Total Volatile Fatty Acid Production Research Articles

Relationship between predicted in vivo and observed in vivo methane production from dairy cows fed a grass-silage based diet with barley, oats, or dehulled oats as a concentrate supplement

The objectives of this study were to compare predicted in vivo methane (CH4) values based on data from an in vitro gas production experiment with observed in vivo values measured by the GreenFeed system, and to investigate the effect of the diet fed to donor animals of rumen inoculum (RI) on predicted in vivo CH4 production. For this purpose, we conducted an in vivo experiment simultaneously with an in vitro gas production experiment. The in vivo experiment (previously published) was a 4 × 4 Latin square design including 16 Nordic Red dairy cows. Cows were fed 60% grass silage and 40% concentrate which consisted of either barley, oats with hulls (hulled oats), dehulled oats, or a 50:50 mixture of hulled and dehulled oats on dry matter basis. The in vitro experiment was a 2 × 4 factorial design replicated in 4 runs. The in vitro diets were incubated for 48 h in two types of RI and formulated according to the diets fed in vivo. The RI was obtained from cows fed either barley (two cows) or hulled oats (two cows) as concentrate in the in vivo experiment. A set of models were applied to the gas and CH4 data obtained from the in vitro system to predict in vivo total gas and CH4 production. For the comparison between predicted in vivo and observed in vivo CH4, two different mean retention times (MRT of 35 and 50 h) in the rumen were used for the predictions. In the in vitro experiment, incubation residues were determined for organic matter digestibility and volatile fatty acids at 48 h of incubation. Assuming a MRT of 35 h in the rumen resulted in a significant relationship (P = 0.04) between predicted in vivo and observed in vivo CH4 yield (g/kg dry matter) with an R-square of 0.91 and a root mean square error of 0.20. Ranking of the diets in terms of their CH4 production was consistent between the in vitro and in vivo experiment. There were no significant interactions between diet and RI for any of the investigated parameters (P = 0.40). Rumen inoculum did not affect organic matter digestibility or total volatile fatty acid production. In conclusion, there was a good agreement between predicted in vivo and observed in vivo CH4 values. In addition, the diet of RI donor animal did not influence the comparison of diets in terms of CH4 production.

Ruminal inocula with distinct fermentation profiles differentially affect the in vitro fermentation pattern of a commercial algal blend.

The in vitro rumen batch technique is widely used for screening novel feed sources; however, it remains unclear to what extent the in vitro fermentability of non-conventional feed sources is affected by non-adapted ruminal inocula. Thus, in this study, we evaluated the effects of distinct ruminal inocula on the in vitro fermentation parameters of a sustainable non-conventional feed, a commercially available algal blend composed of microalgae (Chlorella vulgaris and Nannochloropsis oceanica) and seaweeds (Ulva sp. and Gracilaria gracilis). First, four late-lactation Holstein cows were fed four forage-based diets varying only in the proportions of basal forage (100% corn silage, 70% corn silage and 30% haylage, 30% corn silage and 70% haylage, and 100% haylage) in a 4 × 4 Latin square design with the last square omitted. After 3 weeks of adaptation, haylage-based diets resulted in ruminal fermentation parameters distinct from those promoted by corn silage-based diets, as reflected in increased pH, ammonia-N contents, and acetate proportions. Individual ruminal fluids derived from each of the four diets were further used as inocula in in vitro incubations. Here, a 1:1 mixture of corn silage and haylage was supplemented with 0, 5, 10, or 15% algal blend and incubated with each inoculum for 24 h in a 4 × 4 factorial design. Total gas and methane production decreased with inocula from cows fed haylage-based diets and with increasing algal blend supplementation levels. The fermentation pH increased and the ammonia-N contents decreased with inocula from cows fed haylage-based diets; however, these parameters were not affected by algal blend inclusion levels. The interaction between the ruminal inoculum source and the algal blend supplementation level affected the total volatile fatty acids (VFA) and the proportions of most individual VFA. Total VFA production decreased with increasing algal supplementation levels, particularly with inocula from cows fed 30% corn silage and 70% haylage; the acetate, propionate, and valerate proportions were only affected by algal blend levels under incubation with 100% corn silage inocula. Overall, our findings highlight the importance of the ruminal inoculum source when assessing the fermentability of non-conventional feed as well as the potential of the algal blend as a natural modulator of ruminal fermentation.

