title: Molecular typing, serotyping and cytotoxicity testing of Campylobacter jejuni strains isolated from commercial broilers in Puerto Rico.
abstract: OBJECTIVE
  Thirty Campylobacter jejuni strains isolated from fecal samples (n = 94; 32%) from 13 positive farms (n = 17; 76%) from commercial broiler chickens in Puerto Rico were analysed by molecular methods.
  
  
  RESULTS
  Isolates were identified with multiplex polymerase chain reaction assays, tested for their antimicrobial susceptibility and characterized with pulsed-field gel electrophoresis (PFGE), multilocus sequence typing (MLST), serotyping and bacterial cytotoxicity in mammalian cells. Isolates exhibited high resistance to vancomycin (minimum inhibitory concentration, MIC of >256 microg ml(-1)) and trimethoprim (MIC of >32 microg ml(-1)); few were resistant to clindamycin (MIC(90) 4 microg ml(-1)), erythromycin (MIC(90) 8 microg ml(-1)) and tetracycline (MIC(90) 8 microg ml(-1)); but none was resistant to azithromycin (MIC(90) 4 microg ml(-1)), ciprofloxacin (MIC(90) 1 microg ml(-1)) or gentamycin (MIC(90) 4 microg ml(-1)). Most strains restricted with SmaI, but a combination of SmaI-KpnI digestion was more discriminatory. MLST analysis yielded four sequence types (ST), and ST-2624 was the predominant one. Phylogenetic analysis revealed a high degree of recombination for glnA and pgm genes. The predominant serotypes were O:3 and O:5. Most strains had lowest cytotoxicity potential with Caco-2 cells, medium cytotoxicity with INT-407 and Hep-2 cells and high cytotoxicity with CHO cells.
  
  
  CONCLUSIONS
  A low degree of antimicrobial resistance, 13 PFGE profiles, 4 ST and a large variability in cytotoxicity assays were found for these strains.
  
  
  CONCLUSIONS
  This is the first characterization of C. jejuni strains isolated from broilers in Puerto Rico. The genetic diversity of these strains suggests that several techniques are needed for strain characterization.
authors.affiliation: Department of Poultry Science, Auburn University, Auburn, Alabama, USA. oyarzoa@auburn.edu
authors.forename: O A
authors.initials: OA
authors.lastname: Oyarzabal
authors.forename: S
authors.initials: S
authors.lastname: Backert
authors.forename: L L
authors.initials: LL
authors.lastname: Williams
authors.forename: A J
authors.initials: AJ
authors.lastname: Lastovica
authors.forename: R S
authors.initials: RS
authors.lastname: Miller
authors.forename: S J
authors.initials: SJ
authors.lastname: Pierce
authors.forename: S L
authors.initials: SL
authors.lastname: Vieira
authors.forename: F
authors.initials: F
authors.lastname: Rebollo-Carrato
mesh_terms: D000818:Animals; D018938:Caco-2 Cells; D002169:Campylobacter Infections / Q000382:microbiology* / Q000662:veterinary*; D016123:Campylobacter jejuni / Q000145:classification* / Q000187:drug effects / Q000235:genetics; D002645:Chickens / Q000382:microbiology*; D003601:Cytotoxicity Tests, Immunologic; D052978:Disk Diffusion Antimicrobial Tests; D005516:Food Microbiology; D005838:Genotype; D006801:Humans; D008826:Microbial Sensitivity Tests; D011201:Poultry Diseases / Q000382:microbiology*; D011647:Puerto Rico; D012703:Serotyping; D020713:Vancomycin Resistance