammalian brain, can permeate ruminal epithelia by passive diffusion and enrich in the rumen environment. To explore whether the addition of GABA can regulate rumen fermentation characteristics as well as methane production, a 2 × 6 factorial in vitro rumen batch culture was conducted to determine the supplemental effect of GABA at inclusion levels of 0 (Control), 10, 20, 30, 40 and 50 mg in culture fluids on rumen fermentation of two total mixed rations (HF—a high-fiber ration consisted of 70% corn silage and 30% concentrate; and LF—a low-fiber ration consisted of 30% corn silage and 70% concentrate). After 72 h in vitro incubation of two rations with mixed rumen microoganisms obtained from five rumen-cannulated lactating Holstein dairy cows, increasing GABA addition linearly increased cumulative gas production in the LF group, though in vitro dry matter digestibility was not affected in either the LF or HF group. Kinetic gas production analysis noted that increasing GABA addition mostly decreased the gas production rate (i.e., RmaxG), as well as the ration digestion rate (RmaxS) to reach maximum fermentation. The GABA addition did not affect pH or microbial growth (i.e., MCP). However, total volatile fatty acid production in both LF and HF groups all linearly increased with the increase in GABA addition. Along with the increase in GABA addition in both LF and HF groups, the ratio of non-glucogenic to glucogenic volatile fatty acids both increased, while the molar proportions of propionate and valerate were significantly decreased, and the acetate and butyrate proportions were increased after 72 h in vitro rumen fermentation. The time-course change of fermentation end-products generally showed that carbon dioxide declined from approximately 89% to 74%, and methane increased from approximately 11% to 26%. After 72 h in vitro fermentation, molar methane proportion was greater in the LF than in the HF group, and increasing GABA addition quadratically increased methane production in the LF group while a slight increase occurred in the HF group.

The effects of including wine lees (WL), exhausted olive cake (EOC) and a 1:1 mixture of EOC and tomato pomace (EOCTP) in diets for fattening ruminants on in vitro fermentation parameters and CH4 production were analysed. Ten diets were studied, containing either none of the tested by-products (control), or 6.0, 12.0 or 18.0% of WL, EOC and ECOTP formulated to have similar protein and fiber content. Diets were incubated in vitro with sheep ruminal fluid to measure gas production kinetics and fermentation parameters. Increasing the level of WL, EOC and EOCTP decreased linearly (p ≤ 0.009) the potential gas production, but other gas production parameters were unaffected (p > 0.05), excepting that EOCTP increased the gas production rate. No differences (p ≥ 0.0.05) among diets were observed in total volatile fatty acid (VFA) production at 24 h of incubation for EOC and EOCTP, but NH3-N concentration decreased (p ≤ 0.003). In contrast, WL at 12.0 and 18.0% decreased (p < 0.05) total VFA production and increased the acetate/propionate ratio (p < 0.05). None of the by-products had an effect on CH4 production (p ≥ 0.0.05). Results indicate that EOC and EOCTP could be included up to 18.0% in fattening diets, but lower levels of WL are recommended.

In recent years, animal husbandry has aimed at improving the conditions of livestock animals useful for humans to solve environmental and health problems. The formulation of animal feeds or supplements based on antioxidant plant compounds is considered a valuable approach and an alternative for livestock productivity. Forest biomass materials are an underestimated source of polyphenolic compounds whose sustainable recovery could provide direct benefits to animals and, indirectly, human nutrition. In this context, an alcohol extract from leaves of Fagus sylvatica L. was first investigated through an untargeted ultra-high-performance liquid chromatography–high-resolution tandem mass spectrometry (UHPLC-HRMS/MS) approach. Then, it was fractionated into a fatty acid-rich and a polyphenolic fraction, as evidenced by total lipid, phenol, and flavonoid content assays, with antiradical and reducing activity positively correlated to the latter. When tested in vitro with rumen liquor to evaluate changes in the fermentative parameters, a significant detrimental effect was exerted by the lipid-rich fraction, whereas the flavonoid-rich one positively modulated the production of volatile fatty acids (i.e., acetate, butyrate, propionate, etc.).

