Ruminal microbiota developing in different in vitro simulation systems inoculated with goats’ rumen liquor

Soto, E. C.; Molina-Alcaide, E.; Khelil, Hajer; Yáñez-Ruíz, D. R.

doi:10.1016/j.anifeedsci.2013.06.003

Cited by 15 publications

(13 citation statements)

References 39 publications

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“…After this, the cross inoculated fluid performed most, similarly, to the poorer performing fluid across the remaining consecutive batch cultures. Factors such as bacteriophages and bacterioicins, which are involved in structuring the microbial community (Koskella and Meaden, 2013), the fungal community, and a lack of protozoal survival (Soto et al, 2013;Yáñez-Ruiz et al, 2016) may have prevented the full establishment of the Good community. No effect of cross inoculation was seen in Experiment 2.…”

Section: Discussionmentioning

confidence: 99%

Cross Inoculation of Rumen Fluid to Improve Dry Matter Disappearance and Its Effect on Bacterial Composition Using an in vitro Batch Culture Model

et al. 2020

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Environmental pressures of ruminant production could be reduced by improving digestive efficiency. Previous in vivo attempts to manipulate the rumen microbial community have largely been unsuccessful probably due to the influencing effect of the host. Using an in vitro consecutive batch culture technique, the aim of this study was to determine whether manipulation was possible once the bacterial community was uncoupled from the host. Two cross inoculation experiments were performed. Rumen fluid was collected at time of slaughter from 11 Holstein-Friesian steers from the same herd for Experiment 1, and in Experiment 2 were collected from 11 Charolais cross steers sired by the same bull and raised on a forage only diet on the same farm from birth. The two fluids that differed most in their in vitro dry matter disappearance (IVDMD; "Good," "Bad") were selected for their respective experiment. The fluids were also mixed (1:1, "Mix") and used to inoculate the model. In Experiment 1, the mixed rumen fluid resulted in an IVDMD midway between that of the two rumen fluids from which it was made for the first 24 h batch culture (34, 29, 20 g per 100 g DM for the Good, Mix, and Bad, respectively, P < 0.001) which was reflected in fermentation parameters recorded. No effect of cross inoculation was seen for Experiment 2, where the Mix performed most similarly to the Bad. In both experiments, IVDMD increased with consecutive culturing as the microbial population adapted to the in vitro conditions and differences between the fluids were lost. The improved performance with each consecutive batch culture was associated with reduced bacterial diversity. Increases in the genus Pseudobutyrivibrio were identified, which may be, at least in part, responsible for the improved digestive efficiency observed, whilst Prevotella declined by 50% over the study period. It is likely that along with host factors, there are individual factors within each community that prevent other microbes from establishing. Whilst we were unable to manipulate the bacterial community, uncoupling the microbiota from the host resulted in changes in the community, becoming less diverse with time, likely due to environmental heterogeneity, and more efficient at digesting DM.

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Section: Discussionmentioning

confidence: 99%

Cross Inoculation of Rumen Fluid to Improve Dry Matter Disappearance and Its Effect on Bacterial Composition Using an in vitro Batch Culture Model

et al. 2020

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“…Similar results were observed by Lengowski et al (2016) in the LIQ phase Rusitec fermenters fed silages and inoculated with rumen fluid from cows, but bacterial concentrations in SOL were unchanged over a 13-day incubation period. Studies with continuous-culture fermenters and inocula from goats or cows have observed increased bacterial abundance with time (Soto et al, 2013), no temporal changes (Muetzel etal., 2009;Soto etal., 2012) or decreased abundances (Martínez-Fernández etal., 2015). Differences in the source of the inoculum and its dilution with artificial saliva, the operation conditions of the fermenters, and the type of diet can help to explain the variability of these results.…”

Section: Discussionmentioning

confidence: 99%

“…Both of them are run over periods of several days and quantitative and qualitative changes in the microbial populations through the incubation period are therefore likely to occur. Several studies have monitored the shifts in microbial populations in continuous-culture fermenters (Soto et al, 2012 and2013;Martínez-Fernández et al, 2015), but to our knowledge there is only a recent study on this topic in Rusitec fermenters (Lengowski et al, 2016). Lengowski et al (2016) analyzed the changes in microbial populations over the first 48 h of incubation and on day 13, and reported important changes, recommending that the adaptation phase should last longer than 48 h. However, there is little information on the microbial changes occurring between the day 2 and the end of the incubation, and most is limited to protozoal populations (Martínez et al, 2010a and2011a).…”

Section: Introductionmentioning

confidence: 99%

Shifts in microbial populations in Rusitec fermenters as affected by the type of diet and impact of the method for estimating microbial growth (15N v. microbial DNA)

