1887

Abstract

and are two of the most common causative agents of food-borne gastroenteritis in numerous countries worldwide. In Brazil, campylobacteriosis is underdiagnosed and under-reported, and few studies have molecularly characterized spp. in this country. The current study genotyped 63 strains isolated from humans ( = 12), animals ( = 21), food ( = 10) and the environment ( = 20) between 1995 and 2011 in Brazil. The strains were genotyped using pulsed-field gel electrophoresis (PFGE), sequencing the short variable region (SVR) of the gene (-SVR) and high-resolution melting analysis (HRMA) of the clustered regularly interspaced short palindromic repeat (CRISPR) locus to better understand genotypic diversity and compare the suitability of these three methods for genotyping this species. Additionally, the discrimination index (DI) of each of these methods was assessed. Some strains isolated from clinical and non-clinical origins presented ≥ 80 % genotypic similarity by PFGE and -SVR sequencing. HRMA of the CRISPR locus revealed only four different melting profiles. In total, 22 different A-SVR alleles were detected. Of these, seven alleles, comprising gt1647–gt1653, were classified as novel. The most frequent genotypes were gt30 and gt1647.This distribution reveals the diversity of selected Brazilian isolates in comparison with the alleles described in the PubMLST database. The DIs for PFGE, –SVR sequencing and CRISPR-HRMA were 0.986, 0.916 and 0.550, respectively. PFGE and -SVR sequencing were suitable for subtyping strains, in contrast to CRISPR-HRMA. The high genomic similarity amongst some strains confirms the hypothesis that environmental and food sources potentially lead to human and animal contamination in Brazil.

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2016-01-01
2020-01-25
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References

  1. Abay S., Kayman T., Otlu B., Hizlisoy H., Aydin F., Ertas N.. 2014; Genetic diversity and antibiotic resistance profiles of Campylobacter jejuni isolates from poultry and humans in Turkey. Int J Food Microbiol178:29–38 [CrossRef][PubMed]
    [Google Scholar]
  2. Acke E., McGill K., Lawlor A., Jones B. R., Fanning S., Whyte P.. 2010; Genetic diversity among Campylobacter jejuni isolates from pets in Ireland. Vet Rec166:102–106 [CrossRef][PubMed]
    [Google Scholar]
  3. Anderson J., Horn B. J., Gilpin B. J.. 2012; The prevalence and genetic diversity of Campylobacter spp. in domestic ‘backyard’ poultry in Canterbury, New Zealand. Zoonoses Public Health59:52–60 [CrossRef][PubMed]
    [Google Scholar]
  4. Andrade M. C. R., Gabeira S. C. O., Lopes D. A., Esteves T. C., Vilardo M. C., Thomé J. D. S., Cabello P. H., Filgueiras A. L. L.. 2007; Circulation of Campylobacter spp. in rhesus monkeys (Macaca mulatta) held in captivity: a longitudinal study. Mem Inst Oswaldo Cruz102:53–57[CrossRef]
    [Google Scholar]
  5. Aquino M. H. C., Pacheco A. P. G., Ferreira M. C. S., Tibana A.. 2002; Frequency of isolation and identification of thermophilic campylobacters from animals in Brazil. Vet J164:159–161 [CrossRef][PubMed]
    [Google Scholar]
  6. Aquino M. H. C., Filgueiras A. L. L., Matos R., Santos K. R. N., Ferreira T., Ferreira M. C. S., Teixeira L. M., Tibana A.. 2010; Diversity of Campylobacter jejuni and Campylobacter coli genotypes from human and animal sources from Rio de Janeiro, Brazil. Res Vet Sci88:214–217 [CrossRef][PubMed]
    [Google Scholar]
