scholarly journals In vitro Inoculation of Fresh or Frozen Rumen Fluid Distinguishes Contrasting Microbial Communities and Fermentation Induced by Increasing Forage to Concentrate Ratio

2022 ◽  
Vol 8 ◽  
Author(s):  
Zhi Yuan Ma ◽  
Ju Wang Zhou ◽  
Si Yu Yi ◽  
Min Wang ◽  
Zhi Liang Tan
Keyword(s):  
Bacterial Diversity ◽  
Volatile Fatty Acids ◽  
Rumen Fluid ◽  
Starch Content ◽  
Block Design ◽  
Rumen Fermentation ◽  
Rumen Microorganisms ◽  
Fermentative Bacteria ◽  
In Vitro Inoculation

In vitro rumen batch culture is a technology to simulate rumen fermentation by inoculating microorganisms from rumen fluids. Although inocula (INO) are commonly derived from fresh rumen fluids, frozen rumen fluids are also employed for the advantages of storing, transporting, and preserving rumen microorganisms. The effects of frozen INO on microbial fermentation and community may be interfered with by substrate type, which has not been reported. This study was designed to test whether rumen fluid treatments (i.e., fresh and frozen) could interact with incubated substrates. A complete block design with fractional arrangement treatment was used to investigate the effects of INO (fresh or frozen rumen fluids) and concentrate-to-forage ratios (C/F, 1:4 or 1:1) on rumen fermentation and microbial community. The effects of increasing C/F were typical, including increased dry matter (DM) degradation and total volatile fatty acids (VFA) concentration (P < 0.001), and decreased acetate to propionate ratio (P = 0.01) and bacterial diversity of richness and evenness (P ≤ 0.005) with especially higher fermentative bacteria such as genus Rikenellaceae_RC, F082, Prevotella, Bacteroidales_BS11, Muribaculaceaege, and Christensenellaceae_R-7 (P ≤ 0.04). Although frozen INO decreased (P < 0.001) DM degradation and altered rumen fermentation with lower (P ≤ 0.01) acetate to propionate ratio and molar proportion of butyrate than fresh INO, typical effects of C/F were independent of INO, as indicated by insignificant INO × C/F interaction on substrate degradation, VFA profile and bacterial community (P ≥ 0.20). In summary, the effect of C/F on fermentation and bacterial diversity is not interfered with by INO type, and frozen INO can be used to distinguish the effect of starch content.

scholarly journals Essential oils effect on rumen fermentation and biohydrogenation under in vitro conditions

2014 ◽  
Vol 59 (No. 10) ◽  
pp. 450-459 ◽  
Author(s):  
M. Gunal ◽  
A. Ishlak ◽  
A.A. AbuGhazaleh ◽  
W. Khattab
Keyword(s):  
Fatty Acids ◽  
Essential Oils ◽  
Dose Level ◽  
Volatile Fatty Acids ◽  
Rumen Fluid ◽  
Rumen Fermentation ◽  
Buffer Solution ◽  
Cinnamon Oil ◽  
Tea Tree

The effects of adding essential oils (EO) at different levels (125, 250, 500 mg/l) on rumen fermentation and biohydrogenation were examined in a rumen batch culture study. Treatments were: control without EO (CON), control with anise oil (ANO), cedar wood oil (CWO), cinnamon oil (CNO), eucalyptus oil (EUO), and tea tree oil (TEO). Essential oils, each dissolved in 1 ml of ethanol, were added to the culture flask containing 40 ml of buffer solution, 2 ml of reduction solution, 10 ml of rumen fluid, 25 mg of soybean oil, and 0.5 g of the diet. After 24 h of incubation in a water batch at 39°C, three samples were collected from each flask and analyzed for ammonia-N, volatile fatty acids (VFA), and fatty acids (FA). Expect for CNO, the proportions of acetate, propionate, and acetate to propionate ratios were not affected (P > 0.05) by EO addition. Addition of CWO, CNO, and TEO reduced total VFA concentrations (P < 0.05) regardless of dose level. The ammonia-N concentration was greater in cultures incubated with EO regardless of dose level. Compared with the CON, the concentrations of C18:0 and trans C18:1 were reduced (P < 0.05) with EO addition regardless of dose level. Compared with the CON, the concentration of linoleic acid was greater (P < 0.05) when EO were added at 500 mg/l. EO tested in this study had no effects on VFA profile but significantly reduced the formation of biohydrogenation products (C18:0 and trans C18:1).

scholarly journals Effects of Geraniol and Camphene on in Vitro Rumen Fermentation and Methane Production

2017 ◽  
Vol 48 (2) ◽  
pp. 63-69
Author(s):  
M. Joch ◽  
V. Kudrna ◽  
B. Hučko
Keyword(s):  
Methane Emission ◽  
Methane Production ◽  
Dry Matter ◽  
Volatile Fatty Acids ◽  
Rumen Fluid ◽  
Positive Control ◽  
Rumen Fermentation ◽  
Dry Matter Digestibility

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.

