Prevalence, Antimicrobial Susceptibility, Virulence and Genotyping of Campylobacter jejuni with a Special Reference to the Anti-Virulence Potential of Eugenol and Beta-Resorcylic Acid on Some Multi-Drug Resistant Isolates in Egypt

Ahmed M Ammar; El-Sayed Y El-Naenaeey; Rania M S El-Malt; Attia A El-Gedawy; Eman Khalifa; Shimaa S Elnahriry; Marwa I Abd El-Hamid

doi:10.3390/ani11010003

Prevalence, Antimicrobial Susceptibility, Virulence and Genotyping of Campylobacter jejuni with a Special Reference to the Anti-Virulence Potential of Eugenol and Beta-Resorcylic Acid on Some Multi-Drug Resistant Isolates in Egypt

Animals (Basel). 2020 Dec 22;11(1):3. doi: 10.3390/ani11010003.

Authors

Ahmed M Ammar¹, El-Sayed Y El-Naenaeey¹, Rania M S El-Malt², Attia A El-Gedawy³, Eman Khalifa⁴, Shimaa S Elnahriry⁵, Marwa I Abd El-Hamid¹

Affiliations

¹ Department of Microbiology, Faculty of Veterinary Medicine, Zagazig University, Zagazig 44519, Egypt.
² Department of Microbiology, Animal Health Research Institute, Zagazig 44516, Egypt.
³ Tuberculosis Unit, Department of Bacteriology, Animal Health Research Institute, Giza 12618, Egypt.
⁴ Department of Microbiology, Faculty of Veterinary Medicine, Matrouh University, Matrouh 51511, Egypt.
⁵ Department of Bacteriology, Mycology and Immunology, Faculty of Veterinary Medicine, University of Sadat City, Menofia 32897, Egypt.

Abstract

Campylobacter jejuni is the leading cause of foodborne bacterial gastroenteritis in humans worldwide. Contaminated chickens and their products are the main sources of human campylobacteriosis. Therefore, this study aimed to detect the genotypic and virulence genes' profiles of multi-drug resistant (MDR) C. jejuni isolates and to assess the effects of sub-inhibitory concentrations (SICs) of eugenol and beta-resorcylic acid on the virulence of avian MDR C. jejuni isolates. These isolates were clustered together with the human isolates via enterobacterial repetitive intergenic consensus-PCR (ERIC-PCR) fingerprinting. A total of 345 samples were collected from human stool (100) and different chicken (245) samples in Sharkia Governorate, Egypt. Conventional phenotypic methods identified 113 isolates (32.8%) as C. jejuni, and all C. jejuni isolates were MDR and resistant to erythromycin and ampicillin. The genes virB11, wlaN, and flaA were detected in 52%, 36% and 100% strains, respectively. ERIC-PCR yielded 14 profiles and five main clusters. Interestingly, human and chicken C. jejuni isolates were clustered together in ERIC-PCR clusters II-V, which confirmed the genetic relatedness between the isolates from both origins. Beta-resorcylic acid and eugenol inhibited the invasion of C. jejuni isolates to chicken intestinal cells by 41.66-38.19% and 31.94-29.16%, respectively, and minimized the transcription of flaA, virB11, and wlaN genes in the tested isolates by real-time quantitative reverse transcription PCR (qRT-PCR). In essence, eugenol and beta-resorcylic acid are promising natural antimicrobials for minimizing the virulence of MDR C. jejuni in chickens, thereby managing human campylobacteriosis.

Keywords: Campylobacter jejuni; ERIC-PCR; antimicrobial resistance; beta-resorcylic acid; broiler chickens; cell line; eugenol; invasion; virulence; virulence genes expression.