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      Monitoring of bacterial pathogens at workplaces in power plant using biochemical and molecular methods

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          Abstract

          Purpose

          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 × 10 6cfu/m 3 and 3.8 × 10 4cfu/g, respectively. The number of bacteria in the swab and mask samples also reached a high level of 1.4 × 10 4 cfu/ml and 1.9 × 10 3 cfu/cm 2, 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%.

          Conclusion

          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.

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          Most cited references44

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          The role of the surface environment in healthcare-associated infections.

          This article reviews the evidence demonstrating the importance of contamination of hospital surfaces in the transmission of healthcare-associated pathogens and interventions scientifically demonstrated to reduce the levels of microbial contamination and decrease healthcare-associated infections.
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            Walking-induced particle resuspension in indoor environments

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              Culture-independent methods for studying environmental microorganisms: methods, application, and perspective.

              Since the application of molecular methods, culture-independent methods (CIMs) have been developed to study microbial communities from various environments. In the past 20 years, several methods based on the direct amplification and analyses of the small subunit ribosomal RNA gene have been developed to directly study environmental microorganisms. These methods include denaturing/temperature gradient gel electrophoresis, single-strand-conformation polymorphism, restriction fragment length polymorphism, terminal restriction fragment length polymorphism, and quantitative polymerase chain reaction (PCR). Similarly, non-PCR-based molecular techniques, such as microarray and fluorescence in situ hybridization have also been adopted. In recent years, several novel fields of investigation such as metagenomics, metatranscriptomics, metaproteomics, and single-cell genomics were developed, largely propelled by the innovation and application of next-generation sequencing methods. Several single-cell-based technologies such as Raman microspectroscopy and nano-scale secondary ion mass spectrometry are also increasingly used in the fields of microbial ecology and environmental microbiology. The application of these methods has revolutionized microbiology by allowing scientists to directly analyze natural microbial communities in situ, including their genes, transcripts, proteins, and metabolites and how their interactions impact their distribution patterns. In this review, we present an up-to-date review on different CIMs and their applications, our focuses are on the comparison of different CIMs and their application in the analyses of microbial diversities and communities.
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                Author and article information

                Contributors
                +48226234692 , anlaw@ciop.pl
                Journal
                Int Arch Occup Environ Health
                Int Arch Occup Environ Health
                International Archives of Occupational and Environmental Health
                Springer Berlin Heidelberg (Berlin/Heidelberg )
                0340-0131
                1432-1246
                25 January 2017
                25 January 2017
                2017
                : 90
                : 3
                : 285-295
                Affiliations
                ISNI 0000 0001 2370 2644, GRID grid.460598.6, Biohazard Laboratory, Department of Chemical, Aerosol and Biological Hazards, , Central Institute for Labour Protection–National Research Institute, ; Czerniakowska 16 Street, 00-701 Warsaw, Poland
                Author information
                http://orcid.org/0000-0001-8234-340X
                Article
                1197
                10.1007/s00420-017-1197-z
                5360828
                28124138
                410bb2ce-d2ba-4441-b919-4cf4c7aa4b32
                © The Author(s) 2017

                Open AccessThis article is distributed under the terms of the Creative Commons Attribution 4.0 International License ( http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made.

                History
                : 4 October 2016
                : 2 January 2017
                Funding
                Funded by: the Ministry of Science and Higher Education/National Centre for Research and Development
                Award ID: II.P.16
                Categories
                Original Article
                Custom metadata
                © Springer-Verlag Berlin Heidelberg 2017

                Occupational & Environmental medicine
                molecular typing,rapd-pcr,occupational exposure,bacteria,source tracking

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