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Chronic effects of Campylobacter infection
Abstract
Campylobacter jejuni is one of the most common causes of bacterial gastroenteritis and chronic sequelae, such as reactive arthritis and Guillain-Barré syndrome (GBS), are known to follow uncomplicated infections. While little is known about reactive arthritis following Campylobacter infection, our knowledge on the pathogenesis of Campylobacter-induced GBS is expanding rapidly and is summarized in this review.
... Ich budowa antygenowa jest u niektórych szczepów C. jejuni i C. coli zbli¿ona do budowy antygenów powierzchniowych komórek nerwowych cz³owieka. Zjawisko to okrela siê mianem mimikry antygenowej i prowadzi ono do powa¿nych powik-³añ chorób autoimmunologicznych, takich jak zespó³ Guillain-Barre czy choroba reumatoidalna (2,16,23). ...
... W Holandii, Irlandii, Szwajcarii i krajach Skandynawskich w 2000 r. Campylobacter by³ najczêciej izolowan¹ bakteri¹ od ludzi z przypadków zaka¿eñ pokarmowych (1,2,11,16). ...
... Coraz czêciej powik³aniem wi¹zanym z zaka¿eniem niektórymi szczepami Campylobacter jejuni s¹ choroby o pod³o-¿u autoimmunologicznym, jak np. wspomniany zespó³ Guillain-Barre (2,16). ...
Campylobacter sp. may colonize the intestinal tract of most mammal and bird species. The resultant infections are symptomless or can cause diarrhea. When they concern humans, they are a serious epidemiological problem. In developed countries campylobacteriosis is now one of the most frequent reasons for food-borne diseases and is usually associated with gastroenteritis. In rare cases the infection may lead to complications like septicemia or Gullain-Barre syndrome. Campylobacteriosis is a zoonozis, therefore animal food products are the main source of the infection of Campylobacter for humans. As a zoonozis, in Poland all cases of campylobacteriosis in animals must be registered.
... Similar to H. pylori, C. jejuni AmiA played an important role in septum cleavage during cell division, with a mutant strain producing long chains of unseparated cells. This phenotype is similar to that of an E. coli mutant lacking all three periplasmic amidases, AmiA, AmiB and AmiC (Heidrich et al., 2001;2002). The E. coli ∆ amiA∆ amiB∆ amiB mutant displayed a growth defect, but not to the same extent as that of C. jejuni ∆ amiA. ...
... Coccoid C. jejuni do not activate the intracellular Nod1 and Nod2 receptors of the innate immune system or trigger IL-8 secretion. These coccoid bacteria may serve as a means of escaping the immune system and as a reservoir for the development of chronic C. jejuni infections (Nachamkin, 2002;Sherman et al., 2010;Riddle et al., 2012;Bolton, 2015). As well, the presence of intracellular coccoid bacteria that are immunologically invisible may explain the recrudescence observed in the experimental C. jejuni infection of human volunteers (Baqar et al., 2010;Kirkpatrick and Tribble, 2011;Rimmer et al., 2018). ...
Campylobacter jejuni is a prevalent enteric pathogen that changes morphology from helical to coccoid under unfavorable conditions. Bacterial peptidoglycan maintains cell shape. As C. jejuni transformed from helical to coccoid, peptidoglycan dipeptides increased and tri‐ and tetrapeptides decreased. The DL‐carboxypeptidase Pgp1 important for C. jejuni helical morphology and putative N‐acetylmuramoyl‐L‐alanyl amidase AmiA were both involved in the coccoid transition. Mutants in pgp1 and amiA showed reduced coccoid formation, with ∆pgp1∆amiA producing minimal coccoids. Both ∆amiA and ∆amiA∆pgp1 lacked flagella and formed unseparated chains of cells consistent with a role for AmiA in cell separation. All strains accumulated peptidoglycan dipeptides over time, but only strains capable of becoming coccoid displayed tripeptide changes. C. jejuni helical shape and corresponding peptidoglycan structure are important for pathogenesis‐related attributes. Concomitantly, changing to a coccoid morphology resulted in differences in pathogenic properties; coccoid C. jejuni were non‐motile and non‐infectious, with minimal adherence and invasion of epithelial cells and an inability to stimulate IL‐8. Coccoid peptidoglycan exhibited reduced activation of innate immune receptors Nod1 and Nod2 versus helical peptidoglycan. C. jejuni also transitioned to coccoid within epithelial cells, so the inability of the immune system to detect coccoid C. jejuni may be significant in its pathogenesis.
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... (including C. coli) to human beings [3,4]. Symptoms of campylobacteriosis include mild to severe diarrhea, fever, abdominal cramps, watery to bloody diarrhea, vomiting, and major medical complications like Guillain-Barre syndrome, or chronic and sometimes deadly paralysis [4][5][6]. In addition, myopericarditis has been linked with C. coli enterocolitis [7]. ...
Campylobacter coli resides in the intestine of several commonly consumed animals, as well as water and soil. It leads to campylobacteriosis when humans eat raw/undercooked meat or come into contact with infected animals. A common manifestation of the infection is fever, nausea, headache, and diarrhea. Increasing antibiotic resistance is being observed in this pathogen. The increased incidence of C. coli infection, and post-infection complications like Guillain-Barré syndrome, make it an important pathogen. It is essential to find novel therapeutic targets and drugs against it, especially with the emergence of antibiotic-resistant strains. In the current study, genomes of 89 antibiotic-resistant strains of C. coli were downloaded from the PATRIC database. Potent drug targets (n = 36) were prioritized from the core genome (n = 1,337 genes) of this species. Riboflavin synthase was selected as a drug target and pharmacophore-based virtual screening was performed to predict its inhibitors from the NPASS (n = ~ 30,000 compounds) natural product library. The top three docked compounds (NPC115144, NPC307895, and NPC470462) were selected for dynamics simulation (for 50 ns) and ADMET profiling. These identified compounds appear safe for targeting this pathogen and can be further validated by experimental analysis before clinical trials.
Graphical abstract
... Ever since, campylobacteriosis has been at the top of the list regarding case numbers [1]. Symptoms include watery to hemorrhagic diarrhea and abdominal pain, while post-infection, severe but rare long-term sequelae can occur, such as Guillain-Barré syndrome, reactive arthritis, and erythema nodosum [2][3][4]. ...
Campylobacteriosis is a worldwide-occurring disease and has been the most commonly reported zoonotic gastrointestinal infection in the European Union in recent years. The development of successful phage-based intervention strategies will require a better understanding of phage–bacteria interactions to facilitate advances in phage cocktail design. Therefore, this study aimed to investigate the effects of newly isolated group II and group III phages and their combinations on current Campylobacter field strains. A continuous workflow for host range and efficiency of plating (EOP) value determination was combined with a qPCR-based phage group identification and a liquid-based planktonic killing assay (PKA). An advanced analysis scheme allowed us to evaluate phage cocktails by their efficacy in inhibiting bacterial population growth and the resulting phage concentrations. The results of this study indicate that data obtained from PKAs are more accurate than host range data based on plaque formation (EOP). Planktonic killing assays with Campylobacter appear to be a useful tool for a straightforward cocktail design. Results show that a group II phage vB_CcM-LmqsCP218-2c2 and group III phage vB_CjM-LmqsCP1-1 mixture would be most promising for practical applications against Campylobacter coli and Campylobacter jejuni.
... In addition, C. jejuni constitutes an invasive microorganism that could cause gastroenteritis associated with fever and frequent watery bloody diarrhea, abdominal pains, and occasional nausea (Silva et al., 2011;Bintsis, 2017). It is also linked with post-infection complications such as the immune-mediated neurological disorders of the Guillain-Barré Syndrome (GBS) (Nachamkin, 2002;Alshekhlee et al., 2008;Stojanov et al., 2020), its variant Miller Fisher Syndrome (MFS), or reactive arthritis (Altekruse et al., 1999;Ang et al., 2001). Notably, the infectious dose is thought to be lower than the one for other foodborne pathogens given that merely 500-800 bacteria could trigger human infection (Castano-Rodríguez et al., 2017). ...
Campylobacter jejuni ( C. jejuni ) is one of the major pathogens contributing to the enteritis in humans. Infection can lead to numerous complications, including but not limited to Guillain-Barre syndrome, reactive arthritis, and Reiter’s syndrome. Over the past two decades, joint efforts have been made toward developing a proper strategy of limiting the transmission of C. jejuni to humans. Nevertheless, except for biosecurity measures, no available vaccine has been developed so far. Judging from the research findings, Omp18, AhpC outer membrane protein, and FlgH flagellin subunits of C. jejuni could be adopted as surface protein antigens of C. jejuni for screening dominant epitope thanks to their strong antigenicity, expression of varying strains, and conservative sequence. In this study, bioinformatics technology was adopted to analyze the T-B antigenic epitopes of Omp18, AhpC, and FlgH in C. jejuni strain NCTC11168. Both ELISA and Western Blot methods were adopted to screen the dominant T-B combined epitope. GGS (GGCGGTAGC) sequence was adopted to connect the dominant T-B combined epitope peptides and to construct the prokaryotic expression system of tandem repeats of antigenic epitope peptides. The mouse infection model was adopted to assess the immunoprotective effect imposed by the trivalent T-B combined with antigen epitope peptide based on Omp18/AhpC/FlgH. In this study, a tandem epitope AhpC-2/Omp18-1/FlgH-1 was developed, which was composed of three epitopes and could effectively enhance the stability and antigenicity of the epitope while preserving its structure. The immunization of BALB/c mice with a tandem epitope could induce protective immunity accompanied by the generation of IgG2a antibody response through the in vitro synthesis of IFN-γ cytokines. Judging from the results of immune protection experiments, the colonization of C. jejuni declined to a significant extent, and it was expected that AhpC-2/Omp18-1/FlgH-1 could be adopted as a candidate antigen for genetic engineering vaccine of C. jejuni MAP.
... Symptoms of Campylobacter enteritis include watery to hemorrhagic diarrhea and abdominal pain. Severe long-term sequelae like the Guillain-Barré syndrome, reactive arthritis, and erythema nodosum can occur in rare cases subsequent to the infection (Nachamkin, 2002;Moore et al., 2005;Skarp et al., 2016). ...
Campylobacter spp. are a major cause of bacterial foodborne diarrhea worldwide. While thermophilic Campylobacter species asymptomatically colonize the intestines of chickens, most human infections in industrial countries have been attributed to consumption of chicken meat or cross-contaminated products. Bacteriophages (phages) are natural predators of bacteria and their use at different stages of the food production chain has been shown to reduce the public health burden of human campylobacteriosis. However, regarding regulatory issues, the use of lytic phages in food is still under discussion and evaluation. This study aims to identify lytic phages suitable for reducing Campylobacter bacteria along the food production chain. Therefore, four of 19 recently recovered phages were further characterized in detail for their lytic efficacy against different Campylobacter field strains and their suitability under food production settings at different temperatures and pH values. Based on the results of this study, the phages vB_CjM-LmqsCP1-4 and vB_CjM-LmqsCP1-5 appear to be promising candidates for the reduction of Campylobacter jejuni in food production settings.
