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 / Identification of equine cecal bacteria producing amines in ...
Applied and environmental microbiology2003; 69(4); 2087-2093; doi: 10.1128/AEM.69.4.2087-2093.2003

Identification of equine cecal bacteria producing amines in an in vitro model of carbohydrate overload.

Abstract: Acute laminitis has been associated with the overgrowth of gram-positive bacteria within the equine hindgut, causing the release of factor(s) leading to ischemia-reperfusion of the digits. The products of fermentation which trigger acute laminitis are, as yet, unknown; however, vasoactive amines are possible candidates. The objectives of this study were to use an in vitro model of carbohydrate overload to study the change in populations of cecal streptococci and lactobacilli and to establish whether certain species of these bacteria were capable of producing vasoactive amines from amino acids. Cecal contents from 10 horses were divided into aliquots and incubated anaerobically with either corn starch or inulin (fructan; both at 1 g/100 ml). Samples were taken at 6-h intervals over a 24-h period for enumeration of streptococci, lactobacilli, and gram-negative anaerobes by a dilution method onto standard selective growth media. The effects of the antibiotic virginiamycin (1 mg/100 ml) and calcium hydrogen phosphate (CaHPO(4); 0.3 g/100 ml) were also examined. Fermentation of excess carbohydrate was associated with increases in numbers of streptococci and lactobacilli (2- to 3.5-log unit increases; inhibited by virginiamycin) but numbers of gram-negative anaerobes were not significantly affected. A screening agar technique followed by 16S rRNA gene sequence analysis enabled the identification of 26 different bacterial strains capable of producing one or more vasoactive amines. These included members of the species Streptococcus bovis and five different Lactobacillus spp. These data suggest that certain bacteria, whose overgrowth is associated with carbohydrate fermentation, are capable of producing vasoactive amines which may play a role in the pathogenesis of acute laminitis.
Get Full Text
Publication Date: 2003-04-05 PubMed ID: 12676687PubMed Central: PMC154823DOI: 10.1128/AEM.69.4.2087-2093.2003Google 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 study investigates the role of certain bacteria in the equine (horse) hindgut in relation to acute laminitis, a painful disease affecting horse’s feet. The findings suggest that some of these bacteria, which multiply due to carbohydrate fermentation, produce vasoactive amines, potentially triggering acute laminitis.

Objective of the Study

The research aimed at understanding the role of specific bacteria in the onset of acute laminitis in horses. The researchers hypothesized that certain bacteria in the horse’s hindgut may produce vasoactive amines, a consequence of carbohydrate fermentation, which might play a vital role in triggering laminitis.

Methodology of the Study

  • An in vitro model was created to simulate a condition of carbohydrate overload.
  • Cecal contents from ten horses were further broken down into smaller samples for the experiment.
  • These samples were exposed to an anaerobic environment induced with corn starch or inulin at a concentration of 1g/100ml.
  • Routine samples were collected every six hours, over 24 hours to evaluate the count of streptococci, lactobacilli, and gram-negative anaerobes.
  • The impact of virginiamycin ( an antibiotic) and calcium hydrogen phosphate were also examined in this process.

Results of the Study

  • Fermentation of excess carbohydrate led to an increase in streptococci and lactobacilli. The numbers of gram-negative anaerobes were not significantly affected.
  • The researchers used a screening agar technique, paired with 16S rRNA gene sequence analysis to identify the types of bacteria capable of producing vasoactive amines.
  • 26 different bacterial strains were identified in the process that could give out one or more vasoactive amines.
  • Various species, including Streptococcus bovis and five different species of Lactobacillus, were among the bacteria identified.

Conclusion of the Study

The researchers concluded that specific bacteria, whose proliferation is linked with carbohydrate fermentation, are capable of producing vasoactive amines. This may have significant implications in the development of acute laminitis. These findings contribute to a better understanding of acute laminitis pathogenesis, which could help devise more effective treatment or preventive strategies for the disease.

