Skip to main content
Top
Published in: Gut Pathogens 1/2020

Open Access 01-12-2020 | Research

Characterization of Arcobacter strains isolated from human stool samples: results from the prospective German prevalence study Arcopath

Authors: Vanessa Brückner, Ulrike Fiebiger, Ralf Ignatius, Johannes Friesen, Martin Eisenblätter, Marlies Höck, Thomas Alter, Stefan Bereswill, Markus M. Heimesaat, Greta Gölz

Published in: Gut Pathogens | Issue 1/2020

Login to get access

Abstract

Background

Arcobacter constitute emerging food- and waterborne pathogens causing gastroenteritis in humans, but the underlying mechanisms are only incompletely understood. We therefore characterized Arcobacter isolates derived from human stool samples that had been collected during a prospective prevalence study in Germany in vitro. Thirty-six bacterial isolates belonging to the species A. butzleri (n = 24), A. cryaerophilus (n = 10) and A. lanthieri (n = 2) were genotyped by ERIC-PCR, the presence of 10 putative virulence genes was assessed and cytotoxic effects on the human intestinal cell line HT-29/B6 were analyzed applying the WST-assay.

Results

Genotyping revealed high genetic diversity within the species A. butzleri, A. cryaerophilus and A. lanthieri. Both, A. butzleri and A. lanthieri encoded for a large number of putative virulence genes, while fewer genes were detectable in A. cryaerophilus isolates. Notably, the three cytolethal distending toxin (CDT) genes cdtA, cdtB and cdtC were abundant in both A. lanthieri isolates. Furthermore, all A. butzleri and A. lanthieri, but only one of the A. cryaerophilus isolates exerted cytotoxic effects.

