FREE SHIPPING over $40
OVER 9000 FIVE-STAR REVIEWS
5% OFF ALL SUBSCRIPTIONS
100% HAPPINESS GUARANTEE
Analyze Diet
0
Home / Equine Research Bank / Appl Environ Microbiol / “Bowel on the Bench”: Proof of Concept of a Thre...
Applied and environmental microbiology2019; 86(1); e02093-19; doi: 10.1128/AEM.02093-19

“Bowel on the Bench”: Proof of Concept of a Three-Stage, In Vitro Fermentation Model of the Equine Large Intestine.

Abstract: The intestinal microbiota of the horse, an animal of huge economic and social importance worldwide, is essential to the health of the animal. Understanding the intestinal ecosystem and its dynamic interaction with diet and dietary supplements currently requires the use of experimental animals, with consequent welfare and financial constraints. Here, we describe the development and assessment, using multiple analytical platforms, of a three-vessel, continuous-flow, model of the equine hindgut. After inoculation of the model with fresh horse feces, the bacterial communities established in each vessel had a taxonomic distribution similar to that of the source animal. Short-chain fatty acid (SCFA) and branched-chain fatty acid (BCFA) production within the model at steady state was consistent with the expected bacterial function, although higher concentrations of some SCFA/BCFA relative to those in the gut content were apparent. We demonstrate the intermodel repeatability and the ability of the model to capture some aspects of individual variation in bacterial community profiles. The findings of this proof-of-concept study, including recognition of the limitions of the model, support its future development as a tool for investigating the impact of disease, nutrition, dietary supplementation, and medication on the equine intestinal microbiota. The equine gut model that we have developed and describe has the potential to facilitate the exploration of how the equine gut microbiota is affected by diet, disease, and medication. It is a convenient, cost-effective, and welfare-friendly alternative to research models.
Copyright © 2019 American Society for Microbiology.
Get Full Text
Publication Date: 2019-12-13 PubMed ID: 31676474PubMed Central: PMC6912081DOI: 10.1128/AEM.02093-19Google Scholar: Lookup
The Equine Research Bank provides access to a large database of publicly available scientific literature. Inclusion in the Research Bank does not imply endorsement of study methods or findings by Mad Barn.
LinkedInCopy
  • Journal Article
  • Research Support
  • Non-U.S. Gov't

Summary

This research summary has been generated with artificial intelligence and may contain errors and omissions. Refer to the original study to confirm details provided. Submit correction.

This research article presents the development of a three-stage in vitro model that simulates the horse’s large intestine and its microbial community. The model is designed to allow researchers to study the effects of various factors such as diet, disease, and medication on the equine intestinal microbiota without having to use live animals, making it a cost-effective and welfare-friendly alternative.

Background of the Research

  • The study revolves around the importance of understanding the intestinal ecosystem of horses, particularly the role of the microorganisms living in it. This is crucial for maintaining or improving the health and well-being of these animals.
  • Prior to developing this model, researchers have been using live animals to study these interactions, which posed ethical concerns for animal welfare and also involved significant costs.
  • The researchers sought to create a more ethical, economical and convenient solution for this purpose.

Development of the Model

  • The team developed a three-vessel, continuous-flow model that simulates the horse’s large intestine. They filled this model with fresh horse feces to kickstart the cultivation of the bacterial communities.
  • After some time, the bacterial distribution in each vessel began to resemble the one from the original sample, showing that the model does indeed represent the equine intestinal ecosystem well.
  • The model also produced short-chain fatty acids (SCFA) and branched-chain fatty acids (BCFA), which are products of bacterial metabolism and are consistent with expected bacterial functions.
  • However, some relative concentrations of SCFA and BCFA were higher than those in the actual gut content of horses.

