The dialogue between protozoa and bacteria in a microfluidic device


Autoři: Anna Gaines aff001;  Miranda Ludovice aff001;  Jie Xu aff001;  Marc Zanghi aff001;  Richard J. Meinersmann aff002;  Mark Berrang aff002;  Wayne Daley aff001;  Doug Britton aff001
Působiště autorů: Aerospace, Transportation and Advanced Systems Laboratory, Georgia Tech Research Institute, Georgia Institute of Technology, Atlanta, Georgia, United States of America aff001;  Richard B. Russell Research Center, Agricultural Research Service, United States Department of Agriculture, Athens, Georgia, United States of America aff002
Vyšlo v časopise: PLoS ONE 14(10)
Kategorie: Research Article
doi: 10.1371/journal.pone.0222484

Souhrn

In nature, protozoa play a major role in controlling bacterial populations. This paper proposes a microfluidic device for the study of protozoa behaviors change due to their chemotactic response in the presence of bacterial cells. A three-channel microfluidic device was designed using a nitrocellulose membrane into which channels were cut using a laser cutter. The membrane was sandwiched between two glass slides; a Euglena suspension was then allowed to flow through the central channel. The two side channels were filled with either, 0.1% peptone as a negative control, or a Listeria suspension respectively. The membrane design prevented direct interaction but allowed Euglena cells to detect Listeria cells as secretions diffused through the nitrocellulose membrane. A significant number of Euglena cells migrated toward the chambers near the bacterial cells, indicating a positive chemotactic response of Euglena toward chemical cues released from Listeria cells. Filtrates collected from Listeria suspension with a series of molecular weight cutoffs (3k, 10k and 100k) were examined in Euglena chemotaxis tests. Euglena cells were attracted to all filtrates collected from the membrane filtration with different molecular weight cutoffs, suggesting small molecules from Listeria might be the chemical cues to attract protozoa. Headspace volatile organic compounds (VOC) released from Listeria were collected, spiked to 0.1% peptone and tested as the chemotactic effectors. It was discovered that the Euglena cells responded quickly to Listeria VOCs including decanal, 3,5- dimethylbenzaldehyde, ethyl acetate, indicating bacterial VOCs were used by Euglena to track the location of bacteria.

Klíčová slova:

Flow rate – Chemotaxis – Listeria – Microfluidics – Predation – Protozoans – Peptones – Nitrocellulose


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2019 Číslo 10

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