text:
A human gut microbial gene catalogue established by metagenomic sequencing.
ABSTRACT To understand the impact of gut microbes on human health and well-being it is crucial to assess their genetic potential. Here we describe the Illumina-based metagenomic sequencing, assembly and characterization of 3.3 million non-redundant microbial genes, derived from 576.7 gigabases of sequence, from faecal samples of 124 European individuals. The gene set, approximately 150 times larger than the human gene complement, contains an overwhelming majority of the prevalent (more frequent) microbial genes of the cohort and probably includes a large proportion of the prevalent human intestinal microbial genes. The genes are largely shared among individuals of the cohort. Over 99% of the genes are bacterial, indicating that the entire cohort harbours between 1,000 and 1,150 prevalent bacterial species and each individual at least 160 such species, which are also largely shared. We define and describe the minimal gut metagenome and the minimal gut bacterial genome in terms of functions present in all individuals and most bacteria, respectively.
- SourceAvailable from: sciencedirect.com[Show abstract] [Hide abstract]
ABSTRACT: As our understanding of host gut microbiome increases, research interests in the fields of prebiotics, probiotics and synbiotics are growing rapidly. Currently, the majority of prebiotics in the markets are derived from non-digestible oligosaccharides. Very few investigations have been focused on non-digestible long chain complex carbohydrates/polysaccharides for their potential as novel prebiotics. One of the reasons behind this is the unavailability of non-digestible polysaccharides that can fulfill the criteria as prebiotics. Beta-glucans that exist as non-digestible polysaccharides derived from different food sources have demonstrated not only health promoting effects, but also the potential as a novel source of prebiotics. This review commences with the current trends in the research fields of prebiotics, probiotics and synbiotics. This is followed by a discussion of the potential of long chain beta-glucans to serve as novel prebiotics based on current knowledge on their sources, preparation, fermentation characteristics, and the plausible mechanisms involved in their utilization. Some future research directions with emphasis on new approaches using molecular biology, genetics and genomics are also discussed.Bioactive Carbohydrates and Dietary Fibre. 07/2013; 2(1):45–64.
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ABSTRACT: Bacterial respiratory tract infections, mainly caused by Streptococcus pneumoniae, Haemophilus influenzae and Moraxella catarrhalis are among the leading causes of global mortality and morbidity. Increased resistance of these pathogens to existing antibiotics necessitates the search for novel targets to develop potent antimicrobials.Result: Here, we report a proof of concept study for the reliable identification of potential drug targets in these human respiratory pathogens by combining high-density transposon mutagenesis, high-throughput sequencing, and integrative genomics. Approximately 20% of all genes in these three species were essential for growth and viability, including 128 essential and conserved genes, part of 47 metabolic pathways. By comparing these essential genes to the human genome, and a database of genes from commensal human gut microbiota, we identified and excluded potential drug targets in respiratory tract pathogens that will have off-target effects in the host, or disrupt the natural host microbiota. We propose 249 potential drug targets, 67 of which are targets for 75 FDA-approved antimicrobials and 35 other researched small molecule inhibitors. Two out of four selected novel targets where experimentally validated, proofing the concept.BMC genomics. 11/2014; 15(1):958.
Conference Paper: Gene prediction in metagenomic fragments based on the SVM algorithm.[Show abstract] [Hide abstract]
ABSTRACT: Metagenomic sequencing is becoming a powerful method to explore various environmental organisms without isolation and cultivation. Genomic sequences data generated by this technology is growing explosively while numerous computational methods for analysis are still urgently in need. One of the first and most important processes is exhaustive gene prediction. As short and anonymous DNA fragments, assembly of metagenomic sequences usually has not a fixed end point to obtain complete genomes and moreover is often not available. This situation makes the annotation more complicated than in complete genomes. Here, we present a newly developed SVM-based algorithm which comprises a supervised universal model and a data-specific novel model. It utilizes entropy density profiles of codon usage, translation initiation signal scoring and open read frame length for model training. Tests on fixed-length artificial shotgun sequences of 700 bp showed a sensitivity of 94.7% and a specificity of 94.9% on average, which indicate that our method has the totally higher performance than the best of current gene prediction methods. Thousands of additional genes are predicted when applied to two metagenomic samples from human gut community. Furthermore, compared to other gene predictors, our algorithm predicts the most potential novel genes.4th International Conference on Biomedical Engineering and Informatics, BMEI 2011, Shanghai, China, October 15-17, 2011; 01/2011
Page 1
A human gut microbial gene catalogue
established by metagenomic sequencing
Junjie Qin1*, Ruiqiang Li1*, Jeroen Raes2,3, Manimozhiyan Arumugam2, Kristoffer Solvsten Burgdorf4,
Chaysavanh Manichanh5, Trine Nielsen4, Nicolas Pons6, Florence Levenez6, Takuji Yamada2, Daniel R. Mende2,
Junhua Li1,7, Junming Xu1, Shaochuan Li1, Dongfang Li1,8, JianjunCao1, Bo Wang1, Huiqing Liang1, Huisong Zheng1,
Yinlong Xie1,7, Julien Tap6, Patricia Lepage6, Marcelo Bertalan9, Jean-Michel Batto6, Torben Hansen4, Denis Le
Paslier10, Allan Linneberg11, H. Bjørn Nielsen9, Eric Pelletier10, Pierre Renault6, Thomas Sicheritz-Ponten9,
Keith Turner12, Hongmei Zhu1, Chang Yu1, Shengting Li1, Min Jian1, Yan Zhou1, Yingrui Li1, Xiuqing Zhang1,
Songgang Li1, Nan Qin1, Huanming Yang1, Jian Wang1, Søren Brunak9, Joel Dore ´6, Francisco Guarner5,
Karsten Kristiansen13, Oluf Pedersen4,14, Julian Parkhill12, Jean Weissenbach10, MetaHIT Consortium{, Peer Bork2,
S. Dusko Ehrlich6& Jun Wang1,13
Tounderstand theimpactofgutmicrobesonhumanhealthandwell-beingitiscrucialtoassesstheirgeneticpotential.Here
we describe the Illumina-based metagenomic sequencing, assembly and characterization of 3.3 million non-redundant
microbial genes, derived from 576.7 gigabases of sequence, from faecal samples of 124 European individuals. The gene set,
,150 times larger than the human gene complement, contains an overwhelming majority of the prevalent (more frequent)
genes are largely shared among individuals of the cohort. Over 99% of the genes are bacterial, indicating that the entire
also largely shared. We define and describe the minimal gut metagenome and the minimal gut bacterial genome in terms of
functions present in all individuals and most bacteria, respectively.
It has been estimated that the microbes in our bodies collectively
make up to 100 trillion cells, tenfold the number of human cells,
and suggested that they encode 100-fold more unique genes than
our own genome1. The majority of microbes reside in the gut, have
a profound influence on human physiology and nutrition, and are
energy harvest from food, and changes of gut microbiome may be
associated with bowel diseases or obesity4–8.
To understand and exploit the impact of the gut microbes on
human health and well-being it is necessary to decipher the content,
diversity and functioning of the microbial gut community. 16S ribo-
bacterial divisions, the Bacteroidetes and the Firmicutes, constitute
over 90% of the known phylogenetic categories and dominate the
distal gut microbiota10. Studies also showed substantial diversity of
difference is especially marked among infants12, later in life the gut
microbiome converges to more similar phyla.
Metagenomic sequencing represents a powerful alternative to
Applied to the human gut, such studies have already generated some
3gigabases (Gb) of microbial sequence from faecal samples of 33
individuals from the United States or Japan8,16,17. To get a broader
overview of the human gut microbial genes we used the Illumina
Genome Analyser (GA) technology to carry out deep sequencing of
576.7Gb of sequence, almost 200 times more than in all previous
studies, assembled it into contigs and predicted 3.3 million unique
open reading frames (ORFs). This gene catalogue contains virtually
all of the prevalent gut microbial genes in our cohort, provides a
broad view of the functions important for bacterial life in the gut
and indicates that many bacterial species are shared by different
individuals. Our results also show that short-read metagenomic
sequencing can be used for global characterization of the genetic
potential of ecologically complex environments.
Metagenomic sequencing of gut microbiomes
As part of the MetaHIT (Metagenomics of the Human Intestinal
Tract) project, we collected faecal specimens from 124 healthy, over-
weight and obese individual human adults, as well as inflammatory
tary Table 1). Total DNA was extracted from the faecal specimens18
and an average of 4.5Gb (ranging between 2 and 7.3Gb) of sequence
was generated for each sample, allowing us to capture most of the
*These authors contributed equally to this work.
{Lists of authors and affiliations appear at the end of the paper.
1BGI-Shenzhen, Shenzhen 518083, China.2European Molecular Biology Laboratory, 69117 Heidelberg, Germany.3VIB—Vrije Universiteit Brussel, 1050 Brussels, Belgium.4Hagedorn
Research Institute, DK 2820 Copenhagen, Denmark.5Hospital Universitari Val d'Hebron, Ciberehd, 08035 Barcelona, Spain.6Institut National de la Recherche Agronomique, 78350
Jouy en Josas, France.7School of Software Engineering, South China University of Technology, Guangzhou 510641, China.8Genome Research Institute, Shenzhen University Medical
School, Shenzhen 518000, China.9Center for Biological Sequence Analysis, Technical University of Denmark, DK-2800 Kongens Lyngby, Denmark.10Commissariat a ` l'Energie
Atomique, Genoscope, 91000 Evry, France.11Research Center for Prevention and Health, DK-2600 Glostrup, Denmark.12The Wellcome Trust Sanger Institute, Hinxton, Cambridge
CB101SA,UK.13DepartmentofBiology,UniversityofCopenhagen,DK-2200Copenhagen,Denmark.14Institute ofBiomedicalSciences,UniversityofCopenhagen&FacultyofHealth
Science, University of Aarhus, 8000 Aarhus, Denmark.
