scholarly journals Effect of Acacia purified tannins extract and polyethylene glycol treatment on in vitro ruminal fermentation pattern and methane production

Author(s):  
Khaoula Khelalfa ◽  
Rabah Arhab ◽  
Antonio Ignacio Martín-García ◽  
Nabila Zaabat ◽  
Alejandro Belanche

Condensed tannins are polyphenolic compounds which can exert beneficial effects in ruminants. They have the ability to bind proteins and decrease their degradation. They have also been reported to reduce methanogenesis and improve ruminants performances. The present work aimed to study the effect of purified condensed tannins from Acacia horrida extract on fermentation parameters, gas and methane production. In this context, seven substrates were selected: four Acacia species (Acacia pycnantha, Acacia dealbata, Acacia horrida and Acacia cyanophylla) and three plant by-products (date palm leaves, grenade peel and artichoke stems). Biological activity of tannins was evaluated by the incubation of the substrates in vitro with Polyethylene glycol (PEG). Tannins decreased concentration of all fermentation parameters (gas (p < 0.0001), methane (CH4) (p= 0.0983), ammonia (N-NH3) (p= 0.0382), Volatile Fatty Acids (VFA) (p= 0.0009), acetate (p <0.0001), propionate (p= 0.1024), butyrate (p= 0.0373), isobutyrate (p= 0.0046), valerate (p < 0.0001), isovalerate (p= 0.0032)). Substrates were also characterized by their content in main nutrients (dry matter (DM), organic matter (OM), ash, crude protein (CP), neutral detergent fiber (NDF), acid detergent fiber (ADF) ,acid detergent lignin (ADL)) and in secondary metabolites (total phenols (TP), total tannins (TT) and condensed tannins (CT) ). All substrates presented high CP content except by-products (> 100 g/kg DM) and moderate cell-wall components. CT content was comprised between 120.5 and 680.4 g/kg DM in plants and between 23.1 and 170.4 g/kg DM in plant by-products. A. horrida presented the highest biological activity. Thus, their CT were extracted and purified on a sephadex LH-20 column. Purified tannins from A. horrida were incubated with two different forages alfalfa hay and barley straw at three different concentrations: 50, 100, 150 mg/g DM. Results showed that purified tannins from A. horrida had no effect on fermentation parameters (P > 0.05). The effects of A. horrida tannins may be strongly linked to their structure and their molecular weight more than to their concentration. For this reason, it is interesting that this work may be completed by physical characterization of these tannins.

Author(s):  
Francine M Ciriaco ◽  
Darren D Henry ◽  
Tessa M Schulmeister ◽  
Carla D Sanford ◽  
Luara B Canal ◽  
...  

Abstract To determine the effect of CaO treated Pensacola bahiagrass (Paspalum notatum) hay on intake, ruminal fermentation parameters, and apparent total tract digestibility of nutrients, 9 ruminally cannulated Angus-crossbred steers were used in a triplicated 3 × 3 Latin square design. Steers had ad libitum access to either: 1) untreated dry hay (DH;  n = 8); 2) hay at 50% DM treated with 8.9% CaCO3 [dry matter (DM) basis; CC; n = 9]; or 3) hay at 50% DM treated with 5% CaO (DM basis; CO; n = 8). Water was added to reach 50% DM in the CC and CO diets. Ruminal fluid and blood samples were collected every 3 h for 24 h. Ruminal fluid was analyzed for pH, volatile fatty acids (VFA), and ammonia-nitrogen (NH3-N). Blood was analyzed for plasma urea nitrogen (PUN). Hay and fecal samples were collected for 4 d, 4 times daily for hay and twice daily for feces, to determine apparent total-tract digestibility of nutrients. The hay provided to steers during the digestibility period was analyzed for in vitro organic matter digestibility (IVOMD) for 48h. Data were analyzed as repeated measures for blood and ruminal fermentation parameters. Total DM intake was not affected (P ≥ 0.674) by treatment. A treatment effect (P &lt; 0.001) was observed for average ruminal pH, where steers consuming CO had the greatest pH (P &lt; 0.001). Ruminal concentration of NH3-N tended (P = 0.059) to be reduced in steers consuming CO. There was a treatment × time interaction (P = 0.023) on concentrations of PUN, where at 3 h DH and CO were lesser than CC (P ≤ 0.050) and at 21 h DH was lesser than CC (P = 0.020). Total VFA, acetate, propionate, butyrate, branched-chain VFA, and valerate concentrations were affected by treatment (P ≤ 0.035), where a reduction (P ≤ 0.034) occurred in steers consuming CO. No treatment differences were observed for total-tract digestibility of DM (P = 0.186), organic matter (P = 0.169), or crude protein (P = 0.152); however, steers consuming DH had greater neutral detergent fiber (P = 0.038) than CC and tended to be greater than CO (P = 0.082). The CO hay had greater (P = 0.005) IVOMD compared with DH and tended (P = 0.100) to be greater than CC. Bahiagrass hay treated with CaO may reduce ruminal fermentation, as indicated by decreased total VFA concentration without altering DM intake. The addition of CaO did not improve the digestibility of bahiagrass hay in vivo; however, in vitro results are contradictory and warrant further elucidation.