The objective of this work was to evaluate the pH, chemical composition, minerals, vitamins, and in vitro rumen fermentation characteristics of silage prepared with lucerne, sweet maize stalk (MS), and their mixtures. Freshly chopped lucerne and MS were combined in ratios of 100:0 (M0, control), 80:20 (M20), 60:40 (M40), 40:60 (M60), 20:80 (M80), and 0:100 (M100) on a fresh matter basis. Each treatment was prepared in triplicate, and a total of eighteen silos were fermented for 65 days. After 65 days of fermentation, the pH values in M0, M20, M40, M60, M80, and M100 silages were 5.47, 4.84, 4.23, 4.13, 3.79, and 3.61, respectively. As the MS proportion in the mixtures increased, silage K, Ca, P, Na, Fe, and Cu concentrations linearly decreased (p < 0.001) and so did vitamins B5 and K1 and α-tocopherol. In vitro rumen dry matter and organic matter degradability, pH, ammonia, total volatile fatty acid, and gas production linearly decreased (p < 0.01), while neutral detergent fiber concentration linearly increased (p < 0.001), with increasing proportion of MS. The in vitro dry matter and organic matter degradability rapidly decreased when the MS percentage was ≥60%. In conclusion, the M40 silage is the most suitable for livestock utilization in local forage production considering the balance of silage pH, nutritional quality, and in vitro ruminal fermentation characteristics.

This study investigated the effects of essential oil (EO) from three Korean wormwood (Artemisia Montana) plants on in vitro ruminal digestibility, fermentation, and microbial diversity. Dried (0.5 g) soybean meal (SBM) or bermudagrass hay (BGH) were incubated in buffered rumen fluid (40 mL) for 72 h with or without EO (5 mg/kg) from Ganghwa (GA), Injin (IN), or San (SA) wormwood (Experiment 1). Both SA and IN improved (p < 0.05) dry matter digestibility (DMD) of BGH, while GA reduced (p < 0.05) total short-chain fatty acid of BGH and SBM. Besides, SA increased (p < 0.05) numbers of Ruminococcus albus and Streptococcus bovis in SBM. Experiment 2 examined different doses (0, 0.1, 1, and 10 mg/kg) of SA, the most promising EO from Experiment 1. Applying SA at 10 mg/kg gave the highest DMD (L; p < 0.01) and neutral detergent fiber (Q; p < 0.05) digestibility for BGH. Applying SA at 1 mg/kg gave the highest R. albus population (Q; p < 0.05) in SBM. Therefore, SA was better than GA and IN at improving rumen fermentation, and the 0.1 to 1 and 10 mg/kg doses improved ruminal fermentation and in vitro digestibility of SBM and BGH, respectively.

The study analyzed the characteristics, chemical composition, and in vitro gas production kinetics of Porphyra umbilicalis and Saccharina latissima silages. Each seaweed was ensiled in vacuum bags (three bags/silage) following a 2 × 3 factorial design, with two pre-treatments (unwilted or pre-wilted) and three silage types: unwashed seaweed ensiled without additive; seaweed washed and ensiled without additive; and seaweed washed and ensiled with 4 g of formic acid (FAC) per kg seaweed. Silages were kept for 3 months in darkness at 20 °C. Pre-wilting prevented (p < 0.001) effluent formation and reduced (p ≤ 0.038) the production of NH3-N and volatile fatty acids for both seaweeds. Both pre-wilting and washing increased (p < 0.05) the ruminal degradability of P. umbilicalis silages but not of S. latissima silages. The pH of the FAC-treated silages was below 4.0, but ranged from 4.54 to 6.23 in non FAC-treated silages. DL-lactate concentrations were low (≤23.0 g/kg dry matter) and acetate was the predominant fermentation product, indicating a non-lactic fermentation. The estimated ruminal degradability of the P. umbilicalis and S. latissima silages was as average, 59.9 and 86.1% of that for high-quality rye-grass silages, respectively, indicating a medium-low nutritional value of these seaweed silages for ruminants.