Vol 464|4 March 2010|doi:10.1038/nature08821
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Page 2
novelty (see Methods and Supplementary Table 2). In total, we
obtained 576.7Gb of sequence (Supplementary Table 3).
contigs, which could then be analysed and annotated by standard
designed for assembling very short reads, we performed de novo
assembly for all of the Illumina GA sequence data. Because a high
sample independently (Supplementary Fig. 3). As much as 42.7% of
the Illumina GA reads was assembled into a total of 6.58 million
contigs of a length .500bp, giving a total contig length of 10.3Gb,
with an N50 length of 2.2 kb (Supplementary Fig. 4) and the range of
any one sample could be mapped to contigs from other samples,
indicating the existence of a common sequence core.
the same samples (Supplementary Table 2). A total of 98.7% of the
of the mapped regions. This is comparable to the contigs that were
a control, of which 97.9% were collinear over 99.5% of the mapped
(Mb) of Illumina- and 454-based contigs, respectively (see Methods
and Supplementary Fig. 5), indicating that the short- and long-read-
based assemblies have comparable accuracies.
To complete the contig set we pooled the unassembled reads from
million additional contigs were thus generated, having a length of
370Mb and an N50 length of 939 bp. The total length of our final
contig set was thus 10.7Gb. Some 80% of the 576.7Gb of Illumina
GA sequence could be aligned to the contigs at a threshold of 90%
identity, allowing for accommodation of sequencing errors and
strain variability in the gut (Fig. 1), almost twice the 42.7% of
sequence that was assembled into contigs by SOAPdenovo, because
assembly uses more stringent criteria. This indicates that a vast
majority of the Illumina sequence is represented by our contigs.
To compare the representation of the human gut microbiome in
our contigs with that from previous work, we aligned them to the
reads from the two largest published gut metagenome studies
(1.83Gb of Roche/454 sequencing reads from 18 US adults8, and
0.79Gb of Sanger reads from 13 Japanese adults and infants17), using
the 90% identity threshold. A total of 70.1% and 85.9% of the reads
from the Japanese and US samples, respectively, could be aligned to
our contigs (Fig. 1), showing that the contigs include a high fraction
of sequences from previous studies. In contrast, 85.7% and 69.5% of
our contigs were not covered by the reads from the Japanese and US
samples, respectively, highlighting the novelty we captured.
Only 31.0–48.8% of the reads from the two previous studies and
the present study could be aligned to 194 public human gut bacterial
genomes (Supplementary Table 5), and 7.6–21.2% to the bacterial
genomes deposited in GenBank (Fig. 1). This indicates that the
reference gene set obtained by sequencing genomes of isolated bac-
terial strains is still of a limited scale.
A gene catalogue of the human gut microbiome
To establish a non-redundant human gut microbiome gene set we
first used the MetaGene20program to predict ORFs in our contigs
and found 14,048,045 ORFs longer than 100bp (Supplementary
Table 6). They occupied 86.7% of the contigs, comparable to the
value found for fully sequenced genomes (,86%). Two-thirds of
(N50 of 2.2kb). We next removed the redundant ORFs, by pair-wise
comparison, using a very stringent criterion of 95% identity over
90% of the shorter ORF length, which can fuse orthologues but
avoids inflation of the data set due to possible sequencing errors
(see Methods). Yet, the final non-redundant gene set contained as
many as 3,299,822 ORFs with an average length of 704bp (Sup-
plementary Table 7).
We term the genes of the non-redundant set 'prevalent genes', as
(see Methods). The minimal relative abundance of the prevalent
genes was ,631027, as estimated from the minimum sequence
coverage of the unique genes (close to 3), and the total Illumina
sequence length generated for each individual (on average, 4.5Gb),
assuming the average gene length of 0.85kb (that is, 330.853103/
We mapped the 3.3 million gut ORFs to the 319,812 genes (target
genes) of the 89 frequent reference microbial genomes in the human
gut. At a 90% identity threshold, 80% of the target genes had at least
80% of their length covered by a single gut ORF (Fig. 2b). This
indicates that the gene set includes most of the known human gut
bacterial genes.
We examined the number of prevalent genes identified across all
individuals as a function of the extent of sequencing, demanding at
least two supporting reads for a gene call (Fig. 2a). The incidence-
(the highest number the EstimateS21program could accommodate),
indicates that our catalogue captures 85.3% of the prevalent genes.
Although this is probably an underestimate, it nevertheless indicates
genes of the cohort.
genes (Supplementary Fig. 6b), indicating thatmostof the 3.3 million
gene pool must be shared. However, most of the prevalent genes were
found in only a few individuals: 2,375,655 were present in less than
20%, whereas 294,110 were found in at least 50% of individuals (we
term these 'common' genes). These values depend on the sampling
depth; sequencing of MH0006 and MH0012 revealed more of the
catalogue genes, present at a low abundance (Supplementary Fig. 7).
Nevertheless, even at our routine sampling depth, each individual
harboured 204,05663,603 (mean6s.e.m.) common genes, indi-
cating that about 38% of an individual's total gene pool is shared.
than the individuals not suffering from IBD (Supplementary Fig. 8),
consistent with the observation that the former have lower bacterial
diversity than the latter22.
Common bacterial core
Deep metagenomic sequencing provides the opportunity to explore
contig set
Known human
gut bacteria
Coverage of sequencing reads (%)
Figure 1 | Coverage of human gut microbiome. The three human microbial
sequencing read sets—Illumina GA reads generated from 124 individuals in
thisstudy (black;n5124),Roche/454readsfrom18 humantwins andtheir
mothers (grey; n518) and Sanger reads from 13 Japanese individuals
(white; n513)—were aligned to each of the reference sequence sets. Mean
values6 s.e.m. are plotted.
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Page 3
the cohort. For this purpose, we used a non-redundant set of 650
the Illumina GA reads of each human gut microbial sample onto the
genome set, using a 90% identity threshold, and determined the
proportion of the genomes covered by the reads that aligned onto
only a single position in the set. At a1% coverage, which for atypical
gut bacterial genome corresponds to an average length of about
40kb, some 25-fold more than that of the 16S gene generally used
for species identification, we detected 18 species in all individuals, 57
10% coverage, requiring ,10-fold higher abundance in a sample, we
still found 13 of the above species in$90% of individuals and 35
When the cumulated sequence length increased from 3.96Gb to
8.74Gb and from 4.41Gb to 11.6Gb, for samples MH0006 and
MH0012, respectively, the number of strains common to the two
at the 1% coverage threshold increased by 25%, from 135 to 169.
This indicates the existence of a significantly larger common core
than the one we could observe at the sequence depth routinely used
for each individual.
The variability of abundance of microbial species in individuals
can greatly affect identification of the common core. To visualize
to different genomes across the individuals of our cohort. Even for
the most common 57 species present in$90% of individuals with
variability was between 12- and 2,187-fold (Fig. 3). As expected10,23,
Bacteroidetes and Firmicutes had the highest abundance.
A complex pattern of species relatedness, characterized by clusters
at the genus and family levels, emerges from the analysis of the net-
work based on the pair-wise Pearson correlation coefficients of 155
species present in at least one individual at$1% coverage
abundant gut species, such as members of the Bacteroidetes and
Dorea/Eubacterium/Ruminococcus groups and also bifidobacteria,
Proteobacteria and streptococci/lactobacilli groups. These observa-
different individuals of our cohort, for reasons that remain to be
The above result indicates that the Illumina-based bacterial pro-
filing should reveal differences between the healthy individuals and
patients. To test this hypothesis we compared the IBD patients and
healthy controls (Supplementary Table 1), as it was previously
ponent analysis, based on the same 155 species, clearly separates
patients from healthy individuals and the ulcerative colitis from
the Crohn's disease patients (Fig. 4), confirming our hypothesis.
Functions encoded by the prevalent gene set
We classified the predicted genes by aligning them to the integrated
the KEGG (Kyoto Encyclopedia of Genes and Genomes)24pathways,
and COG (Clusters of Orthologous Groups)25and eggNOG26data-
bases. There were 77.1% genes classified into phylotypes, 57.5% to
eggNOG clusters, 47.0% to KEGG orthology and 18.7% genes
assigned to KEGG pathways, respectively (Supplementary Table 9).
Relative abundance (log10)
Blautia hansenii
Clostridium scindens
Enterococcus faecalis TX0104
Clostridium asparagiforme
Bacteroides fragilis 3_1_12
Bacteroides intestinalis
Ruminococcus gnavus
Anaerotruncus colihominis
Bacteroides pectinophilus
Clostridium nexile
Clostridium sp. L2−50
Parabacteroides johnsonii
Bacteroides finegoldii
Butyrivibrio crossotus
Bacteroides eggerthii
Clostridium sp. M62 1
Coprococcus eutactus
Bacteroides stercoris
Holdemania filiformis
Clostridium leptum
Streptococcus thermophilus LMD−9
Bacteroides capillosus
Subdoligranulum variabile
Ruminococcus obeum A2−162
Bacteroides dorei
Eubacterium ventriosum
Bacteroides sp. D4
Bacteroides sp. D1
Coprococcus comes SL7 1
Bacteriodes xylanisolvens XB1A
Eubacterium rectale M104 1
Bacteroides sp. 2_2_4
Bacteroides sp. 4_3_47FAA
Bacteroides ovatus
Bacteroides sp. 9_1_42FAA
Parabacteroides distasonis ATCC 8503
Eubacterium siraeum 70 3
Bacteroides sp. 2_1_7
Roseburia intestinalis M50 1
Bacteroides vulgatus ATCC 8482
Dorea formicigenerans
Collinsella aerofaciens
Ruminococcus lactaris
Faecalibacterium prausnitzii SL3 3
Ruminococcus sp. SR1 5
Unknown sp. SS3 4
Ruminococcus torques L2−14
Eubacterium hallii
Bacteroides thetaiotaomicron VPI−5482
Clostridium sp. SS2−1
Bacteroides caccae
Ruminococcus bromii L2−63
Dorea longicatena
Parabacteroides merdae
Alistipes putredinis
Bacteroides uniformis
–4–3 –2–1
Figure 3 | Relative abundance of 57 frequent microbial genomes among
individuals of the cohort. See Fig. 2c for definition of box and whisker plot.