Findings and Potentials of the Model

  • The study proved that the model is able to replicate, to a certain extent, the bacterial community profiles from different individuals, demonstrating the repeatability and reliability of the developed model.
  • Though the model cannot perfectly replicate the intricate ecosystem of the equine gut (with some differences observed in the production of certain fatty acids), it provides valuable insights and serves as a starting point for further development.
  • With this model, researchers can easily investigate how the horse’s intestinal microbiota is affected by various factors such as diet, diseases, dietary supplementation, and medication without the need for using live animals. Therefore, it is a cost-effective, welfare-friendly, and easier to manage solution for future studies on equine intestinal health.

Conclusions

  • The team acknowledges the limitations of the current model and recognizes that improvements can be made.
  • Nevertheless, they believe this proof-of-concept study provides a platform for further research into the equine intestinal microbiota and ultimately contributes to the improved well-being of horses.

Cite This Article

APA
Leng J, Walton G, Swann J, Darby A, La Ragione R, Proudman C. (2019). “Bowel on the Bench”: Proof of Concept of a Three-Stage, In Vitro Fermentation Model of the Equine Large Intestine. Appl Environ Microbiol, 86(1), e02093-19. https://doi.org/10.1128/AEM.02093-19

Publication

Applied and environmental microbiology
ISSN: 1098-5336
NlmUniqueID: 7605801
Country: United States
Language: English
Volume: 86
Issue: 1
PII: e02093-19

Researcher Affiliations

Leng, J
  • School of Veterinary Medicine, Faculty of Health and Medical Sciences, University of Surrey, Guildford, Surrey, United Kingdom j.leng@surrey.ac.uk.
Walton, G
  • Department of Food and Nutritional Sciences, University of Reading, Reading, United Kingdom.
Swann, J
  • Division of Integrative Systems Medicine and Digestive Diseases, Department of Surgery and Cancer, Imperial College London, London, United Kingdom.
Darby, A
  • School of Biological Sciences, University of Liverpool, Liverpool, United Kingdom.
La Ragione, R
  • School of Veterinary Medicine, Faculty of Health and Medical Sciences, University of Surrey, Guildford, Surrey, United Kingdom.
Proudman, C
  • School of Veterinary Medicine, Faculty of Health and Medical Sciences, University of Surrey, Guildford, Surrey, United Kingdom.

MeSH Terms

  • Animals
  • Fatty Acids / metabolism
  • Fatty Acids, Volatile / metabolism
  • Feces / microbiology
  • Fermentation / physiology
  • Gastrointestinal Microbiome / physiology
  • Horses
  • In Vitro Techniques / methods
  • Intestine, Large / chemistry
  • Intestine, Large / microbiology
  • Intestine, Large / physiology
  • Models, Biological

