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Objective: This study investigated the impact of Aspergillus niger lysing polysaccharide monooxygenase (AnLPMO) on in vitro rumen microbial fermentation of rice straw. Methods: AnLPMO was heterologously expressed in Escherichia coli. Fourier transform infrared spectrometry and X-ray photoelectron spectroscopy analyzed the surface structure of rice straw after AnLPMO treatment. Two in vitro experiments, coupled with 16S highthroughput sequencing and quantitative real–time polymerase chain reaction techniques, assessed the influence of AnLPMO on rumen microbial fermentation of rice straw. Results: AnLPMO exhibited peak activity at 40°C and pH 6.5, with a preference for rice straw xylan hydrolysis, followed by Avicel. AnLPMO application led to the fractional removal of cellulose and hemicelluloses and a notable reduction in the levels of carbon elements and C–C groups present on the surface of rice straw. Compared to the control (no AnLPMO), supplementing AnLPMO at 1.1 to 2.0 U significantly enhanced in vitro digestibility of dry matter (IVDMD, p<0.01), total gas production (p<0.01), and concentrations of total volatile fatty acids (VFA, p<0.01), acetate (p<0.01), and ammonia-N (p<0.01). Particularly, the 1.4 U AnLPMO group showed a 14.8% increase in IVDMD. In the second experiment, compared to deactivated AnLPMO (1.4 U), supplementing bioactive AnLPMO at 1.4 U increased IVDMD (p = 0.01), total gas production (p = 0.04), and concentrations of total VFA (p<0.01), propionate (p<0.01), and ammonia-N (p<0.01), with a limited 9.6% increase in IVDMD. Supplementing AnLPMO stimulated the growth of ruminal bacterial taxa facilitating fiber degradation, including Proteobacteria, Spirochaetes, Succinivibrio, Rikenellaceae_RC9_ Gut_Group, Prevotelaceae_UCG-003, Desulfovibrio, Fibrobacter succinogenes, Ruminococcus albus, R. flavefaciens, Prevotella bryantii, P. ruminicola, and Treponema bryantii. Conclusion: These findings highlight AnLPMO's potential as a feed additive for improving rice straw utilization in ruminant production. [ABSTRACT FROM AUTHOR]

This study aimed to evaluate the influence of different plant species widespread in alpine pastures on in vitro rumen fermentation parameters and methane kinetic production. A total of 11 plant species were sampled at the beginning of the grazing season and used as substrates in an in vitro batch fermentation system. After 24h of fermentation, plants affected volatile fatty acids profiles, ammonia yield, and dry matter (DM) digestibilities. Carum carvi, Ranunculus. acris and Festuca rubra showed the highest total production of methane per unit of digested DM while Potentilla erecta was the species that produced less methane. In terms of methane as a percentage of the total gas, F. rubra had the highest value (28.9%) while R. acris had the lowest (24.2%). Total gas and methane production were monitored continuously and the percentage of methane in total gas was fitted with the Gompertz model. Plants differed significantly (p < .01) in methane production kinetics, including production rate decline (A), asymptotic methane concentration (B), time to maximum fermentation rate (TMFR), and maximum fermentation rate (MFR). C. carvi, Prunella grandiflora, and R. acris showed high values of MFR and the top values in the production rate decline (A > 0.9). The two grasses (F. rubra and Poa alpina) together with Hypericum maculatum showed an opposite behaviour with low values in MFR, A and a longer TMFR. The results of the methane production kinetics allow for an in-depth evaluation of plant species, adding further information to those registered at the end of fermentation.

To study the effect of hydrated sodium calcium aluminosilicate (HSCAS) in ameliorating adverse effects of aflatoxin on in vitro rumen fermentation, five treatment groups, viz., T1: control (wheat straw; 0.2 g); T2: T1+300 ppb Aflatoxin B1 (AFB1); T3: T2+0.33% HSCAS; T4: T2+0.5% HSCAS and T5: T2+ 1.0% HSCAS were prepared and incubated in vitro. The results revealed that truly degradable dry matter (TDDM), truly degradable organic matter (TDOM), gas production (GP), microbial biomass production (MBP) and partitioning factor (PF) values in aflatoxin contaminated group (T2) were lower (p<0.05) than those of other treatment groups. The TDDM, TDOM, GP, MBP and PF values in control group (T1) were higher than those of other treatment groups, i.e., T2 to T5. These parameters improved with increasing concentration of HSCAS. The total volatile fatty acids (TVFA), acetate (A), propionate (P) and butyrate (B) values in control group (T1) were higher (p<0.05) than those of other treatment groups, i.e., T2 to T5. The TVFA, A, P and B values in aflatoxin contaminated T2 group were lower (p<0.05) than those of other treatment groups. The A:P value among various dietary treatments did not vary significantly. It was concluded that aflatoxin contamination of feed (wheat straw) at 300 ppb level significantly affected the in vitro rumen fermentation in terms of reduced TDDM, TDOM, GP, MBP, PF, TVFA concentration. Inclusion of hydrated sodium calcium aluminosilicate to the aflatoxin contaminated feed partially ameliorated the adverse effects of aflatoxin on in vitro rumen fermentation parameters. [ABSTRACT FROM AUTHOR]

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. [ABSTRACT FROM AUTHOR]

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.