Conclusions

Our study provides evidence for the abundance of putative virulence genes in Arcobacter isolates and prominent cytotoxic effects of A. butzleri and A. lanthieri in vitro. The presence of cdtA, cdtB, cdtC in A. lanthieri points towards CDT secretion as potential mechanism underlying cytotoxicity as opposed to A. butzleri. However, the association of the Arcobacter virulence factors detected and human morbidity should be addressed in future studies.
Literature
1.
go back to reference Perez-Cataluna A, Salas-Masso N, Dieguez AL, Balboa S, Lema A, Romalde JL, et al. Revisiting the taxonomy of the genus Arcobacter: getting order from the chaos. Front Microbiol. 2018;9:2077.CrossRef Perez-Cataluna A, Salas-Masso N, Dieguez AL, Balboa S, Lema A, Romalde JL, et al. Revisiting the taxonomy of the genus Arcobacter: getting order from the chaos. Front Microbiol. 2018;9:2077.CrossRef
2.
go back to reference Vandamme P, Falsen E, Rossau R, Hoste B, Segers P, Tytgat R, et al. Revision of Campylobacter, Helicobacter, and Wolinella taxonomy: emendation of generic descriptions and proposal of Arcobacter gen. nov. Int J Syst Bacteriol. 1991;41(1):88–103.CrossRef Vandamme P, Falsen E, Rossau R, Hoste B, Segers P, Tytgat R, et al. Revision of Campylobacter, Helicobacter, and Wolinella taxonomy: emendation of generic descriptions and proposal of Arcobacter gen. nov. Int J Syst Bacteriol. 1991;41(1):88–103.CrossRef
3.
go back to reference Ferreira S, Queiroz JA, Oleastro M, Domingues FC. Insights in the pathogenesis and resistance of Arcobacter: a review. Crit Rev Microbiol. 2016;42(3):364–83.PubMed Ferreira S, Queiroz JA, Oleastro M, Domingues FC. Insights in the pathogenesis and resistance of Arcobacter: a review. Crit Rev Microbiol. 2016;42(3):364–83.PubMed
4.
go back to reference Hsu TTD, Lee J. Global distribution and prevalence of Arcobacter in food and water. Zoonoses Public Health. 2015;62(8):579–89.CrossRef Hsu TTD, Lee J. Global distribution and prevalence of Arcobacter in food and water. Zoonoses Public Health. 2015;62(8):579–89.CrossRef
5.
go back to reference Ramees TP, Dhama K, Karthik K, Rathore RS, Kumar A, Saminathan M, et al. Arcobacter: an emerging food-borne zoonotic pathogen, its public health concerns and advances in diagnosis and control - a comprehensive review. Vet Q. 2017;37(1):136–61.CrossRef Ramees TP, Dhama K, Karthik K, Rathore RS, Kumar A, Saminathan M, et al. Arcobacter: an emerging food-borne zoonotic pathogen, its public health concerns and advances in diagnosis and control - a comprehensive review. Vet Q. 2017;37(1):136–61.CrossRef
6.
go back to reference Figueras MJ, Levican A, Pujol I, Ballester F, Rabada Quilez MJ, Gomez-Bertomeu F. A severe case of persistent diarrhoea associated with Arcobacter cryaerophilus but attributed to Campylobacter sp. and a review of the clinical incidence of Arcobacter spp. New Microb New Infect. 2014;2(2):31–7.CrossRef Figueras MJ, Levican A, Pujol I, Ballester F, Rabada Quilez MJ, Gomez-Bertomeu F. A severe case of persistent diarrhoea associated with Arcobacter cryaerophilus but attributed to Campylobacter sp. and a review of the clinical incidence of Arcobacter spp. New Microb New Infect. 2014;2(2):31–7.CrossRef
7.
go back to reference Mandisodza O, Burrows E, Nulsen M. Arcobacter species in diarrhoeal faeces from humans in New Zealand. N Z Med J. 2012;125(1353):40–6.PubMed Mandisodza O, Burrows E, Nulsen M. Arcobacter species in diarrhoeal faeces from humans in New Zealand. N Z Med J. 2012;125(1353):40–6.PubMed
8.
go back to reference ICMSF ICoMSfF. In: Tompkin RB (ed), Microbiological testing in food safety management, vol 7. New York: Kluwer Academic/Plenum Publishers; 2002, p. 171 ICMSF ICoMSfF. In: Tompkin RB (ed), Microbiological testing in food safety management, vol 7. New York: Kluwer Academic/Plenum Publishers; 2002, p. 171
9.
