Novel virulence, antibiotic resistance and toxin gene-specific PCR-based assays for rapid pathogenicity assessment of Arcobacter faecis and Arcobacter lanthieri

Zambri, Matthew; Cloutier, Michel; Adam, Zaky; Lapen, David R.; Wilkes, Graham; Sunohara, Mark; Topp, Edward; Talbot, Guylaine; Khan, Izhar U. H.

doi:10.1186/s12866-018-1357-7

Cited by 25 publications

(31 citation statements)

References 54 publications

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“…This is further supported by cytotoxicity of CDT-negative Arcobacter [39]. Notably, we here detected all three CDT toxin genes in both A. lanthieri isolates, which is in concordance with results from Zambri et al [16], who detected these in 8 out of 11 A. lanthieri isolates from various environmental and fecal sources. Thus, while CDT may likely be involved in A. lanthieri-induced cytotoxicity, other factors might contribute to cytotoxic effects of A. butzleri.…”

Section: Cytotoxic Effectssupporting

confidence: 92%

“…isolated from human stool samples. Black: genes detected by primers designed based on A. butzleri sequences (Douidah et al [25], Karadas et al [11], Whiteduck-Leveillee et al [40]; pale grey: genes not detected by both PCR; dark grey: genes detected by primers designed based on A. lanthieri sequences (Zambri et al [16] CDT, i.e., cdtA, cdtB, and cdtC, were present in both A. lanthieri strains (data not shown).…”

Section: Abundance Of Putative Virulence Genesmentioning

confidence: 95%

“…Analyses of respective fragment patterns were performed by using BioNumerics version 7.1 (Applied Maths, Sint-Martens-Latem, Belgium). The similarities between profiles were calculated using Dice coefficient, [25] cadF-R AAA CTA TGC TAA CGC TGG TT Douidah et al [25] irgA-F TGC AGA GGA TGC TTG GAG CGT AAC T 437 Whiteduck-Leveillee et al [40] irgA-R GTA TAA CCC CAT TGA TGA GGA GCA Whiteduck-Leveillee et al [40] hecA-F GTG GAA GTA CAA CGA TAG CAG GCT C 537 Whiteduck-Leveillee et al [40] hecA-R GTC TGT TTT AGT TGC TCT GCA GTC Whiteduck-Leveillee et al [40] hecB-F CTA AAC TCT ACA AAT CGT GC 528 Whiteduck-Leveillee et al [40] hecB-R CTT TTG AGT GTT GAC CTC Whiteduck-Leveillee et al [40] pldA-F TTG ACG AGA CAA TAA GTG CAG C 293 Whiteduck-Leveillee et al [40] pldA-R CGT CTT TAT CTT TGC TTT CAG GGA Whiteduck-Leveillee et al [40] ciaB-F TGG GCA GAT GTG GAT AGA GCT TGG A 284 Whiteduck-Leveillee et al [40] ciaB-R TAG TGC TGG TCG TCC CAC ATA AAG Whiteduck-Leveillee et al [40] cj1349-F CCA GAA ATC ACT GGC TTT TGA G 659 Whiteduck-Leveillee et al [40] cj1349-R GGG CAT AAG TTA GAT GAG GTT CC Whiteduck-Leveillee et al [40] tlyA-F CAA AGT CGA AAC AAA GCG ACT G 230 Whiteduck-Leveillee et al [40] tlyA-R TCC ACC AGT GCT ACT TCC TAT A Whiteduck-Leveillee et al [40] mviN-F TGC ACT TGT TGC AAA ACG GTG 294 Whiteduck-Leveillee et al [40] mviN-R TGC TGA TGG AGC TTT TAC GCA AGC Whiteduck-Leveillee et al [40] iroE-F AAT GGC TAT GAT GTT GTT TAC 415 Karadas et al [11] iroE-R TTG CTG CTA TGA AGT TTT Karadas et al [11] Detection of putative virulence genes with A. lanthieri specific primers AL_cdtB F GCA AAA GGT GAT TGG GCT CC 303 Zambri et al [16] AL_cdtB R TCC TCC AGC TCC TTG AAC AC Zambri et al [16] AL_cadF F TCC AAC TCC AGT TGC TGC TC 243 Zambri et al [16] AL_cadF R TGT CCT TCG ATG TCA GCT TTC Zambri et al [16] AL_irgA F AGA GCT GTT GGT TGG GAT GG 186 Zambri et al [16] AL_irgA R TGC ATT TGC TCT TGT AGG GT Zambri et al [16] AL_cdtC F GAT GAA TCC ACC AGA AAT AGA G 196 Zambri et al [16] AL_cdtC R TTT GGG ATC AAG AGT ATA AAG TTC Zambri et al [16]...…”