Study of the effects of PR toxin, mycophenolic acid and roquefortine C on in vitro gas production parameters and their stability in the rumen environment

2014 ◽  
Vol 153 (1) ◽  
pp. 163-176 ◽  
Author(s):  
A. GALLO ◽  
G. GIUBERTI ◽  
T. BERTUZZI ◽  
M. MOSCHINI ◽  
F. MASOERO
Keyword(s):  
Mycophenolic Acid ◽  
Rumen Fluid ◽  
Rumen Fermentation ◽  
Gas Production ◽  
Farm Animals ◽  
Marginal Effect ◽  
Rumen Microorganisms ◽  
Marginal Effects ◽  
Roquefortine C

SUMMARYMoulds belonging to Penicillium section roqueforti are common contaminants of feedstuffs and produce several mycotoxins that can cause health hazards when ingested by farm animals. Among these, PR toxin (PR), mycophenolic acid (MY) and roquefortine C (RC) have been frequently detected in forages, particularly silages. The aims of the current trials were to study the effects of the presence of pure mycotoxins on in vitro rumen fermentation parameters and to assess their stability in the rumen environment. Two successive in vitro gas production experiments were carried out: a central composite design with four replications of central point (CCD) and a completely randomized design with a fully factorial arrangement of treatments (FFD). In CCD, the effects of PR, MY and RC concentrations in diluted rumen fluid (i.e. 0·01, 0·30, 1·01, 1·71 and 2·00 μg of each mycotoxin/ml) were tested. Gas volume produced after 48 h of incubation (Vf) decreased linearly as concentrations of RC and MY in diluted rumen fluid increased, with marginal effects similar for two mycotoxins, being respectively −14·6 and −13·4 ml/g organic matter (OM) for each 1·0 μg/ml of increment in mycotoxin concentration. Similarly, total volatile fatty acid (VFA) production decreased quadratically as concentrations of RC and MY increased, with marginal effects about two times higher for MY than RC, being −4·22 and −2·62 mmol/l for each 1·0 μg/ml of increment in mycotoxin concentration. With respect to maximum Vf (i.e. 410·6 ml/g OM) and VFA (98·06 mmol/l) values estimated by the model, decreases of 13·6 and 15·2% were obtained when incubating the highest RC and MY concentrations, respectively. The PR did not interfere with rumen fermentation pattern and it was not recovered after 48 h of incubation, whereas the stabilities of MY and RC in rumen fluid were similar and on average equal to about 50%. On the basis of CCD results, a second experiment (FFD) was carried out in which only effects of MY and RC concentrations (i.e. 0, 0·67, 1·33 and 2·00 μg of each mycotoxin/ml of diluted rumen fluid) were tested. Data from FFD showed Vf decreased linearly when concentrations of MY and RC increased, with marginal effect two-folds higher for MY than for RC (−11·1 ml/g OM and −6·7 ml/g OM, respectively). Similar marginal effects of MY and RC in decreasing VFA production were recorded: −2·38 and −2·86 mmol/l for each 1·0 μg/ml of increment in mycotoxin concentration, respectively. At the highest RC and MY tested concentrations, Vf and VFA decreased by 8·7 and 10·7%, respectively, over maximum estimated values. In FFD, the average amounts of MY and RC recovered in rumen fluid after 48 h of incubation were 79·0 and 40·6%, respectively. In conclusion, the MY and RC from standards interfered with rumen microorganisms at relatively low levels and were partially stable in the rumen environment after 48 h of incubation. These findings suggested that MY and RC could interfere with digestive processes and might represent a potential risk for ruminants fed diets containing feeds contaminated by mycotoxins produced by P. roqueforti.

