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Molecular Pathogenesis

Outer Membrane Protein Sequence Variation in Lambs Experimentally Infected with Anaplasma phagocytophilum

Erik G. Granquist, Snorre Stuen, Anna M. Lundgren, Margrethe Bråten, Anthony F. Barbet
Erik G. Granquist
1Department of Production Animal Clinical Sciences, Section of Small Ruminant Research, Norwegian School of Veterinary Science, Sandnes N-4325, Norway
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  • For correspondence: erik.georg.granquist@veths.no
Snorre Stuen
1Department of Production Animal Clinical Sciences, Section of Small Ruminant Research, Norwegian School of Veterinary Science, Sandnes N-4325, Norway
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Anna M. Lundgren
2Department of Infectious Diseases and Pathology, College of Veterinary Medicine, University of Florida, Gainesville, Florida 32611-0880
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Margrethe Bråten
1Department of Production Animal Clinical Sciences, Section of Small Ruminant Research, Norwegian School of Veterinary Science, Sandnes N-4325, Norway
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Anthony F. Barbet
2Department of Infectious Diseases and Pathology, College of Veterinary Medicine, University of Florida, Gainesville, Florida 32611-0880
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DOI: 10.1128/IAI.01206-07
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  • FIG. 1.
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    FIG. 1.

    Clinical parameters of lambs 4203 and 4210 infected with A. phagocytophilum. (A to F) The rickettsemias in lambs 4203 (B) and lamb 4210 (E), presented as relative quantitative data on a logarithmic graph, showed little association with rectal temperature (A and D) or neutropenia (C and F), except during the first phase of infection. The horizontal dotted lines in panels A and D represent the temperature (39.5°C) above which fever was recorded in this study. The horizontal dotted lines in panels B and E illustrate the threshold level (Ct) for positive rickettsemia (more than 35 cycles by real-time PCR). The horizontal dotted lines in panels C and F represent the value (<0.7 × 109 cells/liter) below which significant neutropenia was recorded in this study. The vertical dotted lines show the sampling times that corresponded to days 8, 29, 50, 69, and 93 postinfection for lamb 4203 or days 8, 41, 55, and 79 postinfection for lamb 4210. Hash marks on the x axis indicate a scale change.

  • FIG. 2.
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    FIG. 2.

    Complexity of msp2(p44) expression site clones from each rickettsemic peak by RFLP mapping. (A to E) Sequence diversity among a representation of the different clones of the msp2(p44) expression site from the infection in lamb 4203 only, sampled on days 8 (panel A), 29 (panel B), 50 (panel C), 69 (panel D), and 93 (panel E), respectively, by RFLP analysis. Note the large number of different variants at the initial stage of infection compared to later stages and the apparent increase in sequence diversity between days 50 and 69. The positions of molecular size standards (lane STD) are indicated on the right.

  • FIG. 3.
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    FIG. 3.

    MSP2(P44) hypervariable-region sequences from the predominant variants present in the rickettsemic peaks from sheep 4203 and 4210. Shown is a MUSCLE/CHROMA alignment of the sequence variants present at each sampling time analyzed here (Fig. 1) from lamb 4203 (variants 1) and lamb 4210 (variants 2). That is, variants 1.1 through 1.5 represent the variants present in lamb 4203 at sampling times of 8, 29, 50, 69, and 93 days p.i. and variants 2.1 through 2.4 represent the variants present in lamb 4210 at sampling times of 8, 41, 55, and 79 days p.i. The percentage of a variant in each sampled population is given next to the variant number designation. Identical variants that are present in different populations are indicated by the same superscript letter to the right, except for the two variants indicated by a superscript e. These variants (1.2 and 2.3) are identical except for a short region of difference that is highlighted in red. A consensus sequence for all variants is shown at the bottom, where lowercase letters indicate the following: b, big (EFHIKLMQRWY); c, charged (DEHKR); h, hydrophobic (ACFGHILMTVWY); p, polar (CDEHKNQRST); s, small (ACDGNPSTV); t, tiny (AGS).

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Outer Membrane Protein Sequence Variation in Lambs Experimentally Infected with Anaplasma phagocytophilum
Erik G. Granquist, Snorre Stuen, Anna M. Lundgren, Margrethe Bråten, Anthony F. Barbet
Infection and Immunity Dec 2007, 76 (1) 120-126; DOI: 10.1128/IAI.01206-07

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Outer Membrane Protein Sequence Variation in Lambs Experimentally Infected with Anaplasma phagocytophilum
Erik G. Granquist, Snorre Stuen, Anna M. Lundgren, Margrethe Bråten, Anthony F. Barbet
Infection and Immunity Dec 2007, 76 (1) 120-126; DOI: 10.1128/IAI.01206-07
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KEYWORDS

Anaplasma phagocytophilum
Bacterial Outer Membrane Proteins
Ehrlichiosis
sheep

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