scholarly journals In Vitro Incubations Do Not Reflect In Vivo Differences Based on Ranking of Low and High Methane Emitters in Dairy Cows

Animals ◽  
2021 ◽  
Vol 11 (11) ◽  
pp. 3112
Author(s):  
Edward H. Cabezas-Garcia ◽  
Rebecca Danielsson ◽  
Mohammad Ramin ◽  
Pekka Huhtanen

This study evaluated if ranking dairy cows as low and high CH4 emitters using the GreenFeed system (GF) can be replicated in in vitro conditions using an automated gas system and its possible implications in terms of fermentation balance. Seven pairs of low and high emitters fed the same diet were selected on the basis of residual CH4 production, and rumen fluid taken from each pair incubated separately in the in vitro gas production system. In total, seven in vitro incubations were performed with inoculums taken from low and high CH4 emitting cows incubated in two substrates differing in forage-to-concentrate proportion, each without or with the addition of cashew nutshell liquid (CNSL) as an inhibitor of CH4 production. Except for the aimed differences in CH4 production, no statistical differences were detected among groups of low and high emitters either in in vivo animal performance or rumen fermentation profile prior to the in vitro incubations. The effect of in vivo ranking was poorly replicated in in vitro conditions after 48 h of anaerobic fermentation. Instead, the effects of diet and CNSL were more consistent. The inclusion of 50% barley in the diet (SB) increased both asymptotic gas production by 17.3% and predicted in vivo CH4 by 26.2%, when compared to 100% grass silage (S) substrate, respectively. The SB diet produced on average more propionate (+28 mmol/mol) and consequently less acetate compared to the S diet. Irrespective of CH4 emitter group, CNSL decreased predicted in vivo CH4 (26.7 vs. 11.1 mL/ g of dry matter; DM) and stoichiometric CH4 (CH4VFA; 304 vs. 235 moles/mol VFA), with these being also reflected in decreased total gas production per unit of volatile fatty acids (VFA). Microbial structure was assessed on rumen fluid sampled prior to in vitro incubation, by sequencing of the V4 region of 16S rRNA gene. Principal coordinate analysis (PCoA) on operational taxonomic unit (OTU) did not show any differences between groups. Some differences appeared of relative abundance between groups in some specific OTUs mainly related to Prevotella. Genus Methanobrevibacter represented 93.7 ± 3.33% of the archaeal sequences. There were no clear differences between groups in relative abundance of Methanobrevibacter.

2015 ◽  
pp. 4726-4738 ◽  
Author(s):  
John Ramírez ◽  
Sandra Posada O ◽  
Ricardo Noguera

ABSTRACT Objective. To evaluate the effect of Kikuyu grass (Pennisetum clandestinum) harvested at two different ages and three forage: concentrate supplement ratios (F/C) on methane (CH4) production, dry matter digestibility (DMD), and fermentation profile using the in vitro gas production technique. Materials and methods. six treatments, resulting from the combination of pasture age (30 or 60 days) and F/C (100/0, 75/25, or 50/50) were evaluated using a 2x3 factorial design. The response variables were measured 6, 12, 24 and 48 hours after incubation. A repeated-measure over time design was used to analyze the data, and differences between means were determined with the LSMEANS procedure of SAS. Results. the youngest grass (30 days) was more digestible, produced less CH4 per gram of digestible dry matter (dDM) and more total volatile fatty acids (VFA) compared to the oldest grass (60 days; p <0.05). Reductions of the F/C ratio increased DMD and CH4 production per gram of dDM (p<0.05) but had no significant effect on VFA concentration (p>0.05). Conclusions. under in vitro conditions and pH close to neutrality, the older grass reduces DMD and increases CH4 production per gram of dDM, while a F/C reduction increases DMD and CH4 production per gram of dDM, which differs with reports conducted in vivo.


2014 ◽  
Vol 54 (9) ◽  
pp. 1432 ◽  
Author(s):  
P. Lund ◽  
R. Dahl ◽  
H. J. Yang ◽  
A. L. F. Hellwing ◽  
B. B. Cao ◽  
...  

The effects of a 24-h administration of a ration high in nitrate (20 g/kg DM) on DM intake and enteric gas production in lactating dairy cows as well as the effect of different doses of nitrate on in vitro fermentation were studied. Nitrate reduced in vivo methane (CH4) production by 31%, and CH4 production increased again when nitrate supplementation was stopped. A similar effect was found when relating CH4 to carbon dioxide, and to DM intake. Addition of nitrate was followed by increased hydrogen production, which decreased again when nitrate was stopped. Nitrate addition did not affect in vitro rumen fermentation in terms of DM degradability, pH, ammonia nitrogen, microbial protein and volatile fatty acid production, but it decreased gas production with longer initial delay time before onset of gas production and lower gas production rate. Nitrate added at 7–20 g/kg ration DM significantly decreased net initial (0–12 h) CH4 production by 10–16%, although no further depression was observed afterwards.