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

2020 ◽  
Vol 8 (8) ◽  
pp. 1160 ◽  
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.

scholarly journals Comparison of fermentation characteristics and bacterial diversity in the rumen of sheep and in batch cultures of rumen microorganisms

2015 ◽  
Vol 153 (6) ◽  
pp. 1097-1106 ◽  
Author(s):  
I. MATEOS ◽  
M. J. RANILLA ◽  
C. SARO ◽  
M. D. CARRO
Keyword(s):  
Bacterial Diversity ◽  
Rumen Fluid ◽  
Similarity Index ◽  
Bacterial Strains ◽  
Rumen Microorganisms ◽  
Batch Cultures ◽  
Alfalfa Hay ◽  
Grass Hay ◽  
Forage Type

SUMMARYThe objective of the current study was to assess how closely batch cultures (BC) of rumen microorganisms can mimic the dietary differences in fermentation characteristics found in the rumen, and to analyse changes in bacterial diversity over the in vitro incubation period. Four ruminally and duodenally cannulated sheep were fed four diets having forage : concentrate ratios (FCR) of 70 : 30 or 30 : 70, with either alfalfa hay or grass hay as forage. Rumen fluid from each sheep was used to inoculate BC containing the same diet fed to the donor sheep, and the main rumen fermentation parameters were determined after 24 h of incubation. There were differences between BC and sheep in the magnitude of most measured parameters, but BC detected differences among diets due to forage type similar to those found in sheep. In contrast, BC did not reproduce the dietary differences due to FCR found in sheep for pH, degradability of neutral detergent fibre and total volatile fatty acid (VFA) concentrations. There were differences between systems in the magnitude of most determined parameters and BC showed higher pH values and NH3–N concentrations, but lower fibre degradability and VFA and lactate concentrations compared with sheep. There were significant relationships between in vivo and in vitro values for molar proportions of acetate, propionate and butyrate, and the acetate : propionate ratio. The automated ribosomal intergenic spacer analysis (ARISA) of 16S ribosomal deoxyribonucleic acid showed that FCR had no effect on bacterial diversity either in the sheep rumen fluid used as inoculum (IN) or in BC samples. In contrast, bacterial diversity was greater with alfalfa hay diets than those with grass hay in the IN, but was unaffected by forage type in the BC. Similarity index between the bacterial communities in the inocula and those in the BC ranged from 67·2 to 74·7%, and was unaffected by diet characteristics. Bacterial diversity was lower in BC than in the inocula with 14 peaks out of a total of 181 detected in the ARISA electropherograms never appearing in BC samples, which suggests that incubation conditions in the BC may have caused a selection of some bacterial strains. However, each BC sample showed the highest similarity index with its corresponding rumen IN, which highlights the importance of using rumen fluid from donors fed a diet similar to that being incubated in BC when conducting in vitro experiments.

Influence of acetohydroxamic acid on some activities in vitro of the rumen microbiota

1968 ◽  
Vol 14 (4) ◽  
pp. 409-416 ◽  
Author(s):  
G. A. Jones
Keyword(s):  
Volatile Fatty Acids ◽  
Urease Activity ◽  
Rumen Fluid ◽  
Selective Medium ◽  
Colony Development ◽  
Acetohydroxamic Acid ◽  
Rumen Microorganisms ◽  
Rumen Microbiota

When acetohydroxamic acid was incubated with washed suspensions of bovine rumen microorganisms the urease activity of the suspensions was depressed; activity could not be restored by the addition of divalent cations which, in the absence of acetohydroxamic acid, stimulated the urease activity of the cells. Acetohydroxamic acid was slowly degraded by the rumen microbiota. When the compound was incorporated into a non-selective medium for the enumeration of rumen bacteria it completely prevented visible colony development by some components of the inoculum and retarded the rate of multiplication of others. Acetohydroxamic acid inhibited the production of volatile fatty acids from added cellulose in strained rumen fluid and modified the molar proportions of acetate, propionate, and butyrate produced from the substrate; whereas in the absence of acetohydroxamic acid propionate production was favored at the expense of acetate; in its presence the acetate:propionate ratio remained constant. The effect of acetohydroxamic acid upon rumen microbial activities in vitro was therefore not limited to inhibition of rumen urease. It was impossible, however, to infer from the results obtained whether the potential value of the compound as a urease inhibitor in vivo would be diminished for this reason; this is because the influence of acetohydroxamic acid on the rumen microbiota in vivo is probably subject to modification by factors, such as the composition of the animal's diet, which were not investigated.

scholarly journals Inhibition of in vitro rumen methane production by three statins

2021 ◽  
Vol 0 (0) ◽  
Author(s):  
Miroslav Joch ◽  
Mariana Vadroňová ◽  
Alena Výborná ◽  
Kateřina Jochová
Keyword(s):  
Methane Production ◽  
Volatile Fatty Acids ◽  
Rumen Fluid ◽  
Relative Proportion ◽  
Culture Fluid ◽  
Rumen Fermentation ◽  
Slight Reduction ◽  
Net Production ◽  
Feed Digestibility

