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Food, fibre and pharmaceuticals from animals
RESEARCH ARTICLE

Is polyethylene glycol innocuous to the rumen bacterial community? A preliminary in vitro study

A. Belenguer A C , G. Hervás A , P. G. Toral A , M. Fondevila B and P. Frutos A
+ Author Affiliations
- Author Affiliations

A Instituto de Ganadería de Montaña (IGM, CSIC-Universidad de León), Finca Marzanas, s/n, 24346-Grulleros, León, Spain.

B Departamento de Producción Animal y Ciencia de los Alimentos, Universidad de Zaragoza, Miguel Servet, 177, 50013-Zaragoza, Spain.

C Corresponding author. Email: a.belenguer@csic.es

Animal Production Science 51(11) 990-995 https://doi.org/10.1071/AN11041
Submitted: 27 March 2011  Accepted: 8 August 2011   Published: 21 October 2011

Abstract

Polyethylene glycol (PEG) is a polymer that is widely used in nutritional studies examining the effect of tannins on ruminal fermentation. There is no information however on its potential effect on the structure of the rumen bacterial community. Therefore, the aim herein was to investigate its effect on rumen bacterial profile, using an in vitro batch culture experiment with three substrates (alfalfa hay, maize grain, and a combination of both) to simulate three different rumen environments, treated with or without PEG. Rumen fluid was collected from four cannulated sheep and pooled to inoculate the cultures, which were run at 39°C for 22 h. At the end of the incubation, samples were immediately frozen for microbial DNA extraction. Terminal restriction fragment length polymorphism analysis of 16S rRNA genes revealed that, although there was a high similarity in the fragments detected in the cultures with or without PEG, their relative abundances suggested that PEG might induce some changes in the bacterial community structure when a starch-rich substrate (e.g. maize) is assayed. Furthermore, the relative frequency of some abundant fragments, such as one compatible with bacteria of the phylum Bacteroidetes detected with the enzyme HhaI, and another that may match microorganisms of the genus Ruminococcus obtained with the enzyme MspI, was increased when PEG was added to maize-supplied microbial cultures. These results suggest that the use of PEG in batch cultures may not be as innocuous to rumen bacterial populations as previously described regarding ruminal fermentation, and might be relevant to studies using this polymer to examine the effect of tannins on rumen microbiota.


References

Álvarez del Pino MC, Hervás G, Mantecón AR, Giráldez FJ, Frutos P (2005) Comparison of biological and chemical methods, and internal and external standards, for assaying tannins in shrub species. Journal of the Science of Food and Agriculture 85, 583–590.
Comparison of biological and chemical methods, and internal and external standards, for assaying tannins in shrub species.Crossref | GoogleScholarGoogle Scholar |

Belenguer A, Hervás G, Yáñez-Ruiz DR, Toral PG, Ezquerro C, Frutos P (2010) Preliminary study of the changes in rumen bacterial populations from cattle intoxicated with young oak (Quercus pyrenaica) leaves. Animal Production Science 50, 228–234.
Preliminary study of the changes in rumen bacterial populations from cattle intoxicated with young oak (Quercus pyrenaica) leaves.Crossref | GoogleScholarGoogle Scholar |

Belenguer A, Hervás G, Toral PG, Fondevila M, Frutos P (2011) Effect of polyethilenglycol on the rumen bacterial community: in vitro cultures with inocula from sheep fed only forage or a concentrated-rich diet. In ‘XIV Jornadas sobre Producción Animal AIDA. Tomo II (III Simposio sobre Microbiología Digestiva)’. (Eds A Sanz, I Casasus, M Joy, J Álvarez, JH Calvo, B Panea, P Muñoz, J Balcells) pp. 824–826. (AIDA: Zaragoza)

Cole JR, Wang Q, Cardenas E, Fish J, Chai B, Farris RJ, Kulam-Syed-Mohideen AS, McGarrell DM, Marsh T, Garrity GM, Tiedje JM (2009) The Ribosomal Database Project: improved alignments and new tools for rRNA analysis. Nucleic Acids Research 37, D141–D145.
The Ribosomal Database Project: improved alignments and new tools for rRNA analysis.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BD1cXhsFejtLbF&md5=65c84227c31ea7ac05557d2d0ff7cb9fCAS |

Edwards JE, McEwan NR, Travis AJ, Wallace RJ (2004) 16S rDNA library-based analysis of ruminal bacterial diversity. Antonie van Leeuwenhoek 86, 263–281.
16S rDNA library-based analysis of ruminal bacterial diversity.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BD2cXps1aisLo%3D&md5=4ab6579c0f0fbddbc4537c41de2fddbfCAS |

Frey JC, Pell AN, Berthiaume R, Lapierre H, Lee S, Ha JK, Mendell JE, Angert ER (2009) Comparative studies of microbial populations in the rumen, duodenum, ileum and faeces of lactating dairy cows. Journal of Applied Microbiology 108, 1982–1993.

