Measuring the Prevalence of Campylobacter Species in Diarrheal Samples of Children under 10 Years and Detecting the Ability of Biofilm Production

Background: Campylobacter spp. is one of the most common causes of diarrheal diseases all over the world, with a rapid acquisition of antibiotic resistance. Campylobacter is 1 of 4 key global causes of diarrhoeal diseases.

Objective: To detect the frequency of Campylobacter species in diarrheal stool of children under ten years by conventional and molecular methods, and detect the bacterial ability to produce biofilm.

Methods: A total of 200 children presented with diarrhea were enrolled in this study. Campylobacter was isolated and diagnosed primarily by characteristic features on Gram stain, culture media, biochemical tests, and tested against 13 antibiotics by disc diffusion method. The ability of biofilm production was tested by crystal violet quantitative ELISA microtiter plate assays. Then Campylobacter spp was detected by Multiplex PCR using species specific genes.

Results: The prevalence of Campylobacter spp. was 17% by conventional methods and 15% by molecular method. The results of antibiotic susceptibility test showed that there is complete resistance (100%) to cephalothin, ampicillin, and clindamycin for both species, full resistance (100%) to trimethoprim-sulfamethoxazol and erythromycin by C. coli, and high resistance (92.3%) to trimethoprim-sulfamethoxazol and erythromycin by C. jejuni, while the lowest resistance was to nalidixic acid and amikacin (7.7%) by C. jejuni, and (12.5%) to tetracyclin, amoxicillin, amikacin, chloramphenicol, and ciprofloxacin by C. coli. Statistically, there is no significant difference between biofilm formation and antibiotic resistance in Campylobacter species.

The frequency of biofilm production in all positive Skirrow’s culture was (35.29%) as 12 out of 34 positive isolates, ranging from mild to severe biofilm formation. By PCR assay, 64.7 % (22 of 34) positive Skirrow’s culture were also positive based on hipO gene specific for C. jejuni, while the prevalence of asp gene was (23.5%).

Conclusion: The prevalence of Campylobacter spp. was 17% by conventional methods and 15% by molecular methods. Most Campylobacter isolates were MDR and sensitive to a limited number of antibiotics. Many isolates produced biofilm. A significant correlation was found between hippurate hydrolysis and molecular detection of Campylobacter spp. using hipO and asp genes.