Cite This Article

APA
Bailey SR, Baillon ML, Rycroft AN, Harris PA, Elliott J. (2003). Identification of equine cecal bacteria producing amines in an in vitro model of carbohydrate overload. Appl Environ Microbiol, 69(4), 2087-2093. https://doi.org/10.1128/AEM.69.4.2087-2093.2003

Publication

Applied and environmental microbiology
ISSN: 0099-2240
NlmUniqueID: 7605801
Country: United States
Language: English
Volume: 69
Issue: 4
Pages: 2087-2093

Researcher Affiliations

Bailey, S R
  • Department of Veterinary Basic Sciences, Royal Veterinary College, London, United Kingdom. jelliott@rvc.ac.uk
Baillon, M-L
    Rycroft, A N
      Harris, P A
        Elliott, J

          MeSH Terms

          • Acute Disease
          • Amines / metabolism
          • Animal Feed
          • Animals
          • Bacteria / classification
          • Bacteria / genetics
          • Bacteria / isolation & purification
          • Calcium Phosphates / pharmacology
          • Cecum / microbiology
          • Colony Count, Microbial
          • Dietary Carbohydrates / administration & dosage
          • Gram-Negative Anaerobic Bacteria / classification
          • Gram-Negative Anaerobic Bacteria / genetics
          • Gram-Negative Anaerobic Bacteria / isolation & purification
          • Horse Diseases / etiology
          • Horses
          • Lactobacillus / classification
          • Lactobacillus / genetics
          • Lactobacillus / isolation & purification
          • RNA, Ribosomal, 16S / genetics
          • Reperfusion Injury / etiology
          • Streptococcus / classification
          • Streptococcus / genetics
          • Streptococcus / isolation & purification
          • Virginiamycin / pharmacology