... Among Campylobacter species, C. jejuni and C. coli are the major causes of acute gastroenteritis worldwide (Man, 2011;Kaakoush et al., 2015). In most cases, campylobacteriosis is self-limiting in healthy individuals; however, increasing evidence suggests that Campylobacter infection is associated with the development of autoimmune disorders affecting the nervous system and gastrointestinal tract (Ang et al., 2000;McCarthy and Giesecke, 2001;Nachamkin, 2002;Gradel et al., 2009). Moreover, patients with C. jejuni gastroenteritis are at a high risk of developing IBD (Navarro-Llavat et al., 2009;Antonelli et al., 2012;Arora et al., 2015). ...
Innate lymphoid cells (ILCs) are a heterogeneous group of cytokine-producing lymphocytes which are predominantly located at mucosal barrier surfaces, such as skin, lungs, and gastrointestinal tract. ILCs contribute to tissue homeostasis, regulate microbiota-derived signals, and protect against mucosal pathogens. ILCs are classified into five major groups by their developmental origin and distinct cytokine production. A recently emerged intriguing feature of ILCs is their ability to alter their phenotype and function in response to changing local environmental cues such as pathogen invasion. Once the pathogen crosses host barriers, ILCs quickly activate cytokine production to limit the spread of the pathogen. However, the dysregulated ILC responses can lead to tissue inflammation and damage. Furthermore, the interplay between ILCs and other immune cell types shapes the outcome of the immune response. Recent studies highlighted the important role of ILCs for host defense against intracellular pathogens. Here, we review recent advances in understanding the mechanisms controlling protective and pathogenic ILC responses to intracellular pathogens. This knowledge can help develop new ILC-targeted strategies to control infectious diseases and immunopathology.
... While the majority of cases are self-limiting, they can spread into the bloodstream in immunocompromised individuals and become potentially lethal (Whitehouse et al., 2018). In some instances, affected patients are at risk of Guillain-Barré syndrome (GBS), a severe post-infectious autoimmune disease that occurs weeks or months after the initial infectious gastrointestinal manifestation (Pithadia and Kakadia, 2010), which can also sometimes be life-threatening (Nachamkin, 2002). The high incidence of C. jejuni-associated disease in humans is largely due to its prevalence as a zoonotic agent in animals (Sheppard et al., 2011;Burnham and Hendrixson, 2018). ...
Although campylobacteriosis is a zoonotic foodborne illness, high-risk isolates from animal sources are rarely characterized, and the pathogenic potential of zoonotic strains remains an obstacle to effective intervention against human infection. HS19 has been acknowledged as a maker serotype represented by Campylobacter jejuni ( C. jejuni ) isolates from patients with post-infection Guillain-Barré syndrome (GBS), which is circulation in developed countries. However, a previous serotype epidemiological study of C. jejuni isolates in an animal population revealed that HS19 was also prevalent in isolates from cattle in China. In this study, to investigate the hazardous potential of zoonotic strains, 14 HS19 isolates from cattle were systematically characterized both by genotype and phenotype. The results showed that all of these cattle isolates belonged to the ST-22 complex, a high-risk lineage represented by 77.2% HS19 clinical isolates from patients worldwide in the PubMLST database, indicating that the ST-22 complex is the prominent clonal complex of HS19 isolates, as well as the possibility of clonal spread of HS19 isolates across different regions and hosts. Nevertheless, these cattle strains clustered closely with the HS19 isolates from patients, suggesting a remarkable phylogenetic relatedness and genomic similarity. Importantly, both tetracycline genes tet(O) and gyrA (T86I) reached a higher proportional representation among the cattle isolates than among the human clinical isolates. A worrying level of multidrug resistance (MDR) was observed in all the cattle isolates, and two MDR profiles of the cattle isolates also existed in human clinical isolates. Notably, although shared with the same serotype HS19 and sequence type ST-22, 35.7% of cattle isolates induced severe gastrointestinal pathology in the IL-10 –/– C57BL/6 mice model, indicating that some bacteria could change due to host adaptation to induce a disease epidemic, thus the associated genetic elements deserve further investigation. In this study, HS19 isolates from cattle were first characterized by a systematic evaluation of bacterial genomics and in vitro virulence, which improved our understanding of the potential zoonotic hazard from food animal isolates with high-risk serotypes, and provided critical information for the development of targeted C. jejuni mitigation strategies.
... In 2019, among the eight bacterial pathogens monitored by the Foodborne Diseases Active Surveillance Network (FoodNet), the overall incidence per 100,000 population was highest for Campylobacter (19.5 cases; Tack et al., 2020). While campylobacteriosis is largely a self-limiting illness, it is accompanied by acute gastroenteritis and is a leading antecedent for the severe autoimmune complication Guillain-Barré syndrome (Allos, 1997;Nachamkin, 2002;Scallan Walter et al., 2020). This, together with additional autoimmune sequelae, such as post-infectious irritable bowel syndrome and reactive arthritis, confers a high disease burden to human campylobacteriosis in the United States and worldwide (Batz et al., 2013;Kirk et al., 2015;Scallan et al., 2015;Scallan Walter et al., 2019). ...
Antibiotic-resistant Campylobacter constitutes a serious threat to public health. The clonal expansion of resistant strains and/or the horizontal spread of resistance genes to other strains and species can hinder the clinical effectiveness of antibiotics to treat severe campylobacteriosis. Still, gaps exist in our understanding of the risks of acquisition and spread of antibiotic resistance in Campylobacter. While the in vitro transfer of antimicrobial resistance genes between Campylobacter species via natural transformation has been extensively demonstrated, experimental studies have favored the use of naked DNA to obtain transformants. In this study, we used experimental designs closer to real-world conditions to evaluate the possible transfer of antimicrobial resistance genes between Campylobacter strains of the same or different species (Campylobacter coli or Campylobacter jejuni) and originating from different animal hosts (swine or turkeys). This was evaluated in vitro through co-culture experiments and in vivo with dual-strain inoculation of turkeys, followed by whole genome sequencing of parental and newly emerged strains. In vitro, we observed four independent horizontal gene transfer events leading to the acquisition of resistance to beta-lactams (blaOXA), aminoglycosides [aph(2′′)-If and rpsL] and tetracycline [tet(O)]. Observed events involved the displacement of resistance-associated genes by a mutated version, or the acquisition of genomic islands harboring a resistance determinant by homologous recombination; we did not detect the transfer of resistance-carrying plasmids even though they were present in some strains. In vivo, we recovered a newly emerged strain with dual-resistance pattern and identified the replacement of an existing non-functional tet(O) by a functional tet(O) in the recipient strain. Whole genome comparisons allowed characterization of the events involved in the horizontal spread of resistance genes between Campylobacter following in vitro co-culture and in vivo dual inoculation. Our study also highlights the potential for antimicrobial resistance transfer across Campylobacter species originating from turkeys and swine, which may have implications for farms hosting both species in close proximity.
... With several thousand cases each year, GBS is the most common presentation of acute flaccid paralysis in the United States [18]. Most cases of GBS are believed to follow an infectious disease episode. ...
Campylobacter species are pathogens of both humans and animals, and show a worldwide distribution. Campylobacter jejuni and C. coli are the most common causes of human gastroenteritis. The acute clinical illness ranges from mild watery diarrhea in developed countries to severe inflammatory diarrhea characterized by abdominal pain, fever, nausea, and bloody stool in developing countries. Major medical complications of C. jejuni infections include Guillain-Barre syndrome (GBS) and reactive arthritis. Campylobacter are fastidious organisms, and C. jejuni does not replicate outside a warm-blooded host. However, the infectious dose is low (< 1000 organisms), and transmission occurs through the food chain primarily by eating undercooked poultry or meat. C. jejuni is a natural commensal of chicken, and its presence in chicken flocks is frequent. Contaminated food and subsequent illness are a huge economical burden, and processing control and prevention measures are important.
... Lipooligosaccharides (LOS) and the capsule are involved in immune avoidance (Nachamkin, 2002). The genes that encode for the flagellum, capsule and LOS are found in highly variable regions of the genome, where mutations have been associated with variability in serum resistance, cell adherence and invasion of human intestinal epithelial cell lines (Fry et al., 2000). ...
Bacteria have been evolving and adapting in different hosts and environments for millions of years. Most human infections are zoonotic, occurring mainly in animals but also being transmissible to humans. Humans and animals have been living together for thousands of years and been sharing or retaining their own bacterial populations. Bacteria can adapt in new hosts and respond to different selective pressures but not all bacterial species are found in all hosts and environments. The genetic mechanisms that promote these adaptations are not fully understood. The work presented in this thesis investigates the genomic and phenotypic adaptations that promote colonization/proliferation of bacteria of the genus Campylobacter and explores variation at the species, lineage and gene level. Pangenomic comparative analyses revealed core and accessory gene variation highlighting the importance of gene gain and loss in the evolution of this species and the use of genomics in identifying molecular markers to monitor lineage specific in vivo infection experiments. Campylobacter are highly recombinogenic, thus a focus has been given on quantifying recombination in the genome. A detailed analysis of recombination has shown the proportion of the mobile genetic elements (mobilome) in the Campylobacter genus and pinpointed genes associated with host adaptation. Additionally, analysis of Campylobacter resistomes between species, lineages, hosts and environments revealed multidrug resistant (MDR) genomic islands (GIs) and the involvement of plasmids in horizontal gene transfer (HGT). This work has provided evidence of interspecies recombination between different species that share the same hosts and the genes associated with them. The work in this thesis has broadened understanding of how genomic plasticity can allow these versatile bacterial pathogens to adapt into new niches and environments.
... Campylobacter jejuni is a foodborne pathogen that is one of the most common causes of human gastroenteritis (1,2). Although the acute symptoms are usually self-limiting, long-term post infection complications such as Guillain-Barrésyndrome, Reiter's arthritis (3), and irritable bowel syndrome had been described (4)(5)(6). Association between C. jejuni-driven intestinal pathology and inflammatory bowel disease (IBD) has been also discussed (7,8). ...
Human pathogen Campylobacter jejuni is a significant risk factor for the development of long-term intestinal dysfunction although the cellular and molecular mechanisms remain scantily defined. IL-23 is an emerging therapeutic target for the treatment of inflammatory intestinal diseases, however its role in C. jejuni-driven intestinal pathology is not fully understood. IL-10 deficient mice represent a robust model to study the pathogenesis of C. jejuni infection because C. jejuni infection of mice lacking IL-10 results in symptoms and pathology that resemble human campylobacteriosis. To determine the role of IL-23 in C. jejuni-driven intestinal inflammation, we studied the disease pathogenesis in IL-23-/- mice with inhibited IL-10Rα signaling. These mice exhibited reduced intestinal pathology independent from bacterial clearance. Further, levels of IFNγ, IL-17, IL-22, TNF, and IL-6 were reduced and associated with reduced accumulation of neutrophils, monocytes and macrophages in the colon. Flow cytometry analysis revealed reduced production of IL-17 and IFNγ by group 1 and 3 innate lymphoid cells. Thus, our data suggest that IL-23 contributes to intestinal inflammation in C. jejuni infected mice by promoting IL-17 and IFNγ production by innate lymphoid cells.
... Campylobacteriosis in humans ranges in severity from mild gastroenteritis to acute self-limiting haemorrhagic diarrhoea involving severe inflammation and may lead to long-term sequelae including reactive arthritis and inflammatory neuropathies such as Guillain-Barré Syndrome [8][9][10][11]. Poultry are not generally affected by C. jejuni despite carrying large numbers of bacteria in their gastrointestinal tract [12,13]. However, some studies have reported a decrease in growth performance in chickens harbouring C. jejuni asymptomatically that may be associated with physiological changes in the intestines [14]. ...