See Methods for computation.
Number of individuals sampled
Number of orthologous groups/gene families (×103)
255075100 124
Number of non-redundant
280, 000
1.00.8 0.60.4 0.20
1 2040 6080
OGs + novel gene families
Known + unknown OGs
Known OGs
genes (×106)
Number of target genes covered
Fraction of target genes
Number of samplesFraction of gene length covered
Figure 2 | Predicted ORFs in the human gut microbiome. a, Number of
curve corresponds to the Sobs(Mao Tau) values (number of observed genes),
calculated using EstimateS21(version 8.2.0) onrandomlychosen 100 samples
(due to memory limitation). b, Coverage of genes from 89 frequent gut
by number of samples investigated, based on known (well characterized)
orthologous groups (OGs; bottom), known plus unknown orthologous
groups (including, for example, putative, predicted, conserved hypothetical
functions; middle) and orthologous groups plus novel gene families (.20
proteins) recovered from the metagenome (top). Boxes denote the
interquartilerange (IQR) betweenthe first and thirdquartiles (25thand 75th
percentiles, respectively) and the line inside denotes the median. Whiskers
denote the lowest and highest values within 1.5 times IQR from the first and
third quartiles, respectively. Circles denote outliers beyond the whiskers.
NATURE|Vol 464|4 March 2010
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Page 4
Almost all (99.96%) of the phylogenetically assigned genes belonged
Genes that were not mapped to orthologous groups were clustered
into gene families (see Methods). To investigate the functional con-
tent of the prevalent gene set we computed the total number of
of n individuals (with n52–124; see Fig. 2c). This rarefaction ana-
lysis shows that the 'known' functions (annotated in eggNOG or
sampling any subset of 50 individuals, most have been detected.
However, three-quarters of the prevalent gut functionalities consists
only starts to plateau at the very end, at a much higher level (19,338
groups were detected), confirming that the extensive sampling of a
amount of novel/unknown functionality.
Bacterial functions important for life in the gut
The extensive non-redundant catalogue of the bacterial genes from
the human intestinal tract provides an opportunity to identify bac-
terial functions important for life in this environment. There are
functions necessary for a bacterium to thrive in a gut context (that
is, the 'minimal gut genome') and those involved in the homeostasis
of the whole ecosystem, encoded across many species (the 'minimal
gut metagenome'). Thefirst setof functions is expected to be present
in most or all gut bacterial species; the second set in most or all
individuals' gut samples.
To identify the functions encoded by the minimal gut genome we
use the fact that they should be present in most or all gut bacterial
species and therefore appear in the gene catalogue at a frequency
above that of the functions present in only some of the gut bacterial
species. The relative frequency of different functions can be deduced
from the number of genes recruited to different eggNOG clusters,
after normalization for gene length and copy number (Supplemen-
tary Fig. 10a, b). We ranked all the clusters by gene frequencies and
determined the range that included the clusters specifying well-
mentally for a well-studied firmicute, Bacillus subtilis27, hypothe-
sizing that additional clusters in this range are equally important.
As expected, the range that included most of B. subtilis essential
clusters (86%) was at the very top of the ranking order (Fig. 5).
Some 76% of the clusters with essential genes of Escherichia coli28
were within this range, confirming the validity of our approach.
(Supplementary Table 10; termed 'range clusters' hereafter) specify
functions important for life in the gut.
We found two types of functions among the range clusters: those
required in all bacteria (housekeeping) and those potentially specific
for the gut. Among many examples of the first category are the
functions that are part of main metabolic pathways (for example,
central carbon metabolism, amino acid synthesis), and important
on the KEGG metabolic pathways gives a highly integrated picture of
the global gut cell metabolism (Fig. 6a).
sion to the host proteins (collagen, fibrinogen, fibronectin) or in
harvesting sugars of the globoseries glycolipids, which are carried
on blood and epithelial cells. Furthermore, 15% of range clusters
encode functions that are present in ,10% of the eggNOG genomes
(see Supplementary Fig. 11) and are largely (74.3%) not defined
(Fig. 6b). Detailed studies of these should lead to a deeper compre-
hension of bacterial life in the gut.
To identifythe functionsencoded by theminimal gutmetagenome,
wecomputed theorthologousgroupsthat are shared byindividuals of
one estimated in a previous study8. There are only 2,069 functionally
annotated orthologous groups, showing that they gravely underesti-
fraction of functions (,45%) that are present in ,10% of the
for the gut ecosystem. Eighty per cent of these orthologous groups
containgeneswithatbestpoorly characterizedfunction,underscoring
our limited knowledge of gut functioning.
Of the known fraction, about 5% codes for (pro)phage-related
proteins, implying a universal presence and possible important eco-
logical role of bacteriophages in gut homeostasis. The most striking
secondary metabolism that seems crucial for the minimal metage-
and glycans harvested from the host diet and/or intestinal lining.
Examples include degradation and uptake pathways for pectin
(and its monomer, rhamnose) and sorbitol, sugars which are omni-
as a non-competitive source of energy. Besides these, capacity to
part of the minimal metagenome. Together, these emphasize the
Cluster (%)
1 2,001 4,001 6,0018,00110,001
Cluster rank
Figure 5 | ClustersthatcontaintheB. subtilisessential genes. The clusters
were ranked by the number of genes they contain, normalized by average
lengthandcopy number(seeSupplementaryFig. 10), andtheproportionof
clusters with the essential B. subtilis genes was determined for successive
groups of 100 clusters. Range indicates the part of the cluster distribution
that contains 86% of the B. subtilis essential genes.
Crohn's disease
Ulcerative colitis
P value: 0.031
Figure 4 | Bacterial species abundance differentiates IBD patients and
healthy individuals. Principal component analysis with health status as
instrumental variables, based on the abundance of 155 species with $1%
genome coverage by the Illumina reads in at least 1 individual of the cohort,
colitisand4Crohn's disease)fromSpain(SupplementaryTable1). Twofirst
components (PC1 and PC2) were plotted and represented 7.3% of whole
inertia. Individuals (represented by points) were clustered and centre of
species abundance was assessed using a Monte-Carlo test (999 replicates).
NATURE|Vol 464|4 March 2010
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Page 5
strong dependence of the gut ecosystem on complex sugar degrada-
tion for its functioning.
Functional complementarities of the genome and metagenome
Detailed analysis of the complementarities between the gut metage-
functions in the minimal metagenome and presence/absence of func-
tions in one or the other (Supplementary Table 11). Gut bacteria use
mostly fermentation to generate energy, converting sugars, in part, to
is important for muscle, heart and brain cells31, propionate is used in
hosthepatic neoglucogenic processes,whereas,in addition,butyrate is
amino acids are indispensable to humans33and can be provided by
bacteria34. Similarly, bacteria can contribute certain vitamins3(for
example, biotin,phylloquinone) to thehost. All of the stepsof biosyn-
thesis of these molecules are encoded by the minimal metagenome.
Gut bacteria seem to be able to degrade numerous xenobiotics,
plementary Table 11), even if the steps of most pathways are not part
only. A particularly interestingexample is that of benzoate, which is a
common food supplement, known as E211. Its degradation by the
coenzyme-A ligation pathway, encoded in the minimal metagenome,
leads to pimeloyl-coenzyme-A, which is a precursor of biotin, indi-
catingthat this foodsupplement canhave a potentially beneficialrole
for human health.
Number of individuals sampled
Minimal metagenome size
1 25 5075
Limonene and pinene
biosynthesis II
Penicillins and cephalosporins
and metabolism
Glycan biosynthesis
Amino acid
Biodegradation of
Metabolism of
other amino acids
Metabolism of
cofactors and vitamins
Biosynthesis of
secondary metabolites
0.0% 10.0% 20.0% 30.0% 40.0% 50.0% 60.0% 70.0%
General function - R
Translation - J
Amino acids - E
Unknown - S
Envelope - M
Carbohydrates - G
Energy - C
Transcription - K
Coenzymes - H
Nucleotides - F
Inorganic - P
Protein turnover - O
Lipids - I
Signal transduction - T
Secretion - U
Cell cycle - D
Defence- V
Second metabolites - Q
Cell motility - N
Chromatin - B
Extracellular - W
Nuclear structure - Y
Cytoskeleton - Z
Rare minimal genome
Rare minimal metagenome
Frequent minimal genome
Frequent minimal metagenome
Figure 6 | Characterization of the minimal gut genome and metagenome.
a, Projection of the minimal gut genome on the KEGG pathways using the
iPath tool38. b, Functional composition of the minimal gut genome and
metagenome.Rare and frequentreferto thepresence in sequencedeggNOG
genomes. c, Estimation of the minimal gut metagenome size. Known
orthologous groups (red), known plus unknown orthologous groups (blue)
and orthologous groups plus novel gene families (.20 proteins; grey) are
shown (see Fig. 2c for definition of box and whisker plot). The inset shows
composition of the gut minimal microbiome. Large circle: classification in
the minimal metagenome according to orthologous group occurrence in
(45%) refer to functions that are present in .50%, ,50% but .10%, and
,10% of STRING bacteria genomes, respectively.Small circle: composition
of the rare orthologous groups. Unknown (80%) have no annotation or are
poorly characterized, whereas known bacterial (19%) and phage-related
(1%) orthologous groups have functional description.
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We have used extensive Illumina GA short-read-based sequencing of
total faecal DNA from a cohort of 124 individuals of European
(Nordic and Mediterranean) origin to establish a catalogue of non-
redundant humanintestinal microbialgenes. Thecatalogue contains
3.3 million microbial genes, 150-fold more than the human gene
complement, and includes an overwhelming majority (.86%) of
prevalent genes harboured by our cohort. The catalogue probably
human population, for the following reasons: (1) over 70% of the
metagenomic reads from three previous studies, including American
and Japanese individuals8,16,17, can be mapped on our contigs; (2)
about 80% of the microbial genes from 89 frequent gut reference
genomes are present in our set. This result represents a proof of
principle that short-read sequencing can be used to characterize
complex microbiomes.
