1887
  1. Home
  2. Publications
  3. Journal of Medical Microbiology
  4. Volume 65, Issue 1
  5. Article

f Genotyping of strains isolated in Brazil suggests possible contamination amongst environmental, human, animal and food sources

HTML
106.27 Kb
PDF
331.04 Kb
  • Authors: Carolina N. Gomes1​ ,  Roberto A. Souza1​ ,  Jaqueline Passaglia1​ ,  Sheila S. Duque2​ ,  Marta I. C. Medeiros3​ , Juliana P. Falcão1​
    • VIEW AFFILIATIONS
    • 1​ 1​ Departamento de Análises Clínicas, Toxicológicas e Bromatológicas, Faculdade de Ciências Farmacêuticas de Ribeirão Preto-USP, Av. do Café, s/no, Campus Universitário USP, Ribeirão Preto, SP 14040-903, Brazil 2​ 2​ Fundação Oswaldo Cruz (FIOCRUZ), Instituto Oswaldo Cruz, Pavilhão Rocha Lima, sala 516, Av. Brasil, 4365, Rio de Janeiro, RJ 21040-900, Brazil 3​ 3​ Instituto Adolfo Lutz de Ribeirão Preto, Rua Minas, 877 Ribeirão Preto, SP 14085-410, Brazil
  • Juliana P. Falcão [email protected]
  • First Published Online: 01 January 2016, Journal of Medical Microbiology 65: 80-90, doi: 10.1099/jmm.0.000201
  • Subject: Microbial Epidemiology
  • Received:
  • Accepted:
  • Cover date:
Preview this:
Free Preview thumbnail Magnify
Zoom in
Zoomout

Genotyping of strains isolated in Brazil suggests possible contamination amongst environmental, human, animal and food sources, Page 1 of 1

| /docserver/preview/fulltext/jmm/65/1/80_jmm000201-1.gif

  • 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.

  • Abbreviations:
    DI discrimination index
    HRMA high-resolution melting analysis
    UPGMA unweighted pair group method with arithmetic mean
    SVR short variable region.

