TY - JOUR
T1 - Identification of candidate pathogens of papillomatous digital dermatitis in dairy cattle from quantitative 16S rRNA clonal analysis
AU - Yano, Takahisa
AU - Moe, Kyaw Kyaw
AU - Yamazaki, Kazuko
AU - Ooka, Tadasuke
AU - Hayashi, Tetsuya
AU - Misawa, Naoaki
N1 - Funding Information:
This work was supported by Japan Farriers Association, KAKENHI (Grant-in-Aid for Scientific Research) on Priority Areas “Applied Genomics” from the Ministry of Education, Culture, Sports, Science and Technology of Japan, and Project for Zoonoses Education and Research, University of Miyazaki.
PY - 2010/7
Y1 - 2010/7
N2 - Although it is suspected that papillomatous digital dermatitis (PDD), an infectious foot disease of cattle, is caused by multiple bacteria, it remains unclear precisely which ones are involved in the etiology. To study the bacterial community, we used 16S rRNA gene sequencing of randomly selected clones based on PCR with minimum amplification cycles to search for organisms present in PDD lesions but not in healthy foot skin. The nucleotide sequences of 1525 clones from 5 PDD lesions (836 clones) and 4 samples of healthy foot skin (689 clones) were determined and grouped into 316 operational taxonomic units (OTUs) with a cut-off value of >99% sequence identity. Two OTUs, P-01 (143 clones; 100% nucleotide sequence identity with Treponema phagedenis) and P-02 (112 clones; 86% identity with Bacteroidetes), were detected most frequently in all PDD samples examined. In contrast, OTU N-01 (87 clones), showing 99% nucleotide sequence identity with Moraxella phenylpyruvica, was the most prevalent in the normal samples examined. Spirochaetes were detected in only 1 sample. Phylogenetic analysis showed that T. denticola-like and T. phagedenis-like spirochetes were the predominant groups in the PDD lesions. Detection of multiple treponemes and an unknown bacterium close to Bacteroides sp. at high rates by a culture-independent approach could be evidence of the association of these organisms with PDD.
AB - Although it is suspected that papillomatous digital dermatitis (PDD), an infectious foot disease of cattle, is caused by multiple bacteria, it remains unclear precisely which ones are involved in the etiology. To study the bacterial community, we used 16S rRNA gene sequencing of randomly selected clones based on PCR with minimum amplification cycles to search for organisms present in PDD lesions but not in healthy foot skin. The nucleotide sequences of 1525 clones from 5 PDD lesions (836 clones) and 4 samples of healthy foot skin (689 clones) were determined and grouped into 316 operational taxonomic units (OTUs) with a cut-off value of >99% sequence identity. Two OTUs, P-01 (143 clones; 100% nucleotide sequence identity with Treponema phagedenis) and P-02 (112 clones; 86% identity with Bacteroidetes), were detected most frequently in all PDD samples examined. In contrast, OTU N-01 (87 clones), showing 99% nucleotide sequence identity with Moraxella phenylpyruvica, was the most prevalent in the normal samples examined. Spirochaetes were detected in only 1 sample. Phylogenetic analysis showed that T. denticola-like and T. phagedenis-like spirochetes were the predominant groups in the PDD lesions. Detection of multiple treponemes and an unknown bacterium close to Bacteroides sp. at high rates by a culture-independent approach could be evidence of the association of these organisms with PDD.
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U2 - 10.1016/j.vetmic.2009.12.009
DO - 10.1016/j.vetmic.2009.12.009
M3 - Article
C2 - 20036086
AN - SCOPUS:77953123188
VL - 143
SP - 352
EP - 362
JO - Veterinary Microbiology
JF - Veterinary Microbiology
SN - 0378-1135
IS - 2-4
ER -