The Effect of Saponite Clay on Ruminal Fermentation Parameters during In Vitro Studies.

Reducing the emission of global warming gases currently remains one of the strategic tasks. Therefore, the objective of our work was to determine the effect of saponite clay on fermentation in the rumen of cows. The pH, total gas production, CH4, and volatile fatty acid (VFA) production in ruminal fluid was determined in vitro. Saponite clay from the Tashkiv deposit (Ukraine) has a high content of silicon, iron, aluminum, and magnesium. The addition of 0.15 and 0.25 g of saponite clay to the incubated mixture did not change the pH but reduced the total production (19% and 31%, respectively) and CH4 (24% and 46%, respectively) in the ruminal fluid compared to the control group and had no significant effect on the total VFA levels, but propionic acid increased by 15% and 21% and butyric acid decreased by 39% and 32%, respectively. We observed a decrease in the fermentation rates, with a simultaneous increase in the P:B ratio and an increase in the fermentation efficiency (FE) in the groups fermented with saponite clay, probably a consequence of the high efficiency in the breakdown of starch in the rumen. Therefore, further in vivo studies to determine the effective dose and effect of saponite clay on cow productivity and the reduction of gas emissions are promising and important.

The addition of anaerobic fungi isolates from buffalo rumen to increase fiber digestibility, fermentation, and microbial population in ruminants

Abstract. Agustina S, Wiryawan KG, Suharti S, Meryandini A. 2024. The addition of anaerobic fungi isolates from buffalo rumen to increase fiber digestibility, fermentation, and microbial population in ruminants. Biodiversitas 25: 107-115. Rumen microbes have an important role in the rumen. Anaerobic fungi are microbes needed in the forage digestion process in the rumen. The addition of microbes, particularly anaerobic fungi is essential to increase the digestibility of forage within rumen. Therefore, this study aimed to evaluate the addition of anaerobic fungi isolates from buffalo rumen to increase fiber digestibility, fermentation, and microbial population in sheep rumen. The in vitro tests were carried out using the Tilley and Terry method, using elephant grass and rice straw as tested forage. Piromyces sp. (F1, and F3), Caecomyces sp. (F2, and F5), and Neocallimastix frontalis (F4) isolates from buffalo rumen were used as tested anaerobic fungi. The result showed that the addition of anaerobic fungi isolates from buffalo rumen significantly affected fiber digestibility (Neutral Detergent Fiber (NDF), Acid Detergent Fiber (ADF), cellulose and hemicellulose) except lignin digestibility. The addition of N. frontalis had higher fiber digestibility which is 49.02% NDF digestibility, 42.11% ADF digestibility, 44.28% hemicellulose digestibility, and 38.60% cellulose digestibility. Furthermore, N. frontalis also significantly increased In vitro Dry Matter Digestibility (IVDMD), In vitro Organic Matter Digestibility (IVOMD), ammonium (NH3) production, total Volatile Fatty Acid (VFA) production, and microbial population compared to Piromyces sp., and Caecomyces sp. In conclusion, anaerobic fungus type N. frontalis showed promising potential to be used as a ruminant probiotic due to its superior effect on fiber digestibility, fermentation, and microbial population compared to Caecomyces sp. and Piromyces sp.

Effects of yeast culture and oxalic acid supplementation on in vitro nutrient disappearance, rumen fermentation, and bacterial community composition.

