Gas Production, Digestibility and Efficacy of Stored or Fresh Plant Extracts to Reduce Methane Production on Different Substrates

Natural compounds such as plant secondary metabolites (PSM) can be used to replace antibiotic growth promoters as rumen modifiers. In this study, the effectiveness of stored and freshly extracted Aloe vera (AV), Azadirachta indica (AZ), Moringa oleifera (MO), Jatropha curcas (JA), Tithonia diversifolia (TD) and Carica papaya (CP) crude extract and monensin on in vitro gas and methane production, organic matter digestibility (IVOMD) and volatile fatty acids (VFA) were evaluated using a total mixed ration (TMR), lucerne or Eragrostis curvula substrates. Fresh extracts were processed from the same batch of frozen (−20 °C) plant material a few days before the trial while the stored extracts were extracted and stored at 4 °C for 12 months prior to the study. Extraction was done by solubilising 50 g freeze-dried plant material in 500 mL 100% methanol. Four mL of reconstituted 50 mg crude extract per 1000 mL distilled water was added per incubation vial, which already contained 400 mg substrate and in vitro fermentation, and gas production and IVOMD evaluation were carried out using standard procedures. Results showed that storing plant extracts for 12 months did not affect the activity or stability of metabolites present in the crude extracts, as shown by the lack of differences in total gas production (TGP) and methane produced between fresh or stored extracts across the substrates. In the TMR substrate, plant extracts increased IVOMD but did not affect TGP and methane production, whereas monensin did not have any effect. Plant extracts increased IVOMD of Eragrostis substrate and supressed methane production to a greater extent than monensin (p < 0.05). It can be concluded that storing plant extracts for up to 12 months did not compromise their efficacy. In addition, the use of 50 mg/kg of AV, AZ, MO, JA, TD and CP extract to a forage-based diet will reduce methane production while improving feed digestibility.

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Pomegranate seed oil rich in conjugated linolenic acids reduces in vitro methane production

South African Journal of Animal Science ◽

10.4314/sajas.v46i3.13 ◽

1970 ◽

Vol 46 (3) ◽

pp. 325-335

Author(s):

E. Maleki ◽

G.Y. Meng ◽

M. Faseleh Jahromi ◽

R. Jorfi ◽

A. Khoddami ◽

...

Keyword(s):

Methane Production ◽

Volatile Fatty Acids ◽

Seed Oil ◽

Control Diet ◽

Nutrient Utilization ◽

Gas Production ◽

Metabolizable Energy ◽

Ruminal Fermentation ◽

Ch4 Production

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.

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Effects of Thymol Supplementation on Goat Rumen Fermentation and Rumen Microbiota In Vitro

Microorganisms ◽

10.3390/microorganisms8081160 ◽

2020 ◽

Vol 8 (8) ◽

pp. 1160 ◽

Cited By ~ 1

Author(s):

Jiangkun Yu ◽

Liyuan Cai ◽

Jiacai Zhang ◽

Ao Yang ◽

Yanan Wang ◽

...

Keyword(s):

Methane Production ◽

Volatile Fatty Acids ◽

Rumen Fluid ◽

Optimal Dose ◽

Illumina Miseq ◽

Ammonia Nitrogen ◽

Rumen Fermentation ◽

Gas Production ◽

Rumen Microbiota

This study was performed to explore the predominant responses of rumen microbiota with thymol supplementation as well as effective dose of thymol on rumen fermentation. Thymol at different concentrations, i.e., 0, 100 mg/L, 200 mg/L, and 400 mg/L (four groups × five replications) was applied for 24 h of fermentation in a rumen fluid incubation system. Illumina MiSeq sequencing was applied to investigate the ruminal microbes in addition to the examination of rumen fermentation. Thymol doses reached 200 mg/L and significantly decreased (p < 0.05) total gas production (TGP) and methane production; the production of total volatile fatty acids (VFA), propionate, and ammonia nitrogen, and the digestibility of dry matter and organic matter were apparently decreased (p < 0.05) when the thymol dose reached 400 mg/L. A thymol dose of 200 mg/L significantly affected (p < 0.05) the relative abundance of 14 genera of bacteria, three species of archaea, and two genera of protozoa. Network analysis showed that bacteria, archaea, and protozoa significantly correlated with methane production and VFA production. This study indicates an optimal dose of thymol at 200 mg/L to facilitate rumen fermentation, the critical roles of bacteria in rumen fermentation, and their interactions with the archaea and protozoa.