Saccharina latissima is a brown seaweed that could be used in ruminant feeding, but its fast deteriorating and seasonal growth nature limit their utilisation in the practice. Ensiling could be used as a preservation method, but information of its effects on the nutritional value of the seaweed is limited. This study evaluated the in vitro ruminal fermentation of different S. latissima silages using ruminal inoculum either from goats fed a mixed diet (60:40 oat hay:concentrate) or from sheep fed a high-forage diet (90:10 alfalfa hay:concentrate) to simulate different small ruminant production systems. S. latissima was ensiled in vacuum bags without additives (Control), with formic acid (4 g/kg seaweed; FA), with lactic acid bacteria (LAB) or with LAB after a pre-wilting treatment to reach a seaweed dry matter (DM) content of 30% (30LAB). Ensiling S. latissima decreased (p < 0.05) the content in DM, neutral detergent fibre and total extractable polyphenols, but nitrogen and fat content were unaffected. For both ruminal inoculums, ensiling decreased (p < 0.05) the asymptotic gas production after 120 h of fermentation (excepting for FA silage with goats' inoculum), but the total volatile fatty acid (VFA) production was unaffected. The VFA profile shifted towards greater (p < 0.05) acetate and lower (p < 0.05) propionate proportions in all silages compared with the pre-ensiling S. latissima. When goats inoculum was used, greater (p < 0.05) CH4 production compared with pre-ensiling S. latissima was observed in all silages, except Control one, which led to greater (p < 0.05) CH4/total VFA ratio. In contrast, no differences among samples (p > 0.05) in either CH4 production or CH4/total VFA ratio were observed when sheep' inoculum was used. Fermentation of all samples started earlier with goats' inoculum than with sheep' inoculum, which was attributed to the different diet fed to the animals. These results suggest that ensiling S. latissima with either formic acid or lactic acid bacteria could be a viable conservation method to preserve the nutritive value. [ABSTRACT FROM AUTHOR]

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). [ABSTRACT FROM AUTHOR]

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. [ABSTRACT FROM AUTHOR]

Simple Summary: Despite the importance of legume forage silage in ruminant livestock feeding, its low water-soluble carbohydrate content, great buffering capacity, and urea release to the environment limit its value. Ensilage of artichoke byproducts appears to be an environmentally efficient means of disposing of artichoke crops waste. Hence, this study aimed to investigate the effect of berseem co-ensiling with graded levels of artichoke bracts on silage characteristics. Moreover, the changes in ruminal fermentation characteristics and methane and ammonia production were evaluated using a buffalo inoculum source. The results showed that the co-silage of berseem and artichoke bracts considerably enhanced the silage quality, particularly after 30 days of ensiling at intermediate ratios. Moreover, the in vitro rumen degradation was significantly improved by artichoke bracts concentration of 500 g/kg fresh forage. 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. [ABSTRACT FROM AUTHOR]

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. [ABSTRACT FROM AUTHOR]

This experiment was conducted to investigate the effects of different dosages and types of medium‐chain fatty acids (MCFAs) on rumen fermentation in vitro under low‐ and high‐concentrate diets. For this purpose, two in vitro experiments (Exp.) were conducted. In Exp. 1, the concentrate–roughage ratio of the fermentation substrate [total mixed rations (TMR), dry matter (DM) basis] was 30:70 (low‐concentrate diet), while in Exp. 2, it was 70:30 (high‐concentrate diet). Three types of MCFAs with octanoic acid (C8), capric acid (C10), and lauric acid (C12) were added accounting for 1.5%, 6%, 9%, and 15% of the in vitro fermentation substrate weight (200 mg or 1 g, DM basis) based on control group, respectively. The results showed that the addition of MCFAs all could significantly reduce methane (CH4) production and the number of rumen protozoa, methanogens, and methanobrevibacter under the two diets with the dosages increased (p < 0.05). In addition, MCFAs had a certain degree of improvement on rumen fermentation and influenced in vitro digestibility under low‐ and high‐concentrate diets, and their effects were related to the dosages and types of MCFAs. This study provided a theoretical basis for the selection of types and dosages of MCFAs in ruminants production. [ABSTRACT FROM AUTHOR]

In the present study, we investigated the effects of cellulase and Lactiplantibacillus plantarum (L. plantarum) on the fermentation quality, microbial diversity, gene function prediction, and in vitro rumen fermentation parameters of the Caragana korshinskii (C. korshinskii) silage. The experimental treatment groups included control (CK), cellulase (C), L. plantarum (L), and cellulase + L. plantarum (CL). Triplicate silos were sampled after 14 and 56 days of ensiling. The microbial diversity of C. korshinskii silage at 56 days was analyzed using Illumina MiSeq sequencing technology, and the effects of C. korshinskii silage on rumen fermentation were evaluated by the in vitro artificial rumen gas production method. The results showed that the addition of cellulose and L. plantarum treatments reduced ammonia-nitrogen (NH3-N), dry matter (DM), neutral detergent fiber (NDF), and acid detergent fiber content during ensiling. Compared with the CK group, higher lactic acid content was observed in the silage of the C and L groups, but the CL group had a higher acetic acid (AA) content. Compared with the CK group, the relative abundance of Lactiplantibacillus significantly increased, but that of Enterococcus, Weissella, Enterobacter, and Pediococcus significantly decreased in all other groups except the C group (p < 0.05). The results of gene function prediction were as follows: pyruvate kinase, 1-phosphofructokinase, and fructokinase were lactate production enzymes, which suggested the association of the high AA content in the CL group to the high abundance of 6-phosphate glucose dehydrogenase. The abundance of H + -transporting ATPase and ATP-binding cassette (ABC) transporters in the L and CL groups was higher than that in the CK groups. Metabolism of terpenoids and polyketides, mismatch repair, DNA replication, nucleotide excision repair, and homologous recombination increased in the CL group compared with those in the CK group. Compared with the CK group, NH3-N and microbial protein content and the degradation rates of DM and NDF increased in the L and CL groups. To conclude, the L and CL groups had increased Lactiplantibacillus abundance, improved fermentation quality, and high DM degradation rates compared with the CK group.