Hemicellulose is an important polysaccharide in ruminant nutrition, but it has not been studied as thoroughly as cellulose. Further research is needed to explore supplements that can improve its digestibility and ruminal buffering effects. Our previous research demonstrated the efficacy of oxalic acid (OA) as an essential nutrient in yeast culture (YC) for improving rumen fermentation performance. Consequently, we conducted in vitro rumen digestion experiments to examine the effects of YC and OA on rumen fermentation and bacterial composition. Two diets containing different levels of hemicellulose were formulated: diet 1 with 10.3% and diet 2 with 17% hemicellulose. Three levels of YC (0.00, 0.625, and 1.25 g/kg) and three doses of OA (0.0, 0.4, and 0.8 g/kg, DM) were added into each diet with a 3 × 3 factorial design. A comprehensive assessment was conducted on a total of 18 experimental treatments at fermentation periods of 0, 6, 12, 24, and 48 h. In the first experiment (diet 1), the supplementation of YC, OA, and their interaction significantly increased in vitro DM disappearance (IVDMD) and NDF disappearance (IVNDFD; p < 0.001). In the second experiment (diet 2), the supplementation of OA and the interaction between YC and OA (p < 0.001) increased IVDMD and IVCPD, but had no significant effects on IVNDFD. The interactions of YC and OA significantly increased ammonia nitrogen (p < 0.001). The production of acetic acid, propionic acid, and total volatile fatty acids (TVFA), and pH levels were significantly higher in treatments supplemented with YC and OA (p < 0.001). YC and OA in both diets significantly altered the rumen bacterial community leading to increased Shannon and Simpson diversity indices (p < 0.001). In both diets, OA supplementation significantly increased the relative abundance of the phylum Bacteroidetes and Prevotella genus. The result also showed a positive correlation between the Prevotella and Selenomonas genera with IVDMD, IVNDFD, propionic acid, and TVFA production, suggesting that these dominant bacteria enhanced nutrient disappearance in the rumen. In conclusion, adding YC and OA resulted in modifications to the bacterial community's composition and diversity, and improved nutrient disappearance. These changes indicate improved rumen fermentation efficiency, which is promising for future in vivo studies.

Supplementation of complex natural feed additive containing (C. militaris, probiotics and red ginseng by-product) on rumen-fermentation, growth performance and carcass characteristics in Korean native steers.

This study evaluated the effects of a complex natural feed additive on rumen fermentation, carcass characteristics and growth performance in Korean-native steers. In this study, in vitro and in vivo experiment were conducted. Seven different levels of complex natural feed additive (CA) were added to the buffered rumen fluid using AnkomRF gas production system for 12, 24 and 48 h. All experimental data were analyzed by mixed procedure of SAS. Total gas production increased in the CA groups, with the highest response observed in the 0.06% group at 48 h of incubation (linear, p = 0.02; quadratic, p < 0.01). Regarding rumen fermentation parameters, the total volatile fatty acid (TVFA) tended to increase in all the CA groups (p = 0.07). The concentrations of butyrate, iso-butyrate, and iso-valerate significantly increased in all treatment groups (p < 0.05). In the in vivo experiment, 23 Korean-native steers were allocated to two groups: (1) Control and (2) Treatment; control +0.07% CA (DM basis), in a randomized complete-block design and blocked by body weight (ave. body weight = 641.96 kg ± 62.51 kg, p = 0.80) and feed intake (ave. feed intake = 13.96 kg ± 0.74 kg, p = 0.08) lasted for 252 days. Average daily gain decreased in the treatment group (p < 0.01). Backfat thickness significantly decreased in the CA group (p = 0.03), whereas meat color tended to increase (p = 0.07). In conclusion, in the in vitro experiment, the inclusion of complex natural feed additive decreased methane proportion and tended to increase TVFA production, but supplementation to Korean native steers decreased average daily gain and backfat thickness.

Enteric methane emission reduction potential of natural feed supplements in ewe diets.