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In vitro fermentability and methane production of some alternative forages in Australia

Animal Production Science ◽

10.1071/an15486 ◽

2016 ◽

Vol 56 (3) ◽

pp. 641 ◽

Cited By ~ 4

Author(s):

Z. Durmic ◽

P. J. Moate ◽

J. L. Jacobs ◽

J. Vadhanabhuti ◽

P. E. Vercoe

Keyword(s):

Brassica Napus ◽

Methane Production ◽

Volatile Fatty Acids ◽

Plantago Lanceolata ◽

Ammonia Concentration ◽

Gas Production ◽

Ruminal Fermentation ◽

Brassica Napus L

A study was conducted to examine in vitro ruminal fermentation profiles and methane production of some alternative forage species (n = 10) in Australia. Extent of fermentation was assessed using an in vitro batch fermentation system, where total gas production, methane production, and concentrations in ruminal fluid of volatile fatty acids (VFA) and ammonia were measured. Forages varied in their fermentability, with highest total gas, methane, VFA and ammonia production recorded from selected samples of Brassica napus L. cv. Winfred. Lowest methane production (i.e. 30% less than that formed by the highest-producing one) was observed in Plantago lanceolata L. cv. Tonic and Cichorium intybus L. cv. Choice. Selected plants, including P. lanceolata L. cv. Tonic, Brassica rapa L. cv. Marco, Brassica napus L. cv. Hunter had reduced acetate : propionate ratio and/or ammonia concentration, along with relatively low methane production compared with other species tested, while overall fermentation was not affected. It was concluded that selected novel forages have some advantageous fermentability profiles in the rumen and, in particular, inhibit methane production. However, before these can be recommended as valuable supplementary feedstuffs for ruminants in Australia, further studies are needed to confirm these effects over a range of samples, conditions and in vivo.

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In Vitro Fermentation Characteristics and Methane Mitigation Responded to Flavonoid Extract Levels from Alternanthera sissoo and Dietary Ratios

Fermentation ◽

10.3390/fermentation7030109 ◽

2021 ◽

Vol 7 (3) ◽

pp. 109

Author(s):

Sukruthai Sommai ◽

Anusorn Cherdthong ◽

Chanon Suntara ◽

Sarong So ◽

Metha Wanapat ◽

...

Keyword(s):

Methane Production ◽

Crude Protein ◽

Volatile Fatty Acids ◽

Gas Production ◽

Neutral Detergent Fiber ◽

Ruminal Fermentation ◽

Acid Detergent Fiber ◽

In Vitro Fermentation

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.

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Effects of Microalgae Species on In Vitro Rumen Fermentation Pattern and Methane Production

Annals of Animal Science ◽

10.2478/aoas-2019-0061 ◽

2020 ◽

Vol 20 (1) ◽

pp. 207-218 ◽

Cited By ~ 1

Author(s):

Ekin Sucu

Keyword(s):

Methane Production ◽

Dry Matter ◽

Volatile Fatty Acids ◽

Rumen Fermentation ◽

Gas Production ◽

Corn Silage ◽

Fermentation Pattern ◽

Proximate And Mineral Composition

AbstractThis experiment was conducted to establish the effects of two types of microalgae [Chlorella vulgaris (AI), C. variabilis (AII) and their combination (AI+AII)] with two substrates (wheat and corn silages) on rumen fermentation, gas and methane production. To each substrate, one of 3 algae treatment was supplemented at 0% and 25% of the total incubated dry matter. A series of 5 measurement points (3, 6, 12, 24 and 48 h) were completed and the gas production was monitored. The proximate and mineral composition of microalgae and substrates were examined. At 48 h incubation rumen fermentation variables and CH4 production were also assessed. When compared with wheat silage, corn silage caused an increase in gas production (P<0.05). Ruminal gas production decreased in the algae groups when compared to the controls (0% algae, wheat and corn silages, P<0.05). Among algae, C. vulgaris had the strongest effect, decreasing gas production by 34%. Among algae, the total volatile fatty acids (VFA) and CH4 production were found to be lower in C. variabilis (P<0.001). Ammonia-N increased with the algae inclusion (P<0.05). But, the ruminal gas production, pH, acetate, the total VFA, CH4 and rumen fermentation efficiency were not affected by the substrate and algae interaction (P>0.05). The propionate was the highest (P<0.05) for corn silage when incubated with C. vulgaris. Ruminal butyrate was the lowest for the wheat silage when incubated with the mixture of algae (P<0.05). The NH3-N was the highest in corn silage when incubated with all algae types (P<0.05). Careful selection and combination of substrate and algae may positively manipulate rumen fermentation and may inhibit CH4 production. Further research is needed to validate these results in vivo.