  7. Blaser M. J., Perez G. P., Smith P. F., Patton C., Tenover F. C., Lastovica A. J., Wang W. I.. 1986; Extraintestinal Campylobacter jejuni and Campylobacter coli infections: host factors and strain characteristics. J Infect Dis153:552–559 [CrossRef][PubMed]
    [Google Scholar]
  8. Campioni F., Falcão J. P.. 2014; Genotypic diversity and virulence markers of Yersinia enterocolitica biotype 1A strains isolated from clinical and non-clinical origins. APMIS122:215–222 [CrossRef][PubMed]
    [Google Scholar]
  9. Centers for Disease Control and Prevention (CDC) 2015; Foodborne Diseases Active 378 Surveillance Network (FoodNet): FoodNet Surveillance Report for 2011 Washington, DC: Department of Health and Human Services;
    [Google Scholar]
  10. Centers for Disease Control and Prevention (CDC) 2015; National Center for Emerging and Zoonotic Infectious Diseases Avaliable at: http://www.cdc.gov/nczved/divisions/dfbmd/diseases/campylobacter/
    [Google Scholar]
  11. da Silva Quetz J., Lima I. F. N., Havt A., de Carvalho E. B., Lima N. L., Soares A. M., Mota R. M. S., Guerrant R. L., Lima A. A. M.. 2010; Campylobacter jejuni and Campylobacter coli in children from communities in Northeastern Brazil: molecular detection and relation to nutritional status. Diagn Microbiol Infect Dis67:220–227 [CrossRef][PubMed]
    [Google Scholar]
  12. Denis M., Soumet C., Rivoal K., Ermel G., Blivet D., Salvat G., Colin P.. 1999; Development of a m-PCR assay for simultaneous identification of Campylobacter jejuni and C. coli . Lett Appl Microbiol29:406–410 [CrossRef][PubMed]
    [Google Scholar]
  13. Duarte A., Santos A., Manageiro V., Martins A., Fraqueza M. J., Caniça M., Domingues F. C., Oleastro M.. 2014; Human, food and animal Campylobacter spp. isolated in Portugal: high genetic diversity and antibiotic resistance rates. Int J Antimicrob Agents44:306–313 [CrossRef][PubMed]
    [Google Scholar]
  14. EFSA 2015; The European Union summary report on trends and sources of zoonoses, zoonotic agents and food-borne outbreaks in 2013. EFSA J13:3991
    [Google Scholar]
  15. Ertaş H. B., Cetinkaya B., Muz A., Ongör H.. 2004; Genotyping of broiler-originated Campylobacter jejuni and Campylobacter coli isolates using fla typing and random amplified polymorphic DNA methods. Int J Food Microbiol94:203–209 [CrossRef][PubMed]
    [Google Scholar]
  16. Fica A., Seelmann D., Porte L., Eugenin D., Gallardo R.. 2012; A case of myopericarditis associated to Campylobacter jejuni infection in the southern hemisphere. Braz J Infect Dis16:294–296 [CrossRef][PubMed]
    [Google Scholar]
  17. Franchin P. R., Ogliari P. J., Batista C. R. V.. 2007; Frequency of thermophilic Campylobacter in broiler chickens during industrial processing in a Southern Brazil slaughterhouse. Br Poult Sci48:127–132 [CrossRef][PubMed]
    [Google Scholar]
  18. Friedman C. R., Hoekstra R. M., Samuel M., Marcus R., Bender J., Shiferaw B., Reddy S., Ahuja S. D., Helfrick D. L., other authors. 2004; Risk factors for sporadic Campylobacter infection in the United States: a case-control study in FoodNet sites. Clin Infect Dis38:(Suppl. 3)S285–S296 [CrossRef][PubMed]
    [Google Scholar]
  19. Fry B. N., Wosten M., Wassenaar T. M., van der Zeijst B. A. M.. 2000; Transformation of Campylobacter jejuni . In Electrotransformation of Bacteria pp157–167 Edited by Eynard N., Teissie J.. Berlin: Springer;[CrossRef]
    [Google Scholar]
