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在模拟人类胃肠道环境某些特征的实验条件下,铜绿微囊藻 PCC 7806 的生存、生长和毒性。

Survival, growth and toxicity of Microcystis aeruginosa PCC 7806 in experimental conditions mimicking some features of the human gastro-intestinal environment.

机构信息

Dept. of Environment, Istituto Nazionale per l'Assicurazione contro gli Infortuni sul Lavoro, Via Urbana 167, 00184 Rome, Italy.

Dept. of Environment and Primary Prevention, Istituto Superiore di Sanità, Viale Regina Elena 299, 00161 Rome, Italy.

出版信息

Chem Biol Interact. 2014 May 25;215:54-61. doi: 10.1016/j.cbi.2014.03.006. Epub 2014 Mar 22.

Abstract

Cyanotoxins (CTX) are widely produced by several cyanobacteria (CB), increasingly spreading in most water bodies and terrestrial habitats, and represent a risk for human health. CB are prokaryotes, and although mostly autotrophic, several examples of heterotrophy in symbiotic relationship with different organisms have been described. In addition to the known routes of exposure, it has been hypothesized that CB might 'colonize' human intestine with relevant implications for human health. Colonization is a complex process and requires specific features of the possible invaders. Still, a short-term persistence as living and toxin-producing organisms within the intestinal lumen of the host could represent an 'internal' source of exposure to CTX. In this work we ran microcosm experiments (4-18days), looking at Microcystis aeruginosa PCC7806 resistance and cyanotoxin-producing capabilities in darkness, 37°C, pH 2, and subsequent recovery in a rich medium, in darkness, 37°C, in the presence of enteric bacteria, mimicking few important features of the gastrointestinal environment. We measured cyanobacterial populations and growth, microcystin (MC) production and the presence of mcyB gene. M. aeruginosa could grow in the dark at 37°C up to 17days, and survive at pH 2 at a rate between 30% and 70%, depending on the age and toxicity of the starting culture. Cell lysis resulted in a substantial amounts of MC released, not degraded at gastric pH. Following the acidic passage, still in the dark at 37°C, M. aeruginosa restarted to grow within 24h for the next 3-4days, independently on the presence of intestinal bacteria, maintaining the MC cell quota and mcyB gene. Our results show new features of CB: a significant resistance of M. aeruginosa in conditions far from its optimal one, that is an environment mimicking some of the important characteristics of human gastrointestinal tract, suggesting the possibility of an internal source of exposure to CTX, with implications for the risk assessment.

摘要

蓝藻毒素(CTX)广泛存在于多种蓝藻(CB)中,在大多数水体和陆地生境中不断扩散,对人类健康构成威胁。蓝藻为原核生物,虽然主要为自养生物,但已描述了与不同生物共生的几种异养实例。除了已知的暴露途径外,还有假说认为蓝藻可能会“定植”人类肠道,对人类健康产生相关影响。定植是一个复杂的过程,需要入侵生物的特定特征。然而,作为活体并在宿主肠道腔中产生毒素的生物在短时间内持续存在,可能成为 CTX 的“内部”暴露源。在这项工作中,我们进行了微宇宙实验(4-18 天),研究了铜绿微囊藻 PCC7806 在黑暗、37°C、pH2 下的抗逆性和产毒能力,以及在黑暗、37°C、存在肠道细菌的丰富培养基中的后续恢复能力,模拟了胃肠道环境的几个重要特征。我们测量了蓝藻种群和生长、微囊藻毒素(MC)的产生以及 mcyB 基因的存在。铜绿微囊藻可以在黑暗中 37°C 下生长长达 17 天,并在 pH2 下以 30%-70%的存活率存活,具体取决于起始培养物的年龄和毒性。细胞裂解导致大量 MC 释放,在胃酸性条件下不会降解。在酸性条件下,铜绿微囊藻仍然在黑暗中 37°C 下,在接下来的 3-4 天内,24 小时内重新开始生长,而与肠道细菌的存在无关,维持 MC 细胞配额和 mcyB 基因。我们的研究结果揭示了蓝藻的新特征:铜绿微囊藻在远非最佳条件下具有显著的抗逆性,这一环境模拟了人类胃肠道的一些重要特征,表明 CTX 存在内部暴露源的可能性,对风险评估具有重要意义。

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