2021 ◽  
Vol 51 (3) ◽  
pp. 399-406
Author(s):  
K. Selzer ◽  
A. Hassen ◽  
A.M. Akanmu ◽  
A.Z.M. Salem

Forages play an important role in ruminant animal production worldwide. Unlocking the nutritional potential of poor-quality tropical forages with fibrolytic enzymes would improve forage digestibility and utilization. Using in vitro and in vivo methods this study investigated the effect of pre-treating Smutsfinger hay for 24 hours with a mixture of fibrolytic enzyme (100% cellulase; 75% cellulase: 25% xylanase; 50% cellulase: 50% xylanase; 25% cellulase: 75% xylanase; 100% xylanase and a control with no enzyme) on ruminal fermentation and digestibility of nutrients by sheep. For in vitro fermentation, dry matter, neutral detergent fibre (NDF) degradability and volatile fatty acids (VFA) were determined with standard procedures. The same treatments were used for an in vivo digestibility trial using Merino sheep in a 6 x 6 Latin square design. Feed intake and total tract digestibility were recorded. Rumen fluid samples were collected daily, preserved, and analysed for VFA. The addition of 100% cellulase enzyme to Smutsfinger hay in vitro increased (P <0.05) NDF degradability and gas production compared with the control and inclusion of 100% xylanase enzyme. Both 100% cellulase and xylanase enzymes significantly reduced in vitro end time fermentation pH. A 50:50 mixture of cellulase and xylanase plus enzyme in vivo, increased acetate, total VFA concentration, and higher NDF and ADF digestibility of the test feed compared with the control. Inclusion of a 50-75% mixture of cellulase and 50-25% xylanase enzymes treatment led to higher gas production and butyrate concentration, decreased ruminal pH and improved nutrient digestibility.


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.


2011 ◽  
Vol 11 (2) ◽  
pp. 29-34 ◽  
Author(s):  
Novita Hindratiningrum ◽  
Muhamad Bata ◽  
Setya Agus Santosa

Products of rumen fermentation and protein microbial of dairy cattle feed with rice bran ammonization and some feedstuffs as an energy sourcesABSTRACT. This study aims to examine the energy sources of feed ingredients that can increase the production of Volatile Fatty Acids (VFA), N-NH3, microbial protein synthesis, total gas production and metabolic energy. The material used is as a source of rumen fluid inoculum from Frisian Holstein cows (FH) females, amoniasi rice straw, salt, mineral mix brand "Ultra Minerals' production Eka Farma Semarang, onggok wet and dry, corn, and rice bran. Observed variable is the concentration of (VFA), N-NH3, rumen microbial protein synthesis, and total gas production. Based on the analysis of diversity seen any significant effect (P0.05) on total VFA concentration, N-NH3 and total gas but had no effect (P0.05) on microbial protein synthesis. Conclusion of research is the provision of energy sources with rice bran treatment, onggok wet and dry corn flour can be used as fermentable carbohydrates on feed hay amoniasi in vitro.


1999 ◽  
Vol 1999 ◽  
pp. 151-151 ◽  
Author(s):  
I.C.S. Bueno ◽  
A.L. Abdalla ◽  
S.L.S. Cabral Filho ◽  
D.M.S.S. Vitti ◽  
E. Owen ◽  
...  

The use of small ruminants, such as sheep, in metabolism studies is more convenient as handling problems are reduced and their maintenance costs are lower, in comparison with cattle. However in vivo digestibility estimates obtained at maintenance are known to differ between these two species. With the increased use ofin vitrogas production techniques, to evaluate ruminant feedingstuffs, it is of great importance to identify whether the species from which the rumen fluid inoculum is obtained has a significant influence on the results obtained.Rumen fluid samples were obtained from a non-lactating Holstein cow (C) and six wether sheep (S) offered the same diet (80 % tropical grass and 20 % dairy concentrate) and prepared so as to have similar dry matter (DM) contents and therefore potentially the microbial mass. Nine substrates (two tropical grasses 1-2, tropical alfalfa 3, barley straw 4, and five temperate grasses 5-9) were examined.


