Fiorotti Jéssica, Menna-Barreto Rubem Figueiredo Sadok, Gôlo Patrícia Silva, Coutinho-Rodrigues Caio Junior Balduino, Bitencourt Ricardo Oliveira Barbosa, Spadacci-Morena Diva Denelle, Angelo Isabele da Costa, Bittencourt Vânia Rita Elias Pinheiro
Programa de Pós-Graduação em Ciências Veterinárias, Instituto de Veterinária, Universidade Federal Rural do Rio de Janeiro, Seropédica, Brazil.
Laboratório de Biologia Celular, IOC, Fundação Oswaldo Cruz, Rio de Janeiro, Brazil.
Front Physiol. 2019 May 29;10:654. doi: 10.3389/fphys.2019.00654. eCollection 2019.
is an entomopathogenic fungus widely employed in the biological control of arthropods. Hemocytes present in the hemolymph of invertebrates are the cells involved in the immune response of arthropods. Despite this, knowledge about hemocytes morphological aspects as well as their role in response to the fungal infection is scarce. The present study aimed to analyze the hemocytes of females after infection, using light and electron microscopy approaches associated with the cytotoxicity evaluation. Five types of hemocytes (prohemocytes, spherulocytes, plasmatocytes, granulocytes, and oenocytoids) were described in the hemolymph of uninfected ticks, while only prohemocytes, granulocytes, and plasmatocytes were observed in fungus-infected tick females. Twenty-four hours after the fungal infection, only granulocytes and plasmatocytes were detected in the transmission electron microscopy analysis. Hemocytes from fungus-infected tick females showed several cytoplasmic vacuoles with different electron densities, and lipid droplets in close contact to low electron density vacuoles, as well as the formation of autophagosomes and subcellular material in different stages of degradation could also be observed. propagules were more toxic to tick hemocytes in the highest concentration tested (1.0 × 10 conidia mL). Interestingly, the lowest fungus concentration did not affect significantly the cell viability. Microanalysis showed that cells granules from fungus-infected and uninfected ticks had similar composition. This study addressed the first report of fungal cytotoxicity analyzing ultrastructural effects on hemocytes of infected with entomopathogenic fungi. These results open new perspectives for the comprehension of ticks physiology and pathology, allowing the identification of new targets for the biological control.
是一种广泛用于节肢动物生物防治的昆虫病原真菌。无脊椎动物血淋巴中存在的血细胞是参与节肢动物免疫反应的细胞。尽管如此,关于血细胞形态方面及其在真菌感染反应中的作用的知识却很匮乏。本研究旨在使用与细胞毒性评估相关的光学和电子显微镜方法,分析感染后雌性蜱的血细胞。在未感染蜱的血淋巴中描述了五种血细胞(原血细胞、球血细胞、浆血细胞、粒细胞和oenocytoids),而在真菌感染的蜱雌性中仅观察到原血细胞、粒细胞和浆血细胞。真菌感染24小时后,在透射电子显微镜分析中仅检测到粒细胞和浆血细胞。来自真菌感染蜱雌性的血细胞显示出几个具有不同电子密度的细胞质空泡,以及与低电子密度空泡紧密接触的脂滴,还可观察到自噬体的形成和处于不同降解阶段的亚细胞物质。在所测试的最高浓度(1.0×10分生孢子/毫升)下,繁殖体对蜱血细胞的毒性更大。有趣的是,最低的真菌浓度对细胞活力没有显著影响。微量分析表明,感染真菌和未感染真菌的蜱的细胞颗粒组成相似。本研究首次报道了真菌细胞毒性,分析了昆虫病原真菌感染对蜱血细胞的超微结构影响。这些结果为理解蜱的生理学和病理学开辟了新的视角,有助于确定生物防治的新靶点。