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Anaplasma phagocytophilum-infected ticks, Japan.

Ohashi N, Inayoshi M, Kitamura K, Kawamori F, Kawaguchi D, Nishimura Y, Naitou H, Hiroi M, Masuzawa T - Emerging Infect. Dis. (2005)

Bottom Line: We report Anaplasma phagocytophilum infection of Ixodes persulcatus and I. ovatus ticks in Japan.Unique p44/msp2 paralogs (and/or 16S rRNA genes) were detected in tick tissues, salivary glands, and spleens of experimentally infected mice.These findings indicate the public health threat of anaplasmosis in Japan.

View Article: PubMed Central - PubMed

Affiliation: Laboratory of Experimental Microbiology, Institute for Environmental Sciences, University of Shizuoka, 52-1 Yada, Suruga, Shizuoka 422-8526, Japan. ohashi@u-shizuoka-ken.ac.jp

ABSTRACT
We report Anaplasma phagocytophilum infection of Ixodes persulcatus and I. ovatus ticks in Japan. Unique p44/msp2 paralogs (and/or 16S rRNA genes) were detected in tick tissues, salivary glands, and spleens of experimentally infected mice. These findings indicate the public health threat of anaplasmosis in Japan.

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Phylogram of Anaplasma phagocytophilum p44/msp2 including Japanese paralogs. A) Cluster from Ixodes persulcatus. B) Cluster from I. ovatus, except for Tick41-1. The tree was constructed using the neighbor-joining method. Numbers on the tree indicate bootstrap values for branch points. Japanese p44/msp2 paralogs from I. persulcatus and I. ovatus are underlined and boxed, respectively, in bold. A single star shows p44/msp2 clusters with 99.2%–100% similarities and double stars show a cluster with 85.6% similarity. Two vertical bars and 6 arrows indicate Japanese p44/msp2 clusters and paralogs, respectively, which are distinct from the previously identified p44/msp2 (<73.1% similarity). A horizontal bar indicates percentage of sequence divergence. Accession numbers and location (Japan-Y [Yamanashi], Japan-S [Shizuoka], US [United States], and UK [United Kingdom]) are in parentheses.
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Figure 2: Phylogram of Anaplasma phagocytophilum p44/msp2 including Japanese paralogs. A) Cluster from Ixodes persulcatus. B) Cluster from I. ovatus, except for Tick41-1. The tree was constructed using the neighbor-joining method. Numbers on the tree indicate bootstrap values for branch points. Japanese p44/msp2 paralogs from I. persulcatus and I. ovatus are underlined and boxed, respectively, in bold. A single star shows p44/msp2 clusters with 99.2%–100% similarities and double stars show a cluster with 85.6% similarity. Two vertical bars and 6 arrows indicate Japanese p44/msp2 clusters and paralogs, respectively, which are distinct from the previously identified p44/msp2 (<73.1% similarity). A horizontal bar indicates percentage of sequence divergence. Accession numbers and location (Japan-Y [Yamanashi], Japan-S [Shizuoka], US [United States], and UK [United Kingdom]) are in parentheses.

Mentions: The p44/msp2 amplicons from 8 PCR-positive ticks and 1 PCR-positive mouse were cloned into a pCR2.1 vector with the TA Cloning Kit (Invitrogen, Carlsbad, CA, USA). Recombinant clones were randomly selected and 28 recombinant p44/msp2 clones were sequenced with an ABI 3100-Avant Genetic Analyzer (Applied Biosystems, Foster City, CA, USA). A phylogenetic tree was constructed based on the alignment of Japanese p44/msp2 sequences and the most closely related paralogs (220–400 bp) by using ClustalX (http://www-igbmc.u-strasbg.fr/BioInfo/ClustalX/), followed by the neighbor-joining method with 1,000 bootstrap resamplings (Figure 2). In this tree, the p44/msp2 sequences obtained from I. ovatus were located mostly in clusters different from those where sequences from I. persulcatus were located, except for Tick41-1. This finding suggests that A. phagocytophilum in I. ovatus may encode p44/msp2 paralogs distinct from those of A. phagocytophilum in I. persulcatus. A previous study suggested that the p44/msp2 sequences from the United States and the United Kingdom can be divided into 27 similarity groups based on >90% similarities of DNA sequences, and most sequences from the United Kingdom are distinguishable from those from the United State because of the similarities <79% (15). Of 28 Japanese p44/msp2 sequences in this study, 11 sequences with similarities >85.6% to the previously identified paralogs were probably divided into 8 similarity groups (Figure 2). Of the remaining 17 sequences with similarities <73.1%, 11 members that were grouped into 2 distinctive clusters (Figure 2) and 6 members that were individually located (Figure 2, arrows) were distinguishable from the 8 similarity groups. Thus, some p44/msp2 paralogs of Japanese A. phagocytphilum are unique and distinct from those of A. phgocytophlium in other countries, although multiple copies of p44 in the genome of an organism should be considered (13).


