scholarly journals Characterization and antigenicity of recombinant Campylobacter jejuni flagellar capping protein FliD

2014 ◽  
Vol 63 (4) ◽  
pp. 602-609 ◽  
Author(s):  
Hung-Yueh Yeh ◽  
Kelli L. Hiett ◽  
John E. Line ◽  
Bruce S. Seal
Keyword(s):  
Campylobacter Jejuni ◽  
Broiler Chickens ◽  
Nucleotide Sequencing ◽  
Calculated Molecular Mass ◽  
Ionization Time ◽  
Bacterial Flagella ◽  
Capping Protein ◽  
Flight Mass Spectrometry ◽  
Sds Page ◽  
Bacterial Gastroenteritis

Campylobacter jejuni, a flagellated, spiral-rod, Gram-negative bacterium, is the leading pathogen of human acute bacterial gastroenteritis worldwide, and chickens are regarded as a major reservoir of this micro-organism. Bacterial flagella, composed of more than 35 proteins, play important roles in colonization and adhesion to the mucosal surface of chicken caeca. In this study, the flagellar capping protein, FliD, encoded by the fliD gene, from the Campylobacter jenuni D1-39 isolate was expressed and characterized, and its antigenicity determined. The fliD gene comprised 1929 nt, potentially encoding a 642 aa peptide with a calculated molecular mass of 69.6 kDa. This gene was PCR amplified and overexpressed in Escherichia coli. The recombinant FliD protein was purified by cobalt-chelating affinity chromatography and confirmed by nucleotide sequencing of the expression plasmid, SDS-PAGE analysis, His tag detection and matrix-assisted laser desorption/ionization time of flight mass spectrometry. The immunoblot data showed that the purified recombinant FliD protein reacted strongly to sera from broiler chickens older than 4 weeks, indicating that anti-FliD antibody may be prevalent in the poultry population. These results provide a rationale for further evaluation of the FliD protein as a vaccine candidate for broiler chickens to improve food safety for poultry.

scholarly journals Expression of the Campylobacter jejuni FliD protein and its reaction to chicken sera

2020 ◽  
Vol 367 (14) ◽  
Author(s):  
Xiao-Yan Zhang ◽  
Qian Zhou ◽  
Meng-Jun Tang ◽  
Jun-Hua Pu ◽  
Yan-Feng Fan ◽  
...  
Keyword(s):  
Campylobacter Jejuni ◽  
Broiler Chickens ◽  
Human Infection ◽  
Enzyme Linked Immunosorbent Assay ◽  
Serum Samples ◽  
Capping Protein ◽  
Cultural Status ◽  
Bacterial Gastroenteritis ◽  
Antibody Levels ◽  
Culture Negative

ABSTRACT Campylobacter is a leading causative pathogen of acute bacterial gastroenteritis among humans. Contaminated chicken products are regarded as major sources of human infection. The flagellar capping protein (FliD), which plays important roles in colonization and adhesion to the mucosal surface of chicken ceca, is conserved among Campylobacter jejuni strains. In this study, the recombinant C. jejuni FliD protein was expressed, purified and used as a coated protein to examine the prevalence of C. jejuni antibodies in chickens. The anti-FliD antibody was prevalent among chicken serum samples taken from different farms in the diverse regions of Jiangsu province by using enzyme-linked immunosorbent assay. The Campylobacter antibody was present in culture-negative chickens. No strong dose–response relationships were observed between serum FliD antibody levels and Campylobacter cultural status. These results provide a basis for further evaluating FliD as a vaccine candidate for broiler chickens or for examining host–C. jejuni interactions, with implications for improving food safety.

scholarly journals Identification of a novel mannose-capped lipoarabinomannan from Amycolatopsis sulphurea

2003 ◽  
Vol 372 (3) ◽  
pp. 821-829 ◽  
Author(s):  
Kevin J. C. GIBSON ◽  
Martine GILLERON ◽  
Patricia CONSTANT ◽  
Germain PUZO ◽  
Jérôme NIGOU ◽  
...  
Keyword(s):  
Cell Envelope ◽  
Original Structure ◽  
Average Molecular Mass ◽  
Ionization Time ◽  
Flight Mass Spectrometry ◽  
Sds Page ◽  
Intermediate Size ◽  
Genus Mycobacterium ◽  
Factor Α

