Opportunities and limitations of a standardisation of the rumen simulation technique (RUSITEC) for analyses of ruminal nutrient degradation and fermentation and on microbial community characteristics

Deitmers, Jan-Helge; Gresner, N.; Südekum, Karl‐Heinz

doi:10.1016/j.anifeedsci.2022.115325

Cited by 9 publications

(8 citation statements)

References 97 publications

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“…Many developments designed to enhance and fine-tune the IV procedure have revolved around the technique’s standardization, mechanization, and automation. Likewise, there have been numerous instances in which researchers have noted ethical questions or shortages of labor needed for the maintenance of ruminally fistulated animals ( Mansfield et al, 1995 ; Castillo and Hernández, 2021 ; Deitmers et al, 2022 ), thus necessitating a benchtop technique suitable for feedstuff evaluation. The IVGP technique is a batch culture technique that assesses gas production throughout the incubation period using pressure sensors attached to a needle inserted in the flasks.…”

Section: Yr Later: What Progress Has Been Made Since 1993?mentioning

confidence: 99%

“…This sentiment has continued to the present day. Deitmers et al (2022) indicated that the automated RUSITEC system reduces the animal-to-animal variation present in the IS technique, decreases human error, and decreases the labor costs of sampling. However, Prevot et al (1994) stated that the continuous culture system was not a viable option due to its inability to maintain microbial populations similar to the in vivo conditions, though this is also true for batch culture systems and inside IS bags.…”

Section: Yr Later: What Progress Has Been Made Since 1993?mentioning

confidence: 99%

See 1 more Smart Citation

Forages and pastures symposium: an update on in vitro and in situ experimental techniques for approximation of ruminal fiber degradation

Foster¹,

Smith

Rouquette³

et al. 2023

Journal of Animal Science

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Static quantification measures of chemical components are commonly used to make certain assumptions about forage or feed nutritive value and quality. In order for modern nutrient requirement models to estimate intake and digestibility more accurately, kinetic measures of ruminal fiber degradation are necessary. Compared to in vivo experiments, in vitro (IV) and in situ (IS) experimental techniques are relatively simple and inexpensive methods to determine the extent and rate of ruminal fiber degradation. This paper summarizes limitations of these techniques and statistical analyses of the resulting data, highlights key updates to these techniques in the last 30 yr, and presents opportunities for further improvements to these techniques regarding ruminal fiber degradation. The principle biological component of these techniques, ruminal fluid, is still highly variable because it is influenced by ruminally fistulated animal diet type and timing of feeding, and in the case of the IV technique by collection and transport procedures. Commercialization has contributed to the standardization, mechanization, and automation of the IV true digestibility technique, for example, the well-known DaisyII Incubator. There has been limited commercialization of supplies for the IS technique and several review papers focused on standardization in the last 30 yr; however, the IS experimental technique is not standardized and there remains variation within and among laboratories. Regardless of improved precision resulting from enhancements of these techniques, the accuracy and precision of determining the indigestible fraction are fundamental to modeling digestion kinetics and the use of these estimates in more complex dynamic nutritional modeling. Opportunities for focused research and development are additional commercialization and standardization, methods to improve the precision and accuracy of indigestible fiber fraction, data science applications, and statistical analyses of results, especially for IS data. In situ data is typically fitted to one of a few first-order kinetic models, and parameters are estimated without determining if the selected model has the best fit. Animal experimentation will be fundamental to the future of ruminant nutrition and IV and IS techniques will remain vital to bring together nutritive value with forage quality. It is feasible and important to focus efforts on improving the precision and accuracy of IV and IS results.

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Section: Yr Later: What Progress Has Been Made Since 1993?mentioning

confidence: 99%

Section: Yr Later: What Progress Has Been Made Since 1993?mentioning

confidence: 99%

Forages and pastures symposium: an update on in vitro and in situ experimental techniques for approximation of ruminal fiber degradation

Foster¹,

Smith

Rouquette³

et al. 2023

Journal of Animal Science

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show abstract

“…For these reasons, a more sophisticated approach for testing the promising combinations is the semi-continuously operating rumen simulation technique (Rusitec) [ 21 ]. This simulator allows the evaluation of nutrient degradability and single gas production in a mid-term fermentation period [ 22 ], while meets the need for applying the 3R principles: replace, reduce, refine for experiment purposes [ 23 ]. Even though there are limitations in the use of Rusitec in simulating real in vivo conditions [ 22 , 24 ], it provides useful data in the field of ruminant nutrition and represents a way to better set up in vivo experiments [ 25 ].…”

Section: Introductionmentioning

confidence: 99%

Evaluation of ruminal methane and ammonia formation and microbiota composition as affected by supplements based on mixtures of tannins and essential oils using Rusitec