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In Vitro Estimation of the Effect of Grinding on Rumen Fermentation of Fibrous Feeds

Animals ◽

10.3390/ani10040732 ◽

2020 ◽

Vol 10 (4) ◽

pp. 732

Author(s):

Ignacio Rubén Ortolani ◽

Zahia Amanzougarene ◽

Manuel Fondevila

Keyword(s):

Methane Production ◽

Volatile Fatty Acids ◽

Rumen Fluid ◽

Palm Kernel ◽

Gas Production ◽

Barley Straw ◽

Oat Hulls ◽

Alfalfa Meal ◽

Gas Volume

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.

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Animal grazing selectivity and plant chemistry issues impact on the potential of Rhagodia preissii as an anthelmintic shrub

Parasitology ◽

10.1017/s0031182010001769 ◽

2011 ◽

Vol 138 (5) ◽

pp. 628-637 ◽

Cited By ~ 4

Author(s):

A. C. KOTZE ◽

E. N. ZADOW ◽

P. E. VERCOE ◽

N. PHILLIPS ◽

A. TOOVEY ◽

...

Keyword(s):

Secondary Metabolites ◽

Plant Extracts ◽

Plant Material ◽

Plant Secondary Metabolites ◽

Anthelmintic Activity ◽

Sheep Grazing ◽

Plant Chemistry ◽

Animal Grazing ◽

Selection Of

SUMMARYAs Rhagodia preissii had shown significant in vitro anthelmintic activity in a previous study, we examined the effect of including this shrub in the diet of sheep infected with Trichostrongylus colubriformis. Worm-infected merino wethers were grazed for 7 weeks on either R. preissii or annual pasture, and faecal egg counts (FECs) were conducted weekly. Plant material was collected weekly from eaten and uneaten plants, and analysed for levels of plant secondary metabolites (tannins, oxalates, saponins) and in vitro anthelmintic activity. While mean FECs were consistently lower in sheep grazing R. preissii compared to pasture (reductions of 20–74%), the differences were not significant. There was no relationship between grazing preference (eaten or uneaten) and in vitro anthelmintic activity of plant extracts. The levels of saponins and oxalates did not correlate with grazing preference or in vitro anthelmintic activity, while tannins were not responsible for the anthelmintic activity. While the identity of the grazing deterrent and in vitro anthelmintic compounds remain unknown, the presence of plants which were both highly preferred by the sheep and showed in vitro anthelmintic activity indicates a potential to develop the species as an anthelmintic shrub through selection of shrub populations dominated by such plants.

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Avian (IgY) anti-methanogen antibodies for reducing ruminal methane production: in vitro assessment of their effects

Australian Journal of Experimental Agriculture ◽

10.1071/ea07249 ◽

2008 ◽

Vol 48 (2) ◽

pp. 260 ◽

Cited By ~ 17

Author(s):

S. R. Cook ◽

P. K. Maiti ◽

A. V. Chaves ◽

C. Benchaar ◽

K. A. Beauchemin ◽

...

Keyword(s):

Methane Production ◽

Volatile Fatty Acids ◽

Antibody Activity ◽

Early Lactation ◽

Ch4 Production ◽

Transient Nature ◽

Methanobrevibacter Ruminantium ◽

Freeze Dried ◽

Egg Powder

In vitro dry matter disappearance (IVDMD) and production of methane, volatile fatty acids (VFA) and ammonia from an early lactation diet or from freeze-dried alfalfa were assessed in the presence of anti-methanogen antibody treatments in two in vitro ruminal incubations (experiments 1 and 2). In experiment 1, hens were immunised with crude cell preparations of Methanobrevibacter smithii, Methanobrevibacter ruminantium or Methanosphaera stadtmanae and complete Freund’s adjuvant (CFA). Semipurified egg antibodies (IgY) prepared from the hens’ eggs (α-SMICFA, α-RUMCFA, or α-STADCFA, respectively) were dispensed into 24 replicate vials (400 μL per vial) containing 500 mg of an early lactation total mixed ration (18% crude protein; 33% neutral detergent fibre; DM basis). Vials containing an equal volume of semipurified antibodies from eggs of non-immunised hens were included as a control. In experiment 2, hens were immunised with one of the three antigenic preparations combined with Montanide ISA 70 adjuvant. Triplicate vials per time point included 0.6 g of freeze-dried egg powder (α-SMIMon, α-RUMMon, α-STADMon; 19.0 ± 2.6 mg IgY/g) or a mixture of all three (ComboMon) and 500 mg of freeze-dried alfalfa. Total gas, methane production and pH were measured at intervals over 24 h. After 24 h, samples were analysed for VFA, ammonia and IVDMD. In experiment 1, cumulative CH4 production was similar (P > 0.05) among treatments at each sampling time. At 24 h, average CH4 production across treatments was 27.03 ± 0.205 mg/g DM. In experiment 2, α-SMIMon, α-STADMon and ComboMon reduced methane production at 12 h (P ≤ 0.05) compared with the control, but by 24 h, CH4 levels in all treatments were similar (P > 0.05) to the control. At 24 h, total VFA concentrations were lower (P < 0.05) in α-RUMMon and α-SMIMon than in the control. The transient nature of the inhibition of methane production by the antibodies may have arisen from instability of the antibodies in ruminal fluid, or to the presence of non-culturable methanogens unaffected by the antibody activity that was administered.