Abstract The aim of this study was to evaluate the effects of increasing concentrations of three pure statins on in vitro methane production and rumen fermentation. The effects of atorvastatin, rosuvastatin and simvastatin at three concentrations (1, 10, and 100 mg/L of culture fluid) were evaluated using in vitro 24 h batch incubation of buffered rumen fluid with a 70:30 forage:concentrate substrate. All statins tested demonstrated the ability to reduce methanogenesis. Methane inhibition potential was decreasing in the following order: simvastatin > atorvastatin > rosuvastatin. Methane production was reduced (p < 0.05) by simvastatin at 10 mg/L (by 9.3%) and by atorvastatin at 100 mg/L (by 13.2%) without compromising fermentation and feed digestibility. Simvastatin at 100 mg/L decreased methane production by 26.2%, however, net production of volatile fatty acids (nVFA) was also reduced (p < 0.05). The only effect of rosuvastatin was a slight reduction (p < 0.05) of methane proportion at 10 and 100 mg/L. Simvastatin and atorvastatin at 100 mg/L increased (p < 0.05) relative proportion of propionate at the expense of acetate and butyrate. Ammonia-N concentrations were not affected (p > 0.05) by statins. The current study demonstrated that selected statins could selectively decrease methane production. The effects of statins on methanogenesis and overall rumen fermentation vary depending on statin type and concentration. Hydrophobic statins, such as simvastatin and atorvastatin, seem to be more effective compared to the hydrophilic statins, such as rosuvastatin.

150 Effect of Ergot Alkaloids and a Mycotoxin Deactivating Product on in vitro Rumen Fermentation Using the Rumen Simulation Technique (RUSITEC)

2021 ◽  
Vol 99 (Supplement_3) ◽  
pp. 78-78
Author(s):  
Jenna Sarich ◽  
Kim Stanford ◽  
Karen S Schwartzkopf-Genswein ◽  
Tim A McAllister ◽  
Barry Blakley ◽  
...  
Keyword(s):  
Fixed Effects ◽  
Volatile Fatty Acids ◽  
Block Design ◽  
Control Diet ◽  
Ergot Alkaloids ◽  
Rumen Fermentation ◽  
Simulation Technique ◽  
Neutral Detergent Fiber ◽  
Sample Collection

Abstract A rumen simulation technique (RUSITEC) investigated effects of ergot alkaloids (E) and a mycotoxin deactivating product (Biomin® AA; B) on nutrient digestion, rumen fermentation, total gas and methane emissions. Fermenters received a finishing diet of 90:10 concentrate:barley silage. Using a randomized complete block design, treatments were assigned (4 vessels/treatment) within two RUSITECs in a 2 × 2 factorial. Treatments included: 1) control diet, 2) control + 1 g B, 3) control + 20 ppm E, and 4) control + 20 ppm E + 1 g B. The study had a 14-d experimental period, with 7-d adaptation and 7-d sample collection. Data were analyzed in SAS using PROC mixed including fixed effects of E, B, and E×B interaction. Random effects included RUSITEC apparatus and cow rumen inoculum (n = 4). Ergot decreased dry matter disappearance (DMD) (P &lt; 0.05; 87.9 vs. 87.2%) and organic matter disappearance (OMD) (P &lt; 0.05; 88.8 vs. 88.4%). Adding B increased OMD (P &lt; 0.05; 88.3 vs. 88.9%) and neutral detergent fiber disappearance (NDFD) (P &lt; 0.01); however, an E×B interaction was observed for NDFD (P &lt; 0.01) with B promoting greater increases with E. Ergot decreased acetate proportions (P &lt; 0.01) and increased isovalerate (P &lt; 0.05). Consequently, acetate:propionate was reduced (P &lt; 0.05) with E. Inclusion of B increased total volatile fatty acids (P &lt; 0.01), and proportions of acetate (P &lt; 0.05) and propionate (P &lt; 0.05), and decreased valerate (P &lt; 0.01), isovalerate (P &lt; 0.01), and caproate (P &lt; 0.01). Treatments did not affect (P ≥ 0.17) ammonia-N, total gas, or methane production (mg/d or mg/g of OM fermented). In conclusion, E reduced OMD and acetate production, but these responses were reversed by the addition of B.