Foggi,

Terranova,

Daghio

et al. 2024

J Animal Sci Biotechnol

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Background Dietary supplements based on tannin extracts or essential oil compounds (EOC) have been repeatedly reported as a promising feeding strategy to reduce the environmental impact of ruminant husbandry. A previous batch culture screening of various supplements identified selected mixtures with an enhanced potential to mitigate ruminal methane and ammonia formation. Among these, Q-2 (named after quebracho extract and EOC blend 2, composed of carvacrol, thymol, and eugenol) and C-10 (chestnut extract and EOC blend 10, consisting of oregano and thyme essential oils and limonene) have been investigated in detail in the present study with the semi-continuous rumen simulation technique (Rusitec) in three independent runs. For this purpose, Q-2 and C-10, dosed according to the previous study, were compared with a non-supplemented diet (negative control, NC) and with one supplemented with the commercial EOC-based Agolin® Ruminant (positive control, PC). Results From d 5 to 10 of fermentation incubation liquid was collected and analysed for pH, ammonia, protozoa count, and gas composition. Feed residues were collected for the determination of ruminal degradability. On d 10, samples of incubation liquid were also characterised for bacterial, archaeal and fungal communities by high-throughput sequencing of 16S rRNA and 26S ribosomal large subunit gene amplicons. Regardless of the duration of the fermentation period, Q-2 and C-10 were similarly efficient as PC in mitigating either ammonia (−37% by Q-2, −34% by PC) or methane formation (−12% by C-10, −12% by PC). The PC was also responsible for lower feed degradability and bacterial and fungal richness, whereas Q-2 and C-10 effects, particularly on microbiome diversities, were limited compared to NC. Conclusions All additives showed the potential to mitigate methane or ammonia formation, or both, in vitro over a period of 10 d. However, several differences occurred between PC and Q-2/C-10, indicating different mechanisms of action. The pronounced defaunation caused by PC and its suggested consequences apparently determined at least part of the mitigant effects. Although the depressive effect on NDF degradability caused by Q-2 and C-10 might partially explain their mitigation properties, their mechanisms of action remain mostly to be elucidated.

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“…With the constant inflows and outflows, the rumen simulation technique (RUSITEC) system could serve as a relatively more effective tool in assessing rumen fermentation in vitro in comparison with the traditional method of batch culture ( Deitmers et al, 2022 ). So far, two major types of the RUSITEC system have been separately invented and are commonly applied: the semi-continuous culture system and the continuous culture system ( Hoover et al, 1976 ; Czerkawski and Breckenridge, 1977 ).…”

Section: Introductionmentioning

confidence: 99%

“…So far, two major types of the RUSITEC system have been separately invented and are commonly applied: the semi-continuous culture system and the continuous culture system ( Hoover et al, 1976 ; Czerkawski and Breckenridge, 1977 ). These artificial rumen systems vary from simple vessel sorts with different volumes to complex continuous-flow types with diverse stirrer, and various in- and outflow systems ( Deitmers et al, 2022 ). In our previous investigation on the impacts of rotation speed on rumen fermentation and ruminal microorganisms, a novel dual-flow continuous RUSITEC system was developed and adopted with detailed descriptions ( Adebayo Arowolo et al, 2022 ).…”

Section: Introductionmentioning

confidence: 99%

Evaluation of stirring time through a rumen simulation technique: Influences on rumen fermentation and bacterial community

Zuo

Li²,

Lan³

et al. 2023

Front. Microbiol.

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IntroductionRumen motility is a key element that influences ruminant nutrition, whereas little is known about the effects of rumen contraction duration on rumen fermentation and ruminal microbiome. We previously reported that proper rotation speed of a rumen simulation technique (RUSITEC) system enhanced rumen fermentation and microbial protein (MCP) production. In the present study, different contraction durations and intervals were simulated by setting different stirring times and intervals of the stirrers in a RUSITEC system. The objective of this trial was to evaluate the influences of stirring time on rumen fermentation characteristics, nutrient degradation, and ruminal bacterial microbiota in vitro.MethodsThis experiment was performed in a 3 × 3 Latin square design, with each experimental comprising 4 d for adjustment and 3 d for sample collection. Three stirring time treatments were set: the constant stir (CS), the intermittent stir 1 (each stir for 5 min with an interval of 2 min, IS1), and the intermittent stir 2 (each stir for 4 min with an interval of 3 min, IS2).ResultsThe total volatile fatty acid (TVFA) concentration, valerate molar proportion, ammonia nitrogen level, MCP density, protozoa count, disappearance rates of dry matter, organic matter, crude protein, neutral detergent fiber, and acid detergent fiber, emissions of total gas and methane, and the richness index Chao 1 for the bacterial community were higher (p < 0.05) in the IS1 when compared to those in the CS. The greatest TVFA, MCP, protozoa count, nutrient disappearance rates, gas productions, and bacterial richness indices of Ace and Chao 1 amongst all treatments were observed in the IS2. The relative abundance of the genus Treponema was enriched (p < 0.05) in CS, while the enrichment (p < 0.05) of Agathobacter ruminis and another two less known bacterial genera were identified in IS2.DiscussionIt could be concluded that the proper reduction in the stirring time might help to enhance the feed fermentation, MCP synthesis, gas production, and the relative abundances of specific bacterial taxa.

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Opportunities and limitations of a standardisation of the rumen simulation technique (RUSITEC) for analyses of ruminal nutrient degradation and fermentation and on microbial community characteristics

Cited by 9 publications

References 97 publications

Forages and pastures symposium: an update on in vitro and in situ experimental techniques for approximation of ruminal fiber degradation

Forages and pastures symposium: an update on in vitro and in situ experimental techniques for approximation of ruminal fiber degradation

Evaluation of ruminal methane and ammonia formation and microbiota composition as affected by supplements based on mixtures of tannins and essential oils using Rusitec

Evaluation of stirring time through a rumen simulation technique: Influences on rumen fermentation and bacterial community

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