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Effect of rhubarb (Rheum spp.) root on in vitro and in vivo ruminal methane production and a bacterial community analysis based on 16S rRNA sequence

Animal Production Science ◽

10.1071/an15585 ◽

2016 ◽

Vol 56 (3) ◽

pp. 402 ◽

Cited By ~ 5

Author(s):

Kyoung Hoon Kim ◽

Selvaraj Arokiyaraj ◽

Jinwook Lee ◽

Young Kyoon Oh ◽

Ho Young Chung ◽

...

Keyword(s):

Bacterial Community ◽

Methane Production ◽

Volatile Fatty Acids ◽

Bacterial Community Composition ◽

Gas Production ◽

Feed Additives ◽

Ruminal Fermentation ◽

16S Rna

The objective of this study was to evaluate the anti-methanogenic effect of rhubarb (Rheum spp.) on in vitro, in vivo, and bacterial community composition using Quantitative Insights into Microbial Ecology sequencing. Rhubarb root powder was tested at different concentrations (0, 0.33, 0.67, and 1.33 g/L) in vitro, and all incubations were carried out in triplicate two runs on separate days. Concentrations of 0.67 and 1.33 g/L rhubarb significantly (P < 0.05) reduced methane production and the acetate : propionate ratio compared with those of the Control, without adverse effects on total volatile fatty acids and total gas production. In the second in vivo trial, four Hanwoo (Korean native) steers (live bodyweight, 556 ± 46 kg) with a ruminal cannula were housed individually in metabolic stalls and fed a basal diet twice daily in equal amounts at 0900 hours and 2100 hours. The before rhubarb treatment (before treatment) duration was 24 days for all steers; 14 days were used for diet adaptation and 10 days were used for gas samples collected 1, 2, and 3 h after the morning feeding on Days 3, 5, 7, and 9. We used three syringe needles passed through the ruminal cannula stopper at different time points as a simple and rapid method to sample rumen gas. Thereafter, three mesh bags containing 30 g of sliced rhubarb root each were placed at different depths in the rumen of each steer for 14 days (after treatment), and gas samples were collected on Days 4, 7, 10, 12, and 13. The results showed a significant (P < 0.05) decrease in methane concentration from the rhubarb-treated steers and provide the evidence that this method would be useful for in vivo screening of anti-methanogenic feed additives or plant material. Furthermore, 16s RNA sequencing after treatment showed increases in the numbers of Prevotella, and Lactobacillus, but decreases in Methanobrevibacter. In conclusion, rhubarb had an anti-methanogenic effect in vitro and in vivo, and the increase in the number of Prevotella shifted ruminal fermentation towards propionate production.

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Effects of essence oil additives added to different feeds on methane production

Revista Mvz Córdoba ◽

10.21897/rmvz.1023 ◽

2017 ◽

pp. 5854-5866

Author(s):

Ahmet Tekeli ◽

Gültekin Yıldız ◽

Winfried Drochner ◽

Herbert Steingass

Keyword(s):

Plant Extracts ◽

Methane Production ◽

Positive Control ◽

Gas Production ◽

Production Technique ◽

Control Groups ◽

Production Values ◽

In Vitro Gas Production ◽

Essence Oil

Objective. The aim of the study is to determine the effect of different plant essence oil supplementation to TMR, concentrate and hay on methanogenesis and at 2, 8 and 24 hours (h) using in vitro gas production technique in cattle. Material and methods. Three fistulated Holstein dairy cows were used for rumen fluid collection for application of in vitro gas production technique. Four essence oils (T. vulgaris, O. vulgare, S. aromaticum, Z. officinale) were used as plant extracts. Results. Essence oil supplementation, particularly at increased doses, significantly decreased methane production for TMR, concentrate and hay at 8 and 24 h compared to negative and positive control groups (p<0.05). Among all plant extracts, Syzygium 200 ppm supplementation resulted the lowest methane production values(p<0.05). While methane production at 24 h in positive control groups were recorded respectively as 10.45, 10.75 and 10.07 for TMR, concentrate and hay, the values in Syzygium 200 ppm group were recorded respectively as 1.65, 3.28 and 1.98 for these feed groups. Conclusions. The findings of the study indicate that increased doses of essence oil supplementation significantly decrease methane production in ruminants. Hence, it is suggested that essence oil will be highly beneficial in ruminant nutrition.

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