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. 2015 Jul 28;6:763. doi: 10.3389/fmicb.2015.00763

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Copyright © 2015 Maltz, LeVarge and Graf.

This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) or licensor are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

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Restoration of double mutant (SH-894) using free iron. The leech crop colonization of the double mutant using a chemically manipulated blood meal is shown. One hundred and fifty micrometer EDDA was added to the blood meal along with strains to show minimal amounts of a non-protein-bound iron (free iron) with in the leech crop. SH-894 cannot colonize the leech crop. When 160 μM of ferric chloride (free iron) is added to the blood meal the colonization defect of the double mutant was restored, further linking the inability to obtain iron within the leech crop. Each symbol represents the colonization level in one leech. The statistically significant difference between the parent strain and each mutant was performed using a Mann–Whitney test. Bonferroni corrections were preformed and the critical threshold is 0.025.