go back to reference Carbone M, Maugeri TL, Giannone M, Gugliandolo C, Midiri A, Fera MT. Adherence of environmental Arcobacter butzleri and Vibrio spp. isolates to epithelial cells in vitro. Food Microbiol. 2003;20(5):611–6.CrossRef Carbone M, Maugeri TL, Giannone M, Gugliandolo C, Midiri A, Fera MT. Adherence of environmental Arcobacter butzleri and Vibrio spp. isolates to epithelial cells in vitro. Food Microbiol. 2003;20(5):611–6.CrossRef
10.
go back to reference Karadas G, Bucker R, Sharbati S, Schulzke JD, Alter T, Golz G. Arcobacter butzleri isolates exhibit pathogenic potential in intestinal epithelial cell models. J Appl Microbiol. 2016;120(1):218–25.CrossRef Karadas G, Bucker R, Sharbati S, Schulzke JD, Alter T, Golz G. Arcobacter butzleri isolates exhibit pathogenic potential in intestinal epithelial cell models. J Appl Microbiol. 2016;120(1):218–25.CrossRef
11.
go back to reference Karadas G, Sharbati S, Hanel I, Messelhausser U, Glocker E, Alter T, et al. Presence of virulence genes, adhesion and invasion of Arcobacter butzleri. J Appl Microbiol. 2013;115(2):583–90.CrossRef Karadas G, Sharbati S, Hanel I, Messelhausser U, Glocker E, Alter T, et al. Presence of virulence genes, adhesion and invasion of Arcobacter butzleri. J Appl Microbiol. 2013;115(2):583–90.CrossRef
12.
go back to reference Levican A, Alkeskas A, Gunter C, Forsythe SJ, Figueras MJ. Adherence to and invasion of human intestinal cells by Arcobacter species and their virulence genotypes. Appl Environ Microb. 2013;79(16):4951–7.CrossRef Levican A, Alkeskas A, Gunter C, Forsythe SJ, Figueras MJ. Adherence to and invasion of human intestinal cells by Arcobacter species and their virulence genotypes. Appl Environ Microb. 2013;79(16):4951–7.CrossRef
13.
go back to reference Villarruel-Lopez A, Marquez-Gonzalez M, Garay-Martinez LE, Zepeda H, Castillo A, Mota de la Garza L, et al. Isolation of Arcobacter spp. from retail meats and cytotoxic effects of isolates against vero cells. J Food Prot. 2003;66(8):1374–8.CrossRef Villarruel-Lopez A, Marquez-Gonzalez M, Garay-Martinez LE, Zepeda H, Castillo A, Mota de la Garza L, et al. Isolation of Arcobacter spp. from retail meats and cytotoxic effects of isolates against vero cells. J Food Prot. 2003;66(8):1374–8.CrossRef
14.
go back to reference Bücker R, Troeger H, Kleer J, Fromm M, Schulzke JD. Arcobacter butzleri induces barrier dysfunction in intestinal HT-29/B6 cells. J Infect Dis. 2009;200(5):756–64.CrossRef Bücker R, Troeger H, Kleer J, Fromm M, Schulzke JD. Arcobacter butzleri induces barrier dysfunction in intestinal HT-29/B6 cells. J Infect Dis. 2009;200(5):756–64.CrossRef
15.
go back to reference Miller WG, Parker CT, Rubenfield M, Mendz GL, Wosten MM, Ussery DW, et al. The complete genome sequence and analysis of the epsilon proteobacterium Arcobacter butzleri. PLoS ONE. 2007;2(12):e1358.CrossRef Miller WG, Parker CT, Rubenfield M, Mendz GL, Wosten MM, Ussery DW, et al. The complete genome sequence and analysis of the epsilon proteobacterium Arcobacter butzleri. PLoS ONE. 2007;2(12):e1358.CrossRef
16.
go back to reference Zambri M, Cloutier M, Adam Z, Lapen DR, Wilkes G, Sunohara M, et al. Novel virulence, antibiotic resistance and toxin gene-specific PCR-based assays for rapid pathogenicity assessment of Arcobacter faecis and Arcobacter lanthieri. BMC Microbiol. 2019;19(1):11.CrossRef Zambri M, Cloutier M, Adam Z, Lapen DR, Wilkes G, Sunohara M, et al. Novel virulence, antibiotic resistance and toxin gene-specific PCR-based assays for rapid pathogenicity assessment of Arcobacter faecis and Arcobacter lanthieri. BMC Microbiol. 2019;19(1):11.CrossRef
17.
go back to reference Houf K, Devriese LA, Haesebrouck F, Vandenberg O, Butzler JP, Van Hoof J, et al. Antimicrobial susceptibility patterns of Arcobacter butzleri and Arcobacter cryaerophilus strains isolated from humans and broilers. Microb Drug Resist. 2004;10(3):243–7.CrossRef Houf K, Devriese LA, Haesebrouck F, Vandenberg O, Butzler JP, Van Hoof J, et al. Antimicrobial susceptibility patterns of Arcobacter butzleri and Arcobacter cryaerophilus strains isolated from humans and broilers. Microb Drug Resist. 2004;10(3):243–7.CrossRef