  20. Giacomelli M., Andrighetto C., Rossi F., Lombardi A., Rizzotti L., Martini M., Piccirillo A.. 2012; Molecular characterization and genotypic antimicrobial resistance analysis of Campylobacter jejuni and Campylobacter coli isolated from broiler flocks in northern Italy. Avian Pathol41:579–588 [CrossRef][PubMed]
    [Google Scholar]
  21. Gillespie I. A., O'Brien S. J., Frost J. A., Adak G. K., Horby P., Swan A. V., Painter M. J., Neal K. R., Campylobacter Sentinel Surveillance Scheme Collaborators. 2002; A case–case comparison of Campylobacter coli and Campylobacter jejuni infection: a tool for generating hypotheses. Emerg Infect Dis8:937–942 [CrossRef][PubMed]
    [Google Scholar]
  22. Gomes F. R., Curcio B. R., Ladeira S. R. L., Fernandez H., Meireles M. C. A.. 2006; Campylobacter jejuni occurrence in chicken fecal samples from small properties in Pelotas, southern Brazil. Braz J Infect Dis37:375–378
    [Google Scholar]
  23. Gürtler M., Alter T., Kasimir S., Fehlhaber K.. 2005; The importance of Campylobacter coli in human campylobacteriosis: prevalence and genetic characterization. Epidemiol Infect133:1081–1087 [CrossRef][PubMed]
    [Google Scholar]
  24. Hall G., Kirk M. D., Becker N., Gregory J. E., Unicomb L., Millard G., Stafford R., Lalor K., OzFoodNet Working Group. 2005; Estimating foodborne gastroenteritis, Australia. Emerg Infect Dis11:1257–1264 [CrossRef][PubMed]
    [Google Scholar]
  25. Harrington C. S., Thomson-Carter F. M., Carter P. E.. 1997; Evidence for recombination in the flagellin locus of Campylobacter jejuni: implications for the flagellin gene typing scheme. J Clin Microbiol35:2386–2392[PubMed]
    [Google Scholar]
  26. Health Protection Surveillance Centre 2005; Campylobacteriosis in Ireland. 2005 Annual Report The Health Protection Surveillance Centre; 25–27 Middle Gardiner Street, Dublin 1, Ireland: pp34–36
    [Google Scholar]
  27. Hunter P. R., Gaston M. A.. 1988; Numerical index of the discriminatory ability of typing systems: an application of Simpson's index of diversity. J Clin Microbiol26:2465–2466[PubMed]
    [Google Scholar]
  28. Jansen R., Embden J. D., Gaastra W., Schouls L. M.. 2002; Identification of genes that are associated with DNA repeats in prokaryotes. Mol Microbiol43:1565–1575 [CrossRef][PubMed]
    [Google Scholar]
  29. Lazou T., Houf K., Soultos N., Dovas C., Iossifidou E.. 2014; Campylobacter in small ruminants at slaughter: prevalence, pulsotypes and antibiotic resistance. Int J Food Microbiol173:54–61 [CrossRef][PubMed]
    [Google Scholar]
  30. Lévesque S., Michaud S., Arbeit R. D., Frost E. H.. 2011; High-resolution melting system to perform multilocus sequence typing of Campylobacter jejuni . PLoS One6:e16167 [CrossRef][PubMed]
    [Google Scholar]
  31. Lyhs U., Katzav M., Isohanni P., Heiska H., Maijala R.. 2010; The temporal, PFGE and resistance pattern associations suggest that poultry products are only a minor source of human infections in western Finland. Food Microbiol27:311–315 [CrossRef][PubMed]
    [Google Scholar]
  32. Magnússon S. H., Guðmundsdóttir S., Reynisson E., Rúnarsson A. R., Harðardóttir H., Gunnarson E., Georgsson F., Reiersen J., Marteinsson V. T.. 2011; Comparison of Campylobacter jejuni isolates from human, food, veterinary and environmental sources in Iceland using PFGE, MLST and fla-SVR sequencing. J Appl Microbiol111:971–981 [CrossRef][PubMed]
    [Google Scholar]
  33. Meinersmann R. J., Helsel L. O., Fields P. I., Hiett K. L.. 1997; Discrimination of Campylobacter jejuni isolates by fla gene sequencing. J Clin Microbiol35:2810–2814[PubMed]
    [Google Scholar]