          References

          This article includes 44 references
          1. Al Jassim RAM, Rowe JB. Better understanding of acidosis and its control. Rec. Adv. Anim. Nutr. Aust. 1999 12:91-97.
          2. Allison MJ, Robinson IM, Dougherty RW, Bucklin JA. Grain overload in cattle and sheep: changes in microbial populations in the cecum and rumen.. Am J Vet Res 1975 Feb;36(2):181-5.
            pubmed: 234213
          3. Altschul SF, Madden TL, Schäffer AA, Zhang J, Zhang Z, Miller W, Lipman DJ. Gapped BLAST and PSI-BLAST: a new generation of protein database search programs.. Nucleic Acids Res 1997 Sep 1;25(17):3389-402.
            pmc: PMC146917pubmed: 9254694doi: 10.1093/nar/25.17.3389google scholar: lookup
          4. Bailey SR, Elliott J. Plasma 5-hydroxytryptamine constricts equine digital blood vessels in vitro: implications for pathogenesis of acute laminitis.. Equine Vet J 1998 Mar;30(2):124-30.
            pubmed: 9535068doi: 10.1111/j.2042-3306.1998.tb04471.xgoogle scholar: lookup
          5. Bailey SR, Rycroft A, Elliott J. Production of amines in equine cecal contents in an in vitro model of carbohydrate overload.. J Anim Sci 2002 Oct;80(10):2656-62.
            pubmed: 12413088doi: 10.2527/2002.80102656xgoogle scholar: lookup
          6. Baxter GM, Laskey RE, Tackett RL, Moore JN, Allen D. In vitro reactivity of digital arteries and veins to vasoconstrictive mediators in healthy horses and in horses with early laminitis.. Am J Vet Res 1989 Apr;50(4):508-17.
            pubmed: 2712417
          7. Boeker E, Snell E. Amino acid decarboxylases, p. 217. 1972.
          8. Bover-Cid S, Holzapfel WH. Improved screening procedure for biogenic amine production by lactic acid bacteria.. Int J Food Microbiol 1999 Dec 1;53(1):33-41.
            pubmed: 10598112doi: 10.1016/s0168-1605(99)00152-xgoogle scholar: lookup
          9. Bover-Cid S, Hugas M, Izquierdo-Pulido M, Vidal-Carou MC. Amino acid-decarboxylase activity of bacteria isolated from fermented pork sausages.. Int J Food Microbiol 2001 Jun 15;66(3):185-9.
            pubmed: 11428577doi: 10.1016/s0168-1605(00)00526-2google scholar: lookup
          10. Briggs M. An improved medium for lactobacilli. J. Dairy Res. 1953 20:36-40.
          11. Coe ML, Nagaraja TG, Sun YD, Wallace N, Towne EG, Kemp KE, Hutcheson JP. Effect of virginiamycin on ruminal fermentation in cattle during adaptation to a high concentrate diet and during an induced acidosis.. J Anim Sci 1999 Aug;77(8):2259-68.
            pubmed: 10462007doi: 10.2527/1999.7782259xgoogle scholar: lookup
          12. de Man JC, Rogosa M, Sharpe EM. A medium for the cultivation of lactobacilli. J. Appl. Bacteriol. 1960 23:130-135.
          13. Demeyer DI, Fiedler D, De Graeve KG. Attempted induction of reductive acetogenesis into the rumen fermentation in vitro.. Reprod Nutr Dev 1996;36(3):233-40.
            pubmed: 8766728doi: 10.1051/rnd:19960301google scholar: lookup
          14. Garner HE, Coffman JR, Hahn AW, Hutcheson DP, Tumbleson ME. Equine laminitis of alimentary origin: an experimental model.. Am J Vet Res 1975 Apr;36(4 Pt.1):441-4.
            pubmed: 1124880
          15. Garner HE, Hutcheson DP, Coffman JR, Hahn AW, Salem C. Lactic acidosis: a factor associated with equine laminitis.. J Anim Sci 1977 Nov;45(5):1037-41.
            pubmed: 599094doi: 10.2527/jas1977.4551037xgoogle scholar: lookup
          16. Garner HE, Moore JN, Johnson JH, Clark L, Amend JF, Tritschler LG, Coffmann JR, Sprouse RF, Hutcheson DP, Salem CA. Changes in the caecal flora associated with the onset of laminitis.. Equine Vet J 1978 Oct;10(4):249-52.
