Multi-Omics of Campylobacter jejuni Growth in Chicken Exudate Reveals Molecular Remodelling Associated with Altered Virulence and Survival Phenotypes

Campylobacter jejuni is the leading cause of foodborne human gastroenteritis in the developed world. Infections are largely acquired from poultry produced for human consumption and poor food handling is thus a major risk factor. Chicken exudate (CE) is a liquid produced from defrosted commercial chicken products that facilitates C. jejuni growth. We examined the response of C. jejuni to growth in CE using a multi-omics approach. Changes in the C. jejuni proteome were assessed by label-based liquid chromatography coupled with tandem mass spectrometry (LC-MS/MS). We quantified 1328 and 1304 proteins, respectively, in experiments comparing 5% CE in Mueller–Hinton (MH) medium and 100% CE with MH-only controls. These proteins represent 81.8% and 80.3% of the predicted C. jejuni NCTC11168 proteome. Growth in CE induced profound remodelling of the proteome. These changes were typically conserved between 5% and 100% CE, with a greater magnitude of change observed in 100% CE. We confirmed that CE induced C. jejuni biofilm formation, as well as increasing motility and resistance against oxidative stress, consistent with changes to proteins representing those functions. Assessment of the C. jejuni metabolome showed CE also led to increased intracellular abundances of serine, proline, and lactate that were correlated with the elevated abundances of their respective transporters. Analysis of carbon source uptake showed prolonged culture supernatant retention of proline and succinate in CE-supplemented medium. Metabolomics data provided preliminary evidence for the uptake of chicken-meat-associated dipeptides. C. jejuni exposed to CE showed increased resistance to several antibiotics, including polymyxin B, consistent with changes to tripartite efflux system proteins and those involved in the synthesis of lipid A. The C. jejuni CE proteome was also characterised by very large increases in proteins associated with iron acquisition, while a decrease in proteins containing iron–sulphur clusters was also observed. Our data suggest CE is both oxygen- and iron-limiting and provide evidence of factors required for phenotypic remodelling to enable C. jejuni survival on poultry products.

[1]  Aoxin Guo,et al.  GlcNAc-1,6-anhydro-MurNAc Moiety Affords Unusual Glycosyl Acceptor that Terminates Peptidoglycan Elongation. , 2024, Journal of the American Chemical Society.

[2]  V. DiRita,et al.  Gut metabolite L-lactate supports Campylobacter jejuni population expansion during acute infection , 2024, Proceedings of the National Academy of Sciences of the United States of America.

[3]  S. Cordwell,et al.  Global Acetylomics of Campylobacter jejuni Shows Lysine Acetylation Regulates CadF Adhesin Processing and Human Fibronectin Binding. , 2023, Journal of proteome research.

[4]  W. Vollmer,et al.  Multiple Campylobacter jejuni proteins affecting the peptidoglycan structure and the degree of helical cell curvature , 2023, Frontiers in Microbiology.

[5]  A. Hensel,et al.  Campylobacter jejuni: targeting host cells, adhesion, invasion, and survival , 2023, Applied Microbiology and Biotechnology.

[6]  J. Mcdermott,et al.  The Missing Pieces: The Role of Secretion Systems in Campylobacter jejuni Virulence , 2023, Biomolecules.

[7]  T. Dinh,et al.  Roles of Aerotolerance, Biofilm Formation, and Viable but Non-Culturable State in the Survival of Campylobacter jejuni in Poultry Processing Environments , 2022, Microorganisms.

[8]  R. Shanmugasundaram,et al.  Campylobacter jejuni in Poultry: Pathogenesis and Control Strategies , 2022, Microorganisms.

[9]  V. Korolik,et al.  Extracellular c-di-GMP Plays a Role in Biofilm Formation and Dispersion of Campylobacter jejuni , 2022, Microorganisms.

[10]  C. Szymanski.  Bacterial glycosylation, it’s complicated , 2022, Frontiers in Molecular Biosciences.

