Morphological and functional characterization of the oyster Crassostrea gasar circulating hemocytes: Cell types and phagocytosis activity

Jesarela Merabe Silva Freire , Natanael Dantas Farias , Hélène Hégaret , Patricia Mirella da Silva
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Abstract

Hemocytes are the circulating cells of the hemolymph of oysters and are responsible for numerous physiological functions, including immune defense. The oyster Crassostrea gasar is a native species inhabiting mangrove habitat and is of great commercial interest, cultured throughout the Brazilian coast, mainly in the north and northeast. Despite its commercial importance, little is known about its immunological aspects and defense cells, the hemocytes. This work aimed to morphologically characterize hemocytes of the oyster C. gasar and to study one of the main cellular defense response, phagocytosis, using light microscopy and flow cytometry. The results showed the presence of six hemocyte populations in C. gasar hemolymph. These comprise of large and small granulocytes, large and small hyalinocytes, blast-like cells and a rare type classified as vesicular or serous hemocytes. Hyalinocytes were highly abundant and the most heterogeneous cell population, while small granulocytes, along with vesicular hemocytes were the less abundant population. Hemocytes of C. gasar oysters demonstrated capabilities to phagocytose three different types of particles tested: zymosan A, latex particles and Escherichia coli, indicating a broad defense capacity. The zymosan A were the most engulfed particles, followed by beads, mainly phagocytized by granulocytes, the most phagocytic cells, and finally E. coli, which were the least phagocytized. This study is the first characterization of C. gasar oyster hemocytes and will support future studies that aim to understand the participation of different hemocyte types in defense responses against pathogens and/or environmental changes.

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牡蛎血管循环血细胞的形态和功能特征:细胞类型和吞噬活性
血细胞是牡蛎血淋巴的循环细胞,负责许多生理功能,包括免疫防御。牡蛎Crassostrea gasar是栖息在红树林栖息地的本地物种,具有巨大的商业价值,在整个巴西海岸养殖,主要在北部和东北部。尽管它具有商业重要性,但人们对它的免疫方面和防御细胞——血细胞知之甚少。本工作旨在利用光学显微镜和流式细胞术对牡蛎血细胞进行形态学表征,并研究其主要的细胞防御反应之一吞噬作用。结果表明,在C.gasar血淋巴中存在6个血细胞群。这些细胞包括大小粒细胞、大小透明细胞、成纤维细胞样细胞和一种罕见类型的囊泡或浆液性血细胞。透明质细胞高度丰富,是最异质的细胞群体,而小粒细胞和囊泡血细胞是不太丰富的群体。C.gasar牡蛎的血细胞表现出吞噬三种不同类型颗粒的能力:酵母多糖A、乳胶颗粒和大肠杆菌,表明其具有广泛的防御能力。酵母多糖A是被吞噬最多的颗粒,其次是珠粒,主要被粒细胞吞噬,吞噬细胞最多,最后是被吞噬最少的大肠杆菌。这项研究是对C.gasar牡蛎血细胞的首次表征,将支持未来的研究,旨在了解不同血细胞类型在对抗病原体和/或环境变化的防御反应中的作用。
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