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. 2011 Jan;77(1):330-4.
doi: 10.1128/AEM.02096-10. Epub 2010 Nov 5.

Stability and Tick Transmission Phenotype of Gfp-Transformed Anaplasma Marginale Through a Complete in Vivo Infection Cycle

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Free PMC article

Stability and Tick Transmission Phenotype of Gfp-Transformed Anaplasma Marginale Through a Complete in Vivo Infection Cycle

Susan M Noh et al. Appl Environ Microbiol. .
Free PMC article

Abstract

We tested the stability and tick transmission phenotype of transformed Anaplasma marginale through a complete in vivo infection cycle. Similar to the wild type, the gfp-transformed A. marginale strain established infection in cattle, a natural reservoir host, and persisted in immune competent animals. The tick infection rates for the transformed A. marginale and the wild type were the same. However, there were significantly lower levels of the transformed A. marginale than of the wild type in the tick. Despite the lower levels of replication, ticks transmitted the transformant. Transformants can serve as valuable tools to dissect the molecular requirements of tick colonization and pathogen transmission.

Figures

FIG. 1.
FIG. 1.
Confirmation of infection. PCR to confirm infection with the A. marginale gfp strain (a) and the A. marginale wt (b) in needle-inoculated animals (AF), tick midguts (MG), and salivary glands (SG) and in animals infected by tick transmission feeding (TF). The numbers along the top are individual animal identifiers. Amplification was performed with oligonucleotides that span between the gfp insert and the genome backbone (1) or oligonucleotides specific for A. marginale msp5 (2).
FIG. 2.
FIG. 2.
Green fluorescing colonies in tick tissues. (A) Autofluorescence in a D. andersoni midgut infected with the A. marginale wt. (B) Green fluorescent colonies in a D. andersoni midgut after acquisition feeding on animal 36282 infected with the A. marginale gfp strain. (C) Autofluorescence, which is most intense in the salivary gland ducts, in salivary glands infected with the A. marginale wt. (D) Green fluorescing colonies associated with secondary salivary gland ducts in D. andersoni ticks infected with the A. marginale gfp strain after transmission feeding.
FIG. 3.
FIG. 3.
Green fluorescing colonies in erythrocytes from animal 30122 subinoculated with blood from persistently infected animal 36488.
FIG. 4.
FIG. 4.
PCR demonstrating the stability of transformed A. marginale in acquisition-fed animals (AF), transmission-fed animals (TF), and the animal which was subinoculated (SI) with blood from the persistently infected, transmission-fed animal. Animals 36361 and 36432 were infected with the A. marginale wt. Animals 36282, 36488, and 30122 were infected with the A. marginale gfp strain.

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