The genus Amycolatopsis is a member of the phylogenetic group nocardioform actinomycetes, which also includes the genus Mycobacterium. Members of this group have a characteristic cell envelope structure, dominated by various complex lipids and polysaccharides. Amongst these, lipoglycans are of particular interest since mycobacterial lipoarabinomannans are important immunomodulatory molecules. In this study we report the isolation and structural characterization of Amycolatopsis sulphurea lipoarabinomannan, designated AsuLAM. SDS/PAGE analysis revealed that AsuLAM was of an intermediate size between Mycobacterium tuberculosis lipoarabinomannan and lipomannan, confirmed by matrix-assisted laser-desorption ionization–time-of-flight mass spectrometry that predicted an average molecular mass of 10 kDa. Using a range of chemical degradations, NMR experiments and capillary electrophoresis analysis, AsuLAM was revealed as an original structure. The mannosyl-phosphatidyl-myo-inositol anchor exhibits a single acyl-form, characterized by a diacylated glycerol moiety, and contains, as one of the main fatty acids, 14-methyl-pentadecanoate, a characteristic fatty acid of the Amycolatopsis genus. AsuLAM also contains a short mannan domain; and is dominated by a multi-branched arabinan domain, composed of an (α1→5)-Araf (arabinofuranose) chain substituted, predominately at the O-2 position, by a single β-Araf. The arabinan domain is further elaborated by manno-oligosaccharide caps, with around one per molecule. This is the first description of manno-oligosaccharide caps found in a non-mycobacterial LAM. AsuLAM was unable to induce the production of the pro-inflammatory cytokine tumour necrosis factor α when tested with human or murine macrophage cell lines, reinforcing the paradigm that mannose-capped LAM are poor inducers of pro-inflammatory cytokines.

Urinary protein profiling with surface-enhanced laser desorption/ionization time-of-flight mass spectrometry in ETB receptor-deficient ratsThis article is one of a selection of papers published in the special issue (part 2 of 2) on Forefronts in Endothelin.

2008 ◽  
Vol 86 (8) ◽  
pp. 566-570 ◽  
Author(s):  
Jens Raila ◽  
Philipp Kalk ◽  
Thiemo Pfab ◽  
Christa Thöne-Reineke ◽  
Michael Godes ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Laser Desorption ◽  
Laser Desorption Ionization ◽  
Ionization Time ◽  
Desorption Ionization ◽  
Flight Mass Spectrometry ◽  
Sds Page ◽  
Surface Enhanced ◽  
Urinary Peptide ◽  
Tof Ms

The pathways leading to salt-sensitive hypertension and renal damage in rescued ETB receptor-deficient (ETBRd) rats are still unknown. The objective of the study was therefore to identify modifications of urinary peptide and protein expression in ETBRd rats (n = 9) and wild-type controls (n = 6) using SDS – polyacrylamide gel electrophoresis (SDS-PAGE) and surface-enhanced laser desorption/ionization time-of-flight mass spectrometry (SELDI-TOF-MS) technology. Glomerular filtration rate, glomerulosclerosis, and tubulointerstitial fibrosis did not differ between the groups. ETBRd rats showed slightly higher blood pressure (p < 0.001), media/lumen ratio of intrarenal arteries (p < 0.01), and albuminuria (p < 0.01). SDS-PAGE confirmed albuminuria, but showed no differences in the urinary excretion of low molecular weight proteins (<60 kDa). SELDI-TOF-MS profiling revealed 9 proteomic features at molecular masses (Da) of 2720, 2980, 3130, 3345, 6466, 6682, 8550, 18 729, and 37 492, which were significantly elevated (p < 0.02) in urine of ETBRd rats. The results demonstrate that, independent of structural changes in the kidneys, ETB-receptor deficiency causes specific differences in urinary peptide and protein excretion. SELDI-TOF-MS may be a valuable tool for the characterization of urinary biomarkers helping to uncover the mechanism of ETBR action in the kidney.

scholarly journals Differential Distribution of the wlaN and cgtB Genes, Associated with Guillain-Barré Syndrome, in Campylobacter jejuni Isolates from Humans, Broiler Chickens, and Wild Birds

2020 ◽  
Vol 8 (3) ◽  
pp. 325 ◽  
Author(s):  
Pedro Guirado ◽  
Sonia Paytubi ◽  
Elisenda Miró ◽  
Yaidelis Iglesias-Torrens ◽  
Ferran Navarro ◽  
...  
Keyword(s):  
Campylobacter Jejuni ◽  
Broiler Chickens ◽  
Phase Variation ◽  
Wild Birds ◽  
Guillain Barre Syndrome ◽  
Ecological Niches ◽  
Differential Distribution ◽  
Guillain Barré Syndrome ◽  
Bacterial Gastroenteritis ◽  
Guillain Barré