Research into the potential use of various dietary feed supplements to reduce methane (CH4) production from ruminants has proliferated in recent years. In this study, two 8-wk long experiments were conducted with mature ewes and incorporated the use of a variety of natural dietary feed supplements offered either independently or in combination. Both experiments followed a randomized complete block design. Ewes were offered a basal diet in the form of ad libitum access to grass silage supplemented with 0.5kg concentrates/ewe/d. The entire daily dietary concentrate allocation, incorporating the respective feed supplement, was offered each morning, and this was followed by the daily silage allocation. In experiment 1, the experimental diets contained 1) no supplementation (CON), 2) Ascophyllum nodosum (SW), 3) A. nodosum extract (EX1), 4) a blend of garlic and citrus extracts (GAR), and 5) a blend of essential oils (EO). In experiment 2, the experimental diets contained 1) no supplementation (CON), 2) A. nodosum extract (EX2), 3) soya oil (SO), and 4) a combination of EX2 and SO (EXSO). Twenty ewes per treatment were individually housed during both experiments. Methane was measured using portable accumulation chambers. Rumen fluid was collected at the end of both experiments for subsequent volatile fatty acid (VFA) and ammonia analyses. Data were analyzed using mixed models ANOVA (PROC MIXED, SAS v9.4). Statistically significant differences between treatment means were considered when P < 0.05. Dry matter intake was not affected by diet in either experiment (P > 0.05). Ewes offered EO tended to have an increased feed:gain ratio relative to CON (P < 0.10) and SO tended to increase the average daily gain (P < 0.10) which resulted in animals having a higher final body weight (P < 0.05) than CON. Ewes offered EX1 and SO emitted 9% less CH4 g/d than CON. The only dietary treatment to have an effect on rumen fermentation variables relative to CON was SW, which enhanced total VFA production (P < 0.05). In conclusion, the A. nodosum extract had inconsistent results on CH4 emissions whereby EX1 reduced CH4 g/d while EX2 had no mitigating effect on CH4 production, likely due to the differences in PT content reported for EX1 and EX2. SO was the only dietary feed supplement assessed in the current study that enhanced animal performance whilst mitigating daily CH4 production.

EFFECT OF NITRO COMPOUNDS ON in vitro RUMINAL METHANE, CARBON DIOXIDE, HYDROGEN AND DRY MATTER DEGRADABILITY

This study determined the in vitro effects of ethyl nitroacetate (ENA), ethyl 2-nitropropionate (E-2-NPP) and 3-nitropropionic acid (NPA) on ruminal production of CH4, H2, volatile fatty acids (VFAs) and in vitro dry matter disappearance of ground alfalfa. In vitro incubations were carried out over 24-h succeeding days (3 batches). Total gas and methane production decreased with all nitro compounds (P 0.05) in all the batches. Total production of VFAs was reduced by ENA and E-2-NPP (P 0.05), but not by NPA. DM disappearance was similar between treatments but decreased across batches. Further studies should be aimed at understanding the impact of NPA on ruminal metabolism, as well as its effect on in vivo models.

Enhancing Anaerobic Digestion with an UASB Reactor of the Winery Wastewater for Producing Volatile Fatty Acid Effluent Enriched in Caproic Acid

The production of Volatile Fatty Acids (VFAs) from wastewater holds significant importance in the context of biorefinery concepts due to their potential as valuable precursors for various bio-based processes. Therefore, the primary objective of this research is to investigate the fermentation of Winery Wastewater (WW) in an Upflow Anaerobic Sludge Blanket (UASB) reactor to generate VFAs, with particular emphasis on Caproic Acid (HCa) production and the dynamics of the microbiota, under varying Hydraulic Retention Time (HRT) periods (8, 5, and 2.5 h). The change from an 8 h to a 5 h HRT period resulted in an approximately 20% increase in total VFA production. However, when the HRT was further reduced to 2.5 h, total VFA production decreased by approximately 50%. Concerning the specific production of HCa, expressed in grams of Chemical Oxygen Demand (gCOD), the maximum yield was observed at around 0.9 gCOD/L for a 5-h HRT. Microbial population analysis revealed that Eubacteria outnumbered Archaea across all HRTs. Population dynamics analysis indicated that the Firmicutes Phylum was predominant in all cases. Within this phylum, bacteria such as Clostridium kluyveri and Clostridium sp., known for their ability to produce HCa, were identified. Based on the results obtained, the application of the UASB reactor for WW treatment, within the biorefinery framework, has the potential to provide a practical alternative for HCa production when operated with a 5 h HRT.