Campylobacter jejuni is the leading bacterial cause of human gastroenteritis worldwide and the handling or consumption of contaminated poultry meat is the key source of infection. C. jejuni proteins FlpA and SodB and glycoconjugates containing the C. jejuni N-glycan have been separately reported to be partially protective vaccines in chickens. In this study, two novel glycoproteins generated by protein glycan coupling technology-G-FlpA and G-SodB (with two and three N-glycosylation sites, respectively)-were evaluated for efficacy against intestinal colonisation of chickens by C. jejuni strain M1 relative to their unglycosylated variants. Two independent trials of the same design were performed with either a high challenge dose of 107 colony-forming units (CFU) or a minimum challenge dose of 102 CFU of C. jejuni M1. While antigen-specific serum IgY was detected in both trials, no reduction in caecal colonisation by C. jejuni M1 was observed and glycosylation of vaccine antigens had no effect on the outcome. Our data highlight inconsistencies in the outcome of C. jejuni vaccination trials that may reflect antigen-, challenge strain-, vaccine administration-, adjuvant- and chicken line-specific differences from previously published studies. Refinement of glycoconjugate vaccines by increasing glycosylation levels or using highly immunogenic protein carriers could improve their efficacy.
... In most cases, clinical symptoms such as fever and diarrhea are self-limiting. However, complications may occur, such as reactive arthritis (Hannu et al., 2002) and Guillain-Barr e syndrome (Nachamkin, 2002). Unfortunately, no effective measures to control Campylobacter infection in poultry exist to date (Hermans et al., 2011b). ...
Oral administration of antibodies is a promising strategy against various infectious diseases. Previously, it was demonstrated that passive immunization by providing hyperimmune egg yolk through the feed reduces Campylobacter jejuni colonization in broilers. Campylobacteriosis is the most commonly reported bacterial foodborne zoonosis worldwide, and poultry products are the number one origin of these bacteria for human infection. To date, no effective control measures exist to limit Campylobacter colonization in the chicken's intestinal tract. Here, the effect of lyophilization of hyperimmune egg yolk on protection of broilers against C. jejuni was investigated. During an in vivo trial, broiler chickens were prophylactically given feed with lyophilized hyperimmune or non-immunized egg yolk powder starting from day 1 after hatch. At day 11, broilers were inoculated with C. jejuni according to a seeder model. Five days later, all broilers were euthanized and cecal content was examined for C. jejuni colonization. No decrease in C. jejuni colonization was found. The freeze-drying resulted in a 16-fold decrease of the antibody titer in the yolk powder compared to the fresh yolks, presumably caused by structural changes in the antibodies. In conclusion, applying freeze-dried hyperimmune egg yolk failed to protect broilers against C. jejuni colonization, possibly because lyophilization affected the antibodies' functionality.
... Whereas some patients are even asymptomatic or present with rather mild symptoms, others suffer from abdominal cramps, fever, watery of even bloody and inflammatory diarrhea that usually resolve within 1 week. In rare cases, however, post-infectious sequelae such as Gullain-Barré syndrome, Miller Fisher syndrome, or reactive arthritis may manifest [9][10][11]. The pathogenesis of acute human campylobacteriosis is strongly triggered by the activation of innate immune responses via Toll-like Receptor-4 (TLR-4) mediated sensing of the bacterial lipooligosaccharide (LOS) that is expressed on the surface of C. jejuni [12,13]. ...
Background:
The prevalence of human infections with the zoonotic pathogen Campylobacter jejuni is rising worldwide. Therefore, the identification of compounds with potent anti-pathogenic and anti-inflammatory properties for future therapeutic and/or preventive application to combat campylobacteriosis is of importance for global health. Results of recent studies suggested carvacrol (4-isopropyl-2-methylphenol) as potential candidate molecule for the treatment of campylobacteriosis in humans and for the prevention of Campylobacter colonization in farm animals.
Results:
To address this in a clinical murine infection model of acute campylobacteriosis, secondary abiotic IL-10-/- mice were subjected to synthetic carvacrol via the drinking water starting 4 days before peroral C. jejuni challenge. Whereas at day 6 post-infection placebo treated mice suffered from acute enterocolitis, mice from the carvacrol cohort not only harbored two log orders of magnitude lower pathogen loads in their intestines, but also displayed significantly reduced disease symptoms. Alleviated campylobacteriosis following carvacrol application was accompanied by less distinct intestinal apoptosis and pro-inflammatory immune responses as well as by higher numbers of proliferating colonic epithelial cells. Remarkably, the inflammation-ameliorating effects of carvacrol treatment were not restricted to the intestinal tract, but could also be observed in extra-intestinal organs such as liver, kidneys and lungs and, strikingly, systemically as indicated by lower IFN-γ, TNF, MCP-1 and IL-6 serum concentrations in carvacrol versus placebo treated mice. Furthermore, carvacrol treatment was associated with less frequent translocation of viable C. jejuni originating from the intestines to extra-intestinal compartments.
Conclusion:
The lowered C. jejuni loads and alleviated symptoms observed in the here applied clinical murine model for human campylobacteriosis highlight the application of carvacrol as a promising novel option for both, the treatment of campylobacteriosis and hence, for prevention of post-infectious sequelae in humans, and for the reduction of C. jejuni colonization in the intestines of vertebrate lifestock animals.
... (C. jejuni and C. coli) infection through handling and consuming contaminated water, meat, and raw milk (Nachamkin 2002;Friedman et al. 2004;Zhang et al. 2018). Environmental reservoirs of Campylobacter spp. ...
There is a growing trend to implement biosecurity measures in small commercial broiler flocks and trying to replace ineffective antimicrobial with alternative materials to interevent a strategy for the control of Campylobacter bacteria in these farms. This study was designed to determine the prevalence rate of Campylobacter spp. in broiler flocks and their environment. Thereafter, assess the efficiency of chitosan, zinc oxide nanoparticles (ZnO NPs), and chitosan/ZnO NPs composite against Campylobacter strains to adopt a novel control strategy based on the ability to use those nanocomposites. A total of 220 samples were collected from broiler flocks, their environment, and farm attendants that direct contact with birds. All samples were subjected to microbiological investigation for isolation, then molecular identification of bacteria using PCR. ZnO NPs and chitosan/ZnO NPs composite were synthesized then characterized by transmission electron microscopy (TEM), scanning electron microscopy (SEM), Fourier-transform infrared spectrum (FT-IR), and X-ray diffraction (X-RD). The efficiency of testing compounds was examined against 30 strains of Campylobacter coli (C. coli) to determine the minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC). The highest percentages of C. coli were isolated from the manure storage area, and broiler litter followed by flies, and feeders (66.7, 53.3, 40.0, and 33.3%, respectively). Both chitosan/ZnO NPs and ZnO NPs at a concentration of 0.5 μg/mL and 1.5 μg/mL, respectively showed complete efficiency (100%) against C. coli compared with chitosan compound. In conclusion, manure storage area and broiler litter represented the main reservoir of Campylobacter bacterial contaminant followed by flies in broiler poultry farms. Chitosan/ZnO NPs composite can be used in any biosecurity program of poultry farms as an alternative to ineffective antimicrobial agents.
... In sub-Saharan countries, diarrheal diseases are estimated to contribute up to 25% of all deaths in children under 5 years of age, with Campylobacter among the most common pathogens detected in diarrheic children 20,21 . In addition to gastroenteritis, Campylobacter infections can lead to growth deficits in children, malabsorption, gut inflammation, autoimmune diseases and neurological sequelae 19,21,22 . While Campylobacter jejuni and Campylobacter coli are most frequently associated with gastroenteritis, we could not detect a difference in abundance between treatment groups for these species, which is consistent with recent findings that C. jejuni was not associated with gastroenteritis episodes among infants and children in Mali and Gambia 23 . ...
The MORDOR I trial¹, conducted in Niger, Malawi and Tanzania, demonstrated that mass azithromycin distribution to preschool children reduced childhood mortality¹. However, the large but simple trial design precluded determination of the mechanisms involved. Here we examined the gut microbiome of preschool children from 30 Nigerien communities randomized to either biannual azithromycin or placebo. Gut microbiome γ-diversity was not significantly altered (P = 0.08), but the relative abundances of two Campylobacter species, along with another 33 gut bacteria, were significantly reduced in children treated with azithromycin at the 24-month follow-up. Metagenomic analysis revealed functional differences in gut bacteria between treatment groups. Resistome analysis showed an increase in macrolide resistance gene expression in gut microbiota in communities treated with azithromycin (P = 0.004). These results suggest that prolonged mass azithromycin distribution to reduce childhood mortality reduces certain gut bacteria, including known pathogens, while selecting for antibiotic resistance.
... Campylobacteriosis is one of the most important foodborne bacterial diseases worldwide and has been the most commonly reported zoonosis in the EU since 2005 1 . Clinical symptoms such as fever and diarrhoea are usually self-limiting, although in rare cases complications can occur, leading to reactive arthritis 2 , Guillain-Barré syndrome (GBS) 3 and inflammatory bowel disease (IBD) 4 . The disease is mainly caused by Campylobacter jejuni (C. ...
Campylobacter infections sourced mainly to poultry products, are the most important bacterial foodborne zoonoses worldwide. No effective measures to control these infections in broiler production exist to date. Here, we used passive immunization with hyperimmune egg yolks to confer broad protection of broilers against Campylobacter infection. Two novel vaccines, a bacterin of thirteen Campylobacter jejuni (C. jejuni) and C. coli strains and a subunit vaccine of six immunodominant Campylobacter antigens, were used for the immunization of layers, resulting in high and prolonged levels of specific immunoglobulin Y (IgY) in the hens’ yolks. In the first in vivo trial, yolks (sham, bacterin or subunit vaccine derived) were administered prophylactically in the broiler feed. Both the bacterin- and subunit vaccine-induced IgY significantly reduced the number of Campylobacter-colonized broilers. In the second in vivo trial, the yolks were administered therapeutically during three days before euthanasia. The bacterin IgY resulted in a significant decrease in C. jejuni counts per infected bird. The hyperimmune yolks showed strong reactivity to a broad representation of C. jejuni and C. coli clonal complexes. These results indicate that passive immunization with hyperimmune yolks, especially bacterin derived, offers possibilities to control Campylobacter colonization in poultry.
... Campylobacteriosis is accompanied with clinical manifestations such as abdominal pain, fever, and watery or bloody diarrhea that are mostly selflimiting [1,6,7]. In a minority of cases, severe postinfectious sequelae such as Guillain-Barré syndrome or reactive arthritis can occur [7,8]. ...
Background
Campylobacter jejuni infections constitute serious threats to human health with increasing prevalences worldwide. Our knowledge regarding the molecular mechanisms underlying host–pathogen interactions is still limited. Our group has established a clinical C. jejuni infection model based on abiotic IL-10−/− mice mimicking key features of human campylobacteriosis. In order to further validate this model for unraveling pathogen-host interactions mounting in acute disease, we here surveyed the immunopathological features of the important C. jejuni virulence factors FlaA and FlaB and the major adhesin CadF (Campylobacter adhesin to fibronectin), which play a role in bacterial motility, protein secretion and adhesion, respectively.
