Helicobacter pylori in a poultry slaughterhouse: Prevalence, genotyping and antibiotic resistance pattern

Abstract

Although Helicobacter pylori (H. pylori) is a highly significant pathogen, its source remains unclear. Many people consume chicken daily as a source of animal protein worldwide; thus, hygienic methods of supplying chickens for consumption are critical for public health. Therefore, our study examined the distribution of the glmM (ureC), babA2, vacA and cagA virulence genes in H. pylori strains in chicken meat and giblets (gizzards and livers) and the resistance of the strains to various antibiotics. Ninety chicken meat, gizzard and liver samples were obtained from a semi-automatic abattoir in Sadat City, Egypt, and were cultured and preliminarily analyzed using biochemical tests. The presence of the ureC, babA2, vacA and cagA genotypes was tested for in samples positive for H. pylori by multiplex polymerase chain reaction (Multiplex-PCR). The resistance of H. pylori to various antimicrobial drugs was tested using the disc diffusion method. In total, 7 of the 90 chicken samples were positive for H. pylori (7.78%); in 3/7 (42.85%) samples, the bacteria were found in the chicken liver, while the bacteria were found in the meat in 2/7 (28.57%) and in the gizzard in 2/7 (28.57%) samples. The total prevalence of both the ureC and babA2 genes in the isolated H. pylori strains was 100%, while the prevalence of the vacA and cagA genes was 57.1% and 42.9%, respectively. The resistance of H. pylori to the antibiotics utilized in our study was 100% for streptomycin; 85.7% for amoxicillin and penicillin; 71.4% for oxytetracycline, nalidixic acid and ampicillin; 57.1% for sulfamethoxazole and erythromycin; and 42.9% for neomycin, chloramphenicol and norfloxacin. In conclusion, the chicken meat and giblets were tainted by H. pylori, with a higher occurrence of the ureC, babA2, vacA and cagA genotypes. Future investigations should investigate the resistance of H. pylori to various antimicrobial agents in Egypt.

Keywords: Antibiotic resistance; Chickens; Egypt; Helicobacter pylori; Virulence genes.

Fig. 1

Agarose gel electrophoresis of PCR amplification products using 16S rRNA (375 bp) as a specific primer to identify the Helicobacter species. Lane M: 100-bp ladder as molecular DNA marker; lane C+: positive control for 16S rRNA of Helicobacter species; lane C−: negative control. Lanes 1–7: positive H. pylori; lanes 8–11: positive H. pullorum; lanes 12–13: positive H. cinaedi; lane 15: positive H. bilis; lane 16: positive H. hepaticus; and lane 14: negative Helicobacter species.

Fig. 2

Agarose gel electrophoresis of PCR of the ureC gene (294 bp) for the characterization of the H. pylori strains. Lane M: 100-bp ladder as molecular size DNA marker; lane C+: positive control H. pylori for ureC gene; lane C−: negative control; and lanes 1–7: positive H. pylori strains.

Fig. 3

Agarose gel electrophoresis of Multiplex-PCR of babA2 (259 bp), cagA (350 bp) and vacA (567 bp) as virulence genes of H. pylori strains. Lane M: 100-bp ladder as molecular size DNA marker; lane C+: positive control strain for the babA2, cagA and vacA genes; lane C−: negative control. Lane 1: positive H. pylori strain for the babA2 gene. Lanes 2, 4 and 7: positive H. pylori strains for the babA2 and vacA genes. Lanes 3 and 6: positive H. pylori strains for the babA2 and cagA genes. Lane 5: positive H. pylori strain for the babA2, cagA and vacA genes.