Simple Summary: Exhausted olive cake, tomato pomace and wine lees are agroindustrial by-products extensively produced in the Mediterranean area that could be used in ruminant feeding, but the information on their optimal inclusion level in ruminant diets is scarce. Exhausted olive cake and tomato pomace are fibrous by-products and therefore are more adequate for ruminant feeding, whereas wine lees can have high protein content. The objective of this study was to evaluate the effect of including increasing amounts of these by-products in diets for fattening ruminants on in vitro ruminal fermentation and methane production. We analysed the in vitro fermentation of diets including 0, 6, 12 and 18% of these by-products. Including up to 18% of a 1:1 mixture of exhausted olive cake and tomato pomace had no negative effect on in vitro ruminal fermentation, but decreased ammonia concentrations, probably due to the low crude protein digestibility of both by-products. In contrast, high amounts of wine lees (12 and 18%) had negative effects on in vitro ruminal fermentation, and consequently they could only be included up to 6% to avoid negative effects. The absence of effects of any tested by-product on methane production indicates that they lack antimethanogenic compounds. 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. [ABSTRACT FROM AUTHOR]

Ammoniation of oil palm frond (OPF) with non‐protein nitrogen (N) sources has been shown to improve the nutritional value and digestibility of OPF in ruminants. This study evaluated the effect of treating OPF without (control) or with different urea levels (1%–5%) on chemical composition and in vitro gas production, digestibility and fermentation properties using goat rumen fluids. The results showed that the treated OPF with urea (1%–5%) had significantly lower (p < 0.05) dry matter (DM), organic matter (OM) and ash contents than that of the control. The crude protein (CP) content of treated OPF increased (linear p < 0.05; quadratic p < 0.05) with increasing levels of urea inclusion (1%–5%), whereas the contents of neutral detergent fibre (NDF) and acid detergent lignin (ADL) were significantly (p < 0.05) decreased. The CH4 (ml/500 mg DM incubated) production decreased (linear p < 0.05) with increasing levels of urea inclusion in treated OPF silage. However, in vitro DM and OM degradability were significantly (p < 0.05) increased by higher inclusion levels of urea (4% and 5%). OPF treated with 4% or 5% urea also revealed significantly (p < 0.05) higher total volatile fatty acids and ammonia‐N than the control and OPFs treated with 1%–3% urea. Ammoniation of OPF with urea improved its nutritional value and in vitro rumen fermentation profiles in goats. The impact was more pronounced for 4% or 5% urea‐treated OPF. [ABSTRACT FROM AUTHOR]

Objective: This study investigated the impact of Aspergillus niger lysing polysaccharide monooxygenase (AnLPMO) on in vitro rumen microbial fermentation of rice straw., Methods: AnLPMO was heterologously expressed in Escherichia coli. Fourier transform infrared spectrometry and X-ray photoelectron spectroscopy analyzed the surface structure of rice straw after AnLPMO treatment. Two in vitro experiments, coupled with 16S highthroughput sequencing and quantitative real-time polymerase chain reaction techniques, assessed the influence of AnLPMO on rumen microbial fermentation of rice straw., Results: AnLPMO exhibited peak activity at 40°C and pH 6.5, with a preference for rice straw xylan hydrolysis, followed by Avicel. AnLPMO application led to the fractional removal of cellulose and hemicelluloses and a notable reduction in the levels of carbon elements and C-C groups present on the surface of rice straw. Compared to the control (no AnLPMO), supplementing AnLPMO at 1.1 to 2.0 U significantly enhanced in vitro digestibility of dry matter (IVDMD, p<0.01), total gas production (p<0.01), and concentrations of total volatile fatty acids (VFA, p<0.01), acetate (p<0.01), and ammonia-N (p<0.01). Particularly, the 1.4 U AnLPMO group showed a 14.8% increase in IVDMD. In the second experiment, compared to deactivated AnLPMO (1.4 U), supplementing bioactive AnLPMO at 1.4 U increased IVDMD (p = 0.01), total gas production (p = 0.04), and concentrations of total VFA (p<0.01), propionate (p<0.01), and ammonia-N (p<0.01), with a limited 9.6% increase in IVDMD. Supplementing AnLPMO stimulated the growth of ruminal bacterial taxa facilitating fiber degradation, including Proteobacteria, Spirochaetes, Succinivibrio, Rikenellaceae&#95;RC9&#95; Gut&#95;Group, Prevotelaceae&#95;UCG-003, Desulfovibrio, Fibrobacter succinogenes, Ruminococcus albus, R. flavefaciens, Prevotella bryantii, P. ruminicola, and Treponema bryantii., Conclusion: These findings highlight AnLPMO's potential as a feed additive for improving rice straw utilization in ruminant production.

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.

Some excellent legume forages are difficult to ensile naturally due to their high buffering capacity and low water-soluble carbohydrate content. This may cause serious problems like proteolysis. In the present study, strains of lactic acid bacteria with high acid productivity and high tannin tolerance were screened from different silages and combined with tannic acid (TA) as an addition to ensiling. The screened strains were identified as Lactobacillus plantarum (LP), with four of these strains then selected for their high tannin tolerance. Stylosanthes guianensis and whole-plant soybean (WPS) were ensiled with 1 and 2% (fresh matter basis) TA, four LP strains alone (6 log10 colony forming units per gram of fresh matter), or TA combined with LP strains. Fermentation parameters and in vitro rumen fermentation characteristics were analyzed after 30 days of fermentation. The results showed that TA + LP can be used to reduce pH values (P < 0.01), non-protein nitrogen (P < 0.01), and ammonia-nitrogen (P < 0.01). The in vitro crude protein digestibility of WPS silage was also decreased with the addition of TA + LP (P < 0.01). These results indicate that the addition of TA combined with tannin tolerance LP strains may improve the fermentation quality of legume silage, especially for reducing proteolysis. [ABSTRACT FROM AUTHOR]