The full bacterial gene complement of each individual was not
sampled in our work. Nevertheless, we have detected some 536,000
prevalent unique genes in each, out of the total of 3.3 million carried
by our cohort. Inevitably, the individuals largely share the genes of
thecommonpool.Atthepresentdepthofsequencing,wefound that
almost40% ofthegenes from each individual are shared withatleast
half of the individuals of the cohort. Future studies of world-wide
span, envisaged within the International Human Microbiome
Consortium, will complete, as necessary, our gene catalogue and
establish boundaries to the proportion of shared genes.
yotic and viral origins. The gene catalogue is therefore equivalent to
that of some 1,000 bacterial species with an average-sized genome,
encoding about 3,364 non-redundant genes. We estimate that no
more than 15% of prevalent genes of our cohort may be missing
from the catalogue, and suggest that the cohort harbours no more
sampling. Given the large overlap between microbial sequences in
this and previous studies we suggest that the number of abundant
intestinal bacterial species may be not much higher than that
observed in our cohort. Each individual of our cohort harbours at
least 160 such bacterial species, as estimated by the average prevalent
gene number, and many must thus be shared.
We assigned about 12% of the reference set genes (404,000) to the
194 sequenced intestinal bacterial genomes, and can thus associate
them with bacterial species. Sequencing of at least 1,000 human-
associated bacterial genomes is foreseen within the International
Human Microbiome Consortium, via the Human Microbiome
Project and MetaHIT. This is commensurate with the number of
ment. Nevertheless, we used the presently available sequenced
genomes to explore further the concept of largely shared species
among our cohort and identified 75 species common to .50% of
individuals and 57 species common to .90%. These numbers are
likely to increase with the number of sequenced reference strains and
a deeper sampling. Indeed, a 2–3-fold increase in sequencing depth
raised by 25% the number of species that we could detect as shared
between two individuals. A large number of shared species supports
the view that the prevalent human microbiome is of a finite and not
overly large size.
How can this view be reconciled with that of a considerable inter-
personal diversity of innumerable bacterial species in the gut, arising
from most previous studies using the 16S RNA marker gene4,8,10,11?
Possibly the depth of sampling of these studies was insufficient to
reveal common species when present at low abundance, and empha-
sized the difference in the composition of a relatively few dominant
species. We found a very highvariability of abundance (12- to 2,200-
fold) for the 57 most common species across the individuals of our
cohort. Nevertheless, a recent 16S rRNA-based study concluded that
a common bacterial species 'core', shared among at least 50% of
individuals under study, exists35.
Detailed comparisons of bacterial genes across the individuals of
our cohort will be carried out in the future, within the context of
the ongoing MetaHIT clinical studies of which they are part.
Nevertheless, clustering ofthe genes infamilies allowedus tocapture
a virtually full functional potential of the prevalent gene set and
revealed a considerable novelty, extending the functional categories
by some 30% in regard to previous work8. Similarly, this analysis has
which reflects the full minimal human gut metagenome, encoded
of the gut ecosystem. The size of this minimal metagenome exceeds
several-fold that of the core metagenome reported previously8. It
action, such as degradation of complex polysaccharides, synthesis of
short-chain fatty acids, indispensable amino acids and vitamins.
Finally, we also identified functions that we attribute to a minimal
gut bacterial genome, likely to be required by any bacterium to thrive
in this ecosystem. Besides general housekeeping functions, the
in sequenced genomes and possibly specifically required in the gut.
Beyond providing the global view of the human gut microbiome,
of association of the microbial genes with human phenotypes and,
even more broadly, human living habits, taking into account the
these studies will lead to a much more complete understanding of
human biology than the one we presently have.
by standard methods22. For all 124 individuals, paired-end libraries were con-
structed with different clone insert sizes and subjected to Illumina GA sequen-
cing. All reads were assembled using SOAPdenovo19, with specific parameter
'2M 3' for metagenomics data. MetaGene was used for gene prediction. A
non-redundant gene set was constructed by pair-wise comparison of all genes,
using BLAT36under the criteria of identity .95% and overlap .90%. Gene
taxonomic assignments were made on the basis of BLASTP37search (e-value
,131025) of the NCBI-NR database and 126 known gut bacteria genomes.
with eggNOG and KEGG (v48.2) databases. The total and shared number of
orthologous groups and/or gene families were computed using a random com-
bination of n individuals (with n52 to 124, 100 replicates per bin).
Full Methods and any associated references are available in the online version of
the paper at www.nature.com/nature.
Received 14 August; accepted 23 December 2009.
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Supplementary Information is linked to the online version of the paper at
Acknowledgements We are indebted to the faculty and staff of Beijing Genomics
Institute at Shenzhen, whose names were not included in the author list, but who
contributed to large-scale sequencing of this team work. The research leading to
these results has received funding from the European Community's Seventh
Framework Programme (FP7/2007-2013): MetaHIT, grant agreement
HEALTH-F4-2007-201052, the Ole Rømer grant from the Danish Natural Science
Research Council, the Solexa project (272-07-0196), the Shenzhen Municipal
Government of China, the National Natural Science Foundation of China
(30725008), the International Science and Technology Cooperation Project
(0806), China (CXB200903110066A; ZYC200903240076A), the Danish
Strategic Research Council grant no 2106-07-0021 (Seqnet), and the Lundbeck
Foundation Centre for Applied Medical Genomics in Personalised Disease
Prediction, Prevention and Care. Ciberehd is funded by Instituto de Salud Carlos III
(Spain). We also thank X. Wang from the School of Biosciences and
Bioengineering, South China University of Technology, for his coordination on the
Innovative Program for Undergraduate Students in which J.L. and Y.X. joined.
Author Contributions All authors are members of the Metagenomics of the
Human Intestinal Tract (MetaHIT) Consortium. So.L., H.Y., Je.W., J.D., F.G., K.K.,
O.P., S.B., J.P., Ji.W., S.D.E. and Ju.W. managed the project. T.N., T.H. and K.S.B.
performed clinical analyses; F.L. and C.M. performed DNA extraction. X.Z., B.W.,
J.C., H.L., Hu.Z., K.T., D.L.P., E.P. and M.J. performed sequencing. Ju.W., S.D.E, P.B.,
M.B., H.B.N., T.S.-P., C.Y., She.L., T.Y., N.P., J.-M.B., P.L., D.R.M., S.D.E. and Y.Z.
performed the data analyses. S.D.E., P.B., J.R., J.Q., R.L. and Ju.W. wrote the paper.
J.T., A.L., P.R., Y.L.and N.Q. revised the paper. The MetaHIT Consortium members
contributed to design and execution of the study.
Author Information The raw Illumina read data of all 124 samples has been
available to download from the EMBL (http://www.bork.embl.de/,arumugam/
Qin_et_al_2010/) and BGI (http://gutmeta.genomics.org.cn) websites. Reprints
declare no competing financial interests. This paper is distributed under the terms
freely available to all readers at www.nature.com/nature. Correspondence and
requests for materials should be addressed to Ju.W. (email@example.com) or
S.D.E. (firstname.lastname@example.com).
MetaHIT Consortium (additional members)
Maria Antolin1, Franc ¸ois Artiguenave2, Herve ´ Blottiere3, Natalia Borruel1, Thomas
Bruls2, Francesc Casellas1, Christian Chervaux4, Antonella Cultrone3, Christine
Delorme3, Ge ´rard Denariaz4, Rozenn Dervyn3, Miguel Forte5, Carsten Friss6, Maarten
van de Guchte3, Eric Guedon3, Florence Haimet3, Alexandre Jamet3, Catherine Juste3,
Ghalia Kaci3, Michiel Kleerebezem7, Jan Knol4, Michel Kristensen8, Severine Layec3,
Karine Le Roux3, Marion Leclerc3, Emmanuelle Maguin3, Raquel Melo Minardi2, Raish
Varela1, Willem de Vos7, Yohanan Winogradsky3& Erwin Zoetendal7
1HospitalUniversitariVald'Hebron,Ciberehd,08035Barcelona,Spain.2Commissariat a `
l'Energie Atomique, Genoscope, 91000 Evry, France.3Institut National de la Recherche
5UCB Pharma SA, 28046 Madrid, Spain.6Center for Biological Sequence Analysis,
Technical University of Denmark, DK-2800 Kongens Lyngby, Denmark.7Wageningen
Unviersiteit, 6710BA Ede, The Netherlands.8Hagedorn Research Institute, DK 2820
Copenhagen, Denmark.9Istituto Europeo di Oncologia, 20100 Mila, Italy.
NATURE|Vol 464|4 March 2010
Macmillan Publishers Limited. All rights reserved
Page 8
Human faecal sample collection. Danish individuals were from the Inter-99
cohort40, varying in phenotypes according to BMI and status towards obesity/
diabetes, whereas Spanish individuals were either healthy controls or patients
with chronic inflammatory bowel diseases (Crohn's disease or ulcerative colitis)
in clinical remission.
Patients and healthy controls were asked to provide a frozen stool sample.
Fresh stool samples were obtained at home, and samples were immediately
frozen by storing them in their home freezer. Frozen samples were delivered
stored at 280uC until analysis.
in 250ml of guanidine thiocyanate, 0.1M Tris (pH 7.5) and 40ml of 10%
N-lauroyl sarcosine. Then, DNA extraction was conducted as previously
described22. The DNA concentration and its molecular size were estimated by
nanodrop (Thermo Scientific) and agarose gel electrophoresis.