Anaplasma phagocytophilum-infected ticks, Japan.

Ohashi N, Inayoshi M, Kitamura K, Kawamori F, Kawaguchi D, Nishimura Y, Naitou H, Hiroi M, Masuzawa T - Emerging Infect. Dis. (2005)

Phylogram of Anaplasma phagocytophilum p44/msp2 including Japanese paralogs. A) Cluster from Ixodes persulcatus. B) Cluster from I. ovatus, except for Tick41-1. The tree was constructed using the neighbor-joining method. Numbers on the tree indicate bootstrap values for branch points. Japanese p44/msp2 paralogs from I. persulcatus and I. ovatus are underlined and boxed, respectively, in bold. A single star shows p44/msp2 clusters with 99.2%–100% similarities and double stars show a cluster with 85.6% similarity. Two vertical bars and 6 arrows indicate Japanese p44/msp2 clusters and paralogs, respectively, which are distinct from the previously identified p44/msp2 (<73.1% similarity). A horizontal bar indicates percentage of sequence divergence. Accession numbers and location (Japan-Y [Yamanashi], Japan-S [Shizuoka], US [United States], and UK [United Kingdom]) are in parentheses.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC3367361&req=5

Figure 2: Phylogram of Anaplasma phagocytophilum p44/msp2 including Japanese paralogs. A) Cluster from Ixodes persulcatus. B) Cluster from I. ovatus, except for Tick41-1. The tree was constructed using the neighbor-joining method. Numbers on the tree indicate bootstrap values for branch points. Japanese p44/msp2 paralogs from I. persulcatus and I. ovatus are underlined and boxed, respectively, in bold. A single star shows p44/msp2 clusters with 99.2%–100% similarities and double stars show a cluster with 85.6% similarity. Two vertical bars and 6 arrows indicate Japanese p44/msp2 clusters and paralogs, respectively, which are distinct from the previously identified p44/msp2 (<73.1% similarity). A horizontal bar indicates percentage of sequence divergence. Accession numbers and location (Japan-Y [Yamanashi], Japan-S [Shizuoka], US [United States], and UK [United Kingdom]) are in parentheses.
Mentions: The p44/msp2 amplicons from 8 PCR-positive ticks and 1 PCR-positive mouse were cloned into a pCR2.1 vector with the TA Cloning Kit (Invitrogen, Carlsbad, CA, USA). Recombinant clones were randomly selected and 28 recombinant p44/msp2 clones were sequenced with an ABI 3100-Avant Genetic Analyzer (Applied Biosystems, Foster City, CA, USA). A phylogenetic tree was constructed based on the alignment of Japanese p44/msp2 sequences and the most closely related paralogs (220–400 bp) by using ClustalX (http://www-igbmc.u-strasbg.fr/BioInfo/ClustalX/), followed by the neighbor-joining method with 1,000 bootstrap resamplings (Figure 2). In this tree, the p44/msp2 sequences obtained from I. ovatus were located mostly in clusters different from those where sequences from I. persulcatus were located, except for Tick41-1. This finding suggests that A. phagocytophilum in I. ovatus may encode p44/msp2 paralogs distinct from those of A. phagocytophilum in I. persulcatus. A previous study suggested that the p44/msp2 sequences from the United States and the United Kingdom can be divided into 27 similarity groups based on >90% similarities of DNA sequences, and most sequences from the United Kingdom are distinguishable from those from the United State because of the similarities <79% (15). Of 28 Japanese p44/msp2 sequences in this study, 11 sequences with similarities >85.6% to the previously identified paralogs were probably divided into 8 similarity groups (Figure 2). Of the remaining 17 sequences with similarities <73.1%, 11 members that were grouped into 2 distinctive clusters (Figure 2) and 6 members that were individually located (Figure 2, arrows) were distinguishable from the 8 similarity groups. Thus, some p44/msp2 paralogs of Japanese A. phagocytphilum are unique and distinct from those of A. phgocytophlium in other countries, although multiple copies of p44 in the genome of an organism should be considered (13).

Bottom Line: We report Anaplasma phagocytophilum infection of Ixodes persulcatus and I. ovatus ticks in Japan.Unique p44/msp2 paralogs (and/or 16S rRNA genes) were detected in tick tissues, salivary glands, and spleens of experimentally infected mice.These findings indicate the public health threat of anaplasmosis in Japan.

View Article: PubMed Central - PubMed

Affiliation: Laboratory of Experimental Microbiology, Institute for Environmental Sciences, University of Shizuoka, 52-1 Yada, Suruga, Shizuoka 422-8526, Japan. ohashi@u-shizuoka-ken.ac.jp

ABSTRACT
We report Anaplasma phagocytophilum infection of Ixodes persulcatus and I. ovatus ticks in Japan. Unique p44/msp2 paralogs (and/or 16S rRNA genes) were detected in tick tissues, salivary glands, and spleens of experimentally infected mice. These findings indicate the public health threat of anaplasmosis in Japan.

Show MeSH
Related in: MedlinePlus