  34. Mojica F. J. M., Díez-Villaseñor C., Soria E., Juez G.. 2000; Biological significance of a family of regularly spaced repeats in the genomes of Archaea, Bacteria and mitochondria. Mol Microbiol36:244–246 [CrossRef][PubMed]
    [Google Scholar]
  35. Mojica F. J. M., Díez-Villaseñor C., García-Martínez J., Soria E.. 2005; Intervening sequences of regularly spaced prokaryotic repeats derive from foreign genetic elements. J Mol Evol60:174–182 [CrossRef][PubMed]
    [Google Scholar]
  36. Moore J. E., Corcoran D., Dooley J. S. G., Fanning S., Lucey B., Matsuda M., McDowell D. A., Mégraud F., Millar B. C., other authors. 2005; Campylobacter. Vet Res36:351–382 [CrossRef][PubMed]
    [Google Scholar]
  37. Mughini Gras L., Smid J. H., Wagenaar J. A., Koene M. G. J., Havelaar A. H., Friesema I. H. M., French N. P., Flemming C., Galson J. D., other authors. 2013; Increased risk for Campylobacter jejuni and C. coli infection of pet origin in dog owners and evidence for genetic association between strains causing infection in humans and their pets. Epidemiol Infect141:2526–2535 [CrossRef][PubMed]
    [Google Scholar]
  38. O'Leary A. M., Whyte P., Madden R. H., Cormican M., Moore J. E., Mc Namara E., Mc Gill K., Kelly L., Cowley D., other authors. 2011; Pulsed field gel electrophoresis typing of human and retail foodstuff Campylobacters: an Irish perspective. Food Microbiol28:426–433 [CrossRef][PubMed]
    [Google Scholar]
  39. Olsen S., Fitzgerald C., Swerdlow D.. 2001a; Limitations of pulsed-field gel electrophoresis for the routine surveillance of Campylobacter infections – reply. J Infect Dis184:243–244 [CrossRef]
    [Google Scholar]
  40. Olsen S. J., Hansen G. R., Bartlett L., Fitzgerald C., Sonder A., Manjrekar R., Riggs T., Kim J., Flahart R., other authors. 2001b; An outbreak of Campylobacter jejuni infections associated with food handler contamination: the use of pulsed-field gel electrophoresis. J Infect Dis183:164–167 [CrossRef][PubMed]
    [Google Scholar]
  41. Praakle-Amin K., Roasto M., Korkeala H., Hänninen M. L.. 2007; PFGE genotyping and antimicrobial susceptibility of Campylobacter in retail poultry meat in Estonia. Int J Food Microbiol114:105–112 [CrossRef][PubMed]
    [Google Scholar]
  42. Price E. P., Smith H., Huygens F., Giffard P. M.. 2007; High-resolution DNA melt curve analysis of the clustered, regularly interspaced short-palindromic-repeat locus of Campylobacter jejuni . Appl Environ Microbiol73:3431–3436 [CrossRef][PubMed]
    [Google Scholar]
  43. Quetz J. D., Lima I. F., Havt A., Prata M. M., Cavalcante P. A., Medeiros P. H., Cid D. A., Moraes M. L., Rey L. C., other authors. 2012; Campylobacter jejuni infection and virulence-associated genes in children with moderate to severe diarrhoea admitted to emergency rooms in northeastern Brazil. J Med Microbiol61:507–513[CrossRef]
    [Google Scholar]
  44. Ribot E. M., Fitzgerald C., Kubota K., Swaminathan B., Barrett T. J.. 2001; Rapid pulsed-field gel electrophoresis protocol for subtyping of Campylobacter jejuni . J Clin Microbiol39:1889–1894 [CrossRef][PubMed]
    [Google Scholar]
  45. Ross A. G. P., Olds G. R., Cripps A. W., Farrar J. J., McManus D. P.. 2013; Enteropathogens and chronic illness in returning travelers. N Engl J Med368:1817–1825 [CrossRef][PubMed]
    [Google Scholar]
  46. Roux F., Sproston E., Rotariu O., Macrae M., Sheppard S. K., Bessell P., Smith-Palmer A., Cowden J., Maiden M. C. J., other authors. 2013; Elucidating the aetiology of human Campylobacter coli infections. PLoS One8:e64504 [CrossRef][PubMed]