Objective: This study identified the major lactic acid bacteria (LAB) strains from different fermented total mixed rations (FTMRs) via metataxonomic analysis and evaluated the ability of their standard strain as ensiling inoculants for corn stover silage. Methods: The bacterial composition of eight FTMRs were analyzed by 16S rDNA sequencing. Corn stover was ensiled without LAB inoculation (control) or with 1×106 cfu/g LAB standard strain (Lactobacillus vaginalis, Lactobacillus reuteri, Lactobacillus helveticus, or Lactobacillus paralimentarius) selected from the FTMRs or 10 g/t commercial silage inoculant (CSI) around 25°C for 56 days. For each inoculation, a portion of the silage was sampled to analyze ensiling characteristics at time intervals of 0, 1, 3, 7, 14, 28, and 56 days, gas production (GP), microbial crude protein and volatile fatty acids as the measurements of rumen fermentation characteristics were evaluated in vitro with the silages of 56 days after 72 h incubation. Results: Lactobacillus covered >85% relative abundance of all FTMRs, in which L. pontis, L. vaginalis, L. reuteri, L. helveticus, and L. paralimentarius showed >4% in specific FTMRs. CSI, L. helveticus, and L. paralimentarius accelerated the decline of silage pH. Silage inoculated with L. paralimentarius and CSI produced more lactic acid the early 14 days. Silage inoculated with L. paralimentarius produced less acetic acid and butyric acid. For the in vitro rumen fermentation, silage inoculated with CSI produced more potential GP, isobutyric acid, and isovaleric acid; silage inoculated with L. helveticus produced more potential GP and isovaleric acid, silage inoculated with L. paralimentarius or L. reuteri produced more potential GP only. Conclusion: The standard strain L. paralimentarius (DSM 13238) is a promising ensiling inoculant for corn stover silage. The findings provide clues on strategies to select LAB to improve the quality of silage. [ABSTRACT FROM AUTHOR]

Greenhouse gas (GHG) emissions from livestock are an important consideration in environmental science. Estimating GHG production can be problematic at a farm or animal level, and requires controlled conditions to produce real data. An in vitro gas production technique (IVGPT) was developed to evaluate forage-based total mixed rations in digestion kinetics and GHG production. Two hundred and sixty samples of complete mixed rations (MR), which included a pasture component used in commercial lactating dairy herds, were collected around NZ across three calendar years, 2017-2019. Twenty of the 260 samples were 100% total mixed rations (TMR) with no pasture content. The samples were submitted for proximate analysis as well as IVGPT to generate GHG production figures. The results showed an average total gas production (TGP) of 129.82 ml/g dry matter (DM), 78.6% true digestibility (TDMD), 125.06 mg/g DM microbial biomass (MB), 20.16 g CH4/kg DM, and 12.8 MJME/kg DM. The average nutrient composition was dry matter (DM) 31.55%, crude protein (CP) 21.85%, neutral detergent fibre (NDF) 44.35%, and starch 7.03%. The IVGPT CH4 production was negatively correlated to NDF (r=-0.312), ADF (r=-0.193), TGP (r=-0.216), and was positively correlated with TDMD (r=0.250), apparent digestibility (ADMD) (r=0.614), starch (r=0.117) and volatile fatty acids (r=0.538). The MR diet showed a strong positive relationship with ADMD digestibility (P=0.01) and a negative relationship with fibre content (NDF, P=0.01 and ADF, P=0.01). However, CH4 production reduced linearly with increasing TGP (P=0.01). The results indicated that a greater CH4 production may be related to higher digestibility of mixed ration.

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.

This study characterized the nutritive and microbial profiles and the fermentation characteristics of silage with the following compositions on a dry matter (DM) basis: (1) 100% sorghum, (2) 70% sorghum + 30% carrot or pumpkin, and (3) 40% sorghum + 60% carrot or pumpkin. The treatments were further divided based on the addition or no addition of a probiotic inoculant. After 70 days of ensiling, the silage was incubated for 48 h using the in vitro batch culture technique. Crude protein and non-fiber carbohydrates in the silage increased (P ≤ 0.01) by 5.7 percent point (pp) and 9.6 pp, respectively, with pumpkin at 60% DM. The V4 region of the 16S rRNA gene was sequenced to profile pre-ensiled and ensiled archeal and bacterial communities. Silages containing carrot or pumpkin strongly influenced the microbial structure (PERMANOVA: R 2 = 0.75; P < 0.001), despite the ensiled treatments being dominated by Lactobacillus spp., except for the control, which was dominated by Weissella and Pediococcus spp. (P < 0.01). Linear discriminant analysis indicated that carrot and pumpkin silages were responsible for the increased relative abundance of Lactobacillus and Acinetobacter spp. (log LDA score ≥ 2), respectively. After 48 h of incubation, carrot and pumpkin inclusion increased (P < 0.01) the in vitro DM digestibility by 22.5 and 31.3%, increased the total volatile fatty acids (VFAs) by 16 and 20.6% (P < 0.01), respectively, and showed a tendency (P = 0.07) to increase the gas production. Therefore, this study supports the use of carrot or pumpkin in sorghum silages to maximize feed digestibility and total VFA concentrations. [ABSTRACT FROM AUTHOR]

The experiment studied the effect of different proportions of linseed oil and palm oil in substrate diet on in vitro rumen fermentation of cashmere goats by in vitro method. The experiment adopted a single factor completely randomized design, three Albas white cashmere goats were selected as rumen fluid donors for in vitro culture and divided into five treatment groups, with six replicates in each treatment. The control group (CON group) was only supplemented with 2% linseed oil without palm oil, and the PMO group was only supplemented with 2% palm oil without linseed oil. The linseed oil∶palm oil in the other three groups were 4∶6 (LP1 group), 5∶5 (LP2 group), 6∶4 (LP3 group). The concentrate to forage ratio of substrate was 5∶ 5, and the oil supplemental level in each group was 2%. Rumen fermentation parameters were measured at 3, 6, 9, 12 and 24 h, and multiple combination effect value (MFAEI) was calculated. The results showed that adding different proportions of linseed oil and palm oil decreased the concentration of ammonia nitrogen (NH3-N), protozoa number and gas production, increased the concentration of microbial protein (BCP) and volatile fatty acid (VFA) in rumen medium in vitro. The experiment indicates that when the mixing ratio of linseed oil and palm oil is 6∶4, the effect of promoting the in vitro rumen fermentation function of cashmere goats is better. [ABSTRACT FROM AUTHOR]

