Abstract

White-nose syndrome (WNS) caused by the fungus, Pseudogymnoascus destructans (Pd) has killed millions of North American hibernating bats. Currently, methods to prevent the disease are limited. We conducted two trials to assess potential WNS vaccine candidates in wild-caught Myotis lucifugus. In a pilot study, we immunized bats with one of four vaccine treatments or phosphate-buffered saline (PBS) as a control and challenged them with Pd upon transfer into hibernation chambers. Bats in one vaccine-treated group, that received raccoon poxviruses (RCN) expressing Pd calnexin (CAL) and serine protease (SP), developed WNS at a lower rate (1/10) than other treatments combined (14/23), although samples sizes were small. The results of a second similar trial provided additional support for this observation. Bats vaccinated orally or by injection with RCN-CAL and RCN-SP survived Pd challenge at a significantly higher rate (P = 0.01) than controls. Using RT-PCR and flow cytometry, combined with fluorescent in situ hybridization, we determined that expression of IFN-γ transcripts and the number of CD4 + T-helper cells transcribing this gene were elevated (P < 0.10) in stimulated lymphocytes from surviving vaccinees (n = 15) compared to controls (n = 3). We conclude that vaccination with virally-vectored Pd antigens induced antifungal immunity that could potentially protect bats against WNS.

Highlights

  • Since its discovery in New York in 20061, white-nose syndrome (WNS), caused by the fungus Pseudogymnoascus (Geomyces) destructans (Pd), has killed at least 7 million bats in the US2, causing significant population declines in numerous bat species

  • We first identified a potential CAL-like protein expressed by Pseudogymnoascus destructans (Pd) by using the basic local alignment search tool (BLAST, blast.ncbi.nlm.nih.gov) to search the Pd proteome for proteins with significant homology to the CAL from Paracoccidiodes brasiliensis (Pb), previously shown to be immunogenic and protective[17]

  • All RCNCAL constructs produced an antigen specific band that was not present in cells infected with raccoon poxvirus (RCN) expressing green fluorescent protein (GFP) and corresponded to the major immunoreactive band of 63 kDa from positive control CAL produced in E. coli previously[17]

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Summary

Introduction

Since its discovery in New York in 20061, white-nose syndrome (WNS), caused by the fungus Pseudogymnoascus (Geomyces) destructans (Pd), has killed at least 7 million bats in the US2, causing significant population declines in numerous bat species. Current proposed methods for control of WNS in bats include application of plant – derived compounds or bacteria to hibernating bats to directly inhibit the growth of the fungus[6,7,8,9,10] or inhibition by bacterially produced volatile organic compounds[11]. While results from these studies show promise in the laboratory for reducing the growth of Pd, their practical application may be hindered by logistical factors such as the amount of agent needed www.nature.com/scientificreports/. Because IL-17A is known to recruit and activate phagocytes targeting fungi[18], results suggest that bats are mounting an adaptive immune response to Pd during hibernation[21,22], there is no evidence that this immunity is protective

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