Essential Oil Blends with or without Fumaric Acid Influenced In Vitro Rumen Fermentation, Greenhouse Gas Emission, and Volatile Fatty Acids Production of a Total Mixed Ration

The growing interest in improving rumen fermentation and mitigating methane emissions necessitates the use of essential oil blends (EOB) and fumaric acid (FA). This study evaluated the synergistic effect of four EOB with or without FA supplementation on in vitro dry matter digestibility, greenhouse gas emission, and total volatile fatty acid production using inoculum from three rumen-cannulated Black Angus beef cows. The study was arranged in a 4 × 2 + 1 factorial design to evaluate the effects of the four EOB and two FA levels on a total mixed ration (TMR). The EOB dosage was 100 µL while FA was added at 3% of total mixed ration. The EOB × FA interaction (p &lt; 0.05) influenced the dry matter, neutral detergent fiber, and hemicellulose degradabilities. All the EOB and FA (EFA) treatments decreased (p &lt; 0.001) the dry matter degradability compared to the control (TMR substrate only). The EFA4 treatment reduced the neutral detergent fiber and hemicellulose degradabilities compared to the control. The ruminal pH was influenced (p &lt; 0.001) by both the EOB and FA inclusion, and the EOB × FA interaction was significant. The microbial mass was higher (p &lt; 0.001) in the EFA1, EFA4, and EOB4 compared to the control and the EOB3 treatments. The EFA1 and EOB1 produced less (p &lt; 0.001) gas than the control by 29.1 and 32.1%, respectively. Compared with the control, the EFA1 and EOB1 treatments decreased (p &lt; 0.001) methane gas by 90.8% and 86.4%, respectively, while the carbon dioxide was reduced (p = 0.004) by 65.7 and 57.9%, respectively. The EOB × FA interaction was significant (p &lt; 0.001) for the total and individual volatile fatty acid concentrations. The inclusion of FA increased the propionate concentration by 9.5% and decreased (p = 0.02) the acetate concentration by 4%. In summary, the synergistic effect of the EOB and FA offers an effective way to reduce greenhouse gas emission and enhance total volatile fatty acids.

Microencapsulation of Mitragyna leaf extracts to be used as a bioactive compound source to enhance in vitro fermentation characteristics and microbial dynamics.

Mitragyna speciosa Korth is traditionally used in Thailand. They have a high level of antioxidant capacities and bioactive compounds, the potential to modulate rumen fermentation and decrease methane production. The aim of the study was to investigate the different levels of microencapsulated-Mitragyna leaves extracts (MMLE) supplementation on nutrient degradability, rumen ecology, microbial dynamics, and methane production in an in vitro study. A completely randomized design was used to assign the experimental treatments, MMLE was supplemented at 0%, 4%, 6%, and 8% of the total dry matter (DM) substrate. The addition of MMLE significantly increased in vitro dry matter degradability both at 12, 24, and 48 h, while ammonia-nitrogen (NH3-N) concentration was improved with MMLE supplementation. The MMLE had the greatest propionate and total volatile fatty acid production when added with 6% of total DM substrate, while decreased the methane production (12, 24, and 48 h). Furthermore, the microbial population of cellulolytic bacteria and Butyrivibrio fibrisolvens were increased, whilst Methanobacteriales was decreased with MMLE feeding. The results indicated that MMLE could be a potential alternative plant-based bioactive compound supplement to be used as ruminant feed additives.

PSI-19 A Saccharomyces Cerevisiae-Derived Postbiotic Stimulates in Vitro Volatile Fatty Acid Production and Influences Hindgut Microbial Population Characteristics