Methods and results
Therefore, abiotic IL-10−/− mice were perorally infected with C. jejuni strain 81-176 (WT) or with its isogenic flaA/B (ΔflaA/B) or cadF (ΔcadF) deletion mutants. Cultural analyses revealed that WT and ΔcadF but not ΔflaA/B bacteria stably colonized the stomach, duodenum and ileum, whereas all three strains were present in the colon at comparably high loads on day 6 post-infection. Remarkably, despite high colonic colonization densities, murine infection with the ΔflaA/B strain did not result in overt campylobacteriosis, whereas mice infected with ΔcadF or WT were suffering from acute enterocolitis at day 6 post-infection. These symptoms coincided with pronounced pro-inflammatory immune responses, not only in the intestinal tract, but also in other organs such as the liver and kidneys and were accompanied with systemic inflammatory responses as indicated by increased serum MCP-1 concentrations following C. jejuni ΔcadF or WT, but not ΔflaA/B strain infection.
Conclusion
For the first time, our observations revealed that the C. jejuni flagellins A/B, but not adhesion mediated by CadF, are essential for inducing murine campylobacteriosis. Furthermore, the secondary abiotic IL-10−/− infection model has been proven suitable not only for detailed investigations of immunological aspects of campylobacteriosis, but also for differential analyses of the roles of distinct C. jejuni virulence factors in induction and progression of disease.
Electronic supplementary material
The online version of this article (10.1186/s13099-019-0306-9) contains supplementary material, which is available to authorized users.
... В последние десятилетия в связи с развитием торговых связей отмечается увеличение инфицированности возбудителями кампилобактериоза сельскохозяйственных животных, птиц, в особенности кур [1,2,4]. Наиболее существенными, по мнению специалистов зарубежных стран, природными резервуарами возбудителей кампилобактериоза являются домашние сельскохозяйственные животные и птицы [3,5,6]. Описаны как спорадические случаи кампилобактериозной инфекции, так и вспышки, связанные с употреблением инфицированных пищевых продуктов и воды [7]. ...
The process of campylobacteriosis agent distribution among agricultural poultries was shown to depend upon a number of factors including conditions of maintenance and importation of the agent (with feed and etc.). Unsatisfactory sanitary conditions of poultry farms and high level of infection among hens lead to intensive contamination of different environmental objects with campylobacters. Therefore, the first priority is to select those objects which can really be exposed to contamination with campylobacters and provide circulation of infection among people, animals and poultry due to their functional characteristics.
... In North America, foodborne illness and antimicrobial resistant infections are important causes of morbidity in humans, and may even cause life-threatening infections Tanwar et al., 2014). Additionally, infections with these microorganisms may result in serious long-term health sequelae; for example, previous infection with Campylobacter jejuni has been identified as a risk factor for Guillain-Barré syndrome and reactive arthritis (Nachamkin et al., 2002). ...
This thesis investigated the role of Canada geese as carriers of pathogenic and resistant bacteria in southern Ontario, using samples collected from three sources: hunted birds, diagnostic specimens, and live birds. Based on multi-level logistic regression models, the prevalence of Campylobacter was significantly lower during the nesting period when birds are nonvolant, suggesting that bird mobility outside of the breeding season impacts the carriage of these microorganisms. Antimicrobial resistance in E. coli was significantly associated with source, which suggests the local environment may be an important source of resistant bacteria. Examination of E. coli resistance patterns and Campylobacter subtypes (based on comparative genomic fingerprinting using a 40-gene assay) in different flocks of geese revealed similar resistance profiles and molecular subtypes in birds of the same flock. Canada geese may be a source of these microorganisms, and have the potential to disseminate them in the environment due to their migratory nature.
... Clinical symptoms of campylobacteriosis can range from mild diarrhea to severe, watery to bloody diarrhea, fever and vomiting and can lead to serious medical sequelae, such as Guillain-Barr e syndrome, a debilitating and sometimes fatal paralysis (Nachamkin, 2002;Cameron et al., 2012;Xia et al., 2013). The most important potential transmission routes of Campylobacter spp., specifically Campylobacter coli and Campylobacter jejuni, to humans have been considered to be consumption and handling of improperly prepared poultry meat, activities related to recreational waters, contact with farm animals or pets and consumption of unpasteurized milk (Man, 2011;Silva et al., 2011). ...
... In Canada, annual incidence rates nearing 30 cases per 100,000 individuals have been reported (Galanis, 2007), although statistical models that account for unreported and undiagnosed cases suggest this rate could be as high as 447 cases per 100,000 individuals . While a majority of cases are self-limiting, post-infection complications, such as Guillain-Barré syndrome can be life threatening (Nachamkin et al., 1998;Nachamkin, 2002). Campylobacter jejuni is commonly isolated from the gastrointestinal tract of many different wild and domesticated species, including companion animals and food animals such as poultry and cattle (Lastovica et al., 2014). ...
Campylobacter jejuni is a leading human enteric pathogen worldwide and despite an improved understanding of its biology, ecology, and epidemiology, limited tools exist for identifying strains that are likely to cause disease. In the current study, we used subtyping data in a database representing over 24,000 isolates collected through various surveillance projects in Canada to identify 166 representative genomes from prevalent C. jejuni subtypes for whole genome sequencing. The sequence data was used in a genome-wide association study (GWAS) aimed at identifying accessory gene markers associated with clinically related C. jejuni subtypes. Prospective markers (n = 28) were then validated against a large number (n = 3,902) of clinically associated and non-clinically associated genomes from a variety of sources. A total of 25 genes, including six sets of genetically linked genes, were identified as robust putative diagnostic markers for clinically related C. jejuni subtypes. Although some of the genes identified in this study have been previously shown to play a role in important processes such as iron acquisition and vitamin B5 biosynthesis, others have unknown function or are unique to the current study and warrant further investigation. As few as four of these markers could be used in combination to detect up to 90% of clinically associated isolates in the validation dataset, and such markers could form the basis for a screening assay to rapidly identify strains that pose an increased risk to public health. The results of the current study are consistent with the notion that specific groups of C. jejuni strains of interest are defined by the presence of specific accessory genes.
... It manifests as an acute inflammatory diarrhea with symptoms common to other bacterial enteritidesabdominal pain, fever and watery diarrhea often accompanied with the presence of blood and leukocytes in stool (Blaser and Engberg, 2008). In most cases, campylobacteriosis is selflimiting and does not require specific therapy, however severe autoimmune disorders, such as Guillain-Barré and Miller-Fisher syndromes (Salloway et al., 1996;Nachamkin, 2002), reactive arthritis (Pope et al., 2007), and inflammatory bowel disease (Rodriguez et al., 2006) may appear. These late-onset complications, together with long convalescence time and high occurrence of campylobacteriosis, are the reasons why the disease is ranked as an infection with one of the highest annual burden (Batz et al., 2012;Gibney et al., 2014;Mangen et al., 2015). ...
Campylobacter jejuni has been reported as a major cause of bacterial food-borne enteritides in developed countries during the last decade. Despite its fastidious growth requirements, including low level of oxygen and high level of CO2, this pathogen is able to persist in the environment without permanent loss of its viability and virulence. As C. jejuni is not able to multiply outside a host, the cells spend significant amount of time in stationary phase of growth. The entry into the stationary phase is often correlated to resistance to various stresses in bacteria. The switching between exponential and stationary phases is frequently mediated by the regulator sigma S (RpoS). However, this factor is absent in C. jejuni and molecular mechanisms responsible for transition of cells to the stationary phase remain elusive. In this work, proteomic profiles of cells from exponential and stationary phases were compared using 2-D electrophoresis (2DE) fingerprinting combined with mass spectrometry analysis and qRT-PCR. The identified proteins, whose expression differed between the two phases, are mostly involved in protein biosynthesis, carbon metabolism, stress response and motility. Altered expression was observed also in the pleiotropic regulator CosR that was over-expressed during stationary phase. A shift between transcript and protein level evolution of CosR throughout the growth of C. jejuni was observed using qRT-PCR and (2DE). From these data, we hypothesized that CosR could undergo a negative autoregulation in stationary phase. A consensus sequence resulting from promoter sequence alignment of genes potentially regulated by CosR, including its own upstream region, among C. jejuni strains is proposed. To verify experimentally the potential autoregulation of CosR at the DNA level, electrophoretic mobility shift assay was performed with DNA fragments of CosR promoter region and rCosR. Different migration pattern of the promoter fragments indicates the binding capacity of CosR, suggesting its auto-regulation potential.
... The infection is generally self-limiting and can be treated with rehydration therapy. However, in rare cases chronic sequelae can develop, for example reactive arthritis or Guillain-Barré syndrome (GBS), autoimmune disorders that can lead to temporary or in severe cases permanent paralysis (Nachamkin, 2002). The fact that campylobacters are highly prevalent in the intestinal tracts of farm livestock, such as poultry and pigs, renders it a major foodborne pathogen, and therefore represents a significant risk to consumer health (Boes et al., 2005;McCrea et al., 2005). ...
A screen of bacteriophages infecting a panel of Campylobacter jejuni PT14 gene knock-out mutants identified a role for the minor flagellin encoded by the flaB gene, in the defense of the host against CP8unalikevirus bacteriophage CP_F1 infection. Inactivation of the flaB gene resulted in an increase in the susceptibility of PT14 cultures to infection by CP_F1 and an increase in bacteriophage yields. Infection of wild type PT14 with CP_F1 produces turbid plaques in bacterial lawns, from which 78% of the resistant isolates recovered exhibit either attenuation or complete loss of motility. CP_F1 produces clear plaques on the flaB mutant with no regrowth in the lysis zones. Complementation of the mutant restored overgrowth and the development of resistance at the expense of motility. Further analyses revealed an increase in bacteriophage adsorption constant of nearly two-fold and burst-size three-fold, relative to the wild type. Motility analysis showed no major reduction in swarming motility in the flaB mutant. Thus we propose a new role for FlaB in the defense of campylobacters against bacteriophage infection.
... The infectious dose was estimated to be as low as 500-800 bacteria (Robinson, 1981;Young et al., 2007). In addition, some patients can develop post-infection complications including Guillian Barré Syndrome (GBS) and its variant form Miller Fisher Syndrome (MFS), a chronic and potentially fatal form of paralysis (Nachamkin, 2002;Kuwabara, 2011;Huizinga et al., 2015). ...
Campylobacter jejuni accounts for one of the leading causes of foodborne bacterial enteritis in humans. Despite being considered an obligate microaerobic microorganism, C. jejuni is regularly exposed to oxidative stress. However, its adaptive strategies to survive the atmospheric oxygen level during transmission to humans remain unclear. Recently, the clinical C. jejuni strain Bf was singled out for its unexpected ability to grow under ambient atmosphere. Here, we aimed to understand better the biological mechanisms underlying its atypical aerotolerance trait using two-dimensional protein electrophoresis, gene expression, and enzymatic activities. Forty-seven proteins were identified with a significantly different abundance between cultivation under microaerobic and aerobic conditions. The over-expressed proteins in aerobiosis belonged mainly to the oxidative stress response, enzymes of the tricarboxylic acid cycle, iron uptake, and regulation, and amino acid uptake when compared to microaerobic conditions. The higher abundance of proteins related to oxidative stress was correlated to dramatically higher transcript levels of the corresponding encoding genes in aerobic conditions compared to microaerobic conditions. In addition, a higher catalase-equivalent activity in strain Bf was observed. Despite the restricted catabolic capacities of C. jejuni, this study reveals that strain Bf is equipped to withstand oxidative stress. This ability could contribute to emergence and persistence of particular strains of C. jejuni throughout food processing or macrophage attack during human infection.
