Rumen bacterial community evaluated by 454 pyrosequencing and terminal restriction fragment length polymorphism analyses in dairy sheep fed marine algae.
Developing novel strategies to increase the content of bioactive unsaturated fatty acids (FA) in ruminant-derived products requires a deeper understanding of rumen biohydrogenation and bacteria involved in this process. Although high-throughput pyrosequencing may allow for a great coverage of bacterial diversity, it has hardly been used to investigate the microbiology of ruminal FA metabolism. In this experiment, 454 pyrosequencing and a molecular fingerprinting technique (terminal restriction fragment length polymorphism; T-RFLP) were used concurrently to assess the effect of diet supplementation with marine algae (MA) on the rumen bacterial community of dairy sheep. Eleven lactating ewes were divided in 2 lots and offered a total mixed ration based on alfalfa hay and concentrate (40:60), supplemented with 0 (control) or 8 (MA) g of MA/kg of dry matter. After 54 d on treatments, animals were slaughtered and samples of rumen content and fluid were collected separately for microbial analysis. Pyrosequencing yielded a greater coverage of bacterial diversity than T-RFLP and allowed the identification of low abundant populations. Conversely, both molecular approaches pointed to similar conclusions and showed that relevant changes due to MA addition were observed within the major ruminal phyla, namely Bacteroidetes, Firmicutes, and Proteobacteria. Decreases in the abundance of unclassified Bacteroidales, Porphyromonadaceae, and Ruminococcaceae and increases in as-yet uncultured species of the family Succinivibrionaceae, might be related to a potential role of these groups in different pathways of rumen FA metabolism. Diet supplementation with MA, however, had no effect on the relative abundance of Butyrivibrio and Pseudobutyrivibrio genera. In addition, results from both 454 pyrosequencing and T-RFLP indicate that the effect of MA was rather consistent in rumen content or fluid samples, despite inherent differences between these fractions in their bacterial composition.
Instituto de Ganadería de Montaña, 24346 Grulleros, León, Spain; Institute of Biological, Environmental and Rural Sciences, Aberystwyth University, Aberystwyth SY23 3DA, United Kingdom.
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Castro-Carrera
Instituto de Ganadería de Montaña, 24346 Grulleros, León, Spain.
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Toral
Instituto de Ganadería de Montaña, 24346 Grulleros, León, Spain.
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Frutos
Institute of Biological, Environmental and Rural Sciences, Aberystwyth University, Aberystwyth SY23 3DA, United Kingdom.
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McEwan
Instituto de Ganadería de Montaña, 24346 Grulleros, León, Spain.
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Hervás
Instituto de Nutrición Animal, Estación Experimental del Zaidín, 18008 Granada, Spain.
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Abecia
Institute of Biological, Environmental and Rural Sciences, Aberystwyth University, Aberystwyth SY23 3DA, United Kingdom.
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Pinloche
Institute of Biological, Environmental and Rural Sciences, Aberystwyth University, Aberystwyth SY23 3DA, United Kingdom.
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Girdwood
Instituto de Ganadería de Montaña, 24346 Grulleros, León, Spain. Electronic address: a.belenguer@csic.es.
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Belenguer
D000821:Animal Feed / Q000032:analysis*; D000824:Animal Nutritional Physiological Phenomena; D000818:Animals; D059001:Aquatic Organisms / Q000737:chemistry* / Q000378:metabolism; D001419:Bacteria / Q000145:classification / Q000235:genetics; D018407:Bacterial Physiological Phenomena; D003612:Dairying; D004032:Diet / Q000662:veterinary; D004041:Dietary Fats / Q000008:administration & dosage; D019587:Dietary Supplements / Q000032:analysis; D005260:Female; D005766:Gastrointestinal Contents / Q000382:microbiology; D007774:Lactation; D050356:Lipid Metabolism; D064307:Microbiota / Q000187:drug effects; D010944:Plants / Q000737:chemistry* / Q000378:metabolism; D012150:Polymorphism, Restriction Fragment Length; D011897:Random Allocation; D012417:Rumen / Q000382:microbiology*; D034561:Sheep, Domestic / Q000382:microbiology* / Q000502:physiology*
D004041:Dietary Fats
16S rDNA; ewe; lipid; ruminal microbiota