Antibiotic resistance genes and integrons as indicators of biotic pollution and resistance load of Arctic ecosystems (ATTENTION)

The main aim of this study is the evaluation of impact that water flowing from melting glaciers, permafrost and wastewater discharged directly into fjords in the Arctic region exert on increasing the genes pool of the global aquatic resistome.

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  • marine biology
  • cryosphere

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  • human dimensions / environmental impacts / contaminants

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According to WHO, rapid growth and spread of antibiotic resistance among bacteria is an indisputable global problem and one of the greatest threat to public health in the 21st century. The diversity and spread of antibiotic resistance genes (ARGs) in fragile and fast-changing polar regions is strongly understudied and only basically recognized. Even though remote and with limited human impact, Arctic ecosystems are bound to receive antibiotic resistance through wastewater delivered to the fjords, from melting glaciers and permafrost, which are thought to be repositories of ARGs. Therefore, the main aim of this study is the evaluation of impact that water flowing from melting glaciers, permafrost and wastewater discharged directly into fjords exert on increasing the genes pool of the global aquatic resistome, being the collection of all ARGs in the environment. The proposed research aims at determining the type and frequency of antibiotic-resistant bacteria (ARB), integrase genes (intI1) as a marker of anthropogenic pollution and ARGs in cryospheric resistome and the wastewater in the Svalbard archipelago, which is an example of remote and a relatively pristine polar area. An integrative approach combining a metagenomic and classical culturing methods will be used in the project. In addition, plasmidome will be examined in order to determine the abundance and identify the diversity of ARGs and integrons transmitted via mobile genetic elements (MGEs). The essence of the project is to investigate antibiotic resistance in Spitsbergen characterized by the environments seemingly free from human impact. The study also seeks to recognize the role of WWTPs in the emergence and accelerating the dissemination of ARGs in the highly human-impacted aquatic environments of this Arctic region. Literature data report that wastewater treatment processes do not eliminate ARB, ARGs and integrons. Selection pressure and a horizontal gene transfer in a wastewater treatment plant (WWTP) lead to an increased frequency in the final effluent of bacteria with particularly dangerous mechanisms of resistance, ARGs and integrons, that pose the greatest threat to public health. Moreover, preliminary research has indicated that a glacier in the Svalbard archipelago contains ARB, ARGs and integrons. Taking into account that the role of human impact in this phenomenon as well as sources and the spread of ARGs in Arctic ecosystems are all poorly studied. Each year during polar summer samples will be collected from a WWTP in Ny-Ålesund (raw sewage, final effluent), Kongsfjord (water, sediment), the capital city of Longyearbyen (wastewater stream), Adventfjorden (water, sediment), Larsbreen and Nordenskiöldbreen glaciers (ice), tundra and glacier forefields (permafrost cores). The samples will be collected in the way that to do not influence Svalbard environment. A qualitative analysis of ARGs, integrons and gene cassettes will be performed by conventional PCR. A quantitative analysis of ARG and integrons in the metagenome and plasmidome will be performed using the qPCR and ddPCR methods. The investigation of abundance and diversity of ARGs and variable regions of class 1 integrons in metagenome and plasmidome DNA will be performed by Illumina high-throughput sequencing. To analyze bacterial populations, V3-V4 hypervariable regions of 16S rRNA gene will be sequenced. The idea behind the proposed research is included in the Arctic Monitoring & Assessment Program (AMAP) of the Working Group of the Arctic Council. One of the main goals of AMAP published in a report from 2018 is also to take global action and develop an effective strategy for monitoring and assessing the unknown pathways of the spread of antibiotic resistance in the Arctic that results from human-induced contamination. All these activities are a part of the global holistic "One Health" approach that is to combat the constantly growing and spreading antibiotic resistance in the environment.

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