Distribution of CRISPR Types in Fluoroquinolone-Resistant Campylobacter jejuni Isolates

Mehmet Cemal Adiguzel; Debora Brito Goulart; Zuowei Wu; Jinji Pang; Seyda Cengiz; Qijing Zhang; Orhan Sahin

doi:10.3390/pathogens10030345

Distribution of CRISPR Types in Fluoroquinolone-Resistant Campylobacter jejuni Isolates

Pathogens. 2021 Mar 16;10(3):345. doi: 10.3390/pathogens10030345.

Authors

Mehmet Cemal Adiguzel^{1

2}, Debora Brito Goulart², Zuowei Wu², Jinji Pang², Seyda Cengiz¹, Qijing Zhang², Orhan Sahin³

Affiliations

¹ Department of Microbiology, College of Veterinary Medicine, Ataturk University, Erzurum 25240, Turkey.
² Department of Veterinary Microbiology and Preventive Medicine, College of Veterinary Medicine, Iowa State University, Ames, IA 50011, USA.
³ Department of Veterinary Diagnostic and Production Animal Medicine, College of Veterinary Medicine, Iowa State University, Ames, IA 50011, USA.

Abstract

To aid development of phage therapy against Campylobacter, we investigated the distribution of the clustered regularly interspaced short palindromic repeats (CRISPR) systems in fluoroquinolone (FQ)-resistant Campylobacter jejuni. A total of 100 FQ-resistant C. jejuni strains from different sources were analyzed by PCR and DNA sequencing to determine resistance-conferring mutation in the gyrA gene and the presence of various CRISPR systems. All but one isolate harbored 1-5 point mutations in gyrA, and the most common mutation was the Thr86Ile change. Ninety-five isolates were positive with the CRISPR PCR, and spacer sequences were found in 86 of them. Among the 292 spacer sequences identified in this study, 204 shared 93-100% nucleotide homology to Campylobacter phage D10, 44 showed 100% homology to Campylobacter phage CP39, and 3 had 100% homology with Campylobacter phage CJIE4-5. The remaining 41 spacer sequences did not match with any phages in the database. Based on the results, it was inferred that the FQ-resistant C. jejuni isolates analyzed in this study were potentially resistant to Campylobacter phages D10, CP39, and CJIE4-5 as well as some unidentified phages. These phages should be excluded from cocktails of phages that may be utilized to treat FQ-resistant Campylobacter.

Keywords: CRISPR-Cas system; Campylobacter jejuni; Cas9 gene; fluoroquinolone-resistant bacteria.