[11]  Hyun Cheol Kim,et al.  Using 2D qNMR analysis to distinguish between frozen and frozen/thawed chicken meat and evaluate freshness , 2022, npj Science of Food.

[12]  Vincent T. Lee,et al.  The Campylobacter jejuni Response Regulator and Cyclic-Di-GMP Binding CbrR Is a Novel Regulator of Flagellar Motility , 2021, Microorganisms.

[13]  Wladimir Padilha da Silva,et al.  Virulence factors of foodborne pathogen Campylobacterjejuni. , 2021, Microbial pathogenesis.

[14]  S. Cordwell,et al.  Exploiting pglB Oligosaccharyltransferase-Positive and -Negative Campylobacter jejuni and a Multiprotease Digestion Strategy to Identify Novel Sites Modified by N-Linked Protein Glycosylation. , 2021, Journal of proteome research.

[15]  S. Cordwell,et al.  Integrated mass spectrometry-based multi-omics for elucidating mechanisms of bacterial virulence. , 2021, Biochemical Society transactions.

[16]  V. Kumar,et al.  Turnover Chemistry and Structural Characterization of the Cj0843c Lytic Transglycosylase of Campylobacter jejuni. , 2021, Biochemistry.

[17]  Xiaonan Lu,et al.  Survival and Control of Campylobacter in Poultry Production Environment , 2021, Frontiers in Cellular and Infection Microbiology.

[18]  R. Maia,et al.  Cross-contamination events of Campylobacter spp. in domestic kitchens associated with consumer handling practices of raw poultry. , 2020, International journal of food microbiology.

[19]  B. Wren,et al.  Characterization of Posttranslationally Modified Multidrug Efflux Pumps Reveals an Unexpected Link between Glycosylation and Antimicrobial Resistance , 2020, mBio.

[20]  S. Cordwell,et al.  Proteomics of Campylobacter jejuni Growth in Deoxycholate Reveals Cj0025c as a Cystine Transport Protein Required for Wild-type Human Infection Phenotypes , 2020, Molecular & Cellular Proteomics.

[21]  S. Cordwell,et al.  Identifying the targets and functions of N-linked protein glycosylation in Campylobacter jejuni. , 2020, Molecular omics.

[22]  M. Konkel,et al.  Taking Control: Campylobacter jejuni Binding to Fibronectin Sets the Stage for Cellular Adherence and Invasion , 2020, Frontiers in Microbiology.

[23]  Guoyao Wu,et al.  Composition of amino acids and related nitrogenous nutrients in feedstuffs for animal diets , 2020, Amino Acids.

[24]  Jennifer A. Taylor,et al.  Bent Bacteria: A Comparison of Cell Shape Mechanisms in Proteobacteria. , 2019, Annual review of microbiology.

[25]  V. Korolik.  The role of chemotaxis during Campylobacter jejuni colonisation and pathogenesis. , 2019, Current opinion in microbiology.

[26]  Nichollas E. Scott,et al.  Proteomics Reveals Multiple Phenotypes Associated with N-linked Glycosylation in Campylobacter jejuni* , 2019, Molecular & Cellular Proteomics.

[27]  C. Jobin,et al.  Campylobacter jejuni promotes colorectal tumorigenesis through the action of cytolethal distending toxin , 2018, Gut.

[28]  D. Hendrixson,et al.  FliW controls growth-phase expression of Campylobacter jejuni flagellar and non-flagellar proteins via the post-transcriptional regulator CsrA. , 2018, Microbiology.

[29]  M. Wösten,et al.  Catabolite repression in Campylobacter jejuni correlates with intracellular succinate levels , 2018, Environmental microbiology.

[30]  Xiaonan Lu,et al.  Environmental Stress-Induced Bacterial Lysis and Extracellular DNA Release Contribute to Campylobacter jejuni Biofilm Formation , 2017, Applied and Environmental Microbiology.

[31]  William E. Fondrie,et al.  Identification of the ESKAPE pathogens by mass spectrometric analysis of microbial membrane glycolipids , 2017, Scientific Reports.

