Cuticular Lipids as a First Barrier Defending Ixodid Ticks against Fungal Infection.

Cárita S Ribeiro-Silva, Elen R Muniz, Valesca H Lima, Cíntia C Bernardo, Walquíria Arruda, Rosane N Castro, Patrícia S Gôlo, Isabele C Angelo, Éverton K K Fernandes
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Abstract

The chemical composition of tick cuticles acts as a barrier to pathogens and may limit infection by entomopathogenic fungi. This study characterized the cuticular neutral lipids (NL) and hydrocarbons (HCs) of four ixodid ticks that are widely distributed in Brazil. HC extracts were analyzed by gas chromatography-mass spectrometry and used to challenge Beauveria bassiana IP361 and Metarhizium robertsii IP146; the effect of cuticular extracts in fungal growth were evaluated by disk diffusion and conidial viability assays. In addition, conidial germination on the tick cuticle was evaluated by scanning electron microscopy, and NL from ticks treated with fungi were assessed by thin layer chromatography. Six HCs were exclusively identified in Amblyomma sculptum. Additionally, cuticle extracts from Dermacentor nitens and A. sculptum inhibited the growth of M. robertsii IP146 and reduced conidial germination of B. bassiana IP361 to 70% and 49%, respectively; the same extracts also produced cytotoxic effects, with conidial death above 30% and 60%. Electron micrographs showed a delayed germination of conidia incubated for 48 h or 72 h on D. nitens and A. sculptum. The lipid profile of A. sculptum treated with fungi was not significantly altered; triacylglycerol was not detected in the cuticle extracts of any other tick species. Finally, A. sculptum and D. nitens cuticles have lipid components that may limit the development of M. robertsii.

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表皮脂质作为硬蜱抵抗真菌感染的第一屏障。
蜱虫表皮的化学成分对病原体起屏障作用,并可能限制昆虫病原真菌的感染。本文对巴西广泛分布的4种伊蚊蜱的表皮中性脂(NL)和碳氢化合物(hc)进行了研究。HC提取物经气相色谱-质谱联用分析,攻毒球孢白僵菌IP361和罗伯特绿僵菌IP146;采用圆盘扩散法和分生孢子活力法评价表皮提取物对真菌生长的影响。用扫描电镜观察了蜱表皮上孢子的萌发情况,用薄层色谱法测定了真菌处理过的蜱的NL。6种hcc仅在雕塑性弱视瘤中发现。另外,皮肤革螨和木犀草角质层提取物对robertsii IP146的生长和球孢白僵菌IP361的分生孢子萌发分别有70%和49%的抑制作用;同样的提取物也有细胞毒性作用,分生孢子死亡率分别在30%和60%以上。电镜观察结果显示,两种孢子孵育48 h和72 h后,孢子萌发延迟。真菌处理后木犀草的脂质谱变化不显著;在其他蜱类的角质层提取物中未检出三酰甘油。最后,A. sculptum和D. nitens的角质层含有脂质成分,这可能会限制M. robertsii的发育。
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