Abstract Horses rely on volatile fatty acids (VFA) to provide a source of energy and fermentative end-products for their health and performance. Improvement in VFA production, has been associated in various species with improved feed conversion and availability of nutrients necessary to maintain epithelial cell growth, blood flow, and the normal secretory and absorptive functions of the intestine and positive changes in the microbial ecology of the gut. In this study, we characterized the effect of a dietary Saccharomyces cerevisiae fermentation postbiotic (SCFP; Diamond V, Cedar Rapids, IA) using an in vitro equine intestinal model to estimate the effects on VFA production and microbiome changes in the horse hindgut. Treatments contained inoculated medium containing a predigested blend of alfalfa and timothy hay substrate and a dilution of fresh equine excreta. The test article (SCFP) was evaluated at 150 mg (TEC) and 225 mg (TECP), in addition to a negative control containing inoculated medium alone, and the positive control containing inulin. The full array of treatments and replicates were incubated anaerobically at 37.2 C for 24 h. VFA concentrations were analyzed by gas chromatography and the microbiome was characterized through shotgun sequencing. Both levels of SCFP supported greater (P &amp;lt; 0.05) acetate, propionate, butyrate, and total VFA production versus the negative control. Greater production of acetate, propionate, butyrate and total VFA was observed for TECP compared with the positive control (P &amp;lt; 0.05). Between the two SCFP treatments, TECP promoted more acetate, propionate and total VFA (P &amp;lt; 0.01), and 18.3% more butyrate than TEC. Significant microbiome changes were also observed, with TEC and TECP promoting greater microbiome diversity (P &amp;lt; 0.05) compared with both negative and positive controls. Compared with the negative control, both levels of SCFP influenced an increased abundance (P &amp;lt; 0.05) of lactic acid producers (Veillonella), butyrate producers (Butyricicoccus, Anaerobutyricum), and an increased abundance of multiple Bacteroides species, all known for their enrichment of carbohydrate-degrading enzymes. Interestingly, our data show a distinct effect of inulin in microbial communities by promoting an increased abundance of Streptococcus species. This study provides evidence that a SCFP can stimulate VFA production, and this effect is incremental with the level of administration which could potentially provide gut health benefits in horses.

Cascading Integration of Electrofermentation and Photosynthesis—Low‐Carbon Biorefinery in Closed Loop Approach

AbstractSingle‐chambered electrofermentation (EF) bioreactors are operated using spent wash to evaluate the effect of applied voltage (EF/AV; −0.6 V) against closed‐circuit (EF/CC; 100 Ω) and control (EF/C; no voltage/resistance) operations using deoiled microalgae biomass‐derived biochar electrodes (anode) on increasing acidogenesis rate/efficiency. Higher total volatile fatty acids (VFA) production (mg L−1) is observed in EF/AV (2866), depicting 31% and 57% increments than EF/CC (1984) and EF/C (1224), respectively. The VFA profiles (C2–C4) in EF/AV are higher with acetic/butyric/propionic acids (1922/704/240 mg L−1) and biogas co‐generation [H2 (22%) and CH4 (3%)] than EF/CC and EF/C. The applied voltage (EF/AV) helps in electron flux regulation for increasing substrates conversion and VFA generation. Residual VFA‐rich effluents from EF process is utilized as substrate to mixotrophic microalgae cultivation (nutrient‐ and stress‐phase) that resulted in biomass production with simultaneous wastewater treatment. EF/AV depicts higher biomass growth (0.72 g L−1; 4th day) and substrate removal (63%) than other operations. Integrations of electrofermentation with photosynthetic processes operated with the residual resources in effluents towards product valorization and wastewater polishing, accounts for low‐carbon economy.

Marine Macroalgae in Rabbit Nutrition: In Vitro Digestibility, Caecal Fermentability, and Microbial Inhibitory Activity of Seven Macroalgae Species from Galicia (NW Spain)

The limitation on the prophylactic use of antibiotics in animal feed in Europe has critically challenged the rabbit meat industry, which urgently needs to find solutions. A feasible alternative could be using macroalgae in the diet to improve the gut health. This research studied seven species of marine macroalgae in four formats (dehydrated, enzymatically hydrolyzed, aqueous extract, and aqueous extract of hydrolyzed macroalgae) in order to select the most promising ones for their use in rabbit feed. Chemical composition, in vitro digestibility, in vitro caecal gas, total volatile fatty acid (VFA) production, and minimal inhibitory concentrations (MIC) against common pathogens were studied. All S. latissima products showed high caecal fermentability and VFA production, especially in both types of extracts. The H. elongata aqueous extract was remarkable due to its high in vitro butyrate production, which can be of great interest for improving gut health. The MIC results did not indicate any clear inhibition of the pathogens tested. The macroalgae tested appear to have a potentially prebiotic effect, rather than a direct antimicrobial activity. However, these results must be confirmed in vivo, in order to observe the real benefits of feeding macroalgae during the rabbit weaning period.