            pubmed: 738266doi: 10.1111/j.2042-3306.1978.tb02273.xgoogle scholar: lookup
          17. Ghenghesh KS, Drucker DB. Gas liquid chromatography of amines produced by the Enterobacteriaceae.. Braz J Med Biol Res 1989;22(6):653-65.
            pubmed: 2620174
          18. Golovnya RV, Zhuravleva IL, Kharatyan SG. Gas chromatographic analysis of amines in volatile substances of Streptococcus lactis.. J Chromatogr 1969 Oct 28;44(2):262-8.
            pubmed: 5347346doi: 10.1016/s0021-9673(01)92536-1google scholar: lookup
          19. Hartemink R, Domenech VR, Rombouts RM. LAMVAB—a new selective medium for the isolation of lactobacilli from faeces. J. Microbiol. Methods 1997 29:77-84.
          20. Hartemink R, Rombouts FM. Comparison of media for the detection of bifidobacteria, lactobacilli and total anaerobes from faecal samples.. J Microbiol Methods 1999 Jun;36(3):181-92.
            pubmed: 10379804doi: 10.1016/s0167-7012(99)00031-7google scholar: lookup
          21. Hoffman RM, Wilson JA, Kronfeld DS, Cooper WL, Lawrence LA, Sklan D, Harris PA. Hydrolyzable carbohydrates in pasture, hay, and horse feeds: direct assay and seasonal variation.. J Anim Sci 2001 Feb;79(2):500-6.
            pubmed: 11219461doi: 10.2527/2001.792500xgoogle scholar: lookup
          22. Hood DM, Grosenbaugh DA, Mostafa MB, Morgan SJ, Thomas BC. The role of vascular mechanisms in the development of acute equine laminitis.. J Vet Intern Med 1993 Jul-Aug;7(4):228-34.
            pubmed: 8246212doi: 10.1111/j.1939-1676.1993.tb01012.xgoogle scholar: lookup
          23. Hood DM, Wagner IP, Brumbaugh GW. Evaluation of hoof wall surface temperature as an index of digital vascular perfusion during the prodromal and acute phases of carbohydrate-induced laminitis in horses.. Am J Vet Res 2001 Jul;62(7):1167-72.
            pubmed: 11453497doi: 10.2460/ajvr.2001.62.1167google scholar: lookup
          24. Hurst WJ. A review of HPLC methods for the determination of selected biogenic amines in foods. J. Liquid Chromatogr. 1990 13:1-23.
          25. Julliand V, de Vaux A, Millet L, Fonty G. Identification of Ruminococcus flavefaciens as the predominant cellulolytic bacterial species of the equine cecum.. Appl Environ Microbiol 1999 Aug;65(8):3738-41.
            pmc: PMC91562pubmed: 10427077doi: 10.1128/aem.65.8.3738-3741.1999google scholar: lookup
          26. Kern DL, Slyter LL, Weaver JM, Leffel EC, Samuelson G. Pony cecum vs. steer rumen: the effect of oats and hay on the microbial ecosystem.. J Anim Sci 1973 Aug;37(2):463-9.
            pubmed: 4201259doi: 10.2527/jas1973.372463xgoogle scholar: lookup
          27. Kern DL, Slyter LL, Leffel EC, Weaver JM, Oltjen RR. Ponies vs. steers: microbial and chemical characteristics of intestinal ingesta.. J Anim Sci 1974 Mar;38(3):559-64.
            pubmed: 4856481doi: 10.2527/jas1974.383559xgoogle scholar: lookup
          28. Leuschner RG, Hammes WP. Formation of biogenic amine in mayonnaise, herring and tuna fish salad by lactobacilli.. Int J Food Sci Nutr 1999 May;50(3):159-64.
            pubmed: 10627831doi: 10.1080/096374899101193google scholar: lookup
          29. Marounek M, Skrivanová V, Simůnek J. [The effect of virginiamycin on rumen fermentation in vitro after adaptation of donors to the inoculum].. Vet Med (Praha) 1995 May;40(5):129-32.
            pubmed: 7793012
          30. Miles AA, Misra SS, Irwin JO. The estimation of the bactericidal power of the blood.. J Hyg (Lond) 1938 Nov;38(6):732-49.