Frutos P, Hervás G, Giráldez FJ, Mantecón A (2004) An in vitro study on the ability of polyethylene glycol to inhibit the effect of quebracho tannins and tannic acid on rumen fermentation in sheep, goats, cows, and deer. Australian Journal of Agricultural Research 55, 1125–1132.
An in vitro study on the ability of polyethylene glycol to inhibit the effect of quebracho tannins and tannic acid on rumen fermentation in sheep, goats, cows, and deer.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BD2cXhtVantbnE&md5=945f39db5ec6ad0ab097d7b328e5652fCAS |

Goering MK, Van Soest PJ (1970) ‘Forage fiber analyses (apparatus, reagents, procedures, and some applications). Agriculture Handbook No. 379.’ (Agricultural Research Service, USDA: Washington, DC)

Hervás G, Frutos P, Giráldez FJ, Mora MJ, Fernández B, Mantecón AR (2005) Effect of preservation on fermentative activity of rumen fluid inoculum for in vitro gas production techniques. Animal Feed Science and Technology 123–124, 107–118.
Effect of preservation on fermentative activity of rumen fluid inoculum for in vitro gas production techniques.Crossref | GoogleScholarGoogle Scholar |

Hill TCJ, Walsh KA, Harris JA, Moffett BF (2003) Using ecological diversity measures with bacterial communities. FEMS Microbiology Ecology 43, 1–11.
Using ecological diversity measures with bacterial communities.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BD3sXhtVCqtr8%3D&md5=ed4ca9c7a39f15a18fc1d84e642c7fc4CAS |

Kent AD, Smith DJ, Benson BJ, Triplett EW (2003) Web-based phylogenetic assignment tool for analysis of terminal restriction fragment length polymorphism profiles of microbial communities. Applied and Environmental Microbiology 69, 6768–6776.
Web-based phylogenetic assignment tool for analysis of terminal restriction fragment length polymorphism profiles of microbial communities.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BD3sXptVyktLc%3D&md5=4264ab63c006350e03f897ba40e7a22bCAS |

Kitts CL (2001) Terminal restriction fragment patterns: a tool for comparing microbial communities and assessing community dynamics. Current Issues in Intestinal Microbiology 2, 17–25.

Klieve AV, O’Leary MN, McMillen L, Owerkerk D (2007) Ruminococcus bromii, identification and isolation as a dominant community member in the rumen of cattle fed a barley diet. Journal of Applied Microbiology 103, 2065–2073.
Ruminococcus bromii, identification and isolation as a dominant community member in the rumen of cattle fed a barley diet.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BD1cXnvFantw%3D%3D&md5=9cf01d32fe631d80b579e713926c7373CAS |

Krause DO, Smith WJM, McSweeney CS (2004) Use of community genome arrays (CGAs) to assess the effects of Acacia angustissima on rumen ecology. Microbiology 150, 2899–2909.
Use of community genome arrays (CGAs) to assess the effects of Acacia angustissima on rumen ecology.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BD2cXotF2jsL8%3D&md5=dd445e76c2e9ca78fb779bdf452cb25bCAS |

Li M, Penner GB, Hernandez-Sanabria E, Oba M, Guan LL (2009) Effects of sampling location and time, and host animal on assessment of bacterial diversity and fermentation parameters in the bovine rumen. Journal of Applied Microbiology 107, 1924–1934.
Effects of sampling location and time, and host animal on assessment of bacterial diversity and fermentation parameters in the bovine rumen.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BD1MXhs1Sju73E&md5=fb9097a1ee02180b90fa0eaa41f48d3bCAS |

Makkar HPS (2003) ‘Quantification of tannins in tree and shrub foliage. A laboratory manual.’ (Kluver Academic Publishers: Dordrecht, The Netherlands)

Makkar HPS, Blümmel M, Becker K (1995) Formation of complexes between polyvinyl pyrrolidones or polyethylene glycol and tannins, and their implications in gas production and true digestibility in in vitro techniques. The British Journal of Nutrition 73, 897–913.
Formation of complexes between polyvinyl pyrrolidones or polyethylene glycol and tannins, and their implications in gas production and true digestibility in in vitro techniques.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DyaK2MXmvVyls78%3D&md5=2f09be7a289d43c4873bc2754ba30c33CAS |

McSweeney CS, Palmer B, Bunch R, Krause DO (2001) Effect of the tropical forage Calliandra on microbial protein synthesis and ecology in the rumen. Journal of Applied Microbiology 90, 78–88.
Effect of the tropical forage Calliandra on microbial protein synthesis and ecology in the rumen.Crossref | GoogleScholarGoogle Scholar | 1:STN:280:DC%2BD3M7jtlKksw%3D%3D&md5=180992b45226dc3ce0a38f89adfba410CAS |