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.). [ABSTRACT FROM AUTHOR]

This study characterized the nutritive and microbial profiles and the fermentation characteristics of silage with the following compositions on a dry matter (DM) basis: (1) 100% sorghum, (2) 70% sorghum + 30% carrot or pumpkin, and (3) 40% sorghum + 60% carrot or pumpkin. The treatments were further divided based on the addition or no addition of a probiotic inoculant. After 70 days of ensiling, the silage was incubated for 48 h using the in vitro batch culture technique. Crude protein and non-fiber carbohydrates in the silage increased (P ≤ 0.01) by 5.7 percent point (pp) and 9.6 pp, respectively, with pumpkin at 60% DM. The V4 region of the 16S rRNA gene was sequenced to profile pre-ensiled and ensiled archeal and bacterial communities. Silages containing carrot or pumpkin strongly influenced the microbial structure (PERMANOVA: R2 = 0.75; P < 0.001), despite the ensiled treatments being dominated by Lactobacillus spp., except for the control, which was dominated by Weissella and Pediococcus spp. (P < 0.01). Linear discriminant analysis indicated that carrot and pumpkin silages were responsible for the increased relative abundance of Lactobacillus and Acinetobacter spp. (log LDA score ≥ 2), respectively. After 48 h of incubation, carrot and pumpkin inclusion increased (P < 0.01) the in vitro DM digestibility by 22.5 and 31.3%, increased the total volatile fatty acids (VFAs) by 16 and 20.6% (P < 0.01), respectively, and showed a tendency (P = 0.07) to increase the gas production. Therefore, this study supports the use of carrot or pumpkin in sorghum silages to maximize feed digestibility and total VFA concentrations.

The aim of this study was to determine the effect of grape seed tannin extract (GSTE) in the doses of 0, 25, 50, 75 g/kg dry matter (DM) feed on in vitro gas production (GP), volatile fatty acid (VFA) production and the digestibilities of true dry matter (TDMD) and neutral detergent fiber (NDFD) of alfalfa hay and barley grain. The supplementation of diets with 25 g/kg DM GSTE did not affect (p>0.05) potential gas production, TDMD, NDFD and ruminal total VFA of cultures compared with the diet without GSTE. Compared with the diet without GSTE, the addition of GSTE at > 50 g/kg DM to alfalfa hay diets or at 75 g/kg DM to barley grain diets reduced (p<0.01) potential gas production, TDMD, total VFA. Diet and GSTE had significant interactions (p<0.05) in total VFA and potential gas production. It was concluded that the effects of GSTE on in vitro ruminal fermentation were dose-dependent in both diet, but its effects on in vitro ruminal fermentation were more pronounced with alfalfa digestion than with barley grain. [ABSTRACT FROM AUTHOR]

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.

Abstract Supplementation of appropriate probiotics can improve the health and productivity of ruminants while mitigating environmental methane production. Hence, this study was conducted to determine the effects of Enterococcus faecium SROD on in vitro rumen fermentation, methane concentration, and microbial population structure. Ruminal samples were collected from ruminally cannulated Holstein–Friesian cattle, and 40:60 rice straw to concentrate ratio was used as substrate. Fresh culture of E. faecium SROD at different inclusion rates (0, 0.1%, 0.5%, and 1.0%) were investigated using in vitro rumen fermentation system. Addition of E. faecium SROD had a significant effect on total gas production with the greatest effect observed with 0.1% supplementation; however, there was no significant influence on pH. Supplementation of 0.1% E. faecium SROD resulted in the highest propionate (P = 0.005) but the lowest methane concentration (P = 0.001). In addition, acetate, butyrate, and total VFA concentrations in treatments were comparatively higher than control. Bioinformatics analysis revealed the predominance of the bacterial phyla Bacteroidetes and Firmicutes and the archaeal phylum Euryarchaeota. At the genus level, Prevotella (15–17%) and Methanobrevibacter (96%) dominated the bacterial and archaeal communities of the in vitro rumen fermenta, respectively. Supplementation of 0.1% E. faecium SROD resulted in the highest quantities of total bacteria and Ruminococcus flavefaciens, whereas 1.0% E. faecium SROD resulted in the highest contents of total fungi and Fibrobacter succinogenes. Overall, supplementation of 0.1% E. faecium SROD significantly increased the propionate and total volatile fatty acids concentrations but decreased the methane concentration while changing the microbial community abundance and composition.

Objective: This study was conducted to investigate the effects of molasses and Lactobacillus plantarum on the ensiling quality and in vitro rumen fermentation of sudangrass silage prepared with or without wilting. Methods: The ensiling experiment, measured with 3 replicates, was carried out according to a 2×4 (wilted stages×additives) factorial treatment structure. Dry matter of the fresh (210 g/kg fresh matter) or wilted (305 g/kg fresh matter) sudangrass were ensiled (packed into 5.0-L plastic jars) without additive (control) or with molasses (M), Lactobacillus plantarum (LP), or molasses + Lactobacillus plantarum (M+LP). After 60 days of ensiling, the silages were analyzed for the chemical, fermentation, and in vitro characteristics. Results: After 60 days of ensiling, the fermentation parameters were affected by wilted, the additives and the interactions of wilted with the additives (p<0.05). The M+LP treatment at wilted had higher lactic acid levels and V-score (p<0.05) but lower pH values and butyric acid concentrations than the other treatments. In comparison with sudangrass before ensiling, after ensiling had lower dry matter and higher non-fibrous carbohydrate. The in vitro gas production, in vitro dry matter digestibility, in vitro crude protein digestibility, and in vitro acid fiber detergent digestibility changed under the effects of the additives. Significant interactions were observed between wilted and the additives in terms of in vitro gas production at 48 h, asymptotic gas production, gas production rate, half time, and the average gas production rate. The total volatile fatty acid levels in the additive treatments were higher than those in the control. Conclusion: Wilting and supplementation with molasses and Lactobacillus plantarum had the ability to improve the ensiling quality and in vitro nutrient digestibility of sudangrass silage. The M+LP treatment at wilted exhibited the strongest positive effects on silage quality and in vitro ruminal fermentation characteristics. [ABSTRACT FROM AUTHOR]