            pmc: PMC2199673pubmed: 20475467doi: 10.1017/s002217240001158xgoogle scholar: lookup
          31. Moore JN, Garner HE, Berg JN, Sprouse RF. Intracecal endotoxin and lactate during the onset of equine laminitis: a preliminary report.. Am J Vet Res 1979 May;40(5):722-3.
            pubmed: 38715
          32. Moreno-Arribas V, Torlois S, Joyeux A, Bertrand A, Lonvaud-Funel A. Isolation, properties and behaviour of tyramine-producing lactic acid bacteria from wine.. J Appl Microbiol 2000 Apr;88(4):584-93.
            pubmed: 10792516doi: 10.1046/j.1365-2672.2000.00997.xgoogle scholar: lookup
          33. Mungall BA, Kyaw-Tanner M, Pollitt CC. In vitro evidence for a bacterial pathogenesis of equine laminitis.. Vet Microbiol 2001 Apr 2;79(3):209-23.
            pubmed: 11240100doi: 10.1016/s0378-1135(00)00359-xgoogle scholar: lookup
          34. Nagaraja TG, Taylor MB. Susceptibility and resistance of ruminal bacteria to antimicrobial feed additives.. Appl Environ Microbiol 1987 Jul;53(7):1620-5.
            pmc: PMC203920pubmed: 3116929doi: 10.1128/aem.53.7.1620-1625.1987google scholar: lookup
          35. Potter GD, Arnold FF, Householder DD, Hansen DH, Brown KM. Digestion of starch in the small or large intestine of the equine. Pferdeheilkunde 1992 1:107-111.
          36. Rémésy C, Levrat MA, Gamet L, Demigné C. Cecal fermentations in rats fed oligosaccharides (inulin) are modulated by dietary calcium level.. Am J Physiol 1993 May;264(5 Pt 1):G855-62.
            pubmed: 8388642doi: 10.1152/ajpgi.1993.264.5.g855google scholar: lookup
          37. Rice SL, Koehler PE. Tyrosine and histidine decarboxylase activities of Pediococcus cerevisiae and Lactobacillus species and the production of tyramine in fermented sausages. J. Milk Food Technol. 1976 38:256-258.
          38. ROGOSA M, MITCHELL JA, WISEMAN RF. A selective medium for the isolation and enumeration of oral and fecal lactobacilli.. J Bacteriol 1951 Jul;62(1):132-3.
            pmc: PMC386093pubmed: 14861168doi: 10.1128/jb.62.1.132-133.1951google scholar: lookup
          39. Roos S, Karner F, Axelsson L, Jonsson H. Lactobacillus mucosae sp. nov., a new species with in vitro mucus-binding activity isolated from pig intestine.. Int J Syst Evol Microbiol 2000 Jan;50 Pt 1:251-258.
            pubmed: 10826811doi: 10.1099/00207713-50-1-251google scholar: lookup
          40. Rowe JB, Lees MJ, Pethick DW. Prevention of acidosis and laminitis associated with grain feeding in horses.. J Nutr 1994 Dec;124(12 Suppl):2742S-2744S.
            pubmed: 7996285doi: 10.1093/jn/124.suppl_12.2747sgoogle scholar: lookup
          41. Sanger F, Nicklen S, Coulson AR. DNA sequencing with chain-terminating inhibitors.. Proc Natl Acad Sci U S A 1977 Dec;74(12):5463-7.
            pmc: PMC431765pubmed: 271968doi: 10.1073/pnas.74.12.5463google scholar: lookup
          42. Straub BW, Kicherer M, Schilcher SM, Hammes WP. The formation of biogenic amines by fermentation organisms.. Z Lebensm Unters Forsch 1995 Jul;201(1):79-82.
            pubmed: 7571871doi: 10.1007/bf01193205google scholar: lookup
          43. Whitehead TR, Cotta MA. Development of molecular methods for identification of Streptococcus bovis from human and ruminal origins.. FEMS Microbiol Lett 2000 Jan 15;182(2):237-40.
            pubmed: 10620672doi: 10.1111/j.1574-6968.2000.tb08901.xgoogle scholar: lookup
          44. Wilkins TD, Chalgren S. Medium for use in antibiotic susceptibility testing of anaerobic bacteria.. Antimicrob Agents Chemother 1976 Dec;10(6):926-8.
            pmc: PMC429869pubmed: 1008552doi: 10.1128/aac.10.6.926google scholar: lookup

