Distribution of CRISPR Types in Fluoroquinolone-Resistant Campylobacter jejuni Isolates

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.

Prevalence and antimicrobial resistance of Campylobacter species in shelter‐housed healthy and diarrheic cats and dogs in Turkey

“…Both quinolone and tetracycline resistance can spread rapidly in Campylobacter species and are of particular concern with regards to the potential for future treatment failures (Luangtongkum et al., 2009). In the present study, the most common mutation in the QRDR region of the gyrA gene was Thr‐86‐Ile in the quinolone‐resistant isolates, which has also been reported in previous studies (Adıgüzel et al., 2018; Acke et al., 2009; Adiguzel et al., 2021). In contrast to other studies, the phenotypically tetracycline‐ and gentamicin‐resistant strains did not carry the tetO or aphA‐3 genes, respectively.…”

“…In the present study, the most common mutation in the QRDR region of the gyrA gene was Thr-86-Ile in the quinolone-resistant isolates, which has also been reported in previous studies (Adıgüzel et al, 2018;Acke et al, 2009;Adiguzel et al, 2021). In contrast to other studies, the phenotypically tetracycline-and gentamicin-resistant strains did not carry the tetO or aphA-3 genes, respectively.…”

“…The sequence results were analysed with Chromas software (version 2.6.6 https://technelysium.com.au/wp/ chromas/ accessed 06.04.2023). The data were compared in the NCBI Blast database using C. jejuni NCTC 11168 gyrA gene, DNA gyrase subunit A (NCBI accession number:NC_002163.1) as the reference sequence (Adiguzel et al, 2021).…”

Campylobacter jejuni Developed the Resistance to Bacteriophage CP39 by Phase Variable Expression of 06875 Encoding the CGPTase

“…Both phage CP39 and Los1 belonged to the family Myoviridae with a similar genome size of 130,715 bp and 134,073 bp, respectively, indicating that phage CP39 belonged to Group III phage of the Cp8unalikevirus [17,23,24]. The phylogenetic analysis of CP39 from other studies has also proven our results in that CP39 was cluttered to the group III Campylobacter phage [25,26]. The group III phage was known to exhibit a strong lytic activity, and its successful application could effectively decrease the contamination load C. jejuni in chickens [27].…”

Detection of Campylobacter jejuni diversity by clustered regularly interspaced short palindromic repeats (CRISPR) from an animal farm

“…A database that integrates both MLST sequences and CRISPR sequences and is searchable is greatly in demand. During the revision of this manuscript, a study demonstrates the CRISPR–Cas system is prevalent in the fluoroquinolone‐resistant C. jejuni isolates (Adiguzel et al., 2021 ).…”