With increasing interest in feed-based methane mitigation strategies and regional legal directives aimed at methane production from the agricultural sector, identifying local sources of biological feed additives will be critical for rendering these strategies affordable. In a recent study, the red alga Asparagopsis taxiformis harvested offshore Australia was identified as highly effective for reducing methane production from enteric fermentation. Due to potential difference in methane-reduction potential and the financial burden associated with transporting the harvested seaweed over long distances, we examined locally sourced red seaweed A. taxiformis and brown seaweed Zonaria farlowii for their ability to mitigate methane production when added to feed widely used in the Californian dairy industry. At a dose rate of 5% dry matter (DM), California-sourced A. taxiformis and Z. farlowii reduced methane production by up to 74% (p < 0.05) and 11% (p < 0.05) during in vitro rumen fermentation, respectively. No effect on CO2 production was observed for either seaweed. The measured decrease in methane production induced by A. taxiformis and Z. farlowii amendment, suggest that these local macroalgae are indeed promising candidates for biotic methane mitigation strategies in California, the largest milk producing state in the United States. To determine their real potential as methane mitigating feed supplements in the dairy industry, their effect in vivo will need to be investigated.

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.).

Sorghum-sudangrass hybrid silage has poor fermentation characteristics owing to a high moisture content. Accordingly, a 3 × 4+1 factorial design was applied to investigate the effects of adding different types and amounts of hay (corn stalk, wheat straw, and alfalfa hay at 12.5 kg t-1, 25 kg t-1, 37.5 kg t-1, and 50 kg t-1) on the nutritive value, fermentation quality, 72 h dry matter digestibility, and gas dynamics in vitro to simulate the rumen fermentation of sorghum-sudangrass hybrid silage. Separated silage of sorghum-sudangrass hybrids had a high butyric acid content and a FLIEG’s scores evaluation ranking of only “Fair.” The addition of hay significantly improved the fermentation quality of mixed silage. With respect to hay type, adding wheat straw had the best fermentation quality, alfalfa hay had the best nutritive value, in vitro dry matter digestibility (IVDMD) (662.41 g kg-1), constant fractional rate (C) (0.28 mL h-1), and the average gas production rate (AGPR) (32.70 mL h-1) content. There were no differences in the cumulative gas production at 72 h (GP72h), asymptotic gas production generated at a constant fractional rate (A), and lag time before gas production commenced (lag) among the three hay types. With respect to quantity, 25 kg t-1 hay had the best fermentation quality, 50 kg t-1 hay had the best nutritive value and highest IVDMD content (662.81 g kg-1), 37.5 kg t-1 hay had the highest C (0.28 mL h-1) and AGPR (31.48 mL h-1) contents, 25 kg t-1 hay had the highest Half time (2.20 h), and there were no significant differences in GP72h, A, and lag among the four amounts. Considering both nutritive value and fermentation quality, the best mixed silage mode was 37.5 kg t-1 wheat straw.

Aims: The aim of this research was to study of the effects of mimosa and chestnut tannin on ruminal fermentation. Methods and Results: The effects of inclusion of condensed tannin extract from mimosa and hydrolysed tannin from chestnut in the doses of 0, 25, 50, 75 g/kg dry matter (DM) feed into rumen fluid were investigated on in vitro ruminal fermentation characteristics, gas production, dry matter degradability and microbial protein production. The inclusion of 25 g tannin extract/kg DM into rumen fluid had no effects (P>0.05) on methane (CH4) production and fermentation parameters except for ammonia (NH3) –N concentration. The addition of 75 g tannin extract /kg DM into rumen fluid reduced (P<0.01) gas production from the insoluble fraction, potential gas production, the total volatile fatty acids (VFA), acetic acid, propionic acid, butyric acid and acetic acid/propionic acid ratio and NH3 level. Relative to chestnut tannin extract; gas production from the insoluble fraction, NH3 and CH4 concentration reduced (P<0.05), propionic acid increased (P<0.01) and tended to reduce (P>0.05) with mimosa tannin extract. Conclusions: Chestnut or mimosa tannin extract up to 50 g/kg DM feed can be used in ruminant nutrition but but high doses of these extract may decrease the rumen function and feed efficiency. Mimosa tannin extract has a stronger influence on reduction in NH3 and CH4 production compared to chestnut tannin extract. Significance and Impact of the Study: Depending on the type and concentration used, tannins can reduce microbial fermentation and diet digestibility. However, when supplied at optimal concentrations in ruminant diets, chestnut or mimosa tannin extract have potential to reduce CH4 and NH3 losses without adversely affecting efficiency of ruminal fermentation. [ABSTRACT FROM AUTHOR]