Optimization of Long-Chain Fatty Acid Synthesis from CO 2 Using Response Surface Methodology

A microbial electrosynthesis cell (MES) is an electrochemical technique in which electrolithoautotrophic electroactive microbes fix carbon dioxide (CO2) to longer-chain volatile fatty acids (VFAs). The synthesis of VFAs from CO2 requires optimization to improve the MES performance for industrial feasibility. This work studied the effect of different parameters, such as pH, headspace gas pressure, ethanol concentration, electrolyte, and trace element concentrations, on VFA synthesis from CO2 that used mixed anaerobic consortia in serum bottles. The operational parameters were varied according to a central composite design (CCD) and response surface methodology (RSM). A global optimum for the response variables was determined. The optimum values of different operating factors to maximize VFA production that was obtained from the optimizations were 1.12 × 105 Pa pressure, pH 7.149, ethanol = 2,318.7 mg/L, three times the standard Deutsche Sammlung von Mikroorganismen und Zellkulturen (DSMZ) 300 electrolyte concentration and five times the standard DSMZ 300 trace elements concentration. The experimental study was validated in triplicate, which considered the optimal values. The rate of total VFA production that was obtained from the operational parameters experimental study was 43.7 ± 5.9 mg/L/day under this optimized condition, which agreed with the predicted value of 30.39 mg/L/day. For the media optimization, validation of the experimental study was conducted at the optimal values, and their experimental response was 79.75 ± 19.26 mg/L/day, and the predicted response was 75.89 mg/L/day for total VFA production rates.

Methane reduction by quercetin, tannic and salicylic acids: influence of molecular structures on methane formation and fermentation in vitro

Plant secondary metabolites (PSMs) can potentially reduce ruminal methane formation. However, related to differences in their molecular structures, it is not yet clear what causes an anti-methanogenic effect. In an in vitro system simulating rumen fermentation, we investigated the impact of eight compounds with distinct chemical characteristics (gallic and salicylic acids, tannic acid, catechin, epicatechin, quercetin, rutin, and salicin) when added to a basal feed (maize silage) at a concentration of 12% of the feed dry matter. After 48 h of incubation in buffered rumen fluid, methane production was significantly lowered by quercetin (43%), tannic acid (39%) and salicylic acid (34%) compared to the control (maize silage alone) and without changes in total volatile fatty acid production during fermentation. No other PSM reduced methane formation as compared to control but induced significant differences on total volatile fatty acid production. The observed differences were related to lipophilicity, the presence of double bond and carbonyl group, sugar moieties, and polymerization of the compounds. Our results indicate the importance of distinct molecular structures of PSMs and chemical characteristics for methane lowering properties and volatile fatty acid formation. Further systematic screening studies to establish the structure–function relationship between PSMs and methane reduction are warranted.

Assessment of the Impact of Coffee Waste as an Alternative Feed Supplementation on Rumen Fermentation and Methane Emissions in an In Vitro Study

Spent coffee waste is the most common by-product of coffee processing, and it has the potential to be used as a source of organic compounds for ruminant diets. The objective of this study was to evaluate the optimal inclusion level and method for using spent coffee waste (SCW) as a ruminant feed and investigate its effects on rumen fermentation characteristics and methane (CH4) production. The present in vitro batch culture study was conducted using two different experimental designs. The first experimental design (TRIAL. 1) was performed using a control diet of 500 mg of fresh matter basal diet (60% hay/40% concentrate), with SCW being used as a feed additive at 1%, 10% and 20% of the substrate. The second experimental design was performed using the same control diet, with spent coffee waste replacing either part of the hay (TRIAL. 2) or some of the concentrate mixture (TRIAL. 3) at four different dosages (30:70, 50:50, 70:30 and 100). When SCW was supplemented as a feed additive, there were increases in the production of volatile fatty acids and gas; however, it did not show any suppressive effects on CH4 production. In contrast, when SCW was included as a replacement for hay or concentrate, there were significant reductions in CH4 production with increasing levels of SCW inclusion. These reductions in CH4 production were accompanied by negative effects on nutrient digestibility and total volatile fatty acid production. These findings demonstrate that SCW could potentially be used as a prebiotic feed additive. Additionally, when SCW is used as a replacement for silage at 70:30 and 50:50 dosages appear to be feasible as a substitute for animal feed (hay and concentrate).