Fermentation ◽  
2021 ◽  
Vol 7 (3) ◽  
pp. 109
Author(s):  
Sukruthai Sommai ◽  
Anusorn Cherdthong ◽  
Chanon Suntara ◽  
Sarong So ◽  
Metha Wanapat ◽  
...  

Two experiments were conducted under this study: Experiment 1 was to study production yield, chemical composition, and in vitro degradability of Brazilian spinach (Alternanthera sissoo; BS) leaf and leaf + leaf-stalk at various maturity ages of 15, 30, 45, and 60 days after plantation and regrowth and Experiment 2 was to evaluate the effect of flavonoid extract from BS leaf and leaf + leaf-stalk and dietary ratios on ruminal gas production, fermentation characteristics, and in vitro degradability. Experiment 1 showed that maturity ages after planting and regrowth increased, the yield significantly increased. Increasing maturity ages significantly (p < 0.05) increased neutral detergent fiber and acid detergent fiber content and decreased crude protein content, total flavonoid (TF) content, and degradability for both leaf and leaf + leaf-stalk. Maturity ages from 15 to 30 days after plantation and regrowth resulted (p < 0.05) the highest TF content and degradability for both leaf and leaf + leaf-stalk. Thus, BS leaf and leaf + leaf-stalk samples from 15 to 30 days of age were used for flavonoid extraction and used in the Experiment 2. Experiment 2 was conducted according to a 3 × 5 factorial experiment. Three roughage to concentrate (R:C) ratios at 50:50, 40:60, and 30:70 were used, and five levels of flavonoid extract (FE) at 0, 10, 20, 30, and 40 mg of substrate dry matter (DM) were supplemented. Experiment 2 showed that R:C ratio and FE had an interaction effect only on acetate to propionate ratio. Varying R:C ratios significantly increased (p < 0.05) in vitro DM degradability, total volatile fatty acids (VFA), and propionate (C3) concentration. FE supplementation linearly (p < 0.05) increased total VFA and C3 concentration and decreased methane production and protozoal population. This study could conclude that FE from BS could effectively modulate ruminal fermentation and decrease methane production. However, in vivo study needs to elucidate in order to validate the present results.


Animals ◽  
2020 ◽  
Vol 10 (4) ◽  
pp. 732
Author(s):  
Ignacio Rubén Ortolani ◽  
Zahia Amanzougarene ◽  
Manuel Fondevila

The fermentation patterns of six fiber sources, soybean hulls (SH), sugarbeet pulp (BP), palm kernel cake (PK), oat hulls (OH), dehydrated alfalfa meal (DA), and barley straw (BS) were evaluated for this study on the effect of their presentation form (non-processed, NP and ground, GR). Substrates were tested in a conventional in vitro batch system, using rumen fluid obtained from ewes fed 0.5 alfalfa hay and 0.5 barley straw. All substrates rendered a higher gas production in GR form (p < 0.05) except for BS but ranked similarly irrespective of the presentation form. Among the substrates, when incubated NP, the highest volume of gas was recorded with BP from 8 h onwards (p < 0.05), whereas OH and BS resulted in the lowest gas volume (p < 0.05). During the first half of the incubation period, methane production was higher in GR than NP (p < 0.05). Among substrates, despite NP or GR, methane production with BP was the highest (p < 0.05). Similarly, the presentation form did not qualitatively affect fermentation, as no differences were observed in volatile fatty acids proportions. The effect of particle size of fibrous substrates does not have a major impact on the rate and extent of the rumen microbial fermentation.