Mateos

Ranilla

Saro

et al. 2017

Animal

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Rusitec fermenters are in vitro systems widely used to study ruminal fermentation, but little is known about the microbial populations establishing in them. This study was designed to assess the time evolution of microbial populations in fermenters fed medium- (MC; 50% alfalfa hay : concentrate) and high-concentrate diets (HC; 15 : 85 barley straw : concentrate). Samples from solid (SOL) and liquid (LIQ) content of fermenters were taken immediately before feeding on days 3, 8 and 14 of incubation for quantitative polymerase chain reaction and automated ribosomal intergenic spacer analysis analyses. In SOL, total bacterial DNA concentration and relative abundance of Ruminococcus flavefaciens remained unchanged over the incubation period, but protozoal DNA concentration and abundance of Fibrobacter succinogenes, Ruminococcus albus and fungi decreased and abundance of methanogenic archaea increased. In LIQ, total bacterial DNA concentration increased with time, whereas concentration of protozoal DNA and abundance of methanogens and fungi decreased. Diet×time interactions were observed for bacterial and protozoal DNA and relative abundance of F. succinogenes and R. albus in SOL, as well as for protozoal DNA in LIQ. Bacterial diversity in SOL increased with time, but no changes were observed in LIQ. The incubated diet influenced all microbial populations, with the exception of total bacteria and fungi abundance in LIQ. Bacterial diversity was higher in MC-fed than in HC-fed fermenters in SOL, but no differences were detected in LIQ. Values of pH, daily production of volatile fatty acids and CH4 and isobutyrate proportions remained stable over the incubation period, but other fermentation parameters varied with time. The relationships among microbial populations and fermentation parameters were in well agreement with those previously reported in in vivo studies. Using 15N as a microbial marker or quantifying total microbial DNA for estimating microbial protein synthesis offered similar results for diets comparison, but both methods presented contrasting results for microbial growth in SOL and LIQ phases. The study showed that fermentation parameters remained fairly stable over the commonly used sampling period (days 8 to 14), but shifts in microbial populations were detected. Moreover, microbial populations differed markedly from those in the inocula, which indicates the difficulty of directly transposing results on microbial populations developed in Rusitec fermenters to in vivo conditions.

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“…Removal of soluble particles in the liquid medium may have adverse effects on microbial fermentation, by decreasing the amount of soluble substrate available for microbial growth or may conversely stimulate activity (Roger et al, 1990). Soto et al (2013) reported that the numbers of all quantified microorganisms (total bacteria, protozoa, methanogens, fungi, Fibrobacter succinogenes and Ruminococcus flavefaciens) declined sharply during 24 h to 72 h of incubation. This is likely due to the exhaustion of fermentable substrate and the accumulation of fermentation end products.…”

Section: Incubation Procedures and Ch 4 Measurementsmentioning

confidence: 99%

“…While the use of these techniques is essential when collecting microbial biomass to determine their composition to accurately assess passage of nutrients of microbial origin to the intestine, it is not clear whether a standard protocol for microbial detachment should be applied for in vitro methods. In recent work, Soto et al (2013) studied the development of the microbiota in different in vitro rumen simulation systems inoculated with intact or filtered rumen fluid from goats. Incubation of filtered rumen fluid fraction in batch culture resulted in lower microbial diversity compared with non-filtered rumen fluid inoculum.…”

Section: Preparation Of Inoculum Prior To Incubationmentioning

confidence: 99%

Design, implementation and interpretation of in vitro batch culture experiments to assess enteric methane mitigation in ruminants—a review

Yáñez-Ruíz

Bannink

Dijkstra

et al. 2016

Animal Feed Science and Technology

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a b s t r a c tIn vitro fermentation techniques (IVFT) have been widely used to evaluate the nutritive value of feeds for ruminants and in the last decade to assess the effect of different nutritional strategies on methane (CH4) production. However, many technical factors may influence the results obtained. The present review has been prepared by the 'Global Network' FACCE-JPI international research consortium to provide a critical evaluation of the main factors that need to be considered when designing, conducting and interpreting IVFT experiments that investigate nutritional strategies to mitigate CH 4 emission from ruminants. Given the increasing and wide-scale use of IVFT, there is a need to critically review reports in the literature and establish what criteria are essential to the establishment and implementation of in vitro techniques. Key aspects considered include: i) donor animal species and number of animal used, ii) diet fed to donor animals, iii) collection and processing of rumen fluid as inoculum, iv) choice of substrate and incubation buffer, v) incubation procedures and CH 4 measurements, vi) headspace gas composition and vii) comparability of in vitro and in vivo measurements. Based on an evaluation of experimental evidence, a set of technical recommendations are presented to harmonize IVFT for feed evaluation, assessment of rumen function and CH 4 production.

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Ruminal microbiota developing in different in vitro simulation systems inoculated with goats’ rumen liquor

Cited by 15 publications

References 39 publications

Cross Inoculation of Rumen Fluid to Improve Dry Matter Disappearance and Its Effect on Bacterial Composition Using an in vitro Batch Culture Model

Cross Inoculation of Rumen Fluid to Improve Dry Matter Disappearance and Its Effect on Bacterial Composition Using an in vitro Batch Culture Model

Shifts in microbial populations in Rusitec fermenters as affected by the type of diet and impact of the method for estimating microbial growth (15N v. microbial DNA)

Design, implementation and interpretation of in vitro batch culture experiments to assess enteric methane mitigation in ruminants—a review

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