Ławniczek-Wałczyk Anna, Gołofit-Szymczak Małgorzata, Cyprowski Marcin, Stobnicka Agata, Górny Rafał L
Biohazard Laboratory, Department of Chemical, Aerosol and Biological Hazards, Central Institute for Labour Protection-National Research Institute, Czerniakowska 16 Street, 00-701, Warsaw, Poland.
Int Arch Occup Environ Health. 2017 Apr;90(3):285-295. doi: 10.1007/s00420-017-1197-z. Epub 2017 Jan 25.
The aim of this study was to characterize the ways of spreading of the most common bacterial species isolated from workers as well as from the air and raw materials at the workplaces in power plant utilizing biomass sources. To monitor microbial transmission and identify the source of contamination in the working environment, a combination of molecular and biochemical methods was applied.
The study was carried out at workplaces in power plant utilizes biomass as a main fuel source. At each of the studied workplaces, bioaerosol particles were collected on sterile Teflon filters using personal conical inhalable samplers (CIS), and biomass samples (straw pellets and briquettes, corn briquettes, sunflower pellets and wood chips) were directly taken from their storage places. Simultaneously with that, the swab samples from the hands of ten workers and their used respiratory masks (of FFP2 class) were also collected after the work shift to evaluate individual workers' microbial contamination. In all collected samples, total bacterial concentrations were assessed and the most common microbial isolates were identified to the species level using both biochemical (API tests) and molecular polymerase chain reaction (PCR), followed by random amplification of polymorphic DNA (RAPD) typing methods.
The mean concentrations of culturable bacteria in the air and in biomass samples at the studied workplaces were high, i.e. 1.2 × 10cfu/m and 3.8 × 10cfu/g, respectively. The number of bacteria in the swab and mask samples also reached a high level of 1.4 × 10 cfu/ml and 1.9 × 10 cfu/cm, respectively. Among the most frequently isolated microorganisms from all types of samples were Gram-positive bacteria of the genus Bacillus and Staphylococcus xylosus. 37 bacterial strains belonging to the genus Bacillus (B. licheniformis 8, B. pumilus 15 and B. subtilis 4) and Staphylococcus (10) were genotyped by the RAPD-PCR method. Based on RAPD-PCR analyses, the genomic similarity among 19 Bacillus strains isolated from biomass, air, protective mask and hand samples as well as 6 S. xylosus strains isolated from air, mask and hand samples exceeded 80%.
This study demonstrated that biomass is the primary source of bacteria at power plant workplaces. These results also revealed that biomass-associated bacteria can be easily transferred to workers' hands and mask during their routine activities. To improve health protection at the workplaces, adequate training courses on hand hygiene and how to use and remove respiratory masks correctly for workers should be introduced as a key element of the prevention strategy. From the occupational point of view, the PCR-based methods seem to be an efficient tool for a fast and precise typing of bacterial strains isolated from different sources in the occupational environment. Such methods may help to implement appropriate prophylactic procedures and minimize transmission of infectious agents at workplaces.
本研究旨在描述从利用生物质源的发电厂工作场所的工人、空气和原材料中分离出的最常见细菌种类的传播方式。为监测微生物传播并确定工作环境中的污染源,采用了分子和生化方法相结合的方式。
该研究在以生物质为主要燃料源的发电厂工作场所进行。在每个研究的工作场所,使用个人锥形可吸入采样器(CIS)在无菌聚四氟乙烯滤膜上收集生物气溶胶颗粒,并直接从生物质样本(秸秆颗粒和煤球、玉米煤球、向日葵颗粒和木屑)的储存地点采集样本。与此同时,在轮班后还收集了十名工人手部及其使用过的(FFP2级)呼吸面罩的拭子样本,以评估个体工人的微生物污染情况。在所有收集的样本中,评估了总细菌浓度,并使用生化方法(API测试)和分子聚合酶链反应(PCR)以及随后的随机扩增多态性DNA(RAPD)分型方法将最常见的微生物分离株鉴定到种水平。
在所研究的工作场所,空气中和生物质样本中可培养细菌的平均浓度较高,分别为1.2×10cfu/m和3.8×10cfu/g。拭子和面罩样本中的细菌数量也分别达到了较高水平,即1.4×10 cfu/ml和1.9×10 cfu/cm。在所有类型样本中最常分离出的微生物是芽孢杆菌属和木糖葡萄球菌属的革兰氏阳性细菌。通过RAPD-PCR方法对37株属于芽孢杆菌属(地衣芽孢杆菌8株、短小芽孢杆菌15株和枯草芽孢杆菌4株)和葡萄球菌属(10株)的细菌菌株进行了基因分型。基于RAPD-PCR分析,从生物质、空气、防护面罩和手部样本中分离出的19株芽孢杆菌菌株以及从空气、面罩和手部样本中分离出的6株木糖葡萄球菌菌株之间的基因组相似性超过80%。
本研究表明生物质是发电厂工作场所细菌的主要来源。这些结果还表明,与生物质相关的细菌在工人的日常活动中很容易转移到他们的手上和面罩上。为改善工作场所的健康保护,应引入关于手部卫生以及如何正确使用和摘除呼吸面罩的适当培训课程,作为预防策略的关键要素。从职业角度来看,基于PCR的方法似乎是一种快速准确地对从职业环境中不同来源分离出的细菌菌株进行分型的有效工具。此类方法可能有助于实施适当的预防措施,并最大限度地减少工作场所传染病原体的传播。