Research Article
Gastrointestinal Bacterial and Methanogenic
Archaea Diversity Dynamics Associated with
Condensed Tannin-Containing Pine Bark Diet in
Goats Using 16S rDNA Amplicon Pyrosequencing
Byeng R. Min,
1
Sandra Solaiman,
1
Raymon Shange,
1
and Jong-Su Eun
2
1
Department of Agricultural and Environmental Sciences, Tuskegee University, Tuskegee, AL, USA
2
Department of Animal, Dairy, and Veterinary Sciences, Utah State University, Logan, UT, USA
Correspondence should be addressed to Byeng R. Min; minb@mytu.tuskegee.edu
Received 16 July 2013; Revised 4 September 2013; Accepted 4 September 2013; Published 2 January 2014
Academic Editor: Isabel S´ a-Correia
Copyright © 2014 Byeng R. Min et al. is is an open access article distributed under the Creative Commons Attribution License,
which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
Eighteen Kiko-cross meat goats (=6) were used to collect gastrointestinal (GI) bacteria and methanogenic archaea for diversity
measures when fed condensed tannin-containing pine bark (PB). ree dietary treatments were tested: control diet (0% PB and 30%
wheat straw (WS); 0.17% condensed tannins (CT) dry matter (DM)); 15% PB and 15% WS (1.6% CT DM), and 30% PB and 0% WS
(3.2% CT DM). A 16S rDNA bacterial tag-encoded FLX amplicon pyrosequencing technique was used to characterize and elucidate
changes in GI bacteria and methanogenic archaea diversity among the diets. Proteobacteria was the most dominant phylum in goats
with mean relative abundance values ranging from 39.7 (30% PB) to 46.5% (control) and 47.1% (15% PB). Other phyla individually
accounted for fewer than 25% of the relative abundance observed. Predominant methanogens were Methanobrevibacter (75, 72,
and 49%), Methanosphaera (3.3, 2.3, and 3.4%), and Methanobacteriaceae (1.2, 0.6, and 0.7%) population in control, 15, and 30%
PB, respectively. Among methanogens, Methanobrevibacter was linearly decreased ( = 0.05) with increasing PB supplementation.
ese results indicate that feeding PB selectively altered bacteria and methanogenic archaeal populations in the GI tract of goats.
1. Introduction
Studies on gastrointestinal (GI) microorganisms have tradi-
tionally depended on the use of anaerobic cultivation tech-
niques [1] which only can detect an estimated 11% of the total
bacterial populations present in the rumen [2]. Molecular
methodologies developed over the past decade now enable
researchers to examine the diversity of the gut microflora
independent of cultural methods. Bacterial diversities within
the GI tract of humans [3, 4] and rumen of beef cattle
[5, 6] have been investigated in recent years as a result of
development of 16S rDNA-based analysis, yet similar data
on the microbiomes of bacteria and methanogenic archaea
diversity in the lower GI tract of meat goats are limited. In
addition, there is no clear information on the prevalence of
methanogenic and microbial populations in the hindgut of
animals other than human and swine.
Plant tannins (condensed (CT) and hydrolysable tannins)
are polyphenolic compounds of relatively high molecular
weight with the capacity to form complexes mainly with pro-
teins due to the presence of a large number of phenolic
hydroxyl groups [7]. ey are ubiquitously spread in nutri-
tionally important forage trees, shrubs, legumes, cereals, and
tree barks. e effects of tannins on ruminant production
have been extensively published in the past, and among them
beneficial effects of tannins on animal production system
have been much focused, with particular interest in their
positive effects on protein metabolism, prevention of frothy
bloat, and modification on rumen microbial population [7].
Microbial population changes in the gut when animals were
fed CT-containing diets were reported using a 16S PCR
technique in sheep [8] and rat [9]. In addition, the use of
rapid sequencing technologies combined with molecular
methods is becoming a prevalent standard for evaluating
Hindawi Publishing Corporation
International Journal of Microbiology
Volume 2014, Article ID 141909, 11 pages
http://dx.doi.org/10.1155/2014/141909