... According to a report by the WHO (1997), the use of fluoroquinolones (antibacterials that prevent bacterial DNA from unwinding and duplicating) in poultry has caused a dramatic increase in the incidence of resistant strains of Campylobacter spp. in poultry and subsequently in humans (McDermott et al., 2002; Nachamkin et al., 2000). The first resistant strains of C. jejuni in Europe were discovered during the 1980s (Nachamkin, 2002). The removal of fluoroquinolones from the battery of veterinary medicines has not entirely eliminated the presence of resistant C. jejuni and C. coli in animals and foods of animal origin (Smith and Fratamico, 2010), on the contrary it seems that such resistance is even increasing (Ge et al. 2013; Wimalarathna et al., 2013Fluoroquinolones inhibit the growth of bacteria by binding to bacterial DNA gyrase and DNA topoisomerase IV. ...
... The illness is usually resolved within 2-10 days without the use of antibiotics and without complications. However, C. jejuni infection can result in the development of the rare neuromuscular disease Guillian-Barré syndrome, which occurs at a frequency of about 1 per 1000 infected people (Nachamkin 2002). ...
Helicobacter and Campylobacter species are Gram-negative microaerophilic host-associated heterotrophic bacteria that invade the digestive tract of humans and animals. Campylobacter jejuni is the major worldwide cause of foodborne gastroenteritis in humans, while Helicobacter pylori is ubiquitous in over half of the world's population causing gastric and duodenal ulcers. The colonisation of the gastrointestinal system by Helicobacter and Campylobacter relies on numerous cellular defences to sense the host environment and respond to adverse conditions, including those imposed by the host immunity. An important antimicrobial tool of the mammalian innate immune system is the generation of harmful oxidative and nitrosative stresses to which pathogens are exposed during phagocytosis. This review summarises the regulators, detoxifying enzymes and subversion mechanisms of Helicobacter and Campylobacter that ultimately promote the successful infection of humans.
... Over 90% of human infections are caused by one species, Campylobacter jejuni (Gillespie et al., 2008), the majority of which suffer mild illness manifested as watery or bloody diarrhoea, fever and vomiting. In a minority of cases serious medical sequelae, such as Guillain-Barré syndrome, may occur (Nachamkin, 2002). Campylobacter are common in the intestinal tract of poultry and most campylobacteriosis cases are directly or indirectly associated with broilers (EFSA, 2011). ...
Studies were undertaken to investigate the effect of oxidative stress conditions (exposure to hydrogen peroxide, H2O2) on [1] the expression of 14 Campylobacter jejuni virulence-associated genes associated with motility and/or invasion (flaA, flaB, flhA, flhB, ciaB, iamA), adhesion (cadF), cytotoxin production (cdtA, cdtB, cdtC) as well as some of the regulators of these genes (rpoN, fliA, luxS, cj1000), in 10 C. jejuni strains (5 poultry and 5 human) and [2] the ability of these cells to adhere to and invade Caco-2 cells. Using 16S rRNA as the reference gene (preliminary research demonstrated that this gene was stably expressed), the expression of the 14 virulence associated genes was investigated under normal and oxidative stress conditions using reverse transcription PCR. A Caco-2 cell tissue culture assay was used to examine adhesion and invasion. The response to oxidative stress was strain-dependent. Two strains showed significant (p < 0.05) up or down regulation in 7 of the 14 genes tested, while only 1–2 genes were affected in the remaining strains. Expression of cadF was significantly (p < 0.05) changed in all strains, cdt B in 4 strains and cj1000 in 3 strains. Expression of the remaining genes was either unaffected or significantly altered in 1–2 strains. NCTC 11168 completely lost the ability to adhere to and invade Caco-2 cells. One other strain also demonstrated reduced adherence while two others were unable to invade Caco-2 cells after exposure to oxidative stress conditions. In contrast strain 7, a poultry isolate, showed increased invasion. It was concluded that oxidative stress affects expression of C. jejuni virulence genes in a strain-dependent manner, CadF may have a secondary survival function and the cdtB gene may have a different promoter than cdtA and cdtC.
... La sintomatologia è caratterizzata da manifestazioni gastroenteriche quali forti dolori addominali e diarrea talvolta emorragica; le manifestazioni cliniche normalmente si esauriscono entro CARNE una settimana (Phillips, 1995). Tuttavia, seppur raramente, si possono osservare gravi sequele come l'Artrite Reattiva e la Sindrome di Guillain Barré, che attualmente è considerata la più comune causa di paralisi flaccida acuta dell'uomo (Nachamkin, 2002;Ho et al., 1998). I serbatoi naturali di C. jejuni sono numerosi, come dimostrano gli isolamenti da suini, bovini, pecore, cani e gatti (Blaser et al., 1980;Bacci et al., 2004). ...
A survey was carried out to molecularly characterize isolates of thermophilic campylobacters from raw poultry meat samples marketed at retail level. The isolates were identified to species by multiplex-PCR. PCR-RFLP of the flagellinA gene was performed to genotype the isolated strains. The prevalence of thermophilic campylobacters in the food was high (73%). Interestingly, C. jejuni and C. coli were found to be concomitantly present in 14% of the positive samples analyzed and different genotypes were frequently found in the same sample (27%). This study highlighted that thermophilic campylobacters are commonly present in raw meat poultry meat and the concomitance of several genotypes in the same sample suggests that several sources of contamination are involved.
... The incidence of GBS after C. jejuni infection has been estimated to be about 1.17 per 1000 person-years based on a cohort study in the United Kingdom [40]. Characteristics of the host as well as of the bacteria play a role in immune response, including whether autoantibody proliferation and other features of autoimmune peripheral neuropathy occur [41]. C. jejuni bacteria with specific genetic characteristics and specific lipo-oligosaccharide structures are more strongly associated with cross-reactive antibodies and autoimmune peripheral neuropathy than other types of C. jejuni [42]. ...
Introduction:
Foodborne Campylobacter jejuni infection has been associated with an increased risk of autoimmune peripheral neuropathy, but risks of occupational exposure to C. jejuni have received less attention. This study compared anti-C. jejuni IgA, IgG, and IgM antibody levels, as well as the likelihood of testing positive for any of five anti-ganglioside autoantibodies, between animal farmers and non-farmers. Anti-C. jejuni antibody levels were also compared between farmers with different animal herd or flock sizes. The relationship between anti-C. jejuni antibody levels and detection of anti-ganglioside autoantibodies was also assessed.
Methods:
Serum samples from 129 Agricultural Health Study swine farmers (some of whom also worked with other animals) and 46 non-farmers, all from Iowa, were analyzed for anti-C. jejuni antibodies and anti-ganglioside autoantibodies using ELISA. Information on animal exposures was assessed using questionnaire data. Anti-C. jejuni antibody levels were compared using Mann-Whitney tests and linear regression on log-transformed outcomes. Fisher's Exact Tests and logistic regression were used to compare likelihood of positivity for anti-ganglioside autoantibodies.
Results:
Farmers had significantly higher levels of anti-C. jejuni IgA (p < 0.0001) and IgG (p = 0.02) antibodies compared to non-farmers. There was no consistent pattern of anti-C. jejuni antibody levels based on animal herd or flock size. A higher percentage of farmers (21%) tested positive for anti-ganglioside autoantibodies compared to non-farmers (9%), but this difference was not statistically significant (p = 0.11). There was no significant association between anti-C. jejuni antibody levels and anti-ganglioside autoantibodies.
Conclusions:
The findings provide evidence that farmers who work with animals may be at increased risk of exposure to C. jejuni. Future research should include longitudinal studies of exposures and outcomes, as well as studies of interventions to reduce exposure. Policies to reduce occupational exposure to C. jejuni should be considered.
... Quinolones -According to a report by the WHO (1997), the use of fluoroquinolones (antibacterials that prevent bacterial DNA from unwinding and duplicating) in poultry has caused a dramatic increase in the incidence of resistant strains of Campylobacter spp. in poultry and subsequently in humans (Nachamkin et al., 2000;McDermott et al., 2002). The first resistant strains of C. jejuni in Europe were discovered during the 1980s (Nachamkin, 2002). The removal of fluoroquinolones from the battery of veterinary medicines has not entirely eliminated the presence of resistant C. jejuni and C. coli in animals and foods of animal origin (Smith and Fratamico, 2010), on the contrary it seems that such resistance is even increasing (Ge et al. 2013;Wimalarathna et al., 2013). ...
This review gives an overview on the prevalence of antimicrobial resistance in the food chain
in the European Union. The main emphasis is on two important food pathogens,
Campylobacter spp. and Salmonella spp. Furthermore, antibiotic residues reported in food commodities in the EU during 2008-2012, as well as the current legal framework regarding antibiotic use in the EU are discussed. In addition, the review also presents alternatives for the antibiotic treatment of food of animal origin.
It has been well documented that Campylobacter is the leading cause of foodborne infections and bacterial enteritis in high-income countries. The gastrointestinal tract of most warm-blooded animals, such as mammals and poultry, is prone to this pathogen. Infections caused by this bacterium in humans have usually been associated with the consumption of contaminated poultry meat. The important point about Campylobacter is that this bacterium has adapted to harsh environmental conditions along the food chain (poultry digestive tract to the consumer's plate) and developed an adapted mechanism to those conditions. This study aimed to compare the ability of Campylobacter jejuni and Campylobacter coli strains to form biofilms under aerobic and microaerobic conditions. The presence and expression of flab, FliS, DnaK, luxs, CsrA, Cj0688, and cosR genes involved in biofilm formation were investigated. Finally, the correlation between the biofilm forming ability of Campylobacter isolates and the presence/expression of selected genes has been explored. A significant correlation was observed between the presence and expression of some genes and the degree of biofilm formation in C. jejuni and C. coli isolates. A strong biofilm production was detected in strains harboring all selected genes with greater expression levels. The ability of C. jejuni and C. coli strains in biofilm formation is associated with the coordinated function and convergent expression of the selected genes. Seemingly, stress response- and motility-related genes have the most involvement in biofilm formation of C. jejuni and C. coli strains, while other genes have an accessory role in this phenomenon.
Campylobacter jejuni is the leading bacterial cause of human gastroenteritis worldwide and handling or consumption of contaminated poultry meat is the key source of infection. Glycoconjugate vaccines containing the C. jejuni N-glycan have been reported to be partially protective in chickens. However, our previous studies with subunit vaccines comprising the C. jejuni FlpA or SodB proteins with up to two or three C. jejuni N-glycans, respectively, failed to elicit significant protection. In this study, protein glycan coupling technology was used to add up to ten C. jejuni N-glycans onto a detoxified form of Pseudomonas aeruginosa exotoxin A (ExoA). The glycoprotein, G-ExoA, was evaluated for efficacy against intestinal colonisation of White Leghorn chickens by C. jejuni strains M1 and 11168H relative to unglycosylated ExoA. Chickens were challenged with the minimum dose required for reliable colonisation, which was 10² colony-forming units (CFU) for strain M1 and and 10⁴ CFU for strain 11168H. Vaccine-specific serum IgY was detected in chickens vaccinated with both ExoA and G-ExoA. However, no reduction in caecal colonisation by C. jejuni was observed. While the glycan dose achieved with G-ExoA was higher than FlpA- or SodB-based glycoconjugates that were previously evaluated, it was lower than that of glycoconjugates where protection against C. jejuni has been reported, indicating that protection may be highly sensitive to the amount of glycan presented and/or study-specific variables.
