Summary of Study ST002304

This data is available at the NIH Common Fund's National Metabolomics Data Repository (NMDR) website, the Metabolomics Workbench, https://www.metabolomicsworkbench.org, where it has been assigned Project ID PR001477. The data can be accessed directly via it's Project DOI: 10.21228/M8T40Q This work is supported by NIH grant, U2C- DK119886.

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Study IDST002304
Study TitleWhite-nose syndrome disrupts the splenic lipidome of little brown bats (Myotis lucifugus) at early disease stages
Study SummaryThe fungal disease of bats, white-nose syndrome (WNS), is caused by the pathogen Pseudogymnoascus destructans (Pd). WNS-positive little brown bats (Myotis lucifugus) can exhibit an immune response during infection that include increases in cytokine and pro-inflammatory mediator gene levels. While bioactive lipid mediators (oxylipins) formed by enzymatic oxidation of polyunsaturated fatty acids (PUFAs) can contribute to this type of immune response, their role in WNS pathophysiology have not been investigated. Nonenzymatic conversion of PUFAs can also occur due to reactive oxygen species (ROS), however, these enantiomeric isomers will lack the same signaling properties. In this study, we performed a series of targeted lipidomic approaches on laboratory Pd-inoculated bats to assess changes in their splenic lipidome, including the formation of lipid mediators at early stages of WNS. Hepatic lipids previously identified were also resolved to a higher structural detail. We compared WNS-susceptible M. lucifugus to a WNS-resistant species, the big brown bat (Eptesicus fuscus). Altered splenic lipid levels were only observed in M. lucifugus, with lower total levels of glycerophospholipids (GPs) and free fatty acids (FFAs) in the Pd-inoculated group compared to the sham-inoculated group. Lower concentrations of splenic GPs were observed in lipid compounds containing 18:2 or saturated acyl chains. Differences in splenic FFAs included both omega-3 (including docosahexaenoic acid [DHA]) and omega-6 compounds. Increased levels of an enantiomeric monohydroxy DHA (4-hydroxydocosahexaenoic [HDoHE], 10-HDoHE, and 13-HDoHE) mixture, suggesting nonenzymatic formation, along with 6-keto-PGF1a were found. Changes in previously identified hepatic lipids were confined to omega-3 constituents. Together, these results suggest that increased oxidative stress, but not an inflammatory response, is occurring in bats at early stages of WNS that proceeds fat depletion.
Institute
Georgetown University
Last NamePannkuk
First NameEvan
Address3970 Reservoir Rd, NW New Research Build, washington dc, District of Columbia, 20057, USA
Emailelp44@georgetown.edu
Phone2026875650
Submit Date2022-09-22
Analysis Type DetailLC-MS
Release Date2023-09-22
Release Version1
Evan Pannkuk Evan Pannkuk
https://dx.doi.org/10.21228/M8T40Q
ftp://www.metabolomicsworkbench.org/Studies/ application/zip

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Collection:

Collection ID:CO002383
Collection Summary:Bats were housed within mesh cages (Exo-terra Flexarium©; 22 cm x 35 cm x 43 cm) that were placed within temperature/humidity-controlled incubators (Caron© Environmental Chamber model 6041; 90.1 cm x 84.5 cm x 228.9 cm set to 8°C and 98% humidity) to encourage hibernation. Four cages were used to house the bats, separated by species and inoculation group; M. lucifugus (one sham-treatment: 16 individuals/cage, 10 male and 6 female; one Pd-inoculated; E. fuscus (8 individuals/cage, 4 male and 4 female). Bats were infected with a sham inoculum (20 μl, Phosphate Buffered Saline with Tween®20 (PBS-T)) or a Pd inoculum (20 μl, PBST containing 5x105 conidia) on wing and tail membranes. We monitored arousal frequency and sickness behavior using motion-activated infrared video. During the 71-day period bats found unresponsive or extremely sluggish were humanely euthanized (CO2 inhalation after isoflurane anesthesia and cervical dislocation), and were not included in this experiment. Several M. lucifugus showed signs of morbidity around day 55. We therefore, terminated the experiment 71 – 77 days post-inoculation (mid-March) which coincided with early stages of WNS progression. Disease severity was assessed by fungal load by qPCR33, UV fluorescence indicative of hyphae invasion, survival rate, and behavioral changes 32. E. fuscus was chosen as a WNS-resistant control and we found changes in their splenic lipidome to be minor. Tissue samples were collected, flash frozen, and shipped to Georgetown University Medical Center on dry ice and stored at -80°C until analysis.
Sample Type:Spleen
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