DNA library construction and sequencing. DNA library preparation followed
the manufacturer's instruction (Illumina). We used the same workflow as
described elsewhere to perform cluster generation, template hybridization, iso-
thermal amplification, linearization, blocking and denaturization and hybridi-
zation of the sequencing primers. The base-calling pipeline (version
IlluminaPipeline-0.3) was used to process the raw fluorescent images and call
We constructed one library (clone insert size 200bp) for each of the first 15
To estimate the optimal return between the generation of novel sequence and
sequencingdepth, we alignedthe IlluminaGA reads from samples MH0006and
MH0012 onto 468,335 Sanger reads totalling to 311.7Mb generated from the
same two samples (156.9 and 154.7Mb, respectively, Supplementary Table 2),
using the Short Oligonucleotide Alignment Program (SOAP)41and a match
requirement of 95% sequence identity. With about 4Gb of Illumina sequence,
covered. Further extensive sequencing, to 12.6 and 16.6Gb for MH0006 and
MH0012, respectively, brought only a moderate increase of coverage to about
the Illumina sequences to a very high and uniform level (Supplementary Fig. 2),
a large proportion of Illumina sequences (57% and 74% for M0006 and M0012,
respectively) was novel and could not be mapped onto the Sanger reads. This
fraction was similar at the 4 and 12–16Gb sequencing levels, confirming that
most of the novelty was captured already at 4Gb.
We generated 35.4–97.6 million reads for the remaining 122 samples, with an
average of 62.5 million reads. Sequencing read length of the first batch of 15
samples was 44bp and the second batch was 75bp.
loaded from HMP database (http://www.hmpdacc-resources.org/cgi-bin/hmp_
catalog/main.cgi), GenBank (67 genomes), Washington University in St Louis (85
genomes, version April 2009, http://genome.wustl.edu/pub/organism/Microbes/
version September 2009, http://www.sanger.ac.uk/pathogens/metahit/). The other
sequenced from US individuals8, which was downloaded from NCBI with the
accession SRA002775; (2) human gut metagenomic data from Japanese indivi-
duals17, which was downloaded from P. Bork's group at EMBL (http://www.
bork.embl.de). The integrated NR database we constructed in this study included
NCBI-NR database (version April 2009) and all genes from the known human gut
bacteria genomes.
Illumina GA short reads de novo assembly. High-quality short reads of each
DNA sample were assembled by the SOAPdenovo assembler19. In brief, we first
filtered the low abundant sequences from the assembly according to 17-mer fre-
ing them would significantly reduce memory requirement and make assembly
feasible in an ordinary supercomputer (512GB memory in our institute).
Then the sequences were processed one by one and the de Bruijn graph data
format was used to store the overlap information among the sequences. The
overlap paths supported by a single read were unreliable and removed. Short
low-depth tips and bubbles that were caused by sequencing errors or genetic
variations between microbial strains were trimmed and merged, respectively.
Read paths were used to solve the tiny repeats.
Finally, we broke the connections at repeat boundaries, and outputted the
continuous sequences with unambiguous connections as contigs. The metage-
After de novo assembly for each sample independently, we merged all the
unassembled reads together and performed assembly for them, as to maximize
of all samples together.
Validating Illumina contigs using Sanger reads. We used BLASTN (WU-
BLAST 2.0) to map Sanger reads from samples MH0006 and MH0012
(156.9Mb and154.7Mb,respectively)toIlluminacontigs(singlebest hitlonger
than 75bp and over 95% identity) from the same samples. Each alignment was
scanned for breakage of collinearity where both sequences have at least 50 bases
assembly error in the Illumina contig at the location where collinearity breaks.
exists a Sanger read that agrees with the contig structure for 60bp on both sides
of the error. For comparison, we repeated this on a Newbler2 assembly of 454
number of errors per Mb of assembled Illumina/454 contigs. We estimate 14.12
errors per Mb of contigs for the Illumina assembly, which is comparable to that
of the 454 assembly (20.73 per Mb). 98.7% of Illumina contigs that map at least
one Sanger read were collinear over 99.55% of the mapped regions, which is
comparable to 97.86% of such 454 contigs being collinear over 99.48% of the
mapped regions.
Evaluation of human gut microbiome coverage. The Illumina GA reads were
aligned against the assembled contigs and known bacteria genomes using
SOAP41by allowing at most two mismatches in the first 35-bp region and
90% identity over the read sequence. The Roche/454 and Sanger sequencing
reads were aligned against the same reference using BLASTN with 131028,
were allowed and identity was set 95% over the read sequence when aligned to
Gene prediction and construction of the non-redundant gene set. We use
MetaGene20—which uses di-codon frequencies estimated by the GC content
well as the contigs from the merged assembly.
The predicted ORFs were then aligned to each other using BLAT36. A pair of
genes with greater than 95% identity and aligned length covered over 90% of the
shorter gene was grouped together. The groups sharing genes were then merged,
non-redundant genesetfromallthepredictedgenesby excludingtheredundancy.
into protein sequences using the NCBI Genetic Codes11.
Identification of genes. To make a balance between identifying low-abundance
threshold set for read coverage required to identify a gene in individual micro-
biomes. The number of genes decreased about twice when the number of reads
required for identification was increased from 2 to 6, and changed slowly there-
after (Supplementary Fig. 6a). Nevertheless, to include the rare genes into the
analysis, we selected the threshold of 2 reads.
Gene taxonomic assignment. Taxonomic assignment of predicted genes was
carried out using BLASTP alignment against the integrated NR database.
BLASTP alignment hits with e-values larger than 131025were filtered, and for
taxonomical level of each gene by the lowest common ancestor (LCA)-based
algorithm that was implemented in MEGAN42. The LCA-based algorithm assigns
level of conservation of the gene. For example, if a gene was conserved in many
species, it was assigned to the LCA rather than to a species.
Gene functional classification. We used BLASTP to search the protein
with e-value#131025. The genes were annotated as the function of the NOGs
or KEGG homologues with lowest e-value. The eggNOG database is an integ-
ration of the COG and KOG databases. The genes annotated by COG were
classified into the 25 COG categories, and genes that were annotated by KEGG
were assigned into KEGG pathways.
Macmillan Publishers Limited. All rights reserved
Page 9
genes assignedto the eggNOG clusters was normalized by gene length and cluster
genes was determined, computing the proportion of these clusters among the
successive groups of 100 clusters. Analysis of the range gene clusters involved,
besides iPath projections, use of KEGG and manual verification of the complete-
ness of the pathways and protein machineries they encode.
Determination of total functional complement and minimal metagenome.
We computed the total and shared number of orthologous groups and/or gene
(1) known eggNOG orthologous groups (that is, those with functional annota-
tion, excluding those in which the terms [Uu]ncharacteri[sz]ed, [Uu]nknown,
[Pp]redicted or[Pp]utative occurred); (2) all eggNOG orthologous groups; (3)
all orthologous groupsplus gene families constructed from remaining genesnot
of 60.
Rarefaction analysis. Estimation of total gene richness was done using
EstimateS on 100 randomly picked samples due to memory limitations.
were calculated and the larger estimate of the two (ICE) was used. The estimate
for this sample size was 3,621,646 genes (ICE) whereas Sobs(Mao Tau) was
3,090,575genes, or85.3%.TheICEestimatorcurvedidnotcompletelysaturate,
(data not shown) indicating that additional samples will need to be added to
achieve a final, conclusive estimate.
Common bacterial core. To eliminate the influence of very similar strains and
assess the presence of known microbial species among the individuals of the
by similarity, using a 90% identity cutoff and the similarity over at least 80% of
the length. From each group only the largest genome was used. Illumina reads
from 124 individuals were mapped to the set, for species profiling analysis and
the genomes originating from the same species (by differing in size .20%)
curated by manual inspection and by using the 16S-based clustering when the
sequences were available.
Relative abundance of microbial genomes among individuals. We computed
the genome coverage by uniquely mapping Illumina reads and normalized it to
1Gb of sequence, to correct for different sequencing levels in different indivi-
duals.Thecoveragewassummedoverall speciesofthenon-redundantbacterial
genome set for each individual and the proportion of each species relative to the
sum calculated.
Species co-existencenetwork. For the 155 species that had genome coverage by
the Illumina reads$1% in at least one individual we calculated the pair-wise
inter-species Pearson correlations between sequencing depths (abundance)
throughout the entire cohort of 124 individuals. From the resulting 11,175
inter-species correlations, correlations less than 20.4 or above 0.4 (n5342)
were visualized in a graph using Cytoscape44displaying the average genome
coverage of each species as node size in the graph.
40. Toft, U. et al. The impact of a population-based multi-factorial lifestyle
47, 378–383 (2008).
41. Li, R. et al. SOAP2: an improved ultrafast tool for short read alignment.
Bioinformatics 25, 1966–1967 (2009).
42. Huson,D.H.,Auch,A.F.,Qi,J.&Schuster,S.C.MEGANanalysisofmetagenomic
data. Genome Res. 17, 377–386 (2007).
43. vanDongen,S.GraphClusteringbyFlowSimulation.PhDthesis,Univ.Utrecht(2000).
44. Shannon, P. et al. Cytoscape: a software environment for integrated models of
biomolecular interaction networks. Genome Res. 13, 2498–2504 (2003).
Macmillan Publishers Limited. All rights reserved


text:
Sept. 2, 2014
Dominion is moving forward with plans to build a natural gas pipeline that would cut through Nelson County.
The 550-mile project will start in Harrison County, WV., going through Nelson and Buckingham counties in Virginia and end in Robeson County, NC.
The $5 billion proposal has been met with criticism in Nelson Co. where residents are concerned about the impacts of the pipeline.
The pipe itself would be 42 inches wide, and buried out of sight. However, Dominion would have to remove all trees and other obstructions for 75 feet surrounding the pipeline.
But for residents in Nelson County, the announcement of today's plans was not what they wanted to hear.
'Friends of Nelson', a local group of property owners opposed to the pipeline, said in a press release that this project is not of the public interest.
"All we can say is that we are not happy with the whole big plan it really will have a devastating effect," Kathy Versluys.
Kathy said that herself along with other residents feel like they don't have a voice nor a say in how things are moving forward in their own back yard.
"I mean Nelson County roads are not built for big industry, let's just face it, so these huge trucks with these huge pieces of pipe would be out of place," she said.
Even for small businesses it could have a deeper impact, one that directly hits their pockets. For new business owner Denver Riggleman of Silverback Distillery off of route 151, the pipeline plans could not have come at a more inconvenient time.