Animals ◽  
2019 ◽  
Vol 9 (5) ◽  
pp. 197 ◽  
Author(s):  
Pablo José Rufino-Moya ◽  
Mireia Blanco ◽  
Juan Ramón Bertolín ◽  
Margalida Joy

Alfalfa and sainfoin are high-quality forages with different condensed tannins (CT) content, which can be affected by the stage of maturity. To study the effects of CT on fermentation parameters, three substrates (alfalfa, sainfoin, and sainfoin+PEG) at three stages of maturity were in vitro incubated for 72 h. Sainfoin had greater total polyphenol and CT contents than alfalfa. As maturity advanced, CT contents in sainfoin decreased (p < 0.05), except for the protein-bound CT fraction (p > 0.05). The total gas and methane production was affected neither by the substrate nor by the stage of maturity (p > 0.05). Overall, sainfoin and sainfoin+PEG had greater in vitro organic matter degradability (IVOMD) than alfalfa (p < 0.05). Alfalfa and sainfoin+PEG presented higher ammonia content than sainfoin (p < 0.001). Total volatile fatty acid (VFA) production was only affected by the stage of maturity (p < 0.05), and the individual VFA proportions were affected by the substrate and the stage of maturity (p < 0.001). In conclusion, alfalfa and sainfoin only differed in the IVOMD and the fermentation end products. Moreover, CT reduced ammonia production and the ratio methane: VFA, but the IVOMD was reduced only in the vegetative stage.


Animals ◽  
2021 ◽  
Vol 11 (3) ◽  
pp. 761
Author(s):  
Olinda Guerreiro ◽  
Susana P. Alves ◽  
Mónica Costa ◽  
Maria F. Duarte ◽  
Eliana Jerónimo ◽  
...  

Cistus ladanifer (rockrose) is a perennial shrub quite abundant in the Mediterranean region, and it is a rich source in secondary compounds such as condensed tannins (CTs). Condensed tannins from C. ladanifer were able to change the ruminal biohydrogenation (BH), increasing the t11–18:1 and c9,t11–18:2 production. However, the adequate conditions of the C. ladanifer CTs used to optimize the production of t11–18:1 and c9,t11–18:2 is not yet known. Thus, we tested the effect of increasing the doses of C. ladanifer CT extract (0, 25, 50, 75 and 100 g/kg dry matter (DM)) on in vitro rumen BH. Five in vitro batch incubations replicates were conducted using an oil supplemented high-concentrate substrate, incubated for 24 h with 6 mL of buffered ruminal fluid. Volatile fatty acids (VFAs) and long chain fatty acids (FA) were analyzed at 0 h and 24 h, and BH of c9–18:1, c9, c12–18:2 and c9, c12, c15–18:3, and BH products yield were computed. Increasing doses of C. ladanifer CTs led to a moderate linear decrease (p < 0.001) of the VFA production (a reduction of 27% with the highest dose compared to control). The disappearance of c9–18:1 and c9,c12–18:2 as well as the production of t11–18:1 and c9, t11:18:2 was not affected by increasing doses of C. ladanifer CTs, and only the disappearance of c9, c12, c15–18:3 suffered a mild linear decrease (a reduction of 24% with the highest dose compared to control). Nevertheless, increasing the C. ladanifer CT dose led to a strong depression of microbial odd and branched fatty acids and of dimethyl acetals production (less than 65% with the highest dose compared to control), which indicates that microbial growth was more inhibited than fermentative and biohydrogenation activities, in a possible adaptative response of microbial population to stress induced to CTs and polyunsaturated fatty acids. The ability of C. ladanifer to modulate the ruminal BH was not verified in the current in vitro experimental conditions, emphasizing the inconsistent BH response to CTs and highlighting the need to continue seeking the optimal conditions for using CTs to improve the fatty acid profile of ruminant fat.


