First report of infectious spleen and kidney necrosis virus (ISKNV) in two native cichlids cultured in Brazil

U. P. Pereira, Francisco Eduardo Pereira Rocha, N. A. Ferrari, Leonardo Mantovani Favero, R. M. Mainardi, Mayza Brandão da Silva, A. Alfieri, P. Viadanna, Thomas Waltzek, A. M. Dall Agnol
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Abstract

Peacock bass (syn.: tucunaré, Cichla ocellaris) and the pearl cichlids (syn.: acará, Geophagus brasiliensis) are South American cichlids that are highly valued in both the ornamental and sport fish industries. Since 2017, a number of outbreaks of infectious spleen and kidney necrosis virus (ISKNV) have been reported on Brazilian food and ornamental fish farms. In this study, we detected ISKNV in farmed peacock bass and pearl cichlid by PCR and sequence analysis of the partial major capsid protein (MCP) gene. Moribund peacock bass (n=10) and pearl cichlids (2) from a farm experiencing elevated mortality among juveniles and adults of these species, were submitted for bacteriological and molecular diagnostics. Spleen, liver, brain, and kidney tissues were cultured on 5% sheep blood agar and cystine heart agar with 1% glucose and bovine haemoglobin. No bacteria were isolated from the 12 fish. Additionally, DNA extracts from the liver and spleen of all animals were tested for ISKNV using two conventional polymerase chain reaction (cPCR) assays and two nested PCR (nPCR) assays. ISKNV DNA was amplified in all 12 fish DNA extracts tested, in two or more of the PCR assays. Selected ISKNV amplicons were confirmed by Sanger sequencing. The nucleotide sequences derived from these animals were identical to ISKNV strains previously detected in food (e.g., tilapia and carp) and ornamental species, including strains previously detected in fish from Brazil. To the authors’ knowledge, this is the first report of ISKNV in these native Brazilian cichlids.
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首次报告巴西养殖的两种本地慈鲷感染传染性脾肾坏死病毒(ISKNV)
孔雀鲈(同义词:tucunaré,Cichla ocellaris)和珍珠慈鲷(同义词:acará,Geophagus brasiliensis)是南美洲慈鲷,在观赏鱼和运动鱼行业中都具有很高的价值。自 2017 年以来,巴西食用鱼和观赏鱼养殖场爆发了多起传染性脾肾坏死病毒(ISKNV)疫情。在这项研究中,我们通过 PCR 和部分主囊蛋白(MCP)基因的序列分析,在养殖的孔雀鲈和珍珠慈鲷中检测到了 ISKNV。我们对一个养殖场中病死的孔雀鲈(10 条)和珍珠慈鲷(2 条)幼鱼和成鱼进行了细菌学和分子诊断。脾、肝、脑和肾组织在 5%羊血琼脂和含 1%葡萄糖和牛血红蛋白的胱氨酸心脏琼脂上进行培养。12 条鱼均未分离出细菌。此外,还使用两种传统聚合酶链反应 (cPCR) 方法和两种巢式 PCR (nPCR) 方法对所有动物肝脏和脾脏的 DNA 提取物进行了 ISKNV 检测。在两种或两种以上的 PCR 检测中,所有 12 种鱼类 DNA 提取物都扩增出了 ISKNV DNA。部分 ISKNV 扩增子通过 Sanger 测序得到确认。从这些动物身上获得的核苷酸序列与之前在食用鱼(如罗非鱼和鲤鱼)和观赏鱼身上检测到的 ISKNV 株系相同,包括之前在巴西鱼身上检测到的株系。据作者所知,这是首次报道这些巴西本地慈鲷体内存在 ISKNV。
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