7 research outputs found
D. pulex.
<p><i>D</i>. <i>pulex</i> were exposed to ethylene blue for 15 min, rinsed thoroughly, and imaged using a Maximum Intensity Projection Z Plane Stack (10x, Leica SP5 Scanning Laser Confocal Microscope). The intensity image presented here was used as proof of concept during the characterization of ethylene blue.</p
Synthetic route for the preparation of fluorescently labeled vitamin D<sub>3</sub>.
<p>The carbodiimidazole (CDI) coupling of vitamin D<sub>3</sub> and the fluorescent dye linker.</p
D. pulex.
<p><i>D</i>. <i>pulex</i> were exposed to ethylene blue for 15 min, rinsed thoroughly, and imaged using a Maximum Intensity Projection Z Plane Stack (10x, Leica SP5 Scanning Laser Confocal Microscope). The intensity image presented here was used as proof of concept during the characterization of ethylene blue.</p
<i>D</i>. <i>pulex</i> survival with vitamin D<sub>3</sub>.
<p><i>D</i>. <i>pulex</i> (N = 12) exposed to acute UV-A in the presence of vitamin D<sub>3</sub>. Blue line = 0 mg D<sub>3</sub>; Black line = 5 mg D<sub>3</sub>; Red line = 10 mg D<sub>3</sub>. No reproduction was observed in any individuals during the experimental period. Error = standard error of the mean across 3 trials.</p
Tracking vitamin D<sub>3</sub> in <i>D</i>. <i>pulex</i>.
<p>(A) A live <i>D</i>. <i>pulex</i> was placed in a 1:10 solution of ethylene blue, with no vitamin D<sub>3</sub> (“control”) and images were captured 30 min post exposure (10x, Leica SP5 Scanning Laser Confocal Microscope). Note the presence of the dye in the gut tract (red stain). (B). Live <i>D</i>. <i>pulex</i> were placed in a 1:10 solution of ethylene blue linked vitamin D<sub>3</sub>, rinsed thoroughly, and images were captured 120 min post exposure (10x). Image was compiled from a 90-min time lapse to capture sequestration of vitamin D<sub>3</sub> (red) into the tissues of the <i>D</i>. <i>pulex</i> from the intestine. The high intensity red “dots” in the image are concentrated dye on the outside of the carapace that was not removed during the rinsing stages. The yellow circle indicates a region where D<sub>3</sub> sequestration was detected.</p
UV-Stressed <i>Daphnia pulex</i> Increase Fitness through Uptake of Vitamin D<sub>3</sub>
<div><p>Ultraviolet radiation is known to be highly variable in aquatic ecosystems. It has been suggested that UV-exposed organisms may demonstrate enough phenotypic plasticity to maintain the relative fitness of natural populations. Our long-term objective is to determine the potential photoprotective effect of vitamin D<sub>3</sub> on <i>Daphnia pulex</i> exposed to acute or chronic UV radiation. Herein we report our initial findings in this endeavor. <i>D</i>. <i>pulex</i> survival and reproduction (fitness) was monitored for 5 d as a proof of concept study. Significantly higher fitness was observed in the <i>D</i>. <i>pulex</i> with D<sub>3</sub> than those without (most extreme effects observed were 0% survival in the absence of D<sub>3</sub> and 100% with 10 ppm D<sub>3</sub>). Vitamin D<sub>3</sub> was isolated from the culture media, the algal food (<i>Pseudokirchneriella</i>), and the <i>D</i>. <i>pulex</i> and quantified using high performance liquid chromatography (HPLC). Vitamin D<sub>3</sub> was fluorescently labeled using a phenothiazinium dye and added to cultures of <i>D</i>. <i>pulex</i>. Images demonstrating the uptake of D<sub>3</sub> into the tissues and carapace of the <i>D</i>. <i>pulex</i> were acquired. Our initial findings suggest a positive role for D<sub>3</sub> in ecosystems as both UV-stressed algae and <i>Daphnia</i> sequester D<sub>3</sub>, and <i>D</i>. <i>pulex</i> demonstrate increased fitness in the presence of D<sub>3</sub>.</p></div