2011 ◽  
Vol 77 (8) ◽  
pp. 2634-2639 ◽  
Author(s):  
Zhenming Zhou ◽  
Qingxiang Meng ◽  
Zhongtang Yu

ABSTRACTThe objective of this study was to systematically evaluate and compare the effects of select antimethanogen compounds on methane production, feed digestion and fermentation, and populations of ruminal bacteria and methanogens usingin vitrocultures. Seven compounds, including 2-bromoethanesulphonate (BES), propynoic acid (PA), nitroethane (NE), ethyltrans-2-butenoate (ETB), 2-nitroethanol (2NEOH), sodium nitrate (SN), and ethyl-2-butynote (EB), were tested at a final concentration of 12 mM. Ground alfalfa hay was included as the only substrate to simulate daily forage intake. Compared to no-inhibitor controls, PA, 2NEOH, and SN greatly reduced the production of methane (70 to 99%), volatile fatty acids (VFAs; 46 to 66%), acetate (30 to 60%), and propionate (79 to 82%), with 2NEOH reducing the most. EB reduced methane production by 23% without a significant effect on total VFAs, acetate, or propionate. BES significantly reduced the propionate concentration but not the production of methane, total VFAs, or acetate. ETB or NE had no significant effect on any of the above-mentioned measurements. Specific quantitative-PCR (qPCR) assays showed that none of the inhibitors significantly affected total bacterial populations but that they did reduce theFibrobacter succinogenespopulation. SN reduced theRuminococcus albuspopulation, while PA and 2NEOH increased the populations of bothR. albusandRuminococcus flavefaciens. Archaeon-specific PCR-denaturing gradient gel electrophoresis (DGGE) showed that all the inhibitors affected the methanogen population structure, while archaeon-specific qPCR revealed a significant decrease in methanogen population in all treatments. These results showed that EB, ETB, NE, and BES can effectively reduce the total population of methanogens but that they reduce methane production to a lesser extent. The results may guide futureinvivostudies to develop effective mitigation of methane emission from ruminants.


1970 ◽  
Vol 46 (3) ◽  
pp. 325-335
Author(s):  
E. Maleki ◽  
G.Y. Meng ◽  
M. Faseleh Jahromi ◽  
R. Jorfi ◽  
A. Khoddami ◽  
...  

The objective of this study was to determine the effect of pomegranate (Punica granatum L.) seed oil (PSO) on gas and methane (CH4) production, ruminal fermentation and microbial populations under in vitro conditions. Three treatments consisting of a control diet containing 10 mg tallow (CON); the control diet with 5 mg PSO + 5 mg tallow (MPSO) and the control diet containing 10 mg PSO (HPSO) were compared. Ten mg of the experimental fat/oil samples were inserted into a gas-tight 100 mL plastic syringe containing 30 mL of an incubation inoculum and 250 mg of a basic substrate of a hay/concentrate (1/1, w/w) mixture. In vitro gas production was recorded over 0, 2, 4, 6, 8, 10, 12 and 24 h of incubation. After 24 hours, incubation was stopped, and methane production, pH, volatile fatty acids (VFAs) and microbial counts were measured in the inoculant. Gas production at 4, 6, 8, 10, 12 and 24 h incubation, metabolizable energy and in vitro organic matter disappearance increased linearly and quadratically as level of PSO increased. Furthermore, the 10 mg PSO (HPSO) decreased CH4 production by 21.0% compared with the control (CON) group. There were no significant differences in total and individual VFA concentrations between different levels of PSO, except for butyric acid. After 24 h of incubation, methanogenesis decreased in the HPSO compared with the MPSO and CON treatments. In addition, total bacteria and protozoa counts increased with rising PSO levels, while population methanogenesis declined significantly. These results suggested that PSO could reduce methane emissions, which might be beneficial to nutrient utilization and growth in ruminants.