Campylobacter has been one of the most common causative agent of bacterial food-borne gastroenteritis in humans worldwide. However, in Brazil the campylobacteriosis has been a neglected disease and there is insufficient data to estimate the incidence of this pathogen in the country.
Aims
The current study aimed to determine the phylogenetic relationships among Campylobacter coli strains isolated in Brazil and to compare them with international Campylobacter isolates available in some public databases.
Methods and results
A total of 63 C. coli strains isolated in Brazil were studied. The MLST analysis showed 18 different STs including three STs not yet described in the PubMLST database. The cgMLST allocated the Brazilian strains studied into five main clusters and each cluster comprised groups of strains with nearly identical cgMLST profiles and with significant genetic distance observed among the distinct clusters. The comparison of the Brazilian strains with 3401 isolates from different countries showed a wide distribution of these strains isolated in this country.
Conclusions
The results showed a high similarity among some strains studied and a wide distribution of the Brazilian strains when compared to isolates from different countries, which is an interesting data set since it showed a high genetic diversity of these strains from Brazil in a global context. This study contributed for a better genomic characterization of C. coli strains isolated in Brazil and provides important information about the diversity of this clinically-relevant pathogen.
Campylobacter is the leading cause of the human bacterial foodborne infections in the developed countries. The perception cues from biotic or abiotic environments by the bacteria are often related to bacterial surface and membrane proteins that mediate the cellular response for the adaptation of Campylobacter jejuni to the environment. These proteins function rarely as a unique entity, they are often organized in functional complexes. In C. jejuni, these complexes are not fully identified and some of them remain unknown. To identify putative functional multi-subunit entities at the membrane subproteome level of C. jejuni, a holistic non a priori method was addressed using two-dimensional blue native/Sodium dodecyl sulfate (SDS) polyacrylamide gel electrophoresis (PAGE) in strain C. jejuni 81–176. Couples of acrylamide gradient/migration-time, membrane detergent concentration and hand-made strips were optimized to obtain reproducible extraction and separation of intact membrane protein complexes (MPCs). The MPCs were subsequently denatured using SDS-PAGE and each spot from each MPCs was identified by mass spectrometry. Altogether, 21 MPCs could be detected including multi homo-oligomeric and multi hetero-oligomeric complexes distributed in both inner and outer membranes. The function, the conservation and the regulation of the MPCs across C. jejuni strains were inspected by functional and genomic comparison analyses. In this study, relatedness between subunits of two efflux pumps, CmeABC and MacABputC was observed. In addition, a consensus sequence CosR-binding box in promoter regions of MacABputC was present in C. jejuni but not in Campylobacter coli. The MPCs identified in C. jejuni 81–176 membrane are involved in protein folding, molecule trafficking, oxidative phosphorylation, membrane structuration, peptidoglycan biosynthesis, motility and chemotaxis, stress signaling, efflux pumps and virulence.
Aim:
Following previous research on improving the cleaning of crates used to transport broiler chickens from the farm to the abattoir, a demonstration project was undertaken to investigate improvements in crate washing on a commercial scale.
Methods and results:
The soak tank of a conventional crate washing system was replaced with a high-performance washer fitted with high-volume, high-pressure nozzles. The wash water could be heated, and a greatly improved filtration system ensured that the nozzles did not lose performance or become blocked. Visual cleanliness scores and microbial counts were determined for naturally-contaminated crates which had been randomly assigned to different cleaning protocols.
Conclusions:
When a combination of mechanical energy, heat and chemicals (i.e. detergent and disinfectant) were used, the results showed significant improvements to crate cleaning. Reductions of up to 3·6 and 3·8 log10 CFU per crate base were achieved for Campylobacter and Enterobacteriaceae respectively, along with a marked improvement in visual cleanliness.
Significance and impact of study:
Broiler transport crates may become heavily contaminated with faeces and this may contribute to the spread of disease between farms. The results of this trial may be of use in reducing the spread of zoonotic pathogens in the poultry meat supply chain.
Introduction. Campylobacteriosis is an acute intestinal disease caused by Campylobacter spp., which manifests with symptoms of enterocolitis and gastroenteritis. The causative agents of campylobacteriosis are C. jejuni, C. coli, C. lari, C. upsaliensis, and C. helveticus. The incidence of campylobacteriosis is recorded worldwide as sporadic cases and foodborne or waterborne outbreaks. The industrialization of poultry production has led both to the acceleration of the evolution of commensal pathogens from the genus of Campylobacter, increased contamination of raw products and importance Campylobacter as foodborne pathogens. Material and methods. The main objects were raw poultry products and swabs from the environment of poultry processing enterprises. Cultural, biochemical, immunological methods, methods for the detection of the sensitivity of microorganisms to antibiotics, PCR analysis were used. Results and discussion. The methods of rapid detection of thermophilic campylobacters using combined schemes of bacteriological and molecular genetic analysis are developed. Because C. jejuni makes 85-90% of food isolates of campylobacters, for the purposes of production control the detection of thermophilic campylobacter with a minimum set of cultural and biochemical tests of identification is allowed. This will reduce the duration of analyses up to 3-4 days and decrease their labor-cost. The control critical points of production of poultry products in which it is necessary to control the presence of thermophilic campylobacters are indicated. These are the stages of slaughter, scalding, washing, and processing on the conveyor of carcasses, contact cooling baths, the areas of semi-finished products manufacturing and the packaging. Conclusion. The obtained results were used to develop Guidelines "Methods of rapid determination of bacteria of the genus Campylobacter in food products and evaluation of their antibiotic resistance".
The emergence of nanostructured materials has opened new horizons in the development of next generation biosensors. Being able to control the design of the electrode interface at the nanoscale combined with the intrinsic characteristics of the nanomaterials engenders novel biosensing platforms with improved capabilities. The purpose of this review is to provide a comprehensive and critical overview of the latest trends in emerging nanostructured electrochemical biosensors. A detailed description and discussion of recent approaches to construct label-free electrochemical nanostructured electrodes is given with special focus on pathogen detection for environmental monitoring and food safety. This includes the use of nanoscale materials such as nanotubes, nanowires, nanoparticles and nanosheets, as well as porous nanostructured materials including nanoporous anodic alumina, mesoporous silica, porous silicon and polystyrene nanochannels. These platforms may pave the way towards the development of point-of-care portable electronic devices for applications ranging from environmental analysis to biomedical diagnostics.
Purpose: Guillain Barré syndrome (GBS) is an acute inflammatory, autoimmune disorder of peripheral nervous system (PNS). Interleukin-17 (IL-17) and intercellular adhesion molecule-1 (ICAM-1) polymorphisms with higher expression levels have already been studied in many inflammatory and autoimmune diseases. However, the possible role of IL-17 and ICAM-1 polymorphisms in GBS remains unknown. Therefore, the current study investigated IL-17 (His161Arg, Glu126Gly) and ICAM-1 (Gly241Arg) polymorphisms.
Materials and method: In present study total 80 GBS patients and 75 normal healthy controls were included. IL-17 (His161Arg, Glu126Gly) and ICAM-1 (Gly241Arg) polymorphisms were performed using polymerase chain reaction-restriction fragment length polymorphism analysis. Further, the expression of ICAM-1 and IL-17 were determined by reverse-transcriptase PCR and enzyme-linked immunosorbent assay.
Results: IL-17 (Glu126Gly) mutant and ICAM-1 (Gly241Arg) heterozygous genotypes were strongly associated with increased risk of GBS (p<0.016; OR=3.706, 95% CI=1.28-10.67; p<.001; OR=4.148, 95% CI=2.119-8.119 respectively). IL-17 and ICAM-1 genes showed significantly higher expression in GBS when compared with healthy controls.
Conclusion: IL-17 and ICAM-1 polymorphisms showed significant association with GBS and their enhanced expressions have possible role in GBS development. IL-17 and ICAM-1 polymorphisms could be genetic markers to GBS susceptibility.
Campylobacter is one of the emerging foodborne pathogens, and its worldwide incidence rate is extremely high. This study was undertaken to isolate and identify Campylobacter strains from chicken carcasses in the local markets, and analyze their characteristics regarding oxygen tolerance. They were isolated after aerobic enrichment and identified by biochemical, physiological, and morphological characteristics, PCR, and 16S rDNA sequencing. Their oxygen tolerances were analyzed in terms of the cell surface hydrophobicity, cell fatty acid composition, and oxidoreductase. Five strains of C. jejuni were isolated and identified from 61 isolates from 50 chickens. Among them, C. jejuni IC21 grew well in Brucella broth and commercial milk under aerobic condition. However, in the aerobic exposure, the cell surface hydrophobicity of C. jejuni IC21 was almost the same as the other isolates, even though its morphology changed from the spiral-bacilli form into the coccoid form. Fatty acid analyses showed that all Campylobacter strains had a high composition of C19:1, cyclopropane fatty acid, and that the amount of the other fatty acids were very similar between them. Interestingly, however, only oxidoreductase activities of C. jejuni IC21 increased highly under aerobic exposure even though its activities were almost the same as the other C. jejuni strains just after microaerobic culture. It had 11.8 times higher catalase activity, 4.4 times higher for SOD, and 2.0 times higher for NADH oxidase activities. Therefore, in the case of the aero-adaptive C. jejuni IC21, expression of oxidoreductase significantly increased under oxidative stressed condition, which might allow it to survive for a longer time and grow on food under aerobic exposure. Such new strain might be one of the explanations for the increase of campylobacteriosis.
Campylobacter species are pathogens of both humans and animals and show a worldwide distribution. Campylobacter jejuni and C. coli are the most common causes of human gastroenteritis. The acute clinical illness ranges from mild watery diarrhea in developed countries to severe inflammatory diarrhea characterized by abdominal pain, fever, nausea, and bloody stool in developing countries. Major medical complications of C. jejuni infections include Guillain-Barré syndrome (GBS) and reactive arthritis. Campylobacter are fastidious organisms and C. jejuni does not replicate outside a warm-blooded host. However, the infectious dose is low (<1000 organisms) and transmission occurs through the food chain primarily by eating undercooked poultry or meat. C. jejuni is a natural commensal of chicken and its presence in chicken flocks is frequent. Contaminated food and subsequent illness are a huge economical burden, and processing control and prevention measures are important.
