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成团泛菌:一种亦正亦邪的神秘细菌。第二部分——有害影响:空气中传播的内毒素和过敏原——聚焦谷物粉尘、其他农业粉尘和木尘。

Pantoea agglomerans: a mysterious bacterium of evil and good. Part II--Deleterious effects: Dust-borne endotoxins and allergens--focus on grain dust, other agricultural dusts and wood dust.

作者信息

Dutkiewicz Jacek, Mackiewicz Barbara, Lemieszek Marta Kinga, Golec Marcin, Skórska Czesława, Góra-Florek Anna, Milanowski Janusz

机构信息

Department of Biological Health Hazards and Parasitology, Institute of Rural Health, Lublin, Poland.

Department of Pneumonology, Oncology and Allergology, Medical University of Lublin, Lublin, Poland.

出版信息

Ann Agric Environ Med. 2016;23(1):6-29. doi: 10.5604/12321966.1196848.

Abstract

Pantoea agglomerans, a Gram-negative bacterium developing in a variety of plants as epiphyte or endophyte is particularly common in grain and grain dust, and has been identified by an interdisciplinary group from Lublin, eastern Poland, as a causative agent of work-related diseases associated with exposure to grain dust and other agricultural dusts. The concentration of P. agglomerans in grain as well as in the settled grain and flour dust was found to be high, ranging from 10(4)-10(8) CFU/g, while in the air polluted with grain or flour dust it ranged from 10(3)-10(5) CFU/m(3) and formed 73.2-96% of the total airborne Gram-negative bacteria. The concentration of P. agglomerans was also relatively high in the air of the facilities processing herbs and other plant materials, while it was lower in animal farms and in wood processing facilities. Pantoea agglomerans produces a biologically-potent endotoxin (cell wall lipopolysaccharide, LPS). The significant part of this endotoxin occurs in dusts in the form of virus-sized globular nanoparticles measuring 10-50 nm that could be described as the 'endotoxin super-macromolecules'. A highly significant relationship was found (R=0.804, P=0.000927) between the concentration of the viable P. agglomerans in the air of various agricultural and wood industry settings and the concentration of bacterial endotoxin in the air, as assessed by the Limulus test. Although this result may be interfered by the presence of endotoxin produced by other Gram-negative species, it unequivocally suggests the primary role of the P. agglomerans endotoxin as an adverse agent in the agricultural working environment, causing toxic pneumonitis (ODTS). Numerous experiments by the inhalation exposure of animals to various extracts of P. agglomerans strains isolated from grain dust, including endotoxin isolated with trichloroacetic acid (LPS-TCA), endotoxin nanoparticles isolated in sucrose gradient (VECN), and mixture of proteins and endotoxin obtained by extraction of bacterial mass in saline (CA-S), showed the ability of these extracts to evoke inflammatory and fibrotic changes in the lungs, to stimulate alveolar macrophages to produce superoxide anion (O2(-)), interleukin-1 (IL-1) and chemotactic factors for other macrophages and neutrophils, and to increase the pulmonary concentrations of toll-like receptors and chemokines. The most potent properties showed the CA-S which may be attributed to the allergenic properties of P. agglomerans proteins enhanced by the presence of the autologous endotoxin. The results of these experiments are in accord with the clinical studies which revealed a high reactivity of the agricultural and grain industry workers to allergenic extracts of P. agglomerans, and the presence in these populations of hypersensitivity pneumonitis and asthma cases caused by this bacterium. P. agglomerans has been also identified as a potential causative agent of allergic dermatitis in farmers and of allergic pulmonary disorders in cattle. In conclusion, similar to the cotton industry, also in the grain industry and in agriculture, Pantoea agglomerans should be regarded as one of the major causative agents of work-related diseases, caused by the adverse effects of protein allergens and endotoxin produced by this bacterium.

摘要

成团泛菌是一种革兰氏阴性菌,作为附生菌或内生菌在多种植物中生长,在谷物和谷物粉尘中尤为常见。波兰东部卢布林的一个跨学科小组已将其确定为与接触谷物粉尘和其他农业粉尘相关的职业疾病的病原体。研究发现,谷物以及沉降的谷物和面粉粉尘中成团泛菌的浓度很高,范围为10⁴ - 10⁸ CFU/g,而在被谷物或面粉粉尘污染的空气中,其浓度范围为10³ - 10⁵ CFU/m³,占空气中革兰氏阴性菌总数的73.2 - 96%。在加工草药和其他植物材料的设施空气中,成团泛菌的浓度也相对较高,而在养殖场和木材加工设施中则较低。成团泛菌会产生具有生物活性的内毒素(细胞壁脂多糖,LPS)。这种内毒素的重要部分以直径为10 - 50 nm的病毒大小的球状纳米颗粒形式存在于粉尘中,可被描述为“内毒素超大分子”。通过鲎试剂检测评估发现,在各种农业和木材工业环境的空气中,存活的成团泛菌浓度与空气中细菌内毒素浓度之间存在高度显著的关系(R = 0.804,P = 0.000927)。尽管这一结果可能会受到其他革兰氏阴性菌产生的内毒素的干扰,但它明确表明成团泛菌内毒素在农业工作环境中作为一种有害因子的主要作用,可导致中毒性肺炎(ODTS)。通过将动物吸入暴露于从谷物粉尘中分离出的成团泛菌菌株的各种提取物中进行的大量实验,包括用三氯乙酸分离的内毒素(LPS - TCA)、在蔗糖梯度中分离的内毒素纳米颗粒(VECN)以及通过在盐水中提取细菌团块获得的蛋白质和内毒素混合物(CA - S),结果表明这些提取物能够引起肺部的炎症和纤维化变化,刺激肺泡巨噬细胞产生超氧阴离子(O₂⁻)、白细胞介素 - 1(IL - 1)以及针对其他巨噬细胞和中性粒细胞的趋化因子,并增加肺部Toll样受体和趋化因子的浓度。CA - S表现出最显著的特性,这可能归因于成团泛菌蛋白质的致敏特性因自身内毒素的存在而增强。这些实验结果与临床研究一致,临床研究表明农业和谷物行业工人对成团泛菌的致敏提取物具有高反应性,并且在这些人群中存在由这种细菌引起的过敏性肺炎和哮喘病例。成团泛菌也已被确定为农民过敏性皮炎和牛过敏性肺部疾病的潜在病原体。总之,与棉花行业类似,在谷物行业和农业中,成团泛菌也应被视为由该细菌产生的蛋白质过敏原和内毒素的不利影响所导致的职业疾病的主要病原体之一。

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