Section: Genotyping By Eric-pcrmentioning

confidence: 99%

“…Reaction conditions were 95 °C for 4 min followed by 30 cycles of 95 °C for 30 s, 50 °C for 30 s and 72 °C for 30 s and ended with a final extension step at 72 °C for 5 min. For analysis of A. lanthieri additional primers and a protocol described by Zambri et al [16] were used for the detection of ciaB, mviN, cadF, pldA, tlyA, irgA, cdtA, cdtB and cdtC. Briefly, 25 µl PCR-mixture contained 2 µl template DNA, 1 × PCR buffer, 1.5 mM MgCl 2 , 0.2 mM of each dNTP, 0.5 U Taq polymerase and different concentrations of corresponding primers (0.4 µM of each AL_cdtB, AL_cadF, AL_irgA, and AL_mviN; 0.3 µM of each AL_cdtC and AL_cdtA; 0.2 µM of each AL_ciaB; 0.1 µM of each AL_pldA and AL_tlyA).…”

Section: Presence Of Virulence Associated Genesmentioning

confidence: 99%

“…Furthermore, no toxin, which might be responsible for the cytotoxic effects reported for A. butzleri, has been identified yet. In contrast to A. butzleri and A. cryaerophilus, the cytolethal distending toxin (CDT) encoding genes cdtABC have been detected in several A. lanthieri isolates [16].…”

Section: Introductionmentioning

confidence: 99%

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

et al. 2020

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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.

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Section: Cytotoxic Effectssupporting

confidence: 92%

Section: Abundance Of Putative Virulence Genesmentioning

confidence: 95%

Section: Genotyping By Eric-pcrmentioning

confidence: 99%

Section: Presence Of Virulence Associated Genesmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 3 more Smart Citations

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

et al. 2020

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Virulence genotype and phenotype of two clinical isolates of Arcobacter butzleri obtained from patients with different pathologies

Baztarrika,

Salazar-Sánchez,

Hernaez Crespo

et al. 2023

Arch Microbiol

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The surge in human arcobacteriosis has increased interest in determining the mechanisms involved in the pathogenesis of Arcobacter butzleri. Here, genomic analyses and in vitro Caco-2 infection, motility, urease and antimicrobial susceptibility testing (AST) assays were used to characterise the virulence and antimicrobial resistance (AMR) determinants of strains HC-1, isolated from a patient with travellers’ diarrhoea, and HC-2, isolated from another with pruritus. AMR determinants conferring resistance to tetracycline (tetO, present in both genomes) and to ampicillin and amoxicillin–clavulanic acid (bla3, present in HC-2) were identified. The same determinants associated with flagellum, chemotaxis, adhesion and invasion were detected in both, but HC-1 lacked eight flagellar genes. The urease cluster was only present in HC-1. Motility and urease tests confirmed the genetic differences between strains, but no genetic marker related to the inability of HC-2 to adhere and invade was identified. This inability could be conditioning the patient’s pathology.

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Investigation and characterization of Aliarcobacter spp. isolated from cattle slaughterhouse in Türkiye

Disli,

Hizlisoy,

Gungor

et al. 2024

Int Microbiol

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Aliarcobacter spp. have has been isolated from numerous food products at retail and from animal carcasses and feces at slaughter. The objectives of this study were: i) to isolate Aliarcobacter species (Aliarcobacter butzleri, Aliarcobacter skirrowii, Aliarcobacter cryaerophilus) from different slaughterhouses' samples, ii) to detect genetic diversity, antibiotic resistance, bio lm ability and putative virulence gene pro les of the isolates. The antibiotic susceptibility to antimicrobials was examined using the Kirby-Bauer disc diffusion method. A molecular investigation of antibiotic resistance and virulence factors was also conducted using polymerase chain reaction (PCR).A total of 22 (14.6%) Aliarcobacter spp. isolates were obtained, with varying levels of antibiotic resistance observed. The genes tetO, tetW, and gyrA were detected in 0%, 31.8%, and 27.2% of the isolates, respectively. All isolates were resistant to ampicillin, rifampin, and erythromycin, while tetracycline was found to be the most effective antibiotic, with 81.8% of the isolates showing susceptibility to it. In terms of virulence factors, all isolates (100%) harbored more than one of the nine putative virulence genes tested, with 18.1% of isolates carrying more than three.Regarding bio lm formation, 7 (31.8%) and 4 (18.1%) isolates were found to form strong and moderate bio lms, respectively, while one (4.5%) isolate was classi ed as a weak bio lm producer. ERIC-PCR band patterns suggested that the isolated Aliarcobacter spp. from slaughterhouses had different sources of contamination. These ndings highlight the potential risk posed by pathogenic and multidrug-resistant Aliarcobacter spp. in food, and the need for control measures throughout the food chain to prevent the spread of these strains. The results indicate that foods of animal origin and cattle slaughterhouses are signi cant sources of antimicrobial resistant Aliarcobacters.

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Novel virulence, antibiotic resistance and toxin gene-specific PCR-based assays for rapid pathogenicity assessment of Arcobacter faecis and Arcobacter lanthieri

Cited by 25 publications

References 54 publications

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

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

Virulence genotype and phenotype of two clinical isolates of Arcobacter butzleri obtained from patients with different pathologies

Investigation and characterization of Aliarcobacter spp. isolated from cattle slaughterhouse in Türkiye

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