Campylobacter jejuni causes campylobacteriosis, a bacterial gastroenteritis with high incidence worldwide. Moreover, C. jejuni infection can trigger the polyneuropathic disorder denominated Guillain-Barré syndrome (GBS). The C. jejuni strains that can elicit GBS carry either wlaN or cgtB, coding both genes for a β-1,3-galactosyltransferase enzyme that is required for the production of sialylated lipooligosaccharide (LOSSIAL). We described a differential prevalence of the genes wlaN and cgtB in C. jejuni isolates from three different ecological niches: humans, broiler chickens, and wild birds. The distribution of both genes, which is similar between broiler chicken and human isolates and distinct when compared to the wild bird isolates, suggests a host-dependent distribution. Moreover, the prevalence of the wlaN and cgtB genes seems to be restricted to some clonal complexes. Gene sequencing identified the presence of new variants of the G- homopolymeric tract within the wlaN gene. Furthermore, we detected two variants of a G rich region within the cgtB gene, suggesting that, similarly to wlaN, the G-tract in the cgtB gene mediates the phase variation control of cgtB expression. Caco-2 cell invasion assays indicate that there is no evident correlation between the production of LOSSIAL and the ability to invade eukaryotic cells.

scholarly journals Virulence, Antimicrobial Resistance and Biofilm Production of Escherichia coli Isolates from Healthy Broiler Chickens in Western Algeria

Antibiotics ◽  
2021 ◽  
Vol 10 (10) ◽  
pp. 1157
Author(s):  
Qada Benameur ◽  
Teresa Gervasi ◽  
Filippo Giarratana ◽  
Maria Vitale ◽  
Davide Anzà ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Antimicrobial Resistance ◽  
Broiler Chickens ◽  
Laser Desorption ◽  
Laser Desorption Ionization ◽  
Ionization Time ◽  
E Coli ◽  
Flight Mass Spectrometry ◽  
Biofilm Production ◽  
Western Algeria

The aim of this study was to assess the virulence, antimicrobial resistance and biofilm production of Escherichia coli strains isolated from healthy broiler chickens in Western Algeria. E. coli strains (n = 18) were identified by matrix-assisted laser desorption–ionization time-of-flight mass spectrometry. Susceptibility to 10 antibiotics was determined by standard methods. Virulence and extended-spectrum β-lactamase (ESBL) genes were detected by PCR. The biofilm production was evaluated by microplate assay. All the isolates were negative for the major virulence/toxin genes tested (rfbE, fliC, eaeA, stx1), except one was stx2-positive. However, all were resistant to at least three antibiotics. Ten strains were ESBL-positive. Seven carried the β-lactamase blaTEM gene only and two co-harbored blaTEM and blaCTX-M−1 genes. One carried the blaSHV gene. Among the seven strains harboring blaTEM only, six had putative enteroaggregative genes. Two contained irp2, two contained both irp2 and astA, one contained astA and another contained aggR, astA and irp2 genes. All isolates carrying ESBL genes were non-biofilm producers, except one weak producer. The ESBL-negative isolates were moderate biofilm producers and, among them, two harbored astA, two irp2, and one aggR, astA and irp2 genes. This study highlights the spread of antimicrobial-resistant E. coli strains from healthy broiler chickens in Western Algeria.

Participation of CheR and CheB in the chemosensory response of Campylobacter jejuni

Microbiology ◽  
2011 ◽  
Vol 157 (5) ◽  
pp. 1279-1289 ◽  
Author(s):  
Doungjit Kanungpean ◽  
Tsutomu Kakuda ◽  
Shinji Takai
Keyword(s):  
Campylobacter Jejuni ◽  
Important Determinant ◽  
Commensal Bacterium ◽  
Gradient Sensing ◽  
Sds Page ◽  
Genes Encoding ◽  
Chemotaxis Proteins ◽  
Bacterial Gastroenteritis ◽  
Chemosensory Response

Campylobacter jejuni is a leading cause of bacterial gastroenteritis in humans and a commensal bacterium of the intestinal tracts of animals, especially poultry. Chemotaxis is an important determinant for chicken colonization of C. jejuni. Adaptation has a crucial role in the gradient-sensing mechanism that underlies chemotaxis. The genome sequence of C. jejuni reveals the presence of genes encoding putative adaptation proteins, CheB and CheR. In-frame deletions of cheB, cheR and cheBR were constructed and the chemosensory behaviour of the resultant mutants was examined on swarm plates. CheB and CheR proteins significantly influence chemotaxis but are not essential for this behaviour to occur. Increased mobility of two methyl-accepting chemotaxis proteins (MCPs), DocC and Tlp1, during SDS-PAGE was detected in the mutants lacking functional CheB in the presence of CheR, presumably resulting from stable methylation of receptors. In vitro studies using tissue culture revealed that deletion of cheR resulted in hyperadherent and hyperinvasive phenotypes, while deletion of cheB resulted in nonadherent, noninvasive phenotypes. Furthermore, the ΔcheBR mutant showed significantly reduced ability to colonize chick caeca. Our data suggest that modification of chemoreceptors by the CheBR system is involved in regulation of chemotaxis in C. jejuni although CheB is apparently not controlled by phosphorylation.

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