The nutritive values for ruminants of cauliflower (CAU) and Romanesco (ROM) wastes (leaves, stems and sprouts) were assessed by analyzing their chemical composition, in vitro ruminal fermentation, and in vitro intestinal digestibility. In addition, the in vitro ruminal fermentation of diets containing increasing amounts of CAU was studied. The dry matter (DM) content of leaves, stems and sprouts of both vegetables was lower than 10%, but they contained high crude protein (CP; 19.9 to 33.0%) and sugar (16.3 to 28.7%) levels, and low neutral detergent fiber (21.6 to 32.3%). Stems and sprouts were more rapidly and extensively fermented in the rumen than leaves, but there were only minor differences the fermentation profiles of both vegetables. The estimated metabolizable energy content ranged from 9.3 (leaves) to 10.8 (sprouts) MJ/kg DM. The CP rumen degradability (12-h in situ incubations) was greater than 80.0% for all fractions, and the in vitro intestinal digestibility of CP ranged from 85.7 to 93.2%. The inclusion of up to 24% of dried CAU in the concentrate of a mixed diet (40:60 alfalfa hay:concentrate) increased the in vitro rumen fermentation of the CAU diet, but did not affect methane (CH4) production, indicating the lack of antimethanogenic compounds in CAU.

This study was designed to analyze the chemical composition and in vitro rumen fermentation of eight seaweed species (Brown: Alaria esculenta, Laminaria digitata, Pelvetia canaliculata, Saccharina latissima; Red: Mastocarpus stellatus, Palmaria palmata and Porphyra sp.; Green: Cladophora rupestris) collected in Norway during spring and autumn. Moreover, the in vitro ruminal fermentation of seventeen diets composed of 1:1 oat hay:concentrate, without (control diet) or including seaweeds was studied. The ash and N contents were greater (p < 0.001) in seaweeds collected during spring than in autumn, but autumn-seaweeds had greater total extractable polyphenols. Nitrogen in red and green seaweeds was greater than 2.20 and in brown seaweeds, it was lower than 1.92 g/kg DM. Degradability after 24 h of fermentation was greater in spring seaweeds than in autumn, with Palmaria palmata showing the greatest value and Pelvetia canaliculata the lowest. Seaweeds differed in their fermentation pattern, and autumn Alaria esculenta, Laminaria digitata, Saccharina latissima and Palmaria palmata were similar to high-starch feeds. The inclusion of seaweeds in the concentrate of a diet up to 200 g/kg concentrate produced only subtle effects on in vitro ruminal fermentation.

The objective of this study was to analyze the chemical composition, in vitro ruminal fermentation, and intestinal digestibility of discarded samples of four Brassica vegetables: Brussels sprouts (BS), white cabbage, Savoy cabbage, and red cabbage, and to assess the effects of including increasing amounts of BS in the concentrate of a dairy sheep diet on in vitro fermentation, CH4 production, and in situ degradation of the diets. All cabbages had low dry matter content (DM; 4/total volatile fatty acid ratio. In vivo trials are necessary to confirm these results.

The objective of this study was to analyze the efficacy of a treatment (MAH) of sunflower seed (SS) and meal (SM) with a malic acid solution (1 M; 400 mL/kg) and heating (150 °C, 2 h) to protect protein against rumen degradation and to improve the growth of lambs. Two homogeneous groups of 12 Lacaune lambs each (14.2 ± 0.35 kg body weight) were fed either a concentrate including untreated SS and SM or a concentrate with MAH-treated SS and SM. Lambs were fed concentrate and barley straw ad libitum for 40 days (about 26 kg body weight); feed intake and growth of lambs were recorded; blood samples were taken on days 0, 20, and the slaughter day for analysis of urea-N and amino acid-N; diet digestibility was determined; and ruminal and cecal samples were collected after slaughter. The in vitro incubation of both concentrates with sheep ruminal fluid for 12 h showed that the MAH-treatment tended to reduce NH3-N concentrations and increased propionate production. However, there were no differences (p > 0.05) between groups in any of the tested variables in the in vivo trial.

The objective of this study was to determine the variability in nutritive value for ruminants of tomato pomace (TP) samples and analyze its effect on in vitro fermentation when it was included in a high-concentrate diet. Twelve TP samples were obtained from two processing plants at weekly intervals and analyzed for chemical composition, in vitro rumen fermentation, and intestinal digestibility. The chemical composition of TP did not differ between processing plants and only slight variations were observed among sampling times. Tomato pomace had a low dry matter content (3-N concentrations without affecting CH4. In summary, TP samples showed little variability in nutritive value over sampling time and TP of up to 180 g/kg could be included in high-concentrate diets without negatively affecting rumen fermentation.