We have applied two strategies for the cloning of four genes responsible for the biosynthesis of the GT1a ganglioside mimic
in the lipooligosaccharide (LOS) of a bacterial pathogen,Campylobacter jejuni OH4384, which has been associated with Guillain-Barré syndrome. We first cloned a gene encoding an α-2,3-sialyltransferase
(cst-I) using an activity screening strategy. We then used nucleotide sequence information from the recently completed sequence
from C. jejuni NCTC 11168 to amplify a region involved in LOS biosynthesis from C. jejuni OH4384. The LOS biosynthesis locus from C. jejuni OH4384 is 11.47 kilobase pairs and encodes 13 partial or complete open reading frames, while the corresponding locus in C. jejuni NCTC 11168 spans 13.49 kilobase pairs and contains 15 open reading frames, indicating a different organization between these
two strains. Potential glycosyltransferase genes were cloned individually, expressed in Escherichia coli, and assayed using synthetic fluorescent oligosaccharides as acceptors. We identified genes encoding a β-1,4-N-acetylgalactosaminyl-transferase (cgtA), a β-1,3-galactosyltransferase (cgtB), and a bifunctional sialyltransferase (cst-II), which transfers sialic acid to O-3 of galactose and to O-8 of a sialic acid that is linked α-2,3- to a galactose. The linkage
specificity of each identified glycosyltransferase was confirmed by NMR analysis at 600 MHz on nanomole amounts of model compounds
synthesized in vitro. Using a gradient inverse broadband nano-NMR probe, sequence information could be obtained by detection of3J(C,H) correlations across the glycosidic bond. The role of cgtA and cst-II in the synthesis of the GT1a mimic in C. jejuni OH4384 were confirmed by comparing their sequence and activity with corresponding homologues in two relatedC. jejuni strains that express shorter ganglioside mimics in their LOS.
Lipopolysaccharides (LPS) of four nonencapsulated strains of the human enteric pathogen Campylobacter jejuni were chemically characterized. When applied to two of the strains, extraction by a modified phenol-chloroform-petroleum ether method (H. Brade and C. Galanos, Eur. J. Biochem. 122:233-237, 1982) gave better yields of LPS than did extraction by the conventional hot phenol-water technique. Constituents common to all LPS were D-glucose, D-galactose, L-glycero-D-manno-heptose, 3-deoxy-D-manno-2-octulosonic acid, D-glucuronic acid, D-galactosamine, and phosphorylethanolamine. Phosphate was present in a relatively high amount. In addition, the LPS of three strains contained N-acetylneuraminic acid, whereas the LPS of the strain lacking this component contained 3-amino-3,6-dideoxy-D-glucose. The lipid A component contained phosphate with D-glucosamine and 2,3-diamino-2,3-dideoxy-D-glucose as the major amino sugars. Ethanolamine-phosphate was present also. The major fatty acids were ester- and amide-bound 3-hydroxytetradecanoic and ester-bound hexadecanoic acids, with a minor amount of ester-bound tetradecanoic acid. This is the first report of N-acetylneuraminic acid in the oligosaccharide moiety and diaminoglucose in the lipid A of C. jejuni LPS.
Although infection with Campylobacter jejuni is recognized as a common antecedent of the Guillain-Barré syndrome, the clinical and epidemiologic features of this association are not well understood.
We performed a prospective case-control study in a cohort of patients with Guillain-Barré syndrome (96 patients) or Miller Fisher syndrome (7 patients) who were admitted to hospitals throughout England and Wales between November 1992 and April 1994. Bacteriologic and serologic techniques were used to diagnose preceding C. jejuni infection.
There was evidence of recent C. jejuni infection in 26 percent of the patients with Guillain-Barré or Miller Fisher syndrome, as compared with 2 percent of household controls and 1 percent of age-matched hospital controls (P < 0.001). Of the 27 patients with C. jejuni infection, 19 (70 percent) reported having had a diarrheal illness within 12 weeks before the onset of the neurologic illness. No specific serotypes were associated with Guillain-Barré syndrome. C. jejuni infection was slightly more common in men (P = 0.14) and was more likely to be associated with a pure motor syndrome and a slower recovery (P = 0.03). The patients with preceding C. jejuni infection were more likely to have acute axonal neuropathy or axonal degeneration in association with acute inflammatory demyelinating polyradiculoneuropathy, and they had greater disability after one year (P = 0.02). C. jejuni infection was significantly associated with a poor outcome even after correction for other factors associated with a poor prognosis.
Infection with C. jejuni often precedes the Guillain-Barré syndrome and is associated with axonal degeneration, slow recovery, and severe residual disability.
There is a strong association between Guillain-Barré syndrome (GBS) and Penner's serotype 19 (PEN 19) of Campylobacter jejuni. Sera from patients with GBS after C. jejuni infection have autoantibodies to GM1 ganglioside in the acute phase of the illness. Our previous work has suggested that GBS results from an immune response to cross-reactive antigen between lipopolysaccharide (LPS) of the Gram-negative bacterium and membrane components of peripheral nerves. To clarify the pathogenesis of GBS, we have investigated whether GM1-oligosaccharide structure is present in the LPS of C. jejuni (PEN 19) that was isolated from a GBS patient. After extraction of the LPS, the LPS showing the binding activity of cholera toxin, that specifically recognizes the GM1-oligosaccharide was purified by a silica bead column chromatography. Gas-liquid chromatography-mass spectrometric analysis has shown that the purified LPS contained Gal, GalNAc, and NeuAc, which are sugar components of GM1 ganglioside. 1H NMR methods [Carr-Purcell-Meiboom-Gill (CPMG), total correlation spectroscopy (TOCSY), and nuclear Overhauser effect spectroscopy (NOESY)] have revealed that the oligosaccharide structure [Gal beta 1-3 GalNAc beta 1-4(NeuAc alpha 2-3)Gal beta] protrude from the LPS core. This terminal structure [Gal beta 1-3GalNAc beta 1-4(NeuAc alpha 2-3)Gal beta] is identical to the terminal tetrasaccharide of the GM1 ganglioside. This is the first study to demonstrate the existence of molecular mimicry between nerve tissue and the infectious agent that elicits GBS.
Campylobacter jejuni serotype O19 strains associated with the Guillain-Barré syndrome (GBS) and other strains were examined by restriction fragment
length polymorphism (RFLP) analysis of polymerase chain reaction products of the flaA genes and by random amplified polymorphic DNA (RAPD) analysis. RFLP analysis showed that regardless of LIO serotype, geographic
origins, or association with GBS, the O19 isolates shared an identical digestion pattern by each of four restriction endonucleases,
DdeI, MboI, MseI, and AluI. In contrast, among C. jejuni O1 or O2 strains, RFLP patterns were different even among strains of the same LIO serotype. The results of the RAPD analysis
were consistent with the flaA RFLP data. These data indicate that all of the O19 strains that were tested were closely related to one another whether
they were or were not associated with GBS.
On the basis of the work presented and discussed at the • Serologic assays for diagnosis of C. jejuni infections need to be standardized and validated. workshop, a number of key points can be summarized, and from these, a series of recommendations can be made. These • All patients with acute flaccid paralysis should have their stools cultured for Campylobacter, and all isolated strains points are divided into the following categories: diagnostic aspects, epidemiology and surveillance, microbiology and should be serotyped. • Antibiotic resistance among Campylobacter species needs pathogenesis studies, and information management. to be monitored worldwide.
Guillain-Barré syndrome (GBS) is an autoimmune disease characterized by acute neuromuscular paralysis. Of an estimated annual
number of 2628–9575 US cases, 526–3830 are triggered by Campylobacter infection. Research objectives were to identify the lifetime consequences of GBS and, when possible, to quantify their economic
burden. The cost-of-illness method was used to calculate annual societal resources spent on medical care and lost productivity
due to illness or premature death from Campylobacter-associated GBS. Estimated total costs (in US$) of Campylobacter-associated GBS ($0.2–$1.8 billion) were added to previously estimated costs of campylobacteriosis ($1.3– $6.2 billion) for
a total annual cost from Campylobacter of $1.5–$8.0 billion (1995 dollars). It is concluded that up to $8.0 billion in US human illness costs are spent annually
because of Campylobacter infection. Economic evaluation of the other costs associated with GBS, such as physical and psychological costs, would increase
these estimates.
Lewis rats immunized with P2 protein, a 14.5-kDa protein of the peripheral nerve myelin, develop experimental allergic neuritis, a paralytic disorder
with clinical, histologic, and electrophysiologic features similar to those of human Guillain-Barré syndrome (GBS). T cells
reactive to P2 protein or a peptide corresponding to 53–78 residues of the protein can transfer the disease to naive animals. The mechanisms
by which these T cells induce demyelination are not well understood; however, they may induce inflammation and demyelination
in the nerves by production of Th1 cytokines. Th2 cytokines may lead to suppression of the inflammation and eventual recovery.
There is no conclusive evidence that P2 protein plays a role in the pathogenesis of GBS, with or without association with
Campylobacter jejuni; however, studies of the immunopathogenesis of P2 protein- induced experimental allergic neuritis are important for understanding the pathogenesis of inflammatory demyelination
in the peripheral nerves, the hallmark of GBS.
Over a 20-month period, 3 adult and 6 pediatric patients were diagnosed with Guillain-Barré syndrome (GBS) at Groote Schuur
and Red Cross Hospitals in Cape Town. All 9 GBS patients had Campylobacter jejuni biotype 2, serotype O:41 in their stools. C. jejuni infection was confirmed by ELISA testing of patient sera. Strains of this sero-biotype are rare: Only 12 such strains, including
the GBS-associated strains, were recognized among 776 Campylobacter strains isolated and identified at Red Cross Hospital from March 1994 to October 1995. This is the first known association
of C. jejuni biotype 2, serotype O:41 with GBS. Patients infected with this Campylobacter strain had a particularly severe form of the infection, requiring hospitalization and ventilation much longer than GBS patients
infected with other Campylobacter species and patients with Campylobacter-negative stools. The O:41 Campylobacter isolates from the GBS patients are identical by phenotypic, serologic, and molecular criteria, and they are clonal.
Guillain-Barré syndrome (GBS) is defined clinically as a peripheral neuropathy causing limb weakness that progresses for up
to 4 weeks before reaching a plateau. The symptoms may be caused by inflammatory demyelination, axonal degeneration, or both.
GBS occurs throughout the world, with a median incidence of 1.3 cases/100,000 population (range, 0.4–4.0). Males are more
commonly affected than females, and there are peaks in young adults and the elderly. There is no clear seasonal association
in Western countries, although this may be because the most frequent antecedent events, respiratory and enteric infections,
have opposite seasonality. The most frequently identified cause of GBS is Campylobacter jejuni infection, which has been identified in up to 41% of patients and is associated with more severe disease and prolonged disability.
Summer epidemics of GBS occur among children and young adults in Northern China and are particularly likely to be associated
with C. jejuni infection.
Guillain-Barré syndrome (GBS), a neurologic disease characterized by acute paralysis, is frequently preceded by Campylobacter jejuni infection. Serotype O19 strains are overrepresented among GBS-associated C. jejuni isolates. We previously showed that all O19 strains tested were closely related to one another by randomly amplified polymorphic DNA (RAPD) and restriction fragment length polymorphism analyses. RAPD analysis demonstrated a 1.4-kb band in all O19 strains tested but in no non-O19 strains. We cloned this O19-specific band; nucleotide sequence analysis revealed a truncated open reading frame with significant homology to DNA gyrase subunit B (gyrB) of Helicobacter pylori. PCR using the random primer and a primer specific for gyrB showed that in non-O19 strains, the random primer did not recognize the downstream gyrB binding site. The regions flanking each of the random primer binding sites were amplified by degenerate PCR for further sequencing. Although the random primer had several mismatches with the downstream gyrB binding site, a single nucleotide polymorphism 6 bp upstream from the 3' terminus was found to distinguish O19 and non-O19 strains. PCR using 3'-mismatched primers based on this polymorphism was designed to differentiate O19 strains from non-O19 strains. When a total of 42 (18 O19 and 24 non-O19) strains from five different countries were examined, O19 strains were distinguishable from non-O19 strains in each case. This PCR method should permit identification of O19 C. jejuni strains.