References

This article includes 38 references
  1. Dougal K, Harris PA, Edwards A, Pachebat JA, Blackmore TM, Worgan HJ, Newbold CJ. A comparison of the microbiome and the metabolome of different regions of the equine hindgut.. FEMS Microbiol Ecol 2012 Dec;82(3):642-52.
    doi: 10.1111/j.1574-6941.2012.01441.xpubmed: 22757649google scholar: lookup
  2. Costa MC, Silva G, Ramos RV, Staempfli HR, Arroyo LG, Kim P, Weese JS. Characterization and comparison of the bacterial microbiota in different gastrointestinal tract compartments in horses.. Vet J 2015 Jul;205(1):74-80.
    doi: 10.1016/j.tvjl.2015.03.018pubmed: 25975855google scholar: lookup
  3. Proudman CJ, Hunter JO, Darby AC, Escalona EE, Batty C, Turner C. Characterisation of the faecal metabolome and microbiome of Thoroughbred racehorses.. Equine Vet J 2015 Sep;47(5):580-6.
    doi: 10.1111/evj.12324pubmed: 25041526google scholar: lookup
  4. Costa MC, Stämpfli HR, Allen-Vercoe E, Weese JS. Development of the faecal microbiota in foals.. Equine Vet J 2016 Nov;48(6):681-688.
    doi: 10.1111/evj.12532pubmed: 26518456google scholar: lookup
  5. Ericsson AC, Johnson PJ, Lopes MA, Perry SC, Lanter HR. A Microbiological Map of the Healthy Equine Gastrointestinal Tract.. PLoS One 2016;11(11):e0166523.
    doi: 10.1371/journal.pone.0166523pmc: PMC5112786pubmed: 27846295google scholar: lookup
  6. Costa MC, Arroyo LG, Allen-Vercoe E, Stämpfli HR, Kim PT, Sturgeon A, Weese JS. Comparison of the fecal microbiota of healthy horses and horses with colitis by high throughput sequencing of the V3-V5 region of the 16S rRNA gene.. PLoS One 2012;7(7):e41484.
    doi: 10.1371/journal.pone.0041484pmc: PMC3409227pubmed: 22859989google scholar: lookup
  7. Leng J, Proudman C, Darby A, Blow F, Townsend N, Miller A, Swann J. Exploration of the Fecal Microbiota and Biomarker Discovery in Equine Grass Sickness.. J Proteome Res 2018 Mar 2;17(3):1120-1128.
    doi: 10.1021/acs.jproteome.7b00784pubmed: 29364680google scholar: lookup
  8. Macfarlane GT, Macfarlane S, Gibson GR. Validation of a Three-Stage Compound Continuous Culture System for Investigating the Effect of Retention Time on the Ecology and Metabolism of Bacteria in the Human Colon.. Microb Ecol 1998 Mar;35(2):180-7.
    doi: 10.1007/s002489900072pubmed: 9541554google scholar: lookup
  9. Walton GE, van den Heuvel EG, Kosters MH, Rastall RA, Tuohy KM, Gibson GR. A randomised crossover study investigating the effects of galacto-oligosaccharides on the faecal microbiota in men and women over 50 years of age.. Br J Nutr 2012 May;107(10):1466-75.
    doi: 10.1017/S0007114511004697pubmed: 21910949google scholar: lookup
  10. Grimaldi R, Cela D, Swann JR, Vulevic J, Gibson GR, Tzortzis G, Costabile A. In vitro fermentation of B-GOS: impact on faecal bacterial populations and metabolic activity in autistic and non-autistic children.. FEMS Microbiol Ecol 2017 Feb;93(2).
    doi: 10.1093/femsec/fiw233pmc: PMC5155555pubmed: 27856622google scholar: lookup
  11. Grimaldi R, Gibson GR, Vulevic J, Giallourou N, Castro-Mejía JL, Hansen LH, Leigh Gibson E, Nielsen DS, Costabile A. A prebiotic intervention study in children with autism spectrum disorders (ASDs).. Microbiome 2018 Aug 2;6(1):133.
    doi: 10.1186/s40168-018-0523-3pmc: PMC6091020pubmed: 30071894google scholar: lookup
  12. Lowman RS, Theodorou MK, Hyslop JJ, Dhanoa MS, Cí·¯ord D. Evaluation of an in vitro batch culture technique for estimating the in vivo digestibility and digestible energy content of equine feeds using equine faeces as the source of microbial inoculum. Anim Feed Sci Technol 80:11–27.
    doi: 10.1016/S0377-8401(99)00039-5google scholar: lookup
  13. Desrousseaux G, Santos AS, Pellikaan WF, Van der Poel AFB, Cone JW, Guedes CMV, Ferreira LMM, Rodrigues MAM. Effect of collection time on the fermentative activity of microbes in equine faeces. Anim Feed Sci Technol 178:183–189.
    doi: 10.1016/j.anifeedsci.2012.09.016google scholar: lookup
  14. Biddle AS, Black SJ, Blanchard JL. An in vitro model of the horse gut microbiome enables identification of lactate-utilizing bacteria that differentially respond to starch induction.. PLoS One 2013;8(10):e77599.