18.
go back to reference Houf K, Stephan R. Isolation and characterization of the emerging foodborn pathogen Arcobacter from human stool. J Microbiol Meth. 2007;68(2):408–13.CrossRef Houf K, Stephan R. Isolation and characterization of the emerging foodborn pathogen Arcobacter from human stool. J Microbiol Meth. 2007;68(2):408–13.CrossRef
19.
go back to reference Kayman T, Abay S, Hizlisoy H, Atabay HI, Diker KS, Aydin F. Emerging pathogen Arcobacter spp. in acute gastroenteritis: molecular identification, antibiotic susceptibilities and genotyping of the isolated arcobacters. J Med Microbiol. 2012;61(Pt 10):1439–44.CrossRef Kayman T, Abay S, Hizlisoy H, Atabay HI, Diker KS, Aydin F. Emerging pathogen Arcobacter spp. in acute gastroenteritis: molecular identification, antibiotic susceptibilities and genotyping of the isolated arcobacters. J Med Microbiol. 2012;61(Pt 10):1439–44.CrossRef
20.
go back to reference Perez-Cataluna A, Collado L, Salgado O, Lefinanco V, Figueras MJ. A polyphasic and taxogenomic evaluation uncovers Arcobacter cryaerophilus as a species complex that embraces four genomovars. Front Microbiol. 2018;9:805.CrossRef Perez-Cataluna A, Collado L, Salgado O, Lefinanco V, Figueras MJ. A polyphasic and taxogenomic evaluation uncovers Arcobacter cryaerophilus as a species complex that embraces four genomovars. Front Microbiol. 2018;9:805.CrossRef
21.
go back to reference Houf K, De Zutter L, Van Hoof J, Vandamme P. Assessment of the genetic diversity among arcobacters isolated from poultry products by using two PCR-based typing methods. Appl Environ Microb. 2002;68(5):2172–8.CrossRef Houf K, De Zutter L, Van Hoof J, Vandamme P. Assessment of the genetic diversity among arcobacters isolated from poultry products by using two PCR-based typing methods. Appl Environ Microb. 2002;68(5):2172–8.CrossRef
22.
go back to reference Collado L, Kasimir G, Perez U, Bosch A, Pinto R, Saucedo G, et al. Occurrence and diversity of Arcobacter spp. along the Llobregat River catchment, at sewage effluents and in a drinking water treatment plant. Water Res. 2010;44(12):3696–702.CrossRef Collado L, Kasimir G, Perez U, Bosch A, Pinto R, Saucedo G, et al. Occurrence and diversity of Arcobacter spp. along the Llobregat River catchment, at sewage effluents and in a drinking water treatment plant. Water Res. 2010;44(12):3696–702.CrossRef
23.
go back to reference Lehmann D, Alter T, Lehmann L, Uherkova S, Seidler T, Golz G. Prevalence, virulence gene distribution and genetic diversity of Arcobacter in food samples in Germany. Berl Munch Tierarztl Wochenschr. 2015;128(3–4):163–8.PubMed Lehmann D, Alter T, Lehmann L, Uherkova S, Seidler T, Golz G. Prevalence, virulence gene distribution and genetic diversity of Arcobacter in food samples in Germany. Berl Munch Tierarztl Wochenschr. 2015;128(3–4):163–8.PubMed
24.
go back to reference Sekhar MS, Rao TS, Chinnam BK, Subramanyam KV, Metta M, Sharif NM. Genetic Diversity of Arcobacter Species of Animal and Human Origin in Andhra Pradesh India. Indian J Microbiol. 2017;57(2):250–2.CrossRef Sekhar MS, Rao TS, Chinnam BK, Subramanyam KV, Metta M, Sharif NM. Genetic Diversity of Arcobacter Species of Animal and Human Origin in Andhra Pradesh India. Indian J Microbiol. 2017;57(2):250–2.CrossRef
25.
go back to reference Douidah L, de Zutter L, Bare J, De Vos P, Vandamme P, Vandenberg O, et al. Occurrence of putative virulence genes in arcobacter species isolated from humans and animals. J Clin Microbiol. 2012;50(3):735–41.CrossRef Douidah L, de Zutter L, Bare J, De Vos P, Vandamme P, Vandenberg O, et al. Occurrence of putative virulence genes in arcobacter species isolated from humans and animals. J Clin Microbiol. 2012;50(3):735–41.CrossRef
26.