    [Google Scholar]
  47. Rozynek E., Antos-Bielska M., Dzierzanowska-Fangrat K., Szczepańska B., Trafny E. A.. 2010; Genetic similarity of Campylobacter isolates in humans, food, and water sources in central Poland. Foodborne Pathog Dis7:597–600 [CrossRef][PubMed]
    [Google Scholar]
  48. Sambrook J., Russell W. D.. 2001; Molecular Cloning: a Laboratory Manual, 3rd edn Cold Spring Harbor; NY: Cold Spring Harbor Laboratory
    [Google Scholar]
  49. Scallan E., Hoekstra R. M., Angulo F. J., Tauxe R. V., Widdowson M. A., Roy S. L., Jones J. L., Griffin P. M.. 2011; Foodborne illness acquired in the United States – major pathogens. Emerg Infect Dis17:7–15 [CrossRef][PubMed]
    [Google Scholar]
  50. Scarcelli E., Piatti R. M., Harakava R., Miyashiro S., Fernandes F. M. D., Campos F. R., Francisco W., Genovez M. E., Richtzenhain L. J.. 2005; Molecular subtyping of Campylobacter jejuni subsp jejuni strains isolated from different animal species in the state of Sao Paulo, Brazil. Braz J Infect Dis36:378–382
    [Google Scholar]
  51. Scarcelli E., Piatti R. M., Harakava R., Miyashiro S., Campos F. R., Souza M. C. A., Cardoso M. V., Teixeira S. R., Genovez M. E.. 2009; Use of pcr-rflp of the fla a gene for detection and subtyping of Campylobacter jejuni strains potentially related to Guillain-barré syndrome, isolated from humans and animals. Braz J Microbiol40:952–959 [CrossRef][PubMed]
    [Google Scholar]
  52. Selander B., Rydberg J., Lenner C., Hagerstrand I.. 1993; [Unusual infectious complication in a pregnant woman. Spontaneous abortion caused by Campylobacter coli]. Lakartidningen90:4356–4357 (in Swedish)
    [Google Scholar]
  53. Smith B. L., Lu C. P., Alvarado Bremer J. R.. 2010; High-resolution melting analysis (HRMA): a highly sensitive inexpensive genotyping alternative for population studies. Mol Ecol Resour10:193–196 [CrossRef][PubMed]
    [Google Scholar]
  54. Souza R. A., Falcão J. P.. 2012; A novel high-resolution melting analysis-based method for Yersinia pseudotuberculosis genotyping. J Microbiol Methods91:329–335 [CrossRef][PubMed]
    [Google Scholar]
  55. Wassenaar T. M., Newell D. G.. 2000; Genotyping of Campylobacter spp. Appl Environ Microbiol66:1–9 [CrossRef][PubMed]
    [Google Scholar]
  56. Wassenaar T. M., Geilhausen B., Newell D. G.. 1998; Evidence of genomic instability in Campylobacter jejuni isolated from poultry. Appl Environ Microbiol64:1816–1821[PubMed]
    [Google Scholar]
  57. Wassenaar T. M., Fernández-Astorga A., Alonso R., Marteinsson V. T., Magnússon S. H., Kristoffersen A. B., Hofshagen M.. 2009; Comparison of Campylobacter fla-SVR genotypes isolated from humans and poultry in three European regions. Lett Appl Microbiol49:388–395 [CrossRef][PubMed]
    [Google Scholar]
  58. Wirz S. E., Overesch G., Kuhnert P., Korczak B. M.. 2010; Genotype and antibiotic resistance analyses of Campylobacter isolates from ceca and carcasses of slaughtered broiler flocks. Appl Environ Microbiol76:6377–6386 [CrossRef][PubMed]
    [Google Scholar]
  59. Wittwer C. T.. 2009; High-resolution DNA melting analysis: advancements and limitations. Hum Mutat30:857–859 [CrossRef][PubMed]
    [Google Scholar]
  60. Zhang M., Liu X., Xu X., Gu Y., Tao X., Yang X., Yan G., Zhang J.. 2014; Molecular subtyping and antimicrobial susceptibilities of Campylobacter coli isolates from diarrheal patients and food-producing animals in China. Foodborne Pathog Dis11:610–619 [CrossRef][PubMed]
    [Google Scholar]
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