2020 ◽  
Author(s):  
Hangshu Xin ◽  
Xin Liu ◽  
Xin Jiang ◽  
Chunlong Liu ◽  
Shuzhi Zhang ◽  
...  

Abstract Background: The objectives of this study were to evaluate the profiles of odd- and branched-chain fatty acids (OBCFA; including C15:0, iso-C15:0, anteiso-C15:0, iso-C16:0, C17:0, iso-C17:0 and anteiso-C17:0) during pure carbohydrates incubation in vitro and whether they correlated with ruminal fermentation parameters, microbial crude protein (MCP) synthesis, and bacterial populations. The pure substrates containing five different ratios of fiber and starch (F:S; 0:100, 25:75, 50:50, 75:25 and 100:0) were incubated for 6 h, 12 h, 18 h and 24 h. Results: Except iso-C17:0, OBCFA concentrations were interacted by F:S and incubation time. The highest concentration of total OBCFA was found in the fermented mixture after 24 h of incubation when the F:S = 0:100; while the lowest level was 1.65 mg/g DM produced after 6 h of incubation with F:S = 50:50. The concentrations of total volatile fatty acids (TVFA) and MCP remarkably decreased linearly as the inclusion of fiber in the substrates increased, as expected. The proportions of investigated cellulolytic bacteria in our study were increased linearly (or linearly and quadratically) while those of R. amylophilus and S. bovis were decreased as fiber inclusion increased. The correlation analysis indicated that iso-C16:0 concentration might have potential as a marker of productions of TVFA and MCP with ρ being 0.78 and 0.82 respectively. Compared to starch degrading bacteria, cellulolytic bacteria had more correlations with OBCFA profiles, and the strongest association was found on the population of R. flavefaciens with C15:0 concentration (ρ = 0.70). Conclusions: Our study shows there might be scope for iso-C16:0 to predict rumen productions of VFA and MCP. Notedly, this is the first paper reporting linkage of OBCFA with rumen function based on pure carbohydrate in vitro incubation, which would avoid confounding interference from dietary protein and fat presence. However, more in-depth experiments are needed to substantiate the current findings.


2020 ◽  
Author(s):  
Rafael Muñoz-Tamayo ◽  
Juana C. Chagas ◽  
Mohammad Ramin ◽  
Sophie J. Krizsan

AbstractBackgroundThe red macroalgae Asparagopsis taxiformis is a potent natural supplement for reducing methane production from cattle. A. taxiformis contains several anti-methanogenic compounds including bromoform that inhibits directly methanogenesis. The positive and adverse effects of A. taxiformis on the rumen microbiota are dose-dependent and operate in a dynamic fashion. It is therefore key to characterize the dynamic response of the rumen microbial fermentation for identifying optimal conditions on the use of A. taxiformis as a dietary supplement for methane mitigation. Accordingly, the objective of this work was to model the effect of A. taxiformis supplementation on the rumen microbial fermentation under in vitro conditions. We adapted a published mathematical model of rumen microbial fermentation to account for A. taxiformis supplementation. We modelled the impact of A. taxiformis on the fermentation and methane production by two mechanisms, namely (i) direct inhibition of the growth rate of methanogenesis by bromoform and (ii) hydrogen control on sugars utilization and on the flux distribution towards volatile fatty acids production. We calibrated our model using a multi-experiment estimation approach that integrated experimental data with six macroalgae supplementation levels from a published in vitro study assessing the dose-response impact of A. taxiformis on rumen fermentation.Resultsour model captured satisfactorily the effect of A. taxiformis on the dynamic profile of rumen microbial fermentation for the six supplementation levels of A. taxiformis with an average determination coefficient of 0.88 and an average coefficient of variation of the root mean squared error of 15.2% for acetate, butyrate, propionate, ammonia and methane.Conclusionsour results indicated the potential of our model as prediction tool for assessing the impact of additives such as seaweeds on the rumen microbial fermentation and methane production in vitro. Additional dynamic data on hydrogen and bromoform are required to validate our model structure and look for model structure improvements. We are working on model extensions to account for in vivo conditions. We expect this model development can be useful to help the design of sustainable nutritional strategies promoting healthy rumen function and low environmental footprint.


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