[32]  H. Fernández,et al.  Intrinsic and Extrinsic Aspects on Campylobacter jejuni Biofilms , 2017, Front. Microbiol..

[33]  David A. Dik,et al.  Lytic transglycosylases: concinnity in concision of the bacterial cell wall , 2017, Critical reviews in biochemistry and molecular biology.

[34]  T. J. Johnson,et al.  Current and Potential Treatments for Reducing Campylobacter Colonization in Animal Hosts and Disease in Humans , 2017, Front. Microbiol..

[35]  M. Wösten,et al.  Function and Regulation of the C4-Dicarboxylate Transporters in Campylobacter jejuni , 2017, Front. Microbiol..

[36]  H. Vogel,et al.  Ligand binding specificity of the Escherichia coli periplasmic histidine binding protein, HisJ , 2017, Protein science : a publication of the Protein Society.

[37]  I. Kassem,et al.  Formate simultaneously reduces oxidase activity and enhances respiration in Campylobacter jejuni , 2017, Scientific Reports.

[38]  R. Hancock,et al.  Polymyxin: Alternative Mechanisms of Action and Resistance. , 2016, Cold Spring Harbor perspectives in medicine.

[39]  M. Stahl,et al.  The Helical Shape of Campylobacter jejuni Promotes In Vivo Pathogenesis by Aiding Transit through Intestinal Mucus and Colonization of Crypts , 2016, Infection and Immunity.

[40]  Michael E. Taveirne,et al.  Accumulation of Peptidoglycan O-Acetylation Leads to Altered Cell Wall Biochemistry and Negatively Impacts Pathogenesis Factors of Campylobacter jejuni* , 2016, The Journal of Biological Chemistry.

[41]  R. Reinhardt,et al.  The CsrA-FliW network controls polar localization of the dual-function flagellin mRNA in Campylobacter jejuni , 2016, Nature Communications.

[42]  P. Kiley,et al.  How Is Fe-S Cluster Formation Regulated? , 2015, Annual review of microbiology.

[43]  Euna Oh,et al.  Regulation of oxidative stress resistance in Campylobacter jejuni, a microaerophilic foodborne pathogen , 2015, Front. Microbiol..

[44]  Si Ming Man,et al.  Global Epidemiology of Campylobacter Infection , 2015, Clinical Microbiology Reviews.

[45]  D. Hofreuter.  Defining the metabolic requirements for the growth and colonization capacity of Campylobacter jejuni , 2014, Front. Cell. Infect. Microbiol..

[46]  Kathryn L. Cross,et al.  Chicken Juice Enhances Surface Attachment and Biofilm Formation of Campylobacter jejuni , 2014, Applied and Environmental Microbiology.

[47]  D. Schomburg,et al.  Utilization of host‐derived cysteine‐containing peptides overcomes the restricted sulphur metabolism of Campylobacter jejuni , 2014, Molecular microbiology.

[48]  D. Hendrixson,et al.  Flagellar biosynthesis exerts temporal regulation of secretion of specific Campylobacter jejuni colonization and virulence determinants , 2014, Molecular microbiology.

[49]  S. L. Svensson,et al.  Flagella-Mediated Adhesion and Extracellular DNA Release Contribute to Biofilm Formation and Stress Tolerance of Campylobacter jejuni , 2014, PloS one.

[50]  Jun Lin,et al.  A single nucleotide in the promoter region modulates the expression of the β-lactamase OXA-61 in Campylobacter jejuni. , 2014, The Journal of antimicrobial chemotherapy.

[51]  I. Kassem,et al.  The impairment of methylmenaquinol:fumarate reductase affects hydrogen peroxide susceptibility and accumulation in Campylobacter jejuni , 2014, MicrobiologyOpen.

[52]  B. Imperiali,et al.  The Renaissance of Bacillosamine and Its Derivatives: Pathway Characterization and Implications in Pathogenicity , 2014, Biochemistry.