    doi: 10.1371/journal.pone.0077599pmc: PMC3788102pubmed: 24098591google scholar: lookup
  15. Andoh A, Tsujikawa T, Sasaki M, Mitsuyama K, Suzuki Y, Matsui T, Matsumoto T, Benno Y, Fujiyama Y. Faecal microbiota profile of Crohn's disease determined by terminal restriction fragment length polymorphism analysis.. Aliment Pharmacol Ther 2009 Jan;29(1):75-82.
    doi: 10.1111/j.1365-2036.2008.03860.xpubmed: 18945264google scholar: lookup
  16. Moore-Colyer M, O'Gorman DM, Wakefield K. An in vitro investigation into the effects of a marine-derived, multimineral supplement in simulated equine stomach and hindgut environments. J Equine Vet Sci 34:391–397.
    doi: 10.1016/j.jevs.2013.07.016google scholar: lookup
  17. Murray J, Bice RKT, Moore-Colyer M. The effect of particle size on the in vitro fermentation of different ratios of high-temperature dried lucerne and sugar beet pulp incubated with equine faecal inocula. Anim Feed Sci Technol 162:47–57.
    doi: 10.1016/j.anifeedsci.2010.09.001google scholar: lookup
  18. Murray J, McMullin P, Handel I, Hastie PM. The effect of freezing on the fermentative activity of equine faecal inocula for use in an in vitro gas production technique. Anim Feed Sci Technol 178:175–182.
    doi: 10.1016/j.anifeedsci.2012.09.013google scholar: lookup
  19. Murray JMD, McMullin P, Handel I, Hastie PM. Comparison of intestinal contents from different regions of the equine gastrointestinal tract as inocula for use in an in vitro gas production technique. Anim Feed Sci Technol 187:98–103.
    doi: 10.1016/j.anifeedsci.2013.10.005google scholar: lookup
  20. Murray J, Scott B, Hastie PM. Fermentative capacity of equine faecal inocula obtained from clinically normal horses and those predisposed to laminitis. Anim Feed Sci Technol 151:306–311.
    doi: 10.1016/j.anifeedsci.2009.01.011google scholar: lookup
  21. Abdouli H, Ben Attia S. Evaluation of a two-stage in vitro technique for estimating digestibility of equine feeds using horse faeces as the source of microbial inoculum. Anim Feed Sci Technol 132:155–162.
    doi: 10.1016/j.anifeedsci.2006.03.005google scholar: lookup
  22. Elghandour MMY, Vázquez Chagoyán JC, Salem AZM, Kholif AE, Martínez Castañeda JS, Camacho LM, Buendía G. In vitro fermentative capacity of equine fecal inocula of 9 fibrous forages in the presence of different doses of Saccharomyces cerevisiae. J Equine Vet Sci 34:619–625.
    doi: 10.1016/j.jevs.2013.11.013google scholar: lookup
  23. Hobden MR, Martin-Morales A, Guérin-Deremaux L, Wils D, Costabile A, Walton GE, Rowland I, Kennedy OB, Gibson GR. In vitro fermentation of NUTRIOSE(®) FB06, a wheat dextrin soluble fibre, in a continuous culture human colonic model system.. PLoS One 2013;8(10):e77128.
    doi: 10.1371/journal.pone.0077128pmc: PMC3811981pubmed: 24204753google scholar: lookup
  24. Costabile A, Walton GE, Tzortzis G, Vulevic J, Charalampopoulos D, Gibson GR. Effects of orange juice formulation on prebiotic functionality using an in vitro colonic model system.. PLoS One 2015;10(3):e0121955.
    doi: 10.1371/journal.pone.0121955pmc: PMC4373861pubmed: 25807417google scholar: lookup
  25. Pérez-López E, Cela D, Costabile A, Mateos-Aparicio I, Rupérez P. In vitro fermentability and prebiotic potential of soyabean Okara by human faecal microbiota.. Br J Nutr 2016 Sep;116(6):1116-24.
    doi: 10.1017/S0007114516002816pubmed: 27469454google scholar: lookup
  26. Liu Y, Gibson GR, Walton GE. A three-stage continuous culture approach to study the impact of probiotics, prebiotics and fat intake on faecal microbiota relevant to an over 60 s population. J Funct Foods 32:238–247.
    doi: 10.1016/j.jff.2017.02.035google scholar: lookup
  27. Gharbia SE, Shah HN. Pathways of glutamate catabolism among Fusobacterium species.. J Gen Microbiol 1991 May;137(5):1201-6.
    doi: 10.1099/00221287-137-5-1201pubmed: 1678005google scholar: lookup
