title: Monitoring of bacterial pathogens at workplaces in power plant using biochemical and molecular methods.
abstract: OBJECTIVE
  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.
  
  
  METHODS
  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.
  
  
  RESULTS
  The mean concentrations of culturable bacteria in the air and in biomass samples at the studied workplaces were high, i.e. 1.2 × 106cfu/m3and 3.8 × 104cfu/g, respectively. The number of bacteria in the swab and mask samples also reached a high level of 1.4 × 104cfu/ml and 1.9 × 103cfu/cm2, 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%.
  
  
  CONCLUSIONS
  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.
authors.affiliation: Biohazard Laboratory, Department of Chemical, Aerosol and Biological Hazards, Central Institute for Labour Protection-National Research Institute, Czerniakowska 16 Street, 00-701, Warsaw, Poland. anlaw@ciop.pl.
authors.forename: Anna
authors.identifier: http://orcid.org/0000-0001-8234-340X
authors.initials: A
authors.lastname: Ławniczek-Wałczyk
authors.affiliation: Biohazard Laboratory, Department of Chemical, Aerosol and Biological Hazards, Central Institute for Labour Protection-National Research Institute, Czerniakowska 16 Street, 00-701, Warsaw, Poland.
authors.forename: Małgorzata
authors.initials: M
authors.lastname: Gołofit-Szymczak
authors.affiliation: Biohazard Laboratory, Department of Chemical, Aerosol and Biological Hazards, Central Institute for Labour Protection-National Research Institute, Czerniakowska 16 Street, 00-701, Warsaw, Poland.
authors.forename: Marcin
authors.initials: M
authors.lastname: Cyprowski
authors.affiliation: Biohazard Laboratory, Department of Chemical, Aerosol and Biological Hazards, Central Institute for Labour Protection-National Research Institute, Czerniakowska 16 Street, 00-701, Warsaw, Poland.
authors.forename: Agata
authors.initials: A
authors.lastname: Stobnicka
authors.affiliation: Biohazard Laboratory, Department of Chemical, Aerosol and Biological Hazards, Central Institute for Labour Protection-National Research Institute, Czerniakowska 16 Street, 00-701, Warsaw, Poland.
authors.forename: Rafał L
authors.initials: RL
authors.lastname: Górny
mesh_terms: D000391:Air Microbiology; D016902:Air Pollution, Indoor / Q000032:analysis*; D001419:Bacteria / Q000145:classification / Q000302:isolation & purification*; D018533:Biomass; D015169:Colony Count, Microbial; D004784:Environmental Monitoring / Q000379:methods; D006225:Hand / Q000382:microbiology; D006801:Humans; D016273:Occupational Exposure / Q000032:analysis*; D011044:Poland; D016133:Polymerase Chain Reaction; D011210:Power Plants / Q000706:statistics & numerical data*; D019105:Random Amplified Polymorphic DNA Technique; D012134:Respiratory Protective Devices / Q000382:microbiology; D017132:Workplace / Q000706:statistics & numerical data
keywords: Bacteria; Molecular typing; Occupational exposure; RAPD-PCR; Source tracking