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营养和环境条件对禽致病性大肠杆菌生物膜形成的影响。

Effect of nutritional and environmental conditions on biofilm formation of avian pathogenic Escherichia coli.

机构信息

Engineering Research Center for the Prevention and Control of Animal Original Zoonosis, College of Life Science, Longyan University, Longyan, China.

Shanghai Veterinary Research Institute, The Chinese Academy of Agricultural Sciences (CAAS), Shanghai, China.

出版信息

J Appl Microbiol. 2022 Jun;132(6):4236-4251. doi: 10.1111/jam.15543. Epub 2022 Apr 4.

Abstract

AIMS

To study the effects of environmental stress and nutrient conditions on biofilm formation of avian pathogenic Escherichia coli (APEC).

METHODS AND RESULTS

The APEC strain DE17 was used to study biofilm formation under various conditions of environmental stress (including different temperatures, pH, metal ions, and antibiotics) and nutrient conditions (Luria-Bertani [LB] and M9 media, with the addition of different carbohydrates, if necessary). The DE17 biofilm formation ability was strongest at 25°C in LB medium. Compared to incubation at 37°C, three biofilm-related genes (csgD, dgcC, and pfs) were significantly upregulated and two genes (flhC and flhD) were downregulated at 25°C, which resulted in decreased motility. However, biofilm formation was strongest in M9 medium supplemented with glucose at 37°C, and the number of live bacteria was the highest as determined by confocal laser scanning microscopy. The bacteria in the biofilm were surrounded by a thick extracellular matrix, and honeycomb-like or rough surfaces were observed by scanning electron microscopy. Moreover, biofilm formation of the DE17 strain was remarkably inhibited under acidic conditions, whereas neutral and alkaline conditions were more suitable for biofilm formation. Biofilm formation was also inhibited at specific concentrations of cations (Na , K , Ca , and Mg ) and antibiotics (ampicillin, chloramphenicol, kanamycin, and spectinomycin). The real-time quantitative reverse transcription PCR showed that the transcription levels of biofilm-related genes change under different environmental conditions.

CONCLUSIONS

Nutritional and environmental factors played an important role in DE17 biofilm development. The transcription levels of biofilm-related genes changed under different environmental and nutrient conditions.

SIGNIFICANCE AND IMPACT OF THE STUDY

The findings suggest that nutritional and environmental factors play an important role in APEC biofilm development. Depending on the different conditions involved in this study, it can serve as a guide to treating biofilm-related infections and to eliminating biofilms from the environment.

摘要

目的

研究环境应激和营养条件对禽致病性大肠杆菌(APEC)生物膜形成的影响。

方法和结果

使用 APEC 菌株 DE17 研究各种环境应激条件(包括不同温度、pH 值、金属离子和抗生素)和营养条件(LB 和 M9 培养基,必要时添加不同的碳水化合物)下的生物膜形成。在 LB 培养基中 25°C 时,DE17 生物膜形成能力最强。与 37°C 孵育相比,在 25°C 时,三个生物膜相关基因(csgD、dgcC 和 pfs)显著上调,两个基因(flhC 和 flhD)下调,导致运动性降低。然而,在 37°C 下补充葡萄糖的 M9 培养基中生物膜形成最强,通过共聚焦激光扫描显微镜确定活细菌数量最多。生物膜中的细菌被厚厚的细胞外基质包围,通过扫描电子显微镜观察到蜂窝状或粗糙的表面。此外,在酸性条件下,DE17 菌株的生物膜形成明显受到抑制,而中性和碱性条件更有利于生物膜形成。阳离子(Na+、K+、Ca2+和 Mg2+)和抗生素(氨苄西林、氯霉素、卡那霉素和壮观霉素)的特定浓度也抑制生物膜形成。实时定量反转录 PCR 显示,生物膜相关基因的转录水平在不同环境条件下发生变化。

结论

营养和环境因素对 DE17 生物膜发育起着重要作用。生物膜相关基因的转录水平在不同的环境和营养条件下发生变化。

研究的意义和影响

研究结果表明,营养和环境因素在 APEC 生物膜发育中起着重要作用。根据本研究中涉及的不同条件,可以作为治疗生物膜相关感染和从环境中消除生物膜的指南。

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