  28. Rogers AH, Chen J, Zilm PS, Gully NJ. The behaviour of Fusobacterium nucleatum chemostat-grown in glucose- and amino acid-based chemically defined media.. Anaerobe 1998 Apr;4(2):111-6.
    doi: 10.1006/anae.1997.0140pubmed: 16887630google scholar: lookup
  29. Macrae JC, Armstrong DG. Enzyme method for determination of alpha-linked glucose polymers in biological materials. J Sci Food Agric 19:578.
    doi: 10.1002/jsfa.2740191006google scholar: lookup
  30. Fuller KW. Automated determination of sugars. Autom Anal Chem 2:57–.
  31. Macfarlane GT, Englyst HN. Starch utilization by the human large intestinal microflora.. J Appl Bacteriol 1986 Mar;60(3):195-201.
    doi: 10.1111/j.1365-2672.1986.tb01073.xpubmed: 2423494google scholar: lookup
  32. Zhao G, Nyman M, Jönsson JA. Rapid determination of short-chain fatty acids in colonic contents and faeces of humans and rats by acidified water-extraction and direct-injection gas chromatography.. Biomed Chromatogr 2006 Aug;20(8):674-82.
    doi: 10.1002/bmc.580pubmed: 16206138google scholar: lookup
  33. Escalona EE, Leng J, Dona AC, Merrifield CA, Holmes E, Proudman CJ, Swann JR. Dominant components of the Thoroughbred metabolome characterised by (1) H-nuclear magnetic resonance spectroscopy: A metabolite atlas of common biofluids.. Equine Vet J 2015 Nov;47(6):721-30.
    doi: 10.1111/evj.12333pubmed: 25130591google scholar: lookup
  34. Daims H, Brühl A, Amann R, Schleifer KH, Wagner M. The domain-specific probe EUB338 is insufficient for the detection of all Bacteria: development and evaluation of a more comprehensive probe set.. Syst Appl Microbiol 1999 Sep;22(3):434-44.
    doi: 10.1016/S0723-2020(99)80053-8pubmed: 10553296google scholar: lookup
  35. Caporaso JG, Lauber CL, Walters WA, Berg-Lyons D, Lozupone CA, Turnbaugh PJ, Fierer N, Knight R. Global patterns of 16S rRNA diversity at a depth of millions of sequences per sample.. Proc Natl Acad Sci U S A 2011 Mar 15;108 Suppl 1(Suppl 1):4516-22.
    doi: 10.1073/pnas.1000080107pmc: PMC3063599pubmed: 20534432google scholar: lookup
  36. Bolyen E, Rideout JR, Dillon MR, Bokulich NA, Abnet CC, Al-Ghalith GA, Alexander H, Alm EJ, Arumugam M, Asnicar F, Bai Y, Bisanz JE, Bittinger K, Brejnrod A, Brislawn CJ, Brown CT, Callahan BJ, Caraballo-Rodríguez AM, Chase J, Cope EK, Da Silva R, Diener C, Dorrestein PC, Douglas GM, Durall DM, Duvallet C, Edwardson CF, Ernst M, Estaki M, Fouquier J, Gauglitz JM, Gibbons SM, Gibson DL, Gonzalez A, Gorlick K, Guo J, Hillmann B, Holmes S, Holste H, Huttenhower C, Huttley GA, Janssen S, Jarmusch AK, Jiang L, Kaehler BD, Kang KB, Keefe CR, Keim P, Kelley ST, Knights D, Koester I, Kosciolek T, Kreps J, Langille MGI, Lee J, Ley R, Liu YX, Loftfield E, Lozupone C, Maher M, Marotz C, Martin BD, McDonald D, McIver LJ, Melnik AV, Metcalf JL, Morgan SC, Morton JT, Naimey AT, Navas-Molina JA, Nothias LF, Orchanian SB, Pearson T, Peoples SL, Petras D, Preuss ML, Pruesse E, Rasmussen LB, Rivers A, Robeson MS 2nd, Rosenthal P, Segata N, Shaffer M, Shiffer A, Sinha R, Song SJ, Spear JR, Swafford AD, Thompson LR, Torres PJ, Trinh P, Tripathi A, Turnbaugh PJ, Ul-Hasan S, van der Hooft JJJ, Vargas F, Vázquez-Baeza Y, Vogtmann E, von Hippel M, Walters W, Wan Y, Wang M, Warren J, Weber KC, Williamson CHD, Willis AD, Xu ZZ, Zaneveld JR, Zhang Y, Zhu Q, Knight R, Caporaso JG. Author Correction: Reproducible, interactive, scalable and extensible microbiome data science using QIIME 2.. Nat Biotechnol 2019 Sep;37(9):1091.
    doi: 10.7287/peerj.preprints.27295v2pubmed: 31399723google scholar: lookup
  37. Callahan BJ, McMurdie PJ, Rosen MJ, Han AW, Johnson AJ, Holmes SP. DADA2: High-resolution sample inference from Illumina amplicon data.. Nat Methods 2016 Jul;13(7):581-3.
    doi: 10.1038/nmeth.3869pmc: PMC4927377pubmed: 27214047google scholar: lookup
  38. McDonald D, Price MN, Goodrich J, Nawrocki EP, DeSantis TZ, Probst A, Andersen GL, Knight R, Hugenholtz P. An improved Greengenes taxonomy with explicit ranks for ecological and evolutionary analyses of bacteria and archaea.. ISME J 2012 Mar;6(3):610-8.
    doi: 10.1038/ismej.2011.139pmc: PMC3280142pubmed: 22134646google scholar: lookup