go back to reference Girbau C, Guerra C, Martinez-Malaxetxebarria I, Alonso R, Fernandez-Astorga A. Prevalence of ten putative virulence genes in the emerging foodborne pathogen Arcobacter isolated from food products. Food Microbiol. 2015;52:146–9.CrossRef Girbau C, Guerra C, Martinez-Malaxetxebarria I, Alonso R, Fernandez-Astorga A. Prevalence of ten putative virulence genes in the emerging foodborne pathogen Arcobacter isolated from food products. Food Microbiol. 2015;52:146–9.CrossRef
27.
go back to reference Piva S, Gariano GR, Bonilauri P, Giacometti F, Decastelli L, Florio D, et al. Occurrence of putative virulence genes on Arcobacter butzleri isolated from three different environmental sites throughout the dairy chain. J Appl Microbiol. 2017;122(4):1071–7.CrossRef Piva S, Gariano GR, Bonilauri P, Giacometti F, Decastelli L, Florio D, et al. Occurrence of putative virulence genes on Arcobacter butzleri isolated from three different environmental sites throughout the dairy chain. J Appl Microbiol. 2017;122(4):1071–7.CrossRef
28.
go back to reference Sekhar MS, Tumati SR, Chinnam BK, Kothapalli VS, Sharif NM. Virulence gene profiles of Arcobacter species isolated from animals, foods of animal origin, and humans in Andhra Pradesh India. Vet World. 2017;10(6):716–20.CrossRef Sekhar MS, Tumati SR, Chinnam BK, Kothapalli VS, Sharif NM. Virulence gene profiles of Arcobacter species isolated from animals, foods of animal origin, and humans in Andhra Pradesh India. Vet World. 2017;10(6):716–20.CrossRef
29.
go back to reference Silha D, Vackova B, Silhova L. Occurrence of virulence-associated genes in Arcobacter butzleri and Arcobacter cryaerophilus isolates from foodstuff, water, and clinical samples within the Czech Republic. Folia Microbiol. 2019;64(1):25–31.CrossRef Silha D, Vackova B, Silhova L. Occurrence of virulence-associated genes in Arcobacter butzleri and Arcobacter cryaerophilus isolates from foodstuff, water, and clinical samples within the Czech Republic. Folia Microbiol. 2019;64(1):25–31.CrossRef
30.
go back to reference Tabatabaei M, Shirzad Aski H, Shayegh H, Khoshbakht R. Occurrence of six virulence-associated genes in Arcobacter species isolated from various sources in Shiraz Southern Iran. Microb Pathog. 2014;66:1–4.CrossRef Tabatabaei M, Shirzad Aski H, Shayegh H, Khoshbakht R. Occurrence of six virulence-associated genes in Arcobacter species isolated from various sources in Shiraz Southern Iran. Microb Pathog. 2014;66:1–4.CrossRef
31.
go back to reference Zacharow I, Bystron J, Walecka-Zacharska E, Podkowik M, Bania J. Genetic diversity and incidence of virulence-associated genes of Arcobacter butzleri and Arcobacter cryaerophilus isolates from Pork, Beef, and Chicken Meat in Poland. Biomed Res Int. 2015;2015:956507.CrossRef Zacharow I, Bystron J, Walecka-Zacharska E, Podkowik M, Bania J. Genetic diversity and incidence of virulence-associated genes of Arcobacter butzleri and Arcobacter cryaerophilus isolates from Pork, Beef, and Chicken Meat in Poland. Biomed Res Int. 2015;2015:956507.CrossRef
32.
go back to reference Abdelbaqi K, Menard A, Prouzet-Mauleon V, Bringaud F, Lehours P, Megraud F. Nucleotide sequence of the gyrA gene of Arcobacter species and characterization of human ciprofloxacin-resistant clinical isolates. Fems Immunol Med Microbiol. 2007;49(3):337–45.CrossRef Abdelbaqi K, Menard A, Prouzet-Mauleon V, Bringaud F, Lehours P, Megraud F. Nucleotide sequence of the gyrA gene of Arcobacter species and characterization of human ciprofloxacin-resistant clinical isolates. Fems Immunol Med Microbiol. 2007;49(3):337–45.CrossRef
33.
go back to reference Perez-Cataluna A, Tapiol J, Benavent C, Sarvise C, Gomez F, Martinez B, et al. Antimicrobial susceptibility, virulence potential and sequence types associated with Arcobacter strains recovered from human faeces. J Med Microbiol. 2017;66(12):1736–43.CrossRef Perez-Cataluna A, Tapiol J, Benavent C, Sarvise C, Gomez F, Martinez B, et al. Antimicrobial susceptibility, virulence potential and sequence types associated with Arcobacter strains recovered from human faeces. J Med Microbiol. 2017;66(12):1736–43.CrossRef
34.
