Key Laboratory for Feed Biotechnology of the Ministry of Agriculture, Feed Research Institute, Chinese Academy of Agricultural Sciences, Beijing, People's Republic of China.
PLoS One. 2011;6(12):e29091. doi: 10.1371/journal.pone.0029091. Epub 2011 Dec 19.
The class Myxosporea encompasses about 2,400 species, most of which are parasites of fish and cause serious damage in aquaculture. Due to the concerns about food safety issues and limited knowledge of Myxozoa life cycle and fish immune system, no chemicals, antibiotics or immune modulators are available to control myxozoa infection. Therefore, little can be done once Myxozoa establishment has occurred.
METHODOLOGY/PRINCIPAL FINDINGS: In this paper we isolated Aeromonas veronii CD3 with significant myxospore shell valve-degrading ability from pond sediment. A 3,057-bp full-length chitinase gene was consequently cloned, and the corresponding mature, recombinant chitinase (ChiCD3) produced by Escherichia coli had substantial chitinase activity. The deduced sequence of ChiCD3 contained one catalytic domain, two chitin-binding domains, and one putative signal peptide. ChiCD3 had an optimal activity at 50°C and pH 6.0, and retained more than 50% of its optimal activity under warm water aquaculture conditions (∼30°C and pH ∼7.0). After incubation with ChiCD3, 38.0±4.8% of the myxospores had damaged shell valves, whereas myxospores incubated with commercially available chitinases remained intact.
CONCLUSION/SIGNIFICANCE: This study reveals a new strategy to control myxozoan disease. ChiCD3 that has capacity to damage the shell valve of myxospores can be supplemented into fish feed and used to control Myxozoa-induced diseases specifically.
粘孢子虫纲包含约 2400 种,其中大多数是鱼类寄生虫,在水产养殖中造成严重损害。由于对食品安全问题的关注以及对粘孢子虫生命周期和鱼类免疫系统的了解有限,目前尚无化学品、抗生素或免疫调节剂可用于控制粘孢子虫感染。因此,一旦粘孢子虫建立,就几乎无能为力了。
方法/主要发现:本文从池塘沉积物中分离出具有显著孢子壳瓣降解能力的维氏气单胞菌 CD3。随后克隆了一个 3057bp 的全长几丁质酶基因,并由大肠杆菌产生了相应的成熟重组几丁质酶(ChiCD3),该酶具有显著的几丁质酶活性。ChiCD3 的推导序列包含一个催化结构域、两个几丁质结合结构域和一个假定的信号肽。ChiCD3 的最佳活性在 50°C 和 pH 6.0 下,在水产养殖的温水条件(约 30°C 和 pH 约 7.0)下保留了超过 50%的最佳活性。与 ChiCD3 孵育后,38.0±4.8%的孢子壳瓣受损,而用市售几丁质酶孵育的孢子保持完整。
结论/意义:本研究揭示了一种控制粘孢子虫病的新策略。具有破坏孢子壳瓣能力的 ChiCD3 可以添加到鱼饲料中,专门用于控制粘孢子虫引起的疾病。