Citations

This article has been cited 6 times.
  1. Chaucheyras-Durand F, Sacy A, Karges K, Apper E. Gastro-Intestinal Microbiota in Equines and Its Role in Health and Disease: The Black Box Opens.. Microorganisms 2022 Dec 19;10(12).
    doi: 10.3390/microorganisms10122517pubmed: 36557769google scholar: lookup
  2. Gilroy R, Leng J, Ravi A, Adriaenssens EM, Oren A, Baker D, La Ragione RM, Proudman C, Pallen MJ. Metagenomic investigation of the equine faecal microbiome reveals extensive taxonomic diversity.. PeerJ 2022;10:e13084.
    doi: 10.7717/peerj.13084pubmed: 35345588google scholar: lookup
  3. Goodman-Davis R, Figurska M, Cywinska A. Gut Microbiota Manipulation in Foals-Naturopathic Diarrhea Management, or Unsubstantiated Folly?. Pathogens 2021 Sep 4;10(9).
    doi: 10.3390/pathogens10091137pubmed: 34578169google scholar: lookup
  4. van der Linde C, Barone M, Turroni S, Brigidi P, Keleszade E, Swann JR, Costabile A. An In Vitro Pilot Fermentation Study on the Impact of Chlorella pyrenoidosa on Gut Microbiome Composition and Metabolites in Healthy and Coeliac Subjects.. Molecules 2021 Apr 16;26(8).
    doi: 10.3390/molecules26082330pubmed: 33923841google scholar: lookup
  5. Daniels SP, Leng J, Swann JR, Proudman CJ. Bugs and drugs: a systems biology approach to characterising the effect of moxidectin on the horse's faecal microbiome.. Anim Microbiome 2020 Oct 14;2(1):38.
    doi: 10.1186/s42523-020-00056-2pubmed: 33499996google scholar: lookup
  6. Tassone S, Fortina R, Valle E, Cavallarin L, Raspa F, Boggero S, Bergero D, Giammarino M, Renna M. Comparison of In Vivo and In Vitro Digestibility in Donkeys.. Animals (Basel) 2020 Nov 12;10(11).
    doi: 10.3390/ani10112100pubmed: 33198168google scholar: lookup
Subscribe to our newsletter
Join 99,357 horse owners like you who receive our email updates on horse health and nutrition.