[53]  H. Ingmer,et al.  Campylobacter jejuni carbon starvation protein A (CstA) is involved in peptide utilization, motility and agglutination, and has a role in stimulation of dendritic cells. , 2013, Journal of medical microbiology.

[54]  D. Schomburg,et al.  Contribution of Amino Acid Catabolism to the Tissue Specific Persistence of Campylobacter jejuni in a Murine Colonization Model , 2012, PloS one.

[55]  S. Thompson,et al.  Campylobacter jejuni CsrA complements an Escherichia coli csrA mutation for the regulation of biofilm formation, motility and cellular morphology but not glycogen accumulation , 2012, BMC Microbiology.

[56]  D. Hendrixson,et al.  Identification and analysis of flagellar coexpressed determinants (Feds) of Campylobacter jejuni involved in colonization , 2012, Molecular microbiology.

[57]  K. Strimmer,et al.  MALDIquant: a versatile R package for the analysis of mass spectrometry data , 2012, Bioinform..

[58]  M. Stahl,et al.  Nutrient Acquisition and Metabolism by Campylobacter jejuni , 2012, Front. Cell. Inf. Microbio..

[59]  B. Wren,et al.  Cellular Response of Campylobacter jejuni to Trisodium Phosphate , 2011, Applied and Environmental Microbiology.

[60]  P. Gibbs,et al.  Campylobacter spp. as a Foodborne Pathogen: A Review , 2011, Front. Microbio..

[61]  Xin Liu,et al.  l-Fucose utilization provides Campylobacter jejuni with a competitive advantage , 2011, Proceedings of the National Academy of Sciences.

[62]  B. Wren,et al.  Effect of chicken meat environment on gene expression of Campylobacter jejuni and its relevance to survival in food. , 2011, International journal of food microbiology.

[63]  G. Ermel,et al.  MCLMAN, a new minimal medium for Campylobacter jejuni NCTC 11168. , 2011, Research in microbiology.

[64]  Qijing Zhang,et al.  Phenotypic and Genotypic Evidence for l-Fucose Utilization by Campylobacter jejuni , 2010, Journal of bacteriology.

[65]  Xin Liu,et al.  Campylobacter jejuni free oligosaccharides: function and fate. , 2010, Virulence.

[66]  M. Wösten,et al.  Altered Linkage of Hydroxyacyl Chains in Lipid A of Campylobacter jejuni Reduces TLR4 Activation and Antimicrobial Resistance* , 2010, The Journal of Biological Chemistry.

[67]  N. Shearer,et al.  Reduction of fumarate, mesaconate and crotonate by Mfr, a novel oxygen-regulated periplasmic reductase in Campylobacter jejuni. , 2010, Environmental microbiology.

[68]  B. Pearson,et al.  Biofilm Formation by Campylobacter jejuni Is Increased under Aerobic Conditions , 2010, Applied and Environmental Microbiology.

[69]  L. Dragsted.  Biomarkers of meat intake and the application of nutrigenomics. , 2010, Meat science.

[70]  Carrie Goodson,et al.  The role of respiratory donor enzymes in Campylobacter jejuni host colonization and physiology. , 2009, Microbial pathogenesis.

[71]  Michael E. Taveirne,et al.  The Dual-Functioning Fumarate Reductase Is the Sole Succinate:Quinone Reductase in Campylobacter jejuni and Is Required for Full Host Colonization , 2009, Journal of bacteriology.

[72]  P. Guerry.  Campylobacter flagella: not just for motility. , 2007, Trends in microbiology.

[73]  C. Raetz,et al.  Lipid A modification systems in gram-negative bacteria. , 2007, Annual review of biochemistry.

[74]  John Kelly,et al.  Proteomic Analysis of Campylobacter jejuni 11168 Biofilms Reveals a Role for the Motility Complex in Biofilm Formation , 2006, Journal of bacteriology.

[75]  M. Williamson,et al.  The Campylobacter jejuni PEB1a adhesin is an aspartate/glutamate‐binding protein of an ABC transporter essential for microaerobic growth on dicarboxylic amino acids , 2006, Molecular microbiology.