Objective: The present study aimed to evaluate the effects of γ-aminobutyric acid (GABA)-producing bacteria (GPB) on in vitro rumen fermentation and on the growth performance and meat quality of Hanwoo steers. Methods: The effects of GPB (Lactobacillus brevis YM 3-30)-produced and commercially available GABA were investigated using in vitro rumen fermentation. Using soybean meal as a substrate, either GPB-produced or commercially available GABA were added to the in vitro rumen fermentation bottles, as follows: control, no additive; T1, 2 g/L GPB; T2, 5 g/L GPB; T3, 2 g/L autoclaved GPB; T4, 5 g/L autoclaved GPB; T5, 2 g/L GABA; and T6, 5 g/L GABA. In addition, 27 Hanwoo steers (602.06±10.13 kg) were subjected to a 129-day feeding trial, during which they were fed daily with a commercially available total mixed ration that was supplemented with different amounts of GPB-produced GABA (control, no additive; T1, 2 g/L GPB; T2, 5 g/L GPB). The degree of marbling was assessed using the nine-point beef marbling standard while endotoxin was analyzed using a Chromo-Limulus amebocyte lysate test. Results: In regard to in vitro rumen fermentation, the addition of GPB-produced GABA failed to significantly affect pH or total gas production but did increase the ammonia nitrogen (NH3-N) concentration (p<0.05) and reduce total biogenic amines (p<0.05). Animals fed the GPB-produced GABA diet exhibited significantly lower levels of blood endotoxins than control animals and yielded comparable average daily gain, feed conversion ratio, and beef marbling scores. Conclusion: The addition of GPB improved in vitro fermentation by reducing biogenic amine production and by increasing both antioxidant activity and NH3-N production. Moreover, it also reduced the blood endotoxin levels of Hanwoo steers. [ABSTRACT FROM AUTHOR]

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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.

The aims of this research were to study the nutrient and secondary metabolite contents of solid herbal wastes (SHW) that were preserved by freeze drying, sun drying and silage, as well as to analyze their effects on in vitro fermentation indicators i.e., gas production kinetics and degradability of solid herbal waste. Physical and chemical properties on three forms of SHW (sun dry, freeze dry, and silage) were characterized and then an in vitro gas production experiment was performed to determine the kinetics of gas production, methane production, NH3, microbial protein, and SHW degradability. Polyethylene glycol (PEG) was added to the three treatments to determine the biological activity of tannins. Results showed that all three preparations of SHW still contained high nutrient and plant secondary metabolite contents. Gas production, methane, NH3, microbial protein, in vitro degradability of dry matter (IVDMD) and organic matter (IVDOM) of SHW silage were lower (P

Nutritional evaluation as a fodder and defaunating agent of four multipurpose tree leaves namely Ficus religiosa (Pipal), Ficus bengalensis (Bargad), Mangifera indica (Mango), Enterolobium timoba (Jungle jalebi) and two herbs namely Agave americana (Ramkanta) and Plantago major (Isafghol) was done in vitro. The mean content of OM, CP, EE, NDF, ADF, cellulose and lignin of these tree leaves and herbs were 88.6, 12.6, 2.4, 46.2, 33.5, 25.8 and 7.3% on DM basis, respectively. Enterolobium timoba leaves contained highest amount of CP (22.5%) while highest amount of ADF and lignin content was observed in Ficus bengalensis (41.1% / 12.1%) leaves. Total rumen protozoa as well as Holotrich and spirotrich protozoa number became zero due to inclusion of Agave americana and Enterolobium timoba leaves in the incubation media. Total volatile fatty acids (TVFA) and propionate production was higher where as NH3–N production was lower due to addition of Agave americana leaves in the incubation media. Highest IVTDMD and IVTOMD (61.4% / 64.1%) were observed for the Agave americana followed by Enterolobium timoba (59.8% / 62.5%) and Plantago major (57.5% / 59.2%) leaves. Activity of polysaccharide degrading enzymes like carboxymethyl cellulase and xylanase improved due to addition of Agave americana and Enterolobium timoba leaves in the incubation media. However, activity of β-glucosidase enzyme was similar among all the tested tree leaves and herbs. As a defaunating agent (removal of rumen protozoa / anti ciliate protozoal activity), Agave americana leaves were more effective in comparison to Plantage major leaves. The results indicated that among the tested tree leaves and herbs, Agave americana, Enterolobium timoba and Plantago major were good tree fodder for feeding to the animals and leaves of Agave americana and Enterolobium timoba could be used as defaunating agent for reducing rumen protozoal population to improve animal productivity.

Ensiling vegetables with forage crops is a suggested method of waste diversion and can be directly utilized as a livestock feed. Carrot or pumpkin, ensiled at 0, 20, or 40% dry matter (DM) with crop sorghum, and with or without a second-generation silage inoculant were assessed for nutritive composition, organic acid profiles, aerobic stability and in vitro rumen fermentation characteristics. The study was a completely randomized design, with the fixed effects consisting of vegetable type (carrot vs. pumpkin), level (i.e., the level of vegetables), inoculant (inoculant or non-inoculant) and the interactions, and mini-silos within treatment as the random effect. The experimental unit for sorghum treatments represented by each mini-silo (5 kg capacity). Silage was sampled after 70-days ensiling for nutrient composition, 14-day aerobic stability, organic acid profiles and microbial diversity. After 24 h in vitro incubation, rumen fermentation parameters were assessed, measuring gas and methane (CH4) production, in vitro digestibility and volatile fatty acid concentrations. Sorghum ensiled with carrot or pumpkin at 20% or 40% DM increased crude fat (P ≤ 0.01) and decreased (P ≤ 0.01) silage surface temperature upon aerobic exposure compared to the control. Bacterial communities analyzed through 16S rRNA gene sequencing linearly increased (P ≤ 0.01) in diversity across both vegetables when the vegetable proportion was increased in the silage; dominated by Lactobacillus species. ITS analysis of the fungal microbiota upon silage opening and after 14 days (aerobic stability) identified increased (P ≤ 0.03) fungal diversity with increasing vegetable proportions, predominantly populated by Fusarium denticulatum, Issatchenkia orientalis, Kazachstania humilis, and Monascus purpureus. Upon assessment in vitro, there was an increase (P ≤ 0.04) in in vitro digestibility and some CH4 parameters (% CH4, and mg CH4/g DM), with no effect (P ≥ 0.17) on remaining CH4 parameters (mL CH4/g DM, mg CH4/g digested DM), gas production or pH. However, increasing vegetable amount decreased percentage of acetic acid and increased percentage of propionic acid of the total VFA, decreasing A:P ratio and total VFA concentration as a result (P ≤ 0.01). The results from this study indicate including carrot or pumpkin at 20 or 40% DM in a sorghum silage can produce a highly digestible, microbially diverse and energy-rich livestock feed.