"Well it affects us personally because this is our fifty acres we just finished this distillery on just four days ago and this is our fifth day being open," said Riggleman.
Denver and his wife had plans to retire on their new land and they didn't expect that this plan would impact them so soon.
"An individual that came from Dominion or one of their companies they told me that the pipeline would come right through my land on an angle," he said.
As plans go forward, one can only wonder if the pipeline and the people of Nelson County can co-exist.
"As far as co-existing, you'd have to see the environmental plan and we haven't seen much of anything except that we're going to be affected which is a bit troubling," he said.
The project still has to go through the approval process, and if it's completed it will support 118 jobs per year and an additional $233,000 in state tax revenue.
But only time will tell what true effect this has on not only the people of Nelson, but everyone living along the pipeline.

subset: commoncrawl


text:
A project of People For the American Way dedicated to monitoring and exposing the activities of the right-wing movement. Learn More >
Ted Cruz 'Honored' To Go Hunting With Steve King
Texas senator and likely 2016 presidential candidate Ted Cruz has made a notable friend in Iowa: Rep. Steve King. The Des Moines Register reports that Cruz has accepted King's invitation to go pheasant hunting on the opening day of the hunting season next month, and was "honored to have received the invite."
"Yes, we are confirmed for a hunt with King," Cruz spokeswoman Catherine Frazier said in an email Friday to the Des Moines Register. "The senator has enjoyed getting to know him and work with him on important issues before Congress. He's honored to have received the invite."
Prior to the 2012 Iowa Republican presidential caucuses, King hosted former Pennsylvania Sen. Rick Santorum twice for bird hunting at Iowa game preserves and Texas Gov. Rick Perry on one occasion. The Iowa congressman said in an interview on Thursday he hopes to shape the debate for the 2016 GOP presidential contest by serving as a "guardrail of constitutional conservatism."
Cruz's proud association with King is another sign that the Texan has no plans to moderate his positions in advance of a presidential run. King earned rebukes from his party leadership last month when he insisted that most young undocumented immigrants are drug runners with "calves the size of cantaloupes because they've been hauling 75 pounds of marijuana across the desert." He has also compared immigrants to dogs.
Cruz has been one of the most outspoken opponents of the Senate's bipartisan immigration proposal.
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Additional free tickets have just been released for this summer's sold out Ozzfest!! Tickets can be picked up at the following box offices during the noted hours (WHILE SUPPLIES LAST!!!): * Cricket Pavilion, Phoenix, Arizona - Monday, Wednesday and Friday from 11 a.m. to 5 p.m. * Journal...
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Jul192007CLICK HERE to win a chance to meet Ozzy and have your video premiere on Myspace Music! Ozzy was so happy with the submissions for his last video contest, that he wants to do the same for "Not Going Away...
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Jun222007The Birmingham Walk of Stars, Broad Street, will honor those people who have made a significant contribution in the categories of music, television, film, radio, theatre, sport, business and literacy. The Broad Street area of Birmingham city centre is the Midland's premier, mixed use entertainment...
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See Pamela Anderson's Hot New Baywatch-Inspired "Vogue Brazil" Shoot
Abs, meet under-boob! A little NSFW as under-boob typically is.
Abs, meet under-boob! A little NSFW as under-boob typically is.
Angela Vitello a year ago 25 responses
Commence shame spiral.
Alex Rees a year ago 31 responses
His girly good looks can never be overexposed, quite frankly.
Angela Vitello a year ago 106 responses
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Sexy dwarf who carries a ham and has a cool hat.
Bofur carries a ham.
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India Talks Tough, Seeks Unconditional Apology From US
By IANS - NEW DELHI
Published: 17th December 2013 07:17 PM
Last Updated: 17th December 2013 08:59 PM
India Tuesday sought "unconditional apology" from the US over the arrest and the purported strip search of its ranking diplomat Devyani Khobragade in New York and also took a slew of steps to pare down the privileges of American diplomats in an apparent retaliatory measure.
Parliamentary Affairs Minister Kamal Nath said the US should tender "unconditional apology" to India over the public humiliation of its diplomat, and that "more steps need to be taken to awaken the US that it's a changed world."
The union minister's tough message came after India asked US diplomats in consulates across the country to surrender their Diplomatic Identity Cards issued to them and their families by the Indian government.
India has also withdrawn all airport passes for consulates and import clearances for the embassy. This helped them to import liquor without paying airport fees.
Sources said Delhi Police have also been asked to remove security barricades outside the US embassy. The barricades for years restricted public movement around the embassy and normal vehicular traffic was particularly barred from the slip road in front of the embassy and the visa office at the back of the chancery.
India is also seeking details like salaries paid to Indian staff employed in the US consulates, including those working as domestic helps with the families of American officials because there is suspicion that Indian staff at the US embassy were discriminated against.
Khobragade was charged last week with visa fraud and making false statements.
She was accused by Manhattan's Indian American US Attorney Preet Bharara of visa fraud and exploiting her babysitter and housekeeper. She was handcuffed in public by law enforcement authorities in New York Thursday while she was dropping her daughter at school.
Khobragade was strip searched, confined to a cell with drug addicts and also subjected to DNA swabbing, sources confirmed to IANS.
More stories on the India - US diplomatic row
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The Huntress
The Rose Alley
The second I saw this set I fell in love with it. Goddess of the Hunt by photographer...
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- Attacks a Group or Individuals
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A fun little lifestyle, fashion, and design blog. Full of misadventures, beautiful people, and pretty things!


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Welcome to Starr County, Texas!
If you are looking for local news, events, weather or other local features, you have come to the right place.
Looking for a local business? Click on one of the cities below for business and city area information.Falcon Heights Garciasville Grulla
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North Dansville, New York
|North Dansville, New York
|Coordinates: 42°33′36″N 77°41′44″W / 42.56000°N 77.69556°WCoordinates: 42°33′36″N 77°41′44″W / 42.56000°N 77.69556°W
|United States
|New York
|• Type
|Town Council
|• Town Supervisor
|Dennis P. Mahus (R)
|• Town Council
Members' List
|• Total
|9.8 sq mi (25.5 km2)
|• Land
|9.8 sq mi (25.5 km2)
|• Water
|0.0 sq mi (0.0 km2)
|728 ft (222 m)
|Population (2010)
|• Total
|Time zone
|Eastern (EST) (UTC-5)
|• Summer (DST)
|FIPS code
|GNIS feature ID
North Dansville is a town in Livingston County, New York, United States. The population was 5,538 at the 2010 census.
The Town of North Dansville is in the southeast part of the county.
The first settlers arrived around 1795. The town was organized from the Town of Sparta in 1846, and in 1849 North Dansville was increased by more territory from Sparta. The town was at one time on a branch of the Genesee Valley Canal.
According to the United States Census Bureau, the town has a total area of 9.8 square miles (25 km2), all of it land.
The east and south town lines are the border of Steuben County, New York. The Town of Dansville is in Steuben County, directly south of the Town of North Dansville.
Interstate 390 passes through the town. New York State Route 36, New York State Route 63, and New York State Route 256 pass through the town.
Canaseraga Creek flows northward to the Genesee River.
As of the census[1] of 2000, there were 5,738 people, 2,318 households, and 1,513 families residing in the town. The population density was 583.9 people per square mile (225.4/km²). There were 2,534 housing units at an average density of 257.9 per square mile (99.5/km²). The racial makeup of the town was 96.15% White, 1.06% African American, 0.26% Native American, 0.58% Asian, 1.10% from other races, and 0.85% from two or more races. Hispanic or Latino of any race were 1.78% of the population.
There were 2,318 households out of which 32.4% had children under the age of 18 living with them, 48.1% were married couples living together, 13.2% had a female householder with no husband present, and 34.7% were non-families. 28.9% of all households were made up of individuals and 14.5% had someone living alone who was 65 years of age or older. The average household size was 2.44 and the average family size was 2.98.
In the town the population was spread out with 25.8% under the age of 18, 8.0% from 18 to 24, 27.0% from 25 to 44, 22.3% from 45 to 64, and 16.9% who were 65 years of age or older. The median age was 39 years. For every 100 females there were 89.7 males. For every 100 females age 18 and over, there were 85.8 males.
The median income for a household in the town was $32,519, and the median income for a family was $41,519. Males had a median income of $31,503 versus $25,500 for females. The per capita income for the town was $16,540. About 10.7% of families and 15.4% of the population were below the poverty line, including 16.6% of those under age 18 and 12.5% of those age 65 or over.
Communities and locations in North Dansville[edit]
- Cumminsville – A hamlet on the west town line between Dansville airport and the Interstate.
- Dansville – The Village of Dansville is adjacent to Interstate 390.
- Dansville Municipal Airport (DSV) – A general aviation airport north of Dansville village, lying between NY-36 and NY-63.
- Danville Station – A location northeast of Dansville village.
- ^ "American FactFinder". United States Census Bureau. Retrieved 2008-01-31.
External links[edit]


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A strong quake centred off north-eastern Japan shook buildings as far away as Tokyo last night and triggered a one-metre tsunami in an area devastated by last year's Fukushima disaster, but there were no reports of deaths or serious damage.
The quake had a preliminary magnitude of 7.3, the US Geological Survey said, and thousands of coastal residents were ordered to evacuate to higher ground, but the tsunami warning was lifted two hours after the tremor struck.
The March 2011 earthquake and following tsunami killed nearly 20,000 people and triggered the world's worst nuclear crisis in 25 years when the Fukushima-Daiichi nuclear plant was destroyed, leaking radiation into the sea and air.
Workers at the plant were ordered to move to safety after Friday's quake. Tokyo Electric Power Co, the operator of the Fukushima nuclear plant, reported no irregularities at its nuclear plants.
All but two of Japan's 50 nuclear reactors have been idled since the Fukushima disaster as the government reviews safety.
The quake measured a "lower 5" in Miyagi prefecture on Japan's scale of one to seven, meaning there might be some damage to roads and houses that are less quake resistant.
The scale measures the amount of shaking and in that sense gives a better idea of possible damage than the magnitude. The quake registered a 4 in Tokyo.
