Raudabaugh Daniel B, Miller Andrew N
Department of Plant Biology, University of Illinois, Urbana, Illinois, United States of America ; Illinois Natural History Survey, University of Illinois, Champaign, Illinois, United States of America.
PLoS One. 2013 Oct 21;8(10):e78300. doi: 10.1371/journal.pone.0078300. eCollection 2013.
Pseudogymnoascus destructans, the causal agent of bat white-nose syndrome, has caused nearly six million deaths in North American bats since its introduction into the United States in 2006. Current research has shown that caves can harbor P. destructans even after the infected bats are removed and bats no longer visit or inhabit previously infected caves. Our research focuses on elucidating reservoir requirements by investigating the nutritional capabilities of and substrate suitability requirements for six different P. destructans isolates from various localities including Illinois, Indiana, New York (Type specimen), and Pennsylvania. Enzyme assays implicate that both urease and b-glucosidase appear to be constitutive, lipase and esterase activity were more rapid than proteinase activity on 6% gelatin, gelatin degradation was accompanied by medium alkalinization, the reduction of thiosulfate generated hydrogen sulfide gas, chitinase and manganese dependent peroxidase activity were not visually demonstrated within eight weeks, and keratinase activity was not evident at pH 8 within eight weeks. We demonstrate that all P. destructans isolates are capable of growth and sporulation on dead fish, insect, and mushroom tissues. Sole nitrogen source assays demonstrated that all P. destructans isolates exhibit Class 2 nitrogen utilization and that growth-dependent interactions occur among different pH and nitrogen sources. Substrate suitability assays demonstrated that all isolates could grow and sporulate on media ranging from pH 5-11 and tolerated media supplemented with 2000 mg/L of calcium and 700 mg/L of three separated sulfur compounds: thiosulfate L-cysteine, and sulfite. All isolates were intolerant to PEG-induced matric potential with delayed germination and growth at -2.5 MPa with no visible germination at -5 MPa. Interestingly, decreasing the surface tension with Tween 80 permitted germination and growth of P. destructans in -5 MPa PEG medium within 14 days suggesting a link between substrate suitability and aqueous surface tension altering substances.
毁灭隐球菌是蝙蝠白鼻综合征的病原体,自2006年传入美国以来,已导致近600万只北美蝙蝠死亡。目前的研究表明,即使感染蝙蝠被移除且蝙蝠不再造访或栖息于先前受感染的洞穴,洞穴仍可能存有毁灭隐球菌。我们的研究聚焦于通过调查来自伊利诺伊州、印第安纳州、纽约(模式标本产地)和宾夕法尼亚州等地的六种不同毁灭隐球菌分离株的营养能力和底物适宜性要求,来阐明储存库的条件。酶活性测定表明,脲酶和β-葡萄糖苷酶似乎是组成型的,在6%明胶上,脂肪酶和酯酶活性比蛋白酶活性更快,明胶降解伴随着培养基碱化,硫代硫酸盐的还原产生硫化氢气体,在八周内未观察到几丁质酶和锰依赖性过氧化物酶活性,在pH 8时八周内未观察到角蛋白酶活性。我们证明所有毁灭隐球菌分离株都能够在死鱼、昆虫和蘑菇组织上生长和形成孢子。单一氮源试验表明,所有毁灭隐球菌分离株均表现出2类氮利用,并且在不同pH值和氮源之间存在生长依赖性相互作用。底物适宜性试验表明,所有分离株都能在pH值为5 - 11的培养基上生长和形成孢子,并能耐受添加了2000 mg/L钙和700 mg/L三种不同硫化合物(硫代硫酸盐、L-半胱氨酸和亚硫酸盐)的培养基。所有分离株都不耐受聚乙二醇诱导的基质势,在-2.5 MPa时发芽和生长延迟,在-5 MPa时无可见发芽。有趣的是,用吐温80降低表面张力可使毁灭隐球菌在-5 MPa聚乙二醇培养基中在14天内发芽和生长,这表明底物适宜性与改变水相表面张力的物质之间存在联系。