[76]  O. Oyarzabal.  Reduction of Campylobacter spp. by commercial antimicrobials applied during the processing of broiler chickens: a review from the United States perspective. , 2005, Journal of food protection.

[77]  V. Korolik,et al.  Isolation and Expression of a Novel Molecular Class D β-Lactamase, OXA-61, from Campylobacter jejuni , 2005, Antimicrobial Agents and Chemotherapy.

[78]  M. Vaughn,et al.  Antimicrobial Resistance in Campylobacter , 2005, Emerging Infectious Diseases.

[79]  Brian H. Raphael,et al.  Secretion of Virulence Proteins from Campylobacter jejuni Is Dependent on a Functional Flagellar Export Apparatus , 2004, Journal of bacteriology.

[80]  V. DiRita,et al.  Identification of Campylobacter jejuni genes involved in commensal colonization of the chick gastrointestinal tract , 2004, Molecular microbiology.

[81]  H. Ingmer,et al.  Chicken juice, a food‐based model system suitable to study survival of Campylobacter jejuni , 2004, Letters in applied microbiology.

[82]  P. Barrow,et al.  l-Serine Catabolism via an Oxygen-Labile l-Serine Dehydratase Is Essential for Colonization of the Avian Gut by Campylobacter jejuni , 2004, Infection and Immunity.

[83]  C. Szymanski,et al.  Campylobacter--a tale of two protein glycosylation systems. , 2003, Trends in microbiology.

[84]  S. Hall,et al.  Growth of Campylobacter jejuni Supported by Respiration of Fumarate, Nitrate, Nitrite, Trimethylamine-N-Oxide, or Dimethyl Sulfoxide Requires Oxygen , 2002, Journal of bacteriology.

[85]  Arnoud H M van Vliet,et al.  The role of iron in Campylobacter gene regulation, metabolism and oxidative stress defense. , 2002, FEMS microbiology reviews.

[86]  E. Hani,et al.  JlpA, a novel surface‐exposed lipoprotein specific to Campylobacter jejuni, mediates adherence to host epithelial cells , 2001, Molecular microbiology.

[87]  B. Barrell,et al.  The genome sequence of the food-borne pathogen Campylobacter jejuni reveals hypervariable sequences , 2000, Nature.

[88]  J. Ketley,et al.  Campylobacter jejuni Contains Two Fur Homologs: Characterization of Iron-Responsive Regulation of Peroxide Stress Defense Genes by the PerR Repressor , 1999, Journal of bacteriology.

[89]  P. Coloe,et al.  Survival and growth of Campylobacter jejuni after artificial inoculation onto chicken skin as a function of temperature and packaging conditions. , 1998, Journal of food protection.

[90]  M. Doyle,et al.  Health Risks and Consequences of Salmonella and Campylobacter jejuni in Raw Poultry. , 1995, Journal of food protection.

[91]  M. Blaser,et al.  Experimental Campylobacter jejuni infection in humans. , 1988, The Journal of infectious diseases.

[92]  S. Craven,et al.  Campylobacter jejuni survival in chicken meat as a function of temperature , 1982, Applied and environmental microbiology.

[93]  S. Backert,et al.  Campylobacter Virulence Factors and Molecular Host-Pathogen Interactions. , 2021, Current topics in microbiology and immunology.

[94]  D. Kelly,et al.  The function, biogenesis and regulation of the electron transport chains in Campylobacter jejuni: New insights into the bioenergetics of a major food-borne pathogen. , 2019, Advances in microbial physiology.

[95]  B. Pearson,et al.  Two respiratory enzyme systems in Campylobacter jejuni NCTC 11168 contribute to growth on L-lactate. , 2011, Environmental microbiology.

[96]  Nichollas E. Scott,et al.  Mass spectrometric characterization of the Campylobacter jejuni adherence factor CadF reveals post‐translational processing that removes immunogenicity while retaining fibronectin binding , 2010, Proteomics.