The one-metre tsunami hit at Ishinomaki, in Miyagi, at the centre of the devastation from the March 2011 disaster.
All Miyagi trains halted operations and Sendai airport, which was flooded by the tsunami last year, closed its runway.
Five people in the prefecture were slightly injured.
"I was in the centre of the city the very moment the earthquake struck. I immediately jumped into the car and started running away towards the mountains. I'm still hiding inside the car," said Ishinomaki resident Chikako Iwai.
"...I have the radio on and they say the cars are still stuck in the traffic. I'm planning to stay here for the next couple of hours."
There are vast areas of Ishinomaki that still have not been cleaned up since last year's tsunami. Many houses lie in ruins, full of rubble. Workers by the shore still sort through thousands of cars that were swamped and destroyed. The cars are piled up and being taken apart for parts and scrap.
Narita airport outside Tokyo was back in action after a brief closure for safety checks. There were small tsunamis, measuring in the centimetres, elsewhere near the epicentre.
Last year's quake, which measured 9.0, triggered fuel-rod meltdowns at Fukushima, causing radiation leakage, contamination of food and water and mass evacuations. Much of the area is still deserted.
The government declared in December that the disaster was under control.
"Citizens are now escaping to designated evacuation centres and moving to places on higher ground," office worker Naoki Ara said in Soma, 30 km from the Fukushima-Daiichi plant.
Prime Minister Yoshihiko Noda cancelled campaigning in Tokyo ahead of a December 16 election and was on his way back to his office, but there was no immediate plan to hold a special cabinet meeting.
Public spending on quake-proofing buildings is a big election issue.
Japanese were posting photos of their TV screens with tsunami warnings on Facebook, asking each other whether they're safe, confirming their whereabouts.
"It shook for a long time here in Tokyo, are you guys all all right?" posted Eriko Hamada, enquiring about the safety of her friends.
Phone lines were overloaded and it was difficult to contact residents of Miyagi.
"Owing to the recent earthquake, phone lines are very busy, please try again later," the operator said.
The yen rose against the dollar and the euro on the news, triggering some safe-haven inflows into the Japanese currency.
Japan is one of the world's most earthquake-prone countries, with a tremor occurring at least every five minutes.
Located in the "Ring of Fire" arc of volcanoes and oceanic trenches partly encircling the Pacific Basin, the country accounts for about 20 percent of the world's earthquakes of magnitude 6.0 or greater.
Tokyo, with a population of 12 million, sits on the junction of four tectonic plates: the Eurasian, North American, Philippine and Pacific. The sudden bending or breaking of any plate can trigger an earthquake.
- Reuters
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When someone snorts crack off of a penis.
"She did a white willy vacuum on that guy at the party"
by Sid April 08, 2005
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Book your room at Hotel Lamosa
4-star hotel in Provaglio d'Iseo with a seasonal outdoor pool
Situated in Provaglio d'Iseo, this hotel is close to Guido Berlucchi, Nature Reserve, and Acqua Splash. Local attractions also include Franciacorta Golf Club.
Hotel Features
Hotel Lamosa provides a bar/lounge, complimentary wireless Internet access, a rooftop terrace, and meeting rooms.
Room Amenities
In addition to complimentary wireless Internet access and air conditioning, guestrooms feature climate control along with safes and minibars.Information missing or incorrect? Tell us!
Hotel Lamosa
Hotel Amenities
Hotel Lamosa features a seasonal outdoor pool, a rooftop terrace, and tour/ticket assistance. 3 meeting rooms are available. Public areas are equipped with complimentary high-speed wireless Internet access. A bar/lounge is on site where guests can unwind with a drink. For a surcharge, a roundtrip airport shuttle (available on request) is offered to guests. This Provaglio d'Iseo hotel also offers room service (during limited hours), beach/pool umbrellas, and an elevator (lift). Parking is available onsite for a surcharge.
- 24-hour front desk
- Airport transportation (surcharge)
- Total number of rooms - 68
- Number of floors - 2
- Bar/lounge
- Elevator/lift
- Beach/pool umbrellas
- Tours/ticket assistance
- Free Wi-Fi
- Rooftop terrace
- Outdoor seasonal pool
- Meeting rooms
- Pool sun loungers
- Free parking nearby
- Number of meeting rooms - 3
- Self parking (surcharge)
- Number of buildings/towers - 2
Available in all rooms: Free Wi-Fi
Available in some public areas: Free Wi-Fi
Room Amenities
- Climate control
- Air conditioning
- Minibar
- Daily housekeeping
- Private bathroom
- Complimentary toiletries
- Bidet
- Hair dryer
- Shower only
- In-room safe
- Rollaway/extra beds available
- Free Wi-Fi
- Room service (limited hours)
- Connecting/adjoining rooms available
- Cribs/infant beds (surcharge)
Where to Eat
Room service (during limited hours) is available.
Nearby Things to Do
The recreational activities listed below are available either on site or nearby; fees may apply.
- Windsurfing nearby
- Bicycle rentals nearby
- Hiking/biking trails nearby
- Mountain biking nearby
Hotel Policies
Check-in time starts at 3 PM
Check-out time is 11 AM
Payment types
You need to know
Extra-person charges may apply and vary depending on hotel policy.
Government-issued photo identification and a credit card or cash deposit are required at check-in for incidental charges.
Special requests are subject to availability upon check-in and may incur additional charges. Special requests cannot be guaranteed.
- Self parking fee: EUR 10 per night
- Crib (infant bed) fee: EUR 20 per night
We should mention
Cash transactions at this property cannot exceed EUR 999.99, due to national regulations. For further details, please contact the property using information in the booking confirmation. The property has connecting/adjoining rooms, which are subject to availability and can be requested by contacting the property using the number on the booking confirmation.


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Drag the home icon in this panel and drop it onto the "house icon" in the tool bar for the browser
Select "Yes" from the popup window and you're done!
Daily Times
JACKSON, Miss. — Public schools in Jackson, Mississippi, will no longer handcuff students to poles or other objects and will train staff at its alternative school on better methods of discipline, MSN News reports.
Full story:
Miss. district agrees to stop handcuffing students


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Senator Andrist proposes North Dakota constitutional amendment
April 28, 2009
BISMARCK, North Dakota: John Andrist, a member of the North Dakota State Senate since 1992, says that he wants a proposed constitutional amendment on the November 2010 ballot in North Dakota that would largely overturn the provisions endorsed by about 54% of the state's voters in 2008 when they approved the North Dakota Tobacco Use Prevention and Control Program Initiative.
The basic issue is how the state should use the money that regularly flows into its coffers from a tobacco lawsuit. The 2008 initiative earmarked the tobacco settlement funds for particular purposes, such as prevention.
According to Andrist, the 2008 initiative may have been unconstitutional and thus, subject to lawsuits. He thinks the state's voters should re-consider it.[1]


text:
Infant Diaper Bag Must Haves
What Are Your Top Three Diaper Bag Must Haves For Infants?
by babysugar 9 Reactions 0 Shares Print
Diapers? Check. Wipes? Check. Bottles? Check. Now that we've confirmed the most obvious components to an infant diaper bag, I want to know what other three items you must have with you at all times when bouncing babe on your hip. My list toppers are a toy for distraction, a clean onesie and tube of diaper cream. What essentials do you stash?
Read More Tell MommyBabyDiaper Bags
Lily-Inferno 5 years
Back up outfit, noisy toys and diaper cream. For sure.
roxtarchic 5 years
At 18 mos, i must ALWAYS have pretzels, sippy cup & a small toy. If we will be out for a lil while... i'll throw in the lil diaper holder/wipe holder, just in case. These all get tossed into my huge handbag. No diaper bag.
mstrauss 5 years
It's funny because I always had the diaper rash cream and never EVER used it. My essentials are snacks, a small toy or book, and water.
You-Me-and-Baby 5 years
Back up outfit, baby powder & Hot Sling
Pampire 5 years
Extra outfit, diaper rash cream, hand sanitizer for mom or dad :)
Moms 5 years
Back up outfit, burpers and toy.
bluepuppybites 5 years
Lansinoh cream, a burp cloth and gas drops. My daughter had reflux and was coliky, these helped me when were out and about.
Akpril 5 years
Back up outfit, plastic bag to store wet/dirty clothes, and chapstick for me.
Link Love
- Keep calm when it comes to early puberty — Real Simple
- Halle Berry and ex go to court over daughter's hairdo — Celebrity Baby Scoop
- Prepare for your holiday due date — FitPregnancy
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- Does sunscreen hurt your partner's fertility? — The Bump
- 15 extremely awkward pregnancy photos — BabyCenter Blog
- The things you'll never hear a parent say — HuffPost Parents


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The editors of CONCRETE CONSTRUCTION, MASONRY CONSTRUCTION, THE CONCRETE PRODUCER, and CONCRETE & MASONRY CONSTRUCTION PRODUCTS, all sister publications to PUBLIC WORKS, have joined forces to gather products that comprise the most forward-thinking materials, tools, and equipment on the market.
Higher energy prices will drive concrete prices up 3% to 5%, according to Ken Simonson, chief economist for the Associated General Contractors of America. Wooden roads, anyone?
As infrastructure managers look for ways to economize materials for pavement and water projects, an increasingly cost-effective choice is roller-compacted concrete (RCC).
An attorney from the Milwaukee suburb of River Hills, Wis., was exiting northbound I-43 when he found himself stuck. It wasn't bumper-to-bumper traffic that held him in one place—it was the pavement itself.
This year, the American Concrete Pipe Association (ACPA) is celebrating its 100th anniversary by looking back on the role concrete pipe has played in the country's history.
First introduced nearly 35 years ago, small surface planers (scarifiers) have seen their acceptance and use on municipal maintenance projects grow significantly.
Officials at the New York City Department of Environmental Protection worry that herds of hungry deer munching vegetation around drinking water reservoirs might be threatening the supply.