PSX-B-1 Effect of exposure time and dose of effective microorganisms (EM®) on feed in vitro rumen fermentation and digestibility

2021 ◽  
Vol 99 (Supplement_3) ◽  
pp. 455-457
Author(s):  
Alain Miranda-Figueroa ◽  
Carlos Gutiérrez_Aguilar ◽  
Manuel González-Ronquillo ◽  
Atmir Romero-Pérez ◽  
Claudia Cecilia Márquez-Mota ◽  
...  
Keyword(s):  
Exposure Time ◽  
Volatile Fatty Acids ◽  
Block Design ◽  
Ammonia Nitrogen ◽  
Rumen Fermentation ◽  
Effective Microorganisms ◽  
Time Interaction ◽  
Randomized Block Design ◽  
Quadratic Effects

Abstract The objective was to evaluate the effect of feed inoculation with effective microorganisms (EM®) (mainly containing Lactobacillus spp.,Rhodopseudomona palustrisand Saccharomyces cerevisiae) on rumen fermentation using in vitrogas production technique. We hypothesized that increasing doses and allowing exposure of EM® for up to 48 hours, would improve digestibility and rumen fermentation. The experimental design was a 4×4 completely randomized block design including 4 EM® levels [(0(EM0), 0.5(EM0.5), 1.0 (EM1) and 1.5 (EM1.5) mL EM® / kg DM] and 4 preincubation times [0 (T0), 12(T12), 24(T24), 48 (T48) h], with four repetitions per treatment. Treatments were evaluated using 100ml glass bottles with 0.5g of the diet (20% corn stover, 20% oat hay, 48.8% ground corn, 7% molasses, 1.2% urea, 1% soybean meal, 0.9% mineral premix,1.1% salt, dry matter basis) incubated with sheep ruminal fluid in 3 different occasions. Data were analyzed with PROC MIXED of SAS and orthogonal contrasts to determine the linear and quadratic effects of EM dose and exposure time. Interaction (P &lt; 0.05) of EM x T was observed for in vitrodry matter digestibility (IVDMD), maximum gas volume (Vmax), total volatile fatty acids (VFA), acetate (ACE), propionate (PROP), butyrate (BUT) and ammonia-nitrogen (NH3), IVDMD was higher (P &lt; 0.01, 4.8 and 3.72%) for T48EM1.5 than T12EM0 and T0EM0, PROP was higher (P &lt; 0.05) for T48EM0, T48EM1 and 1.5 than T12EM0. The ACE:PROP ratio was higher (P &lt; 0.05, 17.2%) for T12EM0 than T48EM1.5. IVDMD, PROP and NH3 linearly increased (P &lt; 0.01) with increasing exposure time. EM levels have a quadratic effect (P &lt; 0.01) with maximum response at EM0.5. It was concluded that the addition of 0.5 to 1.5 mL/kg DM of EM® to a sheep diet and increasing preincubation time, up to 48h, improve feed fermentation and digestibility.Project was supported by UNAM, DGAPA, PAPIIT (IT202120).

scholarly journals In vitro rumen fermentation characteristics and microbes of thin tail sheep given sweet potato (Ipomoea batatas L) biomass

2016 ◽  
Vol 21 (2) ◽  
pp. 83
Author(s):  
Asep Sudarman ◽  
M. Hayashida ◽  
M. Miralestari
Keyword(s):  
Organic Matter ◽  
Sweet Potato ◽  
Ipomoea Batatas ◽  
Dry Matter ◽  
Rumen Fluid ◽  
Block Design ◽  
Rumen Fermentation ◽  
Napier Grass ◽  
Sweet Potato Leaves

<p class="abstrak2">Sweet potato plant is produced in Indonesia in large quantities. Sweet potato leaves and stems can be used as a source of protein and the tuber can be used as an energy source. This study was aimed to analyze the effect of feeding of sweet potato biomass on the rumen fermentation characteristics, digestibility of dry matter and organic matter as well as rumen microbial population of thin tail sheep. This study used a randomized block design by applying four feed treatments, i.e., T0 (100% Napier grass), T1 (70% of Napier grass + 30% concentrate), T2 (50% of Napier grass + 50% sweet potato leaves), and T3 (70% sweet potato leaves + 30% sweet potato tuber) with three replicates. Rumen fluid samples were taken with stomach tube method using a vacuum pump.  Results showed that the T3 diet has higher (P&lt;0.05) digestibility of dry matter and organic matter, concentration of NH<sub>3</sub> and VFA, and rumen bacterial population than those of T0 and T1diets but similar to that of T2 diet. Rumen pH and protozoa population was not different among the treatments. It can be concluded that the use of sweet potato biomass can improve the quality of rumen fermentation of sheep.</p>

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