© 2016 The Authors | Published by the Microbiology Society

  • 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 Microbiol 178: 29––38 [CrossRef] [PubMed].
     [Google Scholar]
  • 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 Rec 166: 102––106 [CrossRef] [PubMed].
     [Google Scholar]
  • 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 Health 59: 52––60 [CrossRef] [PubMed].
     [Google Scholar]
  • 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 Cruz 102: 53––57.[CrossRef]
    [Google Scholar]
  • 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 J 164: 159––161 [CrossRef] [PubMed].
     [Google Scholar]
  • 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 Sci 88: 214––217 [CrossRef] [PubMed].
     [Google Scholar]
  • 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 Dis 153: 552––559 [CrossRef] [PubMed].
     [Google Scholar]
  • Campioni F. , Falcão J. P. . ( 2014;). Genotypic diversity and virulence markers of Yersinia enterocolitica biotype 1A strains isolated from clinical and non-clinical origins. APMIS 122: 215––222 [CrossRef] [PubMed].
     [Google Scholar]
  • 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]
  • 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]
  • 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 Dis 67: 220––227 [CrossRef] [PubMed].
     [Google Scholar]
  • 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 Microbiol 29: 406––410 [CrossRef] [PubMed].
     [Google Scholar]
  • 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 Agents 44: 306––313 [CrossRef] [PubMed].
     [Google Scholar]
  • EFSA ( 2015;). The European Union summary report on trends and sources of zoonoses, zoonotic agents and food-borne outbreaks in 2013. EFSA J 13: 3991.
     [Google Scholar]
  • 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 Microbiol 94: 203––209 [CrossRef] [PubMed].
     [Google Scholar]
  • 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 Dis 16: 294––296 [CrossRef] [PubMed].
     [Google Scholar]
  • 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 Sci 48: 127––132 [CrossRef] [PubMed].
     [Google Scholar]
  • 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 Dis 38: (Suppl. 3), S285––S296 [CrossRef] [PubMed].
     [Google Scholar]
  • Fry B. N. , Wosten M. , Wassenaar T. M. , van der Zeijst B. A. M. . ( 2000;). Transformation of Campylobacter jejuni . . In Electrotransformation of Bacteria, pp. 157––167. Edited by Eynard N. , Teissie J. . Berlin: Springer;.[CrossRef]
    [Google Scholar]
  • 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 Pathol 41: 579––588 [CrossRef] [PubMed].
     [Google Scholar]
  • 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 Dis 8: 937––942 [CrossRef] [PubMed].
     [Google Scholar]
  • 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 Dis 37: 375––378.
     [Google Scholar]
  • Gürtler M. , Alter T. , Kasimir S. , Fehlhaber K. . ( 2005;). The importance of Campylobacter coli in human campylobacteriosis: prevalence and genetic characterization. Epidemiol Infect 133: 1081––1087 [CrossRef] [PubMed].
     [Google Scholar]
  • 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 Dis 11: 1257––1264 [CrossRef] [PubMed].
     [Google Scholar]
  • 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 Microbiol 35: 2386––2392 [PubMed].
     [Google Scholar]
  • Health Protection Surveillance Centre ( 2005;). Campylobacteriosis in Ireland. 2005 Annual Report The Health Protection Surveillance Centre; 25–27 Middle Gardiner Street, Dublin 1, Ireland:, pp. 34––36.
     [Google Scholar]
  • 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 Microbiol 26: 2465––2466 [PubMed].
     [Google Scholar]
  • Jansen R. , Embden J. D. , Gaastra W. , Schouls L. M. . ( 2002;). Identification of genes that are associated with DNA repeats in prokaryotes. Mol Microbiol 43: 1565––1575 [CrossRef] [PubMed].
     [Google Scholar]
  • 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 Microbiol 173: 54––61 [CrossRef] [PubMed].
     [Google Scholar]
  • Lévesque S. , Michaud S. , Arbeit R. D. , Frost E. H. . ( 2011;). High-resolution melting system to perform multilocus sequence typing of Campylobacter jejuni . PLoS One 6: e16167 [CrossRef] [PubMed].
     [Google Scholar]
  • 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 Microbiol 27: 311––315 [CrossRef] [PubMed].
     [Google Scholar]
  • 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 Microbiol 111: 971––981 [CrossRef] [PubMed].
     [Google Scholar]
  • Meinersmann R. J. , Helsel L. O. , Fields P. I. , Hiett K. L. . ( 1997;). Discrimination of Campylobacter jejuni isolates by fla gene sequencing. J Clin Microbiol 35: 2810––2814 [PubMed].
     [Google Scholar]
  • 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 Microbiol 36: 244––246 [CrossRef] [PubMed].
     [Google Scholar]
  • 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 Evol 60: 174––182 [CrossRef] [PubMed].
     [Google Scholar]
  • 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 Res 36: 351––382 [CrossRef] [PubMed].
     [Google Scholar]
  • 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 Infect 141: 2526––2535 [CrossRef] [PubMed].
     [Google Scholar]
  • 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 Microbiol 28: 426––433 [CrossRef] [PubMed].
     [Google Scholar]
  • Olsen S. , Fitzgerald C. , Swerdlow D. . ( 2001a;). Limitations of pulsed-field gel electrophoresis for the routine surveillance of Campylobacter infections – reply. J Infect Dis 184: 243––244 [CrossRef].
     [Google Scholar]
  • 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 Dis 183: 164––167 [CrossRef] [PubMed].
     [Google Scholar]
  • 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 Microbiol 114: 105––112 [CrossRef] [PubMed].
     [Google Scholar]
  • 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 Microbiol 73: 3431––3436 [CrossRef] [PubMed].
     [Google Scholar]
  • 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 Microbiol 61: 507––513.[CrossRef]
    [Google Scholar]
  • 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 Microbiol 39: 1889––1894 [CrossRef] [PubMed].
     [Google Scholar]
  • 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 Med 368: 1817––1825 [CrossRef] [PubMed].
     [Google Scholar]
  • 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 One 8: e64504 [CrossRef] [PubMed].
     [Google Scholar]
  • 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 Dis 7: 597––600 [CrossRef] [PubMed].
     [Google Scholar]
  • Sambrook J. , Russell W. D. . ( 2001;). Molecular Cloning: a Laboratory Manual, 3rd edn Cold Spring Harbor; NY: Cold Spring Harbor Laboratory.
     [Google Scholar]
  • 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 Dis 17: 7––15 [CrossRef] [PubMed].
     [Google Scholar]
  • 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 Dis 36: 378––382.
     [Google Scholar]
  • 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 Microbiol 40: 952––959 [CrossRef] [PubMed].
     [Google Scholar]
  • Selander B. , Rydberg J. , Lenner C. , Hagerstrand I. . ( 1993;). [Unusual infectious complication in a pregnant woman. Spontaneous abortion caused by Campylobacter coli]. Lakartidningen 90: 4356––4357 (in Swedish).
     [Google Scholar]
  • 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 Resour 10: 193––196 [CrossRef] [PubMed].
     [Google Scholar]
  • Souza R. A. , Falcão J. P. . ( 2012;). A novel high-resolution melting analysis-based method for Yersinia pseudotuberculosis genotyping. J Microbiol Methods 91: 329––335 [CrossRef] [PubMed].
     [Google Scholar]
  • Wassenaar T. M. , Newell D. G. . ( 2000;). Genotyping of Campylobacter spp. Appl Environ Microbiol 66: 1––9 [CrossRef] [PubMed].
     [Google Scholar]
  • Wassenaar T. M. , Geilhausen B. , Newell D. G. . ( 1998;). Evidence of genomic instability in Campylobacter jejuni isolated from poultry. Appl Environ Microbiol 64: 1816––1821 [PubMed].
     [Google Scholar]
  • 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 Microbiol 49: 388––395 [CrossRef] [PubMed].
     [Google Scholar]
  • 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 Microbiol 76: 6377––6386 [CrossRef] [PubMed].
     [Google Scholar]
  • Wittwer C. T. . ( 2009;). High-resolution DNA melting analysis: advancements and limitations. Hum Mutat 30: 857––859 [CrossRef] [PubMed].
     [Google Scholar]
  • 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 Dis 11: 610––619 [CrossRef] [PubMed].
     [Google Scholar]
http://sgm.metastore.ingenta.com/content/journal/jmm/10.1099/jmm.0.000201
Loading

Supplementary Data

 Data loading....

Loading

Article metrics loading...

/content/journal/jmm/10.1099/jmm.0.000201
2016-01-01
2019-02-19
Loading full text...

Full text loading...

/deliver/fulltext/jmm/65/1/80.html?itemId=/content/journal/jmm/10.1099/jmm.0.000201&mimeType=html&fmt=ahah

Author and Article Information

 Figure data loading....

Footnotes:

Validated antibodies in this article

This is a required field
Please enter a valid email address
Please check the format of the address you have entered.
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error