          Citations

          This article has been cited 15 times.
          1. Ayoub C, Arroyo LG, MacNicol JL, Renaud D, Weese JS, Gomez DE. Fecal microbiota of horses with colitis and its association with laminitis and survival during hospitalization. J Vet Intern Med 2022 Nov;36(6):2213-2223.
            doi: 10.1111/jvim.16562pubmed: 36271677google scholar: lookup
          2. MacNicol JL, Renwick S, Ganobis CM, Allen-Vercoe E, Weese JS, Pearson W. A Comparison of Methods to Maintain the Equine Cecal Microbial Environment In Vitro Utilizing Cecal and Fecal Material. Animals (Basel) 2022 Aug 8;12(15).
            doi: 10.3390/ani12152009pubmed: 35953998google scholar: lookup
          3. Dittoe DK, Olson EG, Ricke SC. Impact of the gastrointestinal microbiome and fermentation metabolites on broiler performance. Poult Sci 2022 May;101(5):101786.
            doi: 10.1016/j.psj.2022.101786pubmed: 35346496google scholar: lookup
          4. Alam MJ, Islam M, Jeon CO, Lee KC, Kim SH, Yang CJ, Kabir ME, Lee SS. In vitro assessment of probiotic potential of selected bacteria isolated from pig faeces with potential application of odour reduction. Int J Vet Sci Med 2021;9(1):22-30.
            doi: 10.1080/23144599.2021.1936962pubmed: 34277842google scholar: lookup
          5. Kauter A, Epping L, Semmler T, Antao EM, Kannapin D, Stoeckle SD, Gehlen H, Lübke-Becker A, Günther S, Wieler LH, Walther B. The gut microbiome of horses: current research on equine enteral microbiota and future perspectives. Anim Microbiome 2019 Nov 13;1(1):14.
            doi: 10.1186/s42523-019-0013-3pubmed: 33499951google scholar: lookup
          6. Curtis L, Burford JH, England GCW, Freeman SL. Risk factors for acute abdominal pain (colic) in the adult horse: A scoping review of risk factors, and a systematic review of the effect of management-related changes. PLoS One 2019;14(7):e0219307.
            doi: 10.1371/journal.pone.0219307pubmed: 31295284google scholar: lookup
          7. Li H, Liu J, Zhu W, Mao S. Intraruminal infusion of oligofructose alters ruminal microbiota and induces acute laminitis in sheep. J Anim Sci 2017 Dec;95(12):5407-5419.
            doi: 10.2527/jas2017.1860pubmed: 29293794google scholar: lookup
          8. Jin W, Xue C, Liu J, Yin Y, Zhu W, Mao S. Effects of Disodium Fumarate on In Vitro Rumen Fermentation, The Production of Lipopolysaccharide and Biogenic Amines, and The Rumen Bacterial Community. Curr Microbiol 2017 Nov;74(11):1337-1342.
            doi: 10.1007/s00284-017-1322-ypubmed: 28761980google scholar: lookup
          9. Bland SD, Venable EB, McPherson JL, Atkinson RL. Effects of liposomal-curcumin on five opportunistic bacterial strains found in the equine hindgut - preliminary study. J Anim Sci Technol 2017;59:15.
            doi: 10.1186/s40781-017-0138-4pubmed: 28638626google scholar: lookup
          10. Harlow BE, Lawrence LM, Hayes SH, Crum A, Flythe MD. Effect of Dietary Starch Source and Concentration on Equine Fecal Microbiota. PLoS One 2016;11(4):e0154037.
            doi: 10.1371/journal.pone.0154037pubmed: 27128793google scholar: lookup
          11. Shepherd ML, Ponder MA, Burk AO, Milton SC, Swecker WS Jr. Fibre digestibility, abundance of faecal bacteria and plasma acetate concentrations in overweight adult mares. J Nutr Sci 2014;3:e10.
            doi: 10.1017/jns.2014.8pubmed: 25191602google scholar: lookup
          12. 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.0077599pubmed: 24098591google scholar: lookup
          13. St-Pierre B, de la Fuente G, O'Neill S, Wright AD, Al Jassim R. Analysis of stomach bacterial communities in Australian feral horses. Mol Biol Rep 2013 Jan;40(1):369-76.
            doi: 10.1007/s11033-012-2070-5pubmed: 23065252google scholar: lookup
          14. Endo A, Futagawa-Endo Y, Dicks LM. Lactobacillus and Bifidobacterium diversity in horse feces, revealed by PCR-DGGE. Curr Microbiol 2009 Dec;59(6):651-5.
            doi: 10.1007/s00284-009-9498-4pubmed: 19730939google scholar: lookup
          15. Bishop RC, Kemper AM, Clark LV, Wilkins PA, McCoy AM. Stability of Gastric Fluid and Fecal Microbial Populations in Healthy Horses under Pasture and Stable Conditions. Animals (Basel) 2024 Oct 16;14(20).
            doi: 10.3390/ani14202979pubmed: 39457909google scholar: lookup
          Subscribe to our newsletter
          Join 99,344 horse owners like you who receive our email updates on horse health and nutrition.