Development and maintenance of a specific pathogen-free (SPF) zebrafish research facility for Pseudoloma neurophilia
Development and maintenance of a specific pathogen-free (SPF) zebrafish research facility for Pseudoloma neurophilia
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DOI：
10.3354/dao02333
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发表时间：
2011-05-24
影响因子：
1.4
通讯作者：
Tanguay, Robert L.
中科院分区：
文献类型：
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作者：
Kent, Michael L.;Buchner, Cari;Tanguay, Robert L.
嗜神经假瘤（微孢子虫）在斑马鱼研究设施中非常常见。一个新的斑马鱼设施已经建立在Sinnhuber水产资源实验室（SARL），俄勒冈州州立大学，科瓦利斯，OR，美国，这是一个机会，建立一个特定的病原体（SPF）的斑马鱼菌落，这种微孢子虫。2007年，在对亲鱼和后代亚群（每个家庭1000条鱼中有258条）进行PCR筛选后，将9条斑马鱼系（n = 2203）的后代最初转移到SARL设施。在设施内对鱼类进行嗜神经原卟啉单胞菌的筛选如下：（1）通过组织学检查濒死或死鱼。(2)每个品系在4个月的轮换中再生，并且在孵化后10天对这些主要繁殖的每个的子样品（60个鱼苗，在10个池中）进行PCR筛选。(3)将来自每条线的成鱼（类似于1岁）安乐死;通过组织学检查20条鱼，并通过PCR筛选另外60条鱼（在5条鱼的池中）的脑。(4)对4个水箱中的哨兵鱼进行了重复筛查，这些水箱接收来自设施中所有水箱的废水（每个水箱20条鱼）。(5)对实验室内进行的毒理学研究中的4个月龄鱼（n = 760）进行组织学检查。到目前为止，我们已经评估了2800鱼PCR和1222鱼组织学没有检测P。因此，我们已经建立了9个斑马鱼SPF系用于嗜神经原卟啉单胞菌。然而，26条鱼表现出分枝杆菌病，组织切片中存在抗酸细菌，另外49条鱼有偶发病变。
Pseudoloma neurophilia (Microsporidia) is very common in zebrafish Danio rerio research facilities. A new zebrafish facility has been established at the Sinnhuber Aquatic Resource Laboratory (SARL), Oregon State University, Corvallis, OR, USA, and this was an opportunity to establish a specific pathogen-free (SPF) colony of zebrafish for this microsporidium. Progeny from 9 zebrafish lines (n = 2203) were initially transferred to the SARL facility in 2007 following PCR screening of broodstock and a subpopulation of progeny (258 of 1000 fish from each family). Screening of fish for P. neurophilia within the facility was conducted as follows: (1) Moribund or dead fish were examined by histology. (2) Each line was regenerated on a 4 mo rotation, and a subsample of each of these major propagations (60 fry, in pools of 10) was PCR-screened at 10 d post hatch. (3) Adult fish (similar to 1 yr old) from each line were euthanized; 20 fish were examined by histology and the brains of another 60 fish (in pools of 5) were screened by PCR. (4) This screening was replicated on sentinel fish held in 4 tanks receiving effluent water from all tanks in the facility (20 fish per tank). (5) Four-month old fish (n = 760) from a toxicology study conducted within the laboratory were examined by histology. To date, we have evaluated 2800 fish by PCR and 1222 fish by histology without detecting P. neurophilia. Thus, we have established 9 lines of zebrafish SPF for P. neurophilia. However, 26 fish exhibited mycobacteriosis, with acid-fast bacteria present in tissue sections, and 49 other fish had incidental lesions.