Metagenomics insights into the effect of co-landfill of incineration fly ash and refuse for bacterial community succession and metabolism pathway of VFAs production

With the development of incineration technologies, incineration has become the most common treatment method of municipal solid waste in China. However, stabilized fly ash may enter landfills during the transition from landfill to incineration, which caused uncertain impact on landfill waste stabilization. Two simulated co-landfill columns were constructed based on different co-landfill methods (layer co-landfill and mixed co-landfill) to investigate the effect of stabilized fly ash co-landfilled municipal solid waste for bacterial community succession and change in metabolic pathways during hydrolysis-acidogenesis stage. The mixed co-landfill method resulted in higher degree of organic matter degradation, and the concentrations of volatile fatty acids (VFA) and chemical oxygen demand (COD) in leachate were higher. The dominant phyla were Firmicutes in the layered co-landfill column and Bacteroidetes in mixed co-landfill column. The dominant genera for the total bacterial composition and VFA production were different, Pseudomonas and Propionibacterium, Proteiniphilum and unclassified Bacteroides were the dominant genera responsible for VFA generation in the layered and mixed co-landfill columns. The genes for butyrate production were enriched in the layered co-landfill column, whereas those related to acetate production were enriched in mixed co-landfill column. However, the layered co-landfill inhibited the microbial metabolic activity at the end of the co-landfill process.

The effect of sainfoin (Onobrychis viciifolia Scop.) drying temperature and pelleting on in vitro fermentation characteristics using equine faecal inocula

This study was conducted to determine the effects of drying temperature and pelleting of the tanniniferous forage legume sainfoin (Onobrychis viciifolia Scop.) on the in vitro fermentation characteristics by equine faecal inocula. The sainfoin used for the experiment was air dried on the land and oven dried at 30, 70 and 130 °C for 2.5 h. Part of the dried sainfoin samples was left as dried hay, another part was pelleted, resulting in 6 different samples. The hay and pellets were tested for particle size distribution and pellets were tested for their pellet quality in terms of hardness, durability and bulk density. The gas production technique was used to simulate fermentation and faecal samples of eight randomly selected horses were used to create four independent inocula mixtures that were examined in two subsequent gas production runs. Per inocula mixture, the hay and pellet samples were incubated in triplicate as is, or in combination with polyethylene glycol to inactivate the tannins. Cumulative gas production, a proxy for the organic matter (OM) fermentation, was measured for 72 h, whereafter fermentation fluids were sampled and analysed for volatile fatty acids (VFA) and pH. The results of the pellet quality measurements showed that the particle sizes of the sainfoin particles in the pellets were decreased by a higher drying temperature or by the pelleting process, indicating a loss of fibre physical effectiveness, a factor that may be indicative for chewing activity and hence saliva production. The gas production results showed a significant (P < 0.05) effect of drying temperature and the addition of polyethylene glycol on the cumulative gas production, the VFA production and the percentage of branched chain VFA produced. Hay dried at 130 °C had a lower (P < 0.05) gas production compared to hay dried at 30 and 70 °C, which indicates a lower OM fermentability. The addition of polyethylene glycol increased the gas and total VFA production, which indicates that tannins in sainfoin decrease the production of gas and VFA in hay pellets compared to the hay product. The pelleting process had no effect on the fermentation characteristics, but pelleting conditions strongly decreased the level of the amino acid lysine and its reactivity. The results indicate the increase in drying temperature to cause a decrease in the OM hindgut fermentation and the total and reactive lysine contents of sainfoin. The positive effects of the tannins (a higher proportion of propionic acid relative to acetic acid and a lower protein fermentation) remain, despite of technological treatments.

Evaluation of different ensiling methods for Saccharina latissima preservation: influence on chemical composition and in vitro ruminal fermentation

ABSTRACT 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.