This study was done to investigate which components of rice bran (RB) are involved in the inhibition of methanogenesis by fractionating the rice bran and adding it to a rumen in vitro culture system. The RB extract obtained using ethanol and water was screened in an in vitro fermentation system. The experimental treatment conditions were as follows: a control group containing a substrate without supplements; substrates with 0.06 g of RB; 0.6 mL of ethanol; 0.6 mL of distilled water (DW); 0.6 mL of ethanol-soluble fraction (ESF); 0.06 g of ethanol-insoluble rice bran (EIRB); 0.6 mL of water-soluble fraction (WSF); and 0.06 g of water-insoluble rice bran (WIRB). Based on the result of the analysis, the addition of ESF significantly decreased CH 4 and CH 4 /g dry matter digested, methanogen population (p < 0.05), while gas and dry matter digestibility (DMD) were comparable with the control group. Total short-chain fatty acid (SCFA), and proportion of propionate were reduced, and the proportion of butyrate was increased by the addition of ethanol and ESF (p < 0.05). This result suggests that the supplementation of 10% ESF can substantially reduce methane production in vitro without a negative effect on substrate digestibility., (© 2024 The Authors. Animal Science Journal published by John Wiley & Sons Australia, Ltd on behalf of Japanese Society of Animal Science.)

Ensiling vegetables with forage crops is a suggested method of waste diversion and can be directly utilized as a livestock feed. Carrot or pumpkin, ensiled at 0, 20, or 40% dry matter (DM) with crop sorghum, and with or without a second-generation silage inoculant were assessed for nutritive composition, organic acid profiles, aerobic stability and in vitro rumen fermentation characteristics. The study was a completely randomized design, with the fixed effects consisting of vegetable type (carrot vs. pumpkin), level (i.e., the level of vegetables), inoculant (inoculant or non-inoculant) and the interactions, and mini-silos within treatment as the random effect. The experimental unit for sorghum treatments represented by each mini-silo (5 kg capacity). Silage was sampled after 70-days ensiling for nutrient composition, 14-day aerobic stability, organic acid profiles and microbial diversity. After 24 h in vitro incubation, rumen fermentation parameters were assessed, measuring gas and methane (CH4) production, in vitro digestibility and volatile fatty acid concentrations. Sorghum ensiled with carrot or pumpkin at 20% or 40% DM increased crude fat (P ≤ 0.01) and decreased (P ≤ 0.01) silage surface temperature upon aerobic exposure compared to the control. Bacterial communities analyzed through 16S rRNA gene sequencing linearly increased (P ≤ 0.01) in diversity across both vegetables when the vegetable proportion was increased in the silage; dominated by Lactobacillus species. ITS analysis of the fungal microbiota upon silage opening and after 14 days (aerobic stability) identified increased (P ≤ 0.03) fungal diversity with increasing vegetable proportions, predominantly populated by Fusarium denticulatum , Issatchenkia orientalis , Kazachstania humilis , and Monascus purpureus. Upon assessment in vitro , there was an increase (P ≤ 0.04) in in vitro digestibility and some CH4 parameters (% CH4, and mg CH4/g DM), with no effect (P ≥ 0.17) on remaining CH4 parameters (mL CH4/g DM, mg CH4/g digested DM), gas production or pH. However, increasing vegetable amount decreased percentage of acetic acid and increased percentage of propionic acid of the total VFA, decreasing A:P ratio and total VFA concentration as a result (P ≤ 0.01). The results from this study indicate including carrot or pumpkin at 20 or 40% DM in a sorghum silage can produce a highly digestible, microbially diverse and energy-rich livestock feed. [ABSTRACT FROM AUTHOR]

Abstract: An in vitro rumen gas production technique was employed to determine the methane production and fermentation characteristics of Leymus chinensis and Medicago ruthenica at differing harvest dates (May 15, May 30, June 30, July 30, August 30 and September 30), which are sequential phases within a single continuous growth of two 10-year-old pastures. To quantify the rate of degradation and compare in vitro rumen fermentation characteristic, a logistic-exponential model, where initial gas volume was zero (LE0), was used to fit gas production and methane output results. Dried, milled forage samples were incubated in vitro for 72 h at 39°C and gas production was recorded intermittently throughout the incubation and gas samples were collected to measure methane production. Results showed that there were significant interactions between species and harvest for all chemical composition variables (P

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.

The aim of the present study was to develop a FTIR method to quantify amounts and proportions of volatile fatty acids (VFA) and ammonia nitrogen (N-NH3) in fermentation fluids collected in vitro using innovative Bayesian models as chemometric technique. A set of 170 fluids, collected before and after 4 in vitro incubations of 8 diets in 5 replication plus 5 blanks, were analysed for VFA, N-NH3 and scanned using the MilcoScan FT2 (Foss Electric, Hillerød, Denmark) in the spectral range between 5000 and 900 cm-1. A Bayes B model was used to calibrate equations for each fermentative trait. The calibration equation predicts well VFA and N-NH3 amounts in calibration and also in validation (R2 VAL ranged from 0.93 to 0.83 for isovaleric and n-butyric acid, respectively). However, the prediction of VFA expressed as proportions of total amount was much less accurate (R2 VAL ranged from 0.81 to 0.52 for iso-valeric and n-butyric acid, respectively). In conclusion, FTIR and Bayesian models can be used as tools to accurately predict VFA amounts in vitro.