Antimicrobial Resistance and in silico Virulence Profiling of Aliarcobacter butzleri Strains From German Water Poultry

Abstract

Aliarcobacter butzleri is an emerging foodborne and zoonotic pathogen that is usually transmitted via contaminated food or water. A. butzleri is not only the most prevalent Aliarcobacter species, it is also closely related to thermophilic Campylobacter, which have shown increasing resistance in recent years. Therefore, it is important to assess its resistance and virulence profiles. In this study, 45 Aliarcobacter butzleri strains from water poultry farms in Thuringia, Germany, were subjected to an antimicrobial susceptibility test using the gradient strip diffusion method and whole-genome sequencing. In the phylogenetic analysis, the genomes of the German strains showed high genetic diversity. Thirty-three isolates formed 11 subgroups containing two to six strains. The antimicrobial susceptibility testing showed that 32 strains were resistant to erythromycin, 26 to doxycycline, and 20 to tetracycline, respectively. Only two strains were resistant to ciprofloxacin, while 39 strains were resistant to streptomycin. The in silico prediction of the antimicrobial resistance profiles identified a large repertoire of potential resistance mechanisms. A strong correlation between a gyrA point mutation (Thr-85-Ile) and ciprofloxacin resistance was found in 11 strains. A partial correlation was observed between the presence of the bla3 gene and ampicillin resistance. In silico virulence profiling revealed a broad spectrum of putative virulence factors, including a complete lipid A cluster in all studied genomes.

Sequence-based detection and typing procedures for Burkholderia mallei: Assessment and prospects

“…WGS would enable the quick detection of genetic manipulation leading to a gain of function, e.g., artificially introduced genes that increase virulence or pathogenicity. Further, the natural gain of resistances by mutations in housekeeping genes and the spread of genes can be detected and monitored by WGS, either based on assemblies or even on sequence reads ( 74 , 79 , 80 ). However, the prerequisite is the availability of a reliable database for quickly screening the WGS data, which has not been established for B. mallei , yet.…”

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

“…Those most frequently reported were detected in both, and were: (1) EP2, comprising only yajR and which being overexpressed can confer multidrug resistance in E. coli (Nishino et al 2001 ); (2) EP3, consisting of four genes including macA1 and macB2 , which encode macrolide export proteins in A. butzleri (Fanelli et al 2019 ); (3) EP4, composed of four genes that include bepD and bepE , responsible for multidrug resistance in Brucella suis (Martin et al 2009 ); (4) EP5 or msbA , coding for a protein that is related to the family of multidrug resistance proteins (Karow et al 1993 ); (5) EP6 or uup , involved in the formation of a soluble ATP-binding cassette (ABC) ATPase that has been implicated in several processes different from transmembrane transport of molecules, such as excision of transposable elements and the deletion of single copies of tandem chromosomal repeats in E. coli (Burgos Zepeda et al 2010 ); (6) EP8 or sugE , encoding a transporter from the small multidrug resistance (SMR) family that also provides resistance to narrow quaternary ammonium compounds in E. coli and Enterobacter cloacae (Chung et al 2002 ; He et al 2011 ); (7) EP9 or fsr, conferring resistance to fosmidomycin in A. butzleri (Fanelli et al 2020 ); (8) EP10, made up of five genes including the macrolide export protein coding macA3 (Fanelli et al 2019 ); (9) EP11; (10) EP15; and (11) EP16, all of which comprise several genes including mexB1 (EP11), mexA1 , mexB2 (EP15), and mexA2 (EP16), allegedly involved in the formation of the MexAB–OprM system responsible for resistance to quinolones, macrolides, novobiocin, chloramphenicol, tetracycline, lincomycin and β-lactams in Pseudomonas aeruginosa (Li et al 1995 ; Masuda et al 2000 ); (12) EP13 or ydhP , that encodes an inner membrane transport protein; and (13) EP14 or bcr1 and (14) EP17 or bcr2 , involved in the resistance to sulphonamides and bicyclomycin in E. coli (Nichols et al 1989 ) and, as far as we know, only to bicyclomycin in A. butzleri (Chieffi et al 2020 ). Studies that have investigated the prevalence of these EP systems in collections of at least 40 strains (Isidro et al 2020 ; Müller et al 2020b ; Uljanovas et al 2023 ) show that their prevalence is generally close to or above 90%, except for EP16 and EP17, which in two studies were detected at rates of less than 70% (Isidro et al 2020 ; Uljanovas et al 2023 ). Based on the three studies cited, the least prevalent systems are EP18 and EP19 (around 2%), followed by EP1 (12–38.8%) and EP7 (21.3–53.1%).…”

“…has been determined by the detection of ten virulence markers: cadF and cj1349 , which encode fibronectin binding proteins that promote the binding of bacteria to intestinal cells (Dasti et al 2010 ; Konkel et al 2005 ); ciaB ( Campylobacter invasive antigen B), which contributes to host cell invasion through a secretion system (Konkel et al 1999 ); hecA , involved in attachment, aggregation and epidermal cell killing (Rojas et al 2002 ); hecB and tlyA , coding for a haemolysin activation protein and a haemolysin, respectively (Miller et al 2007 ; Wren et al 1998 ); the phospholipase encoding gene pldA , also associated with erythrocyte lysis (Istivan et al 2008 ); mviN , coding for a protein essential for peptidoglycan biosynthesis (Inoue et al 2008 ); and irgA and iroE , that code for functional components for iron acquisition and are required for establishing and maintaining infection (Goldberg et al 1990 ). However, whole genome sequencing (WGS) has proven to be a useful technique to further expand knowledge about Arcobacter, and several studies have focused on using it to provide more information (Buzzanca et al 2021 ; Fanelli et al 2019 ; Isidro et al 2020 ; Ma et al 2022 ; Miller et al 2007 ; Müller et al 2020a , b ). It should be noted that although the first genome of A. butzleri was sequenced and analysed several years ago (Miller et al 2007 ), until 2020, no comprehensive genome-scale comparative analysis of A. butzleri has been performed (Buzzanca et al 2021 ; Isidro et al 2020 ; Müller et al 2020a , b ).…”

“…However, whole genome sequencing (WGS) has proven to be a useful technique to further expand knowledge about Arcobacter, and several studies have focused on using it to provide more information (Buzzanca et al 2021 ; Fanelli et al 2019 ; Isidro et al 2020 ; Ma et al 2022 ; Miller et al 2007 ; Müller et al 2020a , b ). It should be noted that although the first genome of A. butzleri was sequenced and analysed several years ago (Miller et al 2007 ), until 2020, no comprehensive genome-scale comparative analysis of A. butzleri has been performed (Buzzanca et al 2021 ; Isidro et al 2020 ; Müller et al 2020a , b ). These latest studies corroborate the high diversity and plasticity of the genome of this emerging human pathogen, highlighting its pathogenic potential and adaptive capacity.…”

Design and Synthesis of Novel Antimicrobial Agents

“…The study emphasizes TQ’s promising effectiveness against multidrug-resistant (MDR) infections, particularly Candida albicans and Gram-positive bacteria. A total of 45 strains of the zoonotic and foodborne pathogen Aliarcobacter butzleri ( A. butzleri ) were examined by Müller et al [ 88 ] utilizing the gradient strip diffusion method and whole-genome sequencing for antibiotic susceptibility testing. Erythromycin, doxycycline, tetracycline, ciprofloxacin, and streptomycin resistance were found in German strains.…”