Isolation, identification, and pathological effects of beach sand bacterial extract on human skin keratinocytes in vitro

Fazli Subhan; Raheem Shahzad; Isfahan Tauseef; Kashif Syed Haleem; Atta-Ur Rehman; Sajid Mahmood; In-Jung Lee

doi:10.7717/peerj.4245

Isolation, identification, and pathological effects of beach sand bacterial extract on human skin keratinocytes in vitro

PeerJ. 2018 Jan 12:6:e4245. doi: 10.7717/peerj.4245. eCollection 2018.

Authors

Fazli Subhan^#¹, Raheem Shahzad^#², Isfahan Tauseef¹, Kashif Syed Haleem¹, Atta-Ur Rehman³, Sajid Mahmood³, In-Jung Lee^{2

4}

Affiliations

¹ Department of Microbiology, Hazara University, Mansehra, Pakistan.
² School of Applied Biosciences, Kyungpook National University, Daegu, Republic of Korea.
³ Department of Zoology, Hazara University, Mansehra, Pakistan.
⁴ Research Institute for Dok-do and Ulleung-do Island, Kyungpook National University, Daegu, Republic of Korea.

^# Contributed equally.

Abstract

Background: Beaches are recreational spots for people. However, beach sand contains harmful microbes that affect human health, and there are no established methods for either sampling and identifying beach-borne pathogens or managing the quality of beach sand.

Method: This study was conducted with the aim of improving human safety at beaches and augmenting the quality of the beach experience. Beach sand was used as a resource to isolate bacteria due to its distinctive features and the biodiversity of the beach sand biota. A selected bacterial isolate termed FSRS was identified as Pseudomonas stutzeri using 16S rRNA sequencing and phylogenetic analysis, and the sequence was deposited in the NCBI GenBank database under the accession number MF599548. The isolated P. stutzeri bacterium was cultured in Luria-Bertani growth medium, and a crude extract was prepared using ethyl acetate to examine the potential pathogenic effect of P. stutzeri on human skin. A human skin keratinocyte cell line (HaCaT) was used to assess cell adhesion, cell viability, and cell proliferation using a morphological analysis and a WST-1 assay.

Result: The crude P. stutzeri extract inhibited cell adhesion and decreased cell viability in HaCaT cells. We concluded that the crude extract of P. stutzeri FSRS had a strong pathological effect on human skin cells.

Discussion: Beach visitors frequently get skin infections, but the exact cause of the infections is yet to be determined. The beach sand bacterium P. stutzeri may, therefore, be responsible for some of the dermatological problems experienced by people visiting the beach.

Keywords: Bacteria; Beach sand; Pathology; Sequencing; Skin.

Grants and funding

This research was supported by the Basic Science Research Program through the National Research Foundation of Korea (NRF) funded by the Ministry of Education (2016R1A6A1A05011910). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.