2019 ◽  
Vol 64 (No. 8) ◽  
pp. 352-360
Author(s):  
Jiu Yuan ◽  
Xinjie Wan

The associative effects (AE) between concentrate (C), peanut shell (P) and alfalfa (A) were investigated by means of an automated gas production (GP) system. The C, P and A were incubated alone or as 40 : 60 : 0, 40 : 45 : 15, 40 : 30 : 30, 40 : 15 : 45, 40 : 0 : 60 and 30 : 70 : 0, 30 : 55 : 15, 30 : 40 : 30, 30 : 25 : 45, 30 : 10 : 60, 30 : 0 : 70 mixtures where the C : roughage (R) ratios were 40 : 60 and 30 : 70. Samples (0.2000 ± 0.0010 g) of single feeds or mixtures were incubated for 96 h in individual bottles (100 ml) with 30 ml of buffered rumen fluid. GP parameters were analysed using a single exponential equation. After incubation, the residues were used to determine pH, dry matter digestibility (DMD), organic matter digestibility (OMD), volatile fatty acids (VFA) and ammonia nitrogen (NH<sub>3</sub>-N) of the incubation fluid, and their single factor AE indices (SFAEI) and multiple-factors AE indices (MFAEI) were determined. The results showed that group of 30 peanut shell had higher SFAEI of GP<sub>48 h</sub>, DMD, OMD and total volatile fatty acids (p &lt; 0.05) and MFAEI (p &lt; 0.05) than groups 60, 45 and 0 when C : R was 40 : 60. The group of 10 peanut shell showed higher SFAEI of GP<sub>48 h</sub>, DMD and OMD (p &lt; 0.05) than groups 70, 55 and 40 and MFAEI (p &lt; 0.01) when C : R was 30 : 70. It is concluded that optimal SFAEI and MFAEI were obtained when the C : P : A ratios were 40 : 30 : 30 and 30 : 10 : 60.


1998 ◽  
Vol 1998 ◽  
pp. 168-168 ◽  
Author(s):  
A. Hussain ◽  
E. L. Miller

Inclusion of lactose in dairy cow rations increases dry matter intake (DMI) and milk yield (Garnsworthy 1996). This may be due to the relatively slow rate of lactose fermentation ( Hussain and Miller, 1998) sustaining better regulation of rumen pH and also possible consequence for microbial protein synthesis (Chamberlain et al., 1993).This experiment was conducted to study the changes in rumen environment over the adaptation period and effect of these changes on the fermentation of lactose itself.Three Suffolk wethers (b.wt 56± 7.36 kg) maintained on hay and concentrate (600:400) were offered 50g lactose per day for 16 days. Rumen liquor collected on dayO (before offering lactose), 4, 8, 12 and 16 was used to measure gas production from sucrose and lactose ( Menke et al., 1979). On these days rumen samples were collected at 0, 1, 2, 4, 6 and 8 hrs after the morning feed. Rumen pH, ammonia N (NH3N) and volatile fatty acids (VFA) were measured. At 8 hrs time rumen samples were also taken for protozoa enumeration. Data obtained were analysed using ANOVA procedure of Genstat 5.


2017 ◽  
Vol 48 (2) ◽  
pp. 63-69
Author(s):  
M. Joch ◽  
V. Kudrna ◽  
B. Hučko

AbstractThe objective of this study was to determine the effects of geraniol and camphene at three dosages (300, 600, and 900 mg l-1) on rumen microbial fermentation and methane emission in in vitro batch culture of rumen fluid supplied with a 60 : 40 forage : concentrate substrate (16.2% crude protein, 33.1% neutral detergent fibre). The ionophore antibiotic monensin (8 mg/l) was used as positive control. Compared to control, geraniol significantly (P < 0.05) reduced methane production with increasing doses, with reductions by 10.2, 66.9, and 97.9%. However, total volatile fatty acids (VFA) production and in vitro dry matter digestibility were also reduced (P < 0.05) by all doses of geraniol. Camphene demonstrated weak and unpromising effects on rumen fermentation. Camphene did not decrease (P > 0.05) methane production and slightly decreased (P < 0.05) VFA production. Due to the strong antimethanogenic effect of geraniol a careful selection of dose and combination with other antimethanogenic compounds may be effective in mitigating methane emission from ruminants. However, if a reduction in total VFA production and dry matter digestibility persisted in vivo, geraniol would have a negative effect on animal productivity.


2010 ◽  
Vol 2010 ◽  
pp. 1-7 ◽  
Author(s):  
M. A. Abdl-Rahman ◽  
F. A. R. Sawiress ◽  
A. M. Abd El-Aty

The aim of the current study was to evaluate the effect of sodium lauryl sulfate-fumaric acid coupled addition onin vitromethangenesis and rumen fermentation. Evaluation was carried out usingin vitrogas production technique. Ruminal contents were collected from five steers immediately after slaughtering and used for preparation of inoculums of mixed rumen microorganisms. Rumen fluid was then mixed with the basal diet of steers and used to generate four treatments, negative control (no additives), sodium lauryl sulfate (SLS) treated, fumaric acid treated, and SLS-fumaric acid coupled addition treated. The results revealed that, relative to control, efficiency in reduction of methanogenesis was as follows: coupled addition > SLS-addition > fumaric acid addition. Both SLS-addition and SLS-fumaric acid coupled addition demonstrated a decremental effect on ammonia nitrogen (–), total short chain volatile fatty acids (SCVFAs) concentrations and the amount of substrate degraded, and an increment effect on microbial mass and microbial yield (). Nevertheless, fumaric acid did not alter any of the previously mentioned parameters but induced a decremental effect on –. Furthermore, both fumaric acid and SLS-fumaric acid coupled addition increased propionate at the expense of acetate and butyrate, while, defaunation increased acetate at the expense of propionate and butyrate. The pH value was decreased by all treatments relative to control, while, cellulase activity did not differ by different treatments. The current study can be promising strategies for suppressing ruminal methane emissions and improving ruminants feed efficiency.


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