Nelson KE, Zinder SH, Hance I, Burr P, Odongo D, Wasawo D, Odenyo A, Bishop R (2003) Phylogenetic analysis of the microbial populations in the wild herbivore gastrointestinal tract: insights into an unexplored niche. Environmental Microbiology 5, 1212–1220.
Phylogenetic analysis of the microbial populations in the wild herbivore gastrointestinal tract: insights into an unexplored niche.Crossref | GoogleScholarGoogle Scholar |

Newbold CJ, Lopez S, Nelson N, Ouda JO, Wallace RJ, Moss AJ (2005) Propionate precursors and other metabolic intermediates as possible alternative electron acceptors to methanogenesis in ruminal fermentation in vitro. The British Journal of Nutrition 94, 27–35.
Propionate precursors and other metabolic intermediates as possible alternative electron acceptors to methanogenesis in ruminal fermentation in vitro.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BD2MXhtFantr7J&md5=6b59949daabc77f3f3b26047dfc7a0e1CAS |

Odenyo AA, Bishop R, Asefa G, Jamnadass R, Odongo D, Osuji P (2001) Characterization of tannin-tolerant bacterial isolates from East African ruminants. Anaerobe 7, 5–15.
Characterization of tannin-tolerant bacterial isolates from East African ruminants.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BD3MXhslyit70%3D&md5=c1567bd8130ca7f65552b4f33c566deaCAS |

Prates A, de Oliveira JA, Abecia L, Fondevila M (2010) Effects of preservation procedures of rumen inoculum on in vitro microbial diversity and fermentation. Animal Feed Science and Technology 155, 186–193.
Effects of preservation procedures of rumen inoculum on in vitro microbial diversity and fermentation.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BC3cXhtVGqtbs%3D&md5=d642ac2c5c8d2f1c5c8dea0a96ab1442CAS |

Rees GN, Baldwin DS, Watson GO, Perryman S, Nielsen DL (2004) Ordination and significance testing of microbial community composition derived from terminal restriction fragment length polymorphisms: application of multivariate statistics. Antonie van Leeuwenhoek 86, 339–347.
Ordination and significance testing of microbial community composition derived from terminal restriction fragment length polymorphisms: application of multivariate statistics.Crossref | GoogleScholarGoogle Scholar |

Rodríguez R, Mota M, Castrillo C, Fondevila M (2010) In vitro rumen fermentation of the tropical grass Pennisetum purpureum and mixtures with browse legumes: effects of tannin contents. Journal of Animal Physiology and Animal Nutrition 94, 696–705.
In vitro rumen fermentation of the tropical grass Pennisetum purpureum and mixtures with browse legumes: effects of tannin contents.Crossref | GoogleScholarGoogle Scholar |

Sanabria C, Barahona R, Rodriguez DA, Martin E, Rodriguez F (2007) Effect of tanniniferous forages Leucaena and Desmodium on rumen microbial populations. In ‘Tropical grasses and legumes: optimizing genetic diversity for multipurpose use: 2006 Annual Report of Project IP-5’ pp. 7–8. (Centro Internacional de Agricultura Tropical: Cali, Colombia)

Smith AH, Zoetendal E, Mackie RI (2005) Bacterial mechanisms to overcome inhibitory effects of dietary tannins. Microbial Ecology 50, 197–205.
Bacterial mechanisms to overcome inhibitory effects of dietary tannins.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BD2MXht1Gru7vO&md5=7ac12d953aa3f6bdf97eb4024a7a185bCAS |

Stewart CS, Flint HJ, Bryant MP (1997) The rumen bacteria. In ‘The rumen microbial ecosystem’. (Eds PN Hobson, CS Stewart) pp. 10–72. (Chapman & Hall: London)

Tiemann TT, Avila P, Ramírez G, Lascano CE, Kreuzer M, Hess HD (2008) In vitro ruminal fermentation of tanniniferous tropical plants: plant-specific tannin effects and counteracting efficiency of PEG. Animal Feed Science and Technology 146, 222–241.
In vitro ruminal fermentation of tanniniferous tropical plants: plant-specific tannin effects and counteracting efficiency of PEG.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BD1cXhtFCku7fK&md5=3a5ca9900308213e36c37af17b83b5d7CAS |

Wang Y, Alxander TW, McAllister TA (2009) In vitro effects of phlorotannins from Ascophyllum nodosum (brown seaweed) on rumen bacterial populations and fermentation. Journal of the Science of Food and Agriculture 89, 2252–2260.
In vitro effects of phlorotannins from Ascophyllum nodosum (brown seaweed) on rumen bacterial populations and fermentation.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BD1MXhtV2htbjI&md5=f61d6be541ea9310959cd1847466a83cCAS |