Copyright ©
2014 Hanley Wood Media, Inc. All rights reserved.


text:
BOSTON – Raytheon Company, a Waltham-based defense contractor, has agreed to pay $350,000 to resolve allegations that it charged the Department of Defense for meal expenses that were ineligible for reimbursement. Specifically, the government alleged that, from 2007 to 2009, Raytheon submitted overhead expenses for group meals, business group meals, and group meals at conference events that were ineligible for reimbursement under the Federal Acquisition Regulation.
This matter was investigated by Leo Lamont, Special Agent in Charge of the Naval Criminal Investigative Service, Northeast Field Office, and Michael Conner, Resident Agent in Charge of the U.S. Army Criminal Investigation Division, Hartford Fraud Resident Agency, with the assistance of the Defense Contract Audit Agency's Investigative Support Division and the Defense Contract Management Agency. The matter was handled by Assistant United States Attorney Jennifer Cardello or Ortiz's Civil Division.
Receive information regarding those charged in the Boston Marathon bombing cases.
Making sure that victims of federal crimes are treated with compassion, fairness and respect.
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BBQ Pitmasters Season Finale, Plus Episode 5 Recap
The finale for Season 3 of BBQ Pitmasters is tonight. It was filmed in Lexington, NC at the BBQ Capital Cook Off, and the contestants in this episode are cooking pork shoulder and spare ribs. This episode will feature the winners from the previous 5 episodes: … Continue reading


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Radio Birdman
53 Shouts - 712,791 Scrobbles
Read More...
Radio Birdman are a punk band that formed in Sydney, Australia in 1974. The band's best known lineup consisted of Deniz Tek (vocals, guitar), Rob Younger (vocals, guitar), Chris Masuak (guitar), Philip "Pip" Hoyle (keyboards), Warwick Gilbert (bass) and Ron Keeley (drums) Radio Birdman formed in 1974 and began recording in 1976 when the single Burn My Eye was released. They took their name from a misheard lyric in The Stooges song, 1970.
Read more about Radio Birdman on Last.fm.
User-contributed text is available under the Creative Commons By-SA License and may also be available under the GNU FDL.
Top SongsTotal plays on Last.fm over the last 6 months
- LyricsMoving to the sound of the midnight masses
Fighting off terminal seventies stasis
Cruising down Woodward gotta find me some action
Looking for a lover with a power reaction
Murder city nights, oh yeah
LyricsGot to get a line to Danno
- Download This Track
- Amazon MP30.89 USD
- 7digital
- iTunes0.99 USD
- Ringtone
Got to pick up his gun
Get out an APB
Chinn is going to tell you why
Government says it's high priority
- New Race - (2:42) - 1,410 plays
- LyricsTook a ride on the ocean, started swimming out
Lost sight of land, time ran out
Arms getting heavy, exhaustion's setting in
Waves getting bigger, life's getting thin
My brain was screaming
- Love Kills - (3:44) - 1,104 plays
- Anglo Girl Desire - (3:06) - 1,140 plays
- Do The Pop - (2:32) - 992 plays
- Hand Of Law - (5:59) - 1,048 plays
- What Gives? - (2:25) - 795 plays
- Man With Golden Helmet - (5:39) - 716 plays


text:
Find records of Stuart Lucas
Find birth, death, marriage, military, and other records of Stuart Lucas at Ancestry.com.Search for records
Biography & Family History Add details
Stuart Lucas (1914 - 1986) was born on August 18, 1914. He was born into the Lucas family.
There are no known marriages or children on file for Stuart. Stuart died in October 1986 at 72 years old.
Stuart Lucas's last known residence is at Troy, Montgomery County, North Carolina.
The data on this page comes from the official record for Stuart Lucas in the SSDI (United States Social Security Master Death Index). What is the SSDI?
This profile of Stuart Lucas's life is a collaborative effort by AncientFaces users Share any information you have to reach the people who know him. Some common things we would like to know about Stuart's genealogy:
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- Given name
- Stuart
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- Last Known Residence
- Troy, Montgomery County, North Carolina
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- Male
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Possible Relatives
- Lankie Lucas
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- Richard Lucas
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- Perry M Lucas
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- Helen Marie Wood Lucas
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- Robert B Lucas
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- Marjorie Lucas
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- Cathy Lucas
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- Elsie P Lucas
- ·
- George Lucas
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- John Lucas
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- Leon Lucas
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- Sharon M Lucas
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- Burnell Lucas
- ·
- William R Lucas
- ·
- Cecil L Lucas
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- Wilma Lucas
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- Annie A Lucas
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text:
COVER STORY: Shovel Ready Sites Are Growing From The Ground Up
Many locations are gaining an advantage in the battle for new projects with pre-certification programs for available sites. From the September/October 2013 issue. ...Read more ›
This section includes articles about the economic development and site selection coverage of the storage and distribution of goods.
Many locations are gaining an advantage in the battle for new projects with pre-certification programs for available sites. From the September/October 2013 issue. ...Read more ›
The report cards are in for the Superstorm Sandy recovery and the grade is "incomplete" as misery continues for thousands who have lost their homes or businesses. ...Read more ›
HairUWear will invest $7.1 million in its move to the new facility, which is located in Lenexa. ...Read more ›
Global consumer goods giant to invest $108.7 Million and add 428 new jobs. ...Read more ›
Jones Lang LaSalle's 2012 Life Sciences Cluster Report shows greater Boston, San Diego, Philadelphia and Raleigh-Durham rising in importance as industry hubs. ...Read more ›
The recession of 2008 left Florida overcast economically for years. Looks like it may be time to break out the shades. From the July/August 2013 issue. ...Read more ›
The expanded anodize facility will be operational in a matter of weeks. ...Read more ›
eBay Enterprise, an eBay Inc. company, plans to expand operations in Louisville, creating more than 150 new, full-time jobs and investing up to $45 million in the Commonwealth. ...Read more ›
Business Facilities mourns the loss of a widely respected consultant, colleague and friend, a perennial speaker at our LiveXchange event. ...Read more ›
European company's first U.S. steel facility will result in more than 2,200 direct and indirect jobs in Northwest Louisiana. ...Read more ›
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The Thanksgiving season is upon us, which for many means celebrations chock-full of time with family, friends and, of course, shopping for those loved ones. But what happens after we give thanks on Thursday, flock to stores on Black Friday and surf through the best deals on Cyber Monday? It's the perfect time to give back in honor of all this year's blessings while finishing your holiday gift "to-do" list!
You can participate in #GivingTuesday and support Meals from the Heartland in several ways, including purchasing our hunger-fighting T-shirts as gifts for family and friends.
Click here for the red T-shirt, here for gray and here for blue.
And for those hard-to-shop-for people in your life, you can give a Hunger Fighter Sponsorship Card. These cards are available for $25 (funds 125 meals), $50 (funds 250 meals) or $100 (funds 500 meals) and can be presented as a gift in a loved one's honor that provides meals for hungry children and families in Iowa, around the nation and across the world.
What are we thankful for this year? Our many committed and generous donors and volunteers that keep the fight against hunger going strong. We're thankful for YOU.


text: ESPN Feed: cam newton http://assets.espn.go.com/i/tvlistings/tv_espn_original.gif ESPN en-us 10 ESPN Inc. http://espn.go.com/ Copyright 2010 ESPN Inc. ESPN draft analyst Mel Kiper explains why Ohio State quarterback Terrelle Pryor will be hurt by not going through the usual draft process of playing i...]]> June 09, 2011 06:59:34 PDT A couple days ago, we took a gander at whether an AFC East club would be interested in grabbing besieged Ohio State quarterback Terrelle Pryor in the ...]]> June 03, 2011 08:02:23 PDT ESPN analyst Tedy Bruschi discussed the importance of informal lockout practices for getting rookie quarterbacks ready to play in 2011. Bruschi sai...]]> June 01, 2011 09:41:56 PDT At some point, the unlikely collection of NFL teammates struck somebody as unusual, and they began to sort out who the highest draft pick was. Toge...]]> May 18, 2011 08:00:27 PDT For six weeks leading into the draft, I compiled your votes for an AFC East reader mock that would be measured against ESPN experts Mel Kiper and Todd...]]> May 02, 2011 05:49:11 PDT The 2011 draft was short on sensationalism for the AFC East. Even the sexiest pick was easy to rationalize. The New England Patriots took Arkansas ...]]> April 30, 2011 15:31:34 PDT ORCHARD PARK, N.Y. -- Buffalo Bills general manager Buddy Nix said "there were some shockers" when it came to first-round quarterbacks. Those surpr...]]> April 30, 2011 07:59:57 PDT We'll find out what Ryan Mallett is all about. No setting would have provided a truer test than New England, where the coach runs a tight operation...]]> April 29, 2011 21:01:51 PDT ORCHARD PARK, N.Y. -- As expected, a famous quarterback was the first pick in Thursday night's NFL draft. Then things got a little strange. Another...]]> April 28, 2011 21:31:27 PDT ORCHARD PARK, N.Y. -- With the Buffalo Bills drafting first in the AFC East (again), I'll be reporting Thursday night from their facility. Who do y...]]> April 28, 2011 16:43:05 PDT U.S. Department of Labor analysts have confirmed Mel Kiper is the second-busiest man in the country this week and would have been first had the presid...]]> April 27, 2011 18:15:32 PDT With the NFL draft one day away, Scouts Inc. analysts Kevin Weidl and Steve Muench tap into the latest buzz. They discuss the possibility quarterba...]]> April 27, 2011 15:50:34 PDT Mel Kiper's final Big Board was posted Wednesday afternoon at ESPN Insider, and Louisiana State cornerback Patrick Peterson remained in the top spot. ...]]> April 27, 2011 11:33:23 PDT There's a belief that if the Carolina Panthers don't select Auburn quarterback Cam Newton first overall Thursday night, then the Buffalo Bills will ha...]]> April 27, 2011 10:17:15 PDT With the 32nd pick in the 2011 AFC East reader mock draft, the Green Bay Packers select ... Akeem Ayers, outside linebacker, UCLA. We wrapped up th...]]> April 27, 2011 06:57:06 PDT