Limited work has previously been done using classical microbiology to identify organisms found in the rumen of moose . One male moose from Alaska was shot in August of 1985, and bacteria which were isolated and characterized consisted of Streptococcus bovis (21 strains), Butyrivibrio fibrisolvens (9 strains), Lachnospira multiparus (7 strains), and Selenomonas ruminantium (2 strains) . For the present study, the second generation (G2) PhyloChip (PhyloTech Inc., California) was used to survey rumen and colon samples for the presence and presumptive identification of bacteria. The G2 PhyloChip uses 16S rRNA gene sequences to rapidly type bacteria
and methanogens in a mixed microbial sample without the use of cloning or sequencing [15, 16]. The PhyloChip contains approximately 500,000 probes on its surface, representing over 8,400 species of bacteria and roughly 300 species of archaea . There Selleckchem GDC 973 PI3K inhibitor are 11, 25mer, probes that are designed to hybridize to each specific taxon, allowing for specificity in determining taxa present . Depending on what the probes are designed to target, the PhyloChip can be used to differentiate between different serotypes of Escherichia coli, or determine the presence of a species regardless of strain. It is already a popular bacterial screening method for air , water , and soil [19, 20], and has recently gained favor for digestive tract
samples [21, 22]. Due to their specificity and sensitivity, DNA microarrays have also been used to categorize diseased and CHIR-99021 datasheet healthy states [22, 23]. The major objectives of the present study were to type the bacteria present in rumen and colonic samples, and to compare these findings with other studies of ruminants and herbivores. Given that moose are large browsing herbivores , it was hypothesized that the bacterial populations in the browse-fed wild moose would be more closely related to bacterial populations
found in other browse/forage fed animals. This study reports on the bacteria found in the rumen and colon of the North American moose, as well as how these environments relate to other studies of the gut microbiome in various species. Results Quantitative Real-Time PCR Mean bacteria cell densities were calculated for each HSP90 rumen sample using standard curves generated by Bio-Rad’s CFX96 software. Based on a regression line created using the bacterial standards (R2 = 0.997), estimated cell density ranged from 8.46 × 1011 to 2.77 × 1012 copies of 16S rRNA/g in the rumen (Table 1). Table 1 Estimated densities (16S rRNA copy numbers per gram wet weight) of bacteria in the rumen (R) of the moose in October, 2010, Vermont Sample Bacterial copies of 16S rRNA/g (SEM) 1R 8.46 x 1011 2R 1.61 x 1012 3R 2.57 x 1012 4R 2.02 x 1012 5R 9.36 x 1011 6R 1.21 x 1012 7R 2.77 x 1012 8R 1.34 x 1012 Mean (SEM) 1.66 x 1012 (7.