go back to reference Johnson WM, Lior H. A new heat-labile cytolethal distending toxin (CLDT) produced by Campylobacter spp. Microb Pathog. 1988;4(2):115–26.CrossRef Johnson WM, Lior H. A new heat-labile cytolethal distending toxin (CLDT) produced by Campylobacter spp. Microb Pathog. 1988;4(2):115–26.CrossRef
35.
go back to reference Pickett CL, Pesci EC, Cottle DL, Russell G, Erdem AN, Zeytin H. Prevalence of cytolethal distending toxin production in Campylobacter jejuni and relatedness of Campsssylobacter sp. cdtB gene. Infect Immun. 1996;64(6):2070–8.CrossRef Pickett CL, Pesci EC, Cottle DL, Russell G, Erdem AN, Zeytin H. Prevalence of cytolethal distending toxin production in Campylobacter jejuni and relatedness of Campsssylobacter sp. cdtB gene. Infect Immun. 1996;64(6):2070–8.CrossRef
36.
go back to reference Zheng J, Meng J, Zhao S, Singh R, Song W. Campylobacter-induced interleukin-8 secretion in polarized human intestinal epithelial cells requires Campylobacter-secreted cytolethal distending toxin- and Toll-like receptor-mediated activation of NF-kappaB. Infect Immun. 2008;76(10):4498–508.CrossRef Zheng J, Meng J, Zhao S, Singh R, Song W. Campylobacter-induced interleukin-8 secretion in polarized human intestinal epithelial cells requires Campylobacter-secreted cytolethal distending toxin- and Toll-like receptor-mediated activation of NF-kappaB. Infect Immun. 2008;76(10):4498–508.CrossRef
37.
go back to reference Gugliandolo C, Irrera GP, Lentini V, Maugeri TL. Pathogenic Vibrio, Aeromonas and Arcobacter spp. associated with copepods in the Straits of Messina (Italy). Mar Pollut Bull. 2008;56(3):600–6.CrossRef Gugliandolo C, Irrera GP, Lentini V, Maugeri TL. Pathogenic Vibrio, Aeromonas and Arcobacter spp. associated with copepods in the Straits of Messina (Italy). Mar Pollut Bull. 2008;56(3):600–6.CrossRef
38.
go back to reference Musmanno RA, Russi M, Lior H, Figura N. In vitro virulence factors of Arcobacter butzleri strains isolated from superficial water samples. New Microbiol. 1997;20(1):63–8.PubMed Musmanno RA, Russi M, Lior H, Figura N. In vitro virulence factors of Arcobacter butzleri strains isolated from superficial water samples. New Microbiol. 1997;20(1):63–8.PubMed
39.
go back to reference Johnson LG, Murano EA. Lack of a cytolethal distending toxin among Arcobacter isolates from various sources. J Food Protect. 2002;65(11):1789–95.CrossRef Johnson LG, Murano EA. Lack of a cytolethal distending toxin among Arcobacter isolates from various sources. J Food Protect. 2002;65(11):1789–95.CrossRef
40.
go back to reference Whiteduck-Leveillee J, Cloutier M, Topp E, Lapen DR, Talbot G, Villemur R, et al. Development and evaluation of multiplex PCR assays for rapid detection of virulence-associated genes in Arcobacter species. J Microbiol Methods. 2016;121:59–655.CrossRef Whiteduck-Leveillee J, Cloutier M, Topp E, Lapen DR, Talbot G, Villemur R, et al. Development and evaluation of multiplex PCR assays for rapid detection of virulence-associated genes in Arcobacter species. J Microbiol Methods. 2016;121:59–655.CrossRef
41.
go back to reference Kreusel KM, Fromm M, Schulzke JD, Hegel U. Cl- secretion in epithelial monolayers of mucus-forming human colon cells (HT-29/B6). Am J Physiol. 1991;261(4 Pt 1):C574–C582582.CrossRef Kreusel KM, Fromm M, Schulzke JD, Hegel U. Cl- secretion in epithelial monolayers of mucus-forming human colon cells (HT-29/B6). Am J Physiol. 1991;261(4 Pt 1):C574–C582582.CrossRef
Metadata
Title
Characterization of Arcobacter strains isolated from human stool samples: results from the prospective German prevalence study Arcopath
Authors
Vanessa Brückner
Ulrike Fiebiger
Ralf Ignatius
Johannes Friesen
Martin Eisenblätter
Marlies Höck
Thomas Alter
Stefan Bereswill
Markus M. Heimesaat
Greta Gölz
Publication date
01-12-2020
Publisher
BioMed Central
Published in
Gut Pathogens / Issue 1/2020
Electronic ISSN: 1757-4749
DOI
https://doi.org/10.1186/s13099-019-0344-3

Other articles of this Issue 1/2020

Gut Pathogens 1/2020 Go to the issue