To determine whether GM1-like epitopes in Campylobacter species are specific to O serotypes associated with Guillain-Barré syndrome (GBS) or whether they are frequent among random
Campylobacter isolates causing enteritis, 275 random enteritis-associated isolates of Campylobacter jejuni were analyzed. The isolates were collected in the United States using a cholera toxin—binding assay. Overall, 26.2% of the
isolates were positive for the GM1-like epitope. Of the 36 different O serotypes in the sample, 21 (58.3%) contained no strains
positive for GM1, whereas in 6 serotypes (16.7%), >50% of isolates were positive for GM1. GBS-associated serotypes were more
likely to contain strains positive for GM1 than were non—GBS-associated serotypes (37.8% vs. 15.1%, P = .0116). The results suggest that humans are frequently exposed to strains exhibiting GM1-like mimicry and, while certain
serotypes may be more likely to possess GM1-like epitopes, the presence of GM1-like epitopes on Campylobacter strains does not itself trigger GBS.
Guillain-Barré syndrome and its variant, Miller-Fisher syndrome, are acute, postinfectious, autoimmune neuropathies that frequently follow Campylobacter jejuni enteritis. The pathogenesis is believed to involve molecular mimicry between sialylated epitopes on C. jejuni LPSs and neural gangliosides. More than 90% of Miller-Fisher syndrome cases have serum anti-GQ1b and anti-GT1a ganglioside antibodies that may also react with other disialylated gangliosides including GD3 and GD1b. Structural studies on LPS from neuropathy-associated C. jejuni strains have revealed GT1a-like and GD3-like core oligosaccharides. To determine whether this structural mimicry results in pathogenic autoantibodies, we immunized mice with GT1a/GD3-like C. jejuni LPS and then cloned mAb's that reacted with both the immunizing LPS and GQ1b/GT1a/GD3 gangliosides. Immunohistology demonstrated antibody binding to ganglioside-rich sites including motor nerve terminals. In ex vivo electrophysiological studies of nerve terminal function, application of antibodies either ex vivo or in vivo via passive immunization induced massive quantal release of acetylcholine, followed by neurotransmission block. This effect was complement-dependent and associated with extensive deposits of IgM and C3c at nerve terminals. These data provide strong support for the molecular mimicry hypothesis as a mechanism for the induction of cross-reactive pathogenic anti-ganglioside/LPS antibodies in postinfectious neuropathies.
Campylobacter jejuni O:41 strains are found in association with Guillain-Barré syndrome in South Africa. Strains of this serotype collected over 17 years were characterized by amplified fragment length polymorphism and flagellin typing to determine their clonal nature. Despite minor variation in GM1 expression, all of the strains were genetically indistinguishable, indicating that they are representative of a genetically stable clone.
Campylobacter jejuni, from the delta-epsilon group of proteobacteria, is a microaerophilic, Gram-negative, flagellate, spiral bacterium-properties it shares with the related gastric pathogen Helicobacter pylori. It is the leading cause of bacterial food-borne diarrhoeal disease throughout the world. In addition, infection with C. jejuni is the most frequent antecedent to a form of neuromuscular paralysis known as Guillain-Barré syndrome. Here we report the genome sequence of C. jejuni NCTC11168. C. jejuni has a circular chromosome of 1,641,481 base pairs (30.6% G+C) which is predicted to encode 1,654 proteins and 54 stable RNA species. The genome is unusual in that there are virtually no insertion sequences or phage-associated sequences and very few repeat sequences. One of the most striking findings in the genome was the presence of hypervariable sequences. These short homopolymeric runs of nucleotides were commonly found in genes encoding the biosynthesis or modification of surface structures, or in closely linked genes of unknown function. The apparently high rate of variation of these homopolymeric tracts may be important in the survival strategy of C. jejuni.
Campylobacter jejuni has been identified as the predominant cause of antecedent infection in Guillain-Barré syndrome (GBS) and Miller Fisher syndrome (MFS). The risk of developing GBS or MFS may be higher after infection with specific C. jejuni types. To investigate the putative clonality, 18 GBS- or MFS-related C. jejuni strains from The Netherlands and Belgium and 17 control strains were analyzed by serotyping (Penner and Lior), restriction fragment length polymorphism analysis of PCR products of the flaA gene, amplified fragment length polymorphism analysis, pulsed-field gel electrophoresis, and randomly amplified polymorphic DNA analysis. Serotyping revealed 10 different O serotypes and 7 different Lior serotypes, thereby indicating a lack of serotype clustering. Two new O serotypes, O:35 and O:13/65, not previously associated with GBS or MFS were found. Serotype O:19 was encountered in 2 of 18 strains, and none was of serotype O:41. The results of all genotypic methods also demonstrated substantial heterogeneity. No clustering of GBS- or MFS-related strains occurred and no molecular marker capable of separating pathogenic GBS or MFS from non-GBS- or non-MFS-related enteritis strains could be identified in this study. Sialic-acid-containing lipopolysaccharides (LPS) are thought to be involved in the triggering of GBS or MFS through molecular mimicry with gangliosides in human peripheral nerves. Therefore, further characterization of GBS- or MFS-related C. jejuni should target the genes involved in the synthesis of LPS and the incorporation of sialic acid.
Infection with Campylobacter jejuni serotype HS:19 is associated with the development of Guillain-Barré syndrome (GBS). To determine whether a particular HS:19
clone is associated with GBS, multilocus enzyme electrophoresis (MLEE) was used to analyze a worldwide collection of isolates.
There were 34 electropherotypes (ETs) in 3 phylogenetic clusters among 83 C. jejuni isolates. Cluster I contained all HS:19 strains, and a single ET (ET4) accounted for most HS:19 strains. HS:19 strains did
not occur in any of the other clusters. ET4 contained isolates from different geographic locations, indicating global spread
of this clone. Furthermore, ET4 contained isolates from patients with uncomplicated enteritis and GBS, as well as isolates
from animal sources. The results of this study show that HS:19 strains comprise a clonal, although not monomorphic, population,
which is distinct from non-HS:19 strains within C. jejuni. A unique clone associated with GBS was not identified by use of MLEE
Guillain-Barré syndrome (GBS), a neurologic disease characterized by acute paralysis, is frequently preceded by Campylobacter jejuni infection. Serotype O19 strains are overrepresented among GBS-associated C. jejuni isolates. We previously showed that all O19 strains tested were closely related to one another by randomly amplified polymorphic DNA (RAPD) and restriction fragment length polymorphism analyses. RAPD analysis demonstrated a 1.4-kb band in all O19 strains tested but in no non-O19 strains. We cloned this O19-specific band; nucleotide sequence analysis revealed a truncated open reading frame with significant homology to DNA gyrase subunit B ( gyrB ) of Helicobacter pylori . PCR using the random primer and a primer specific for gyrB showed that in non-O19 strains, the random primer did not recognize the downstream gyrB binding site. The regions flanking each of the random primer binding sites were amplified by degenerate PCR for further sequencing. Although the random primer had several mismatches with the downstream gyrB binding site, a single nucleotide polymorphism 6 bp upstream from the 3′ terminus was found to distinguish O19 and non-O19 strains. PCR using 3′-mismatched primers based on this polymorphism was designed to differentiate O19 strains from non-O19 strains. When a total of 42 (18 O19 and 24 non-O19) strains from five different countries were examined, O19 strains were distinguishable from non-O19 strains in each case. This PCR method should permit identification of O19 C. jejuni strains.
GM1 ganglioside has been implicated as a target of immune attack in some diseases of the peripheral nervous system. Anti-GM1 ganglioside antibodies are associated with certain acquired immune-mediated neuropathies. It is not clear how anti-GM1 antibodies cause nerve dysfunction and injury; however, sodium and/or potassium ion channel dysfunction at the node of Ranvier has been implicated. To gain insight into the pathogenesis of these neuropathies, we examined the distribution of GM1 ganglioside and Gal(β1–3)GalNAc moieties in nerve fibres and their relationship to voltage-gated sodium and potassium (Kv1.1, 1.5) channels at the nodes of Ranvier in peripheral nerves from human, rat and dystrophic mice. Gal(β1–3)GalNAc moieties were localized via the binding of cholera toxin and peanut agglutinin. As a control for the specificity of these findings, we compared the distribution of GM1 moieties to that of the ganglioside GT1b. Our study provides definitive evidence for the presence of Gal(β1–3)GalNAc bearing moieties on the axolemmal surface of mature myelinated fibres and on Schwann cells. Gal(β1–3)GalNAc binding sites did not have an obligatory co-localization with voltage-gated sodium channels or the potassium ion channels Kv1.1 and Kv1.5 and are thus not likely carried by these ion channels. In contrast with Gal(β1–3)GalNAc, GT1b-like moieties are restricted to the axolemma.
Using Penner's method and Lior's scheme, we serotyped 31 isolates from patients with Guillain-Barré syndrome, 7 isolates from those with Fisher's syndrome, and isolates from patients with sporadic enteritis. PEN 19 of Campylobacter jejuni was isolated more frequently from the Guillain-Barré syndrome patients (16/31 isolates, 52%) than from the sporadic enteritis patients (5%). LIO 7 of C. jejuni also was isolated more frequently from the Gillain-Barré syndrome patients (14/31 isolates, 45%) than from the enteritis patients (3%). One reason why Guillain-Barré syndrome is rare, despite the high incidence of C. jejuni enteritis, may be the low frequencies of PEN 19 and LIO 7. The frequency of positive anti-GM 1 antibody titers in the Guillain-Barré syndrome patients with PEN 19 isolates was higher than that in the Guillain-Barré syndrome and Fisher's syndrome patients without PEN 19 isolates. We speculate that the serotypic determinant of PEN 19 aids in the production of anti-GM 1 antibody by a GM 1−like lipopolysaccharide. In contrast, 5 of the 7 isolates from the Fisher's syndrome patients belonged to PEN 2:LIO 4. The IgG anti-GQ 1b antibody was associated with PEN 2 and LIO 4. These serotypic determinants may aid in the production of IgG anti-GQ 1b antibody by a GQ 1b−like lipopolysaccharide.
Campylobactor jejuni was isolacted from stool cultures from 14(30%) of 46 partients with Guillain-Barré syndrome and from 6(1.2%) of healthy persons, and the difference was highly significant (P<0.0001). In addition, serological evidence of recent. C. Jejuni infection. Ten of 12(83%) isolated from partients with Guillain-Barré syndrome belonged to Penner serogroup 19, which is a rare serogroup in sporadic patients with C. Jejuni enterties. In the lection typing stude, all serogroup 19 strains from patients with Guillain-Barré syndrome patients with Guillain-Barré syndrome were shown to contain terminal b̃-N-acetylglucosamine residues on their cell surface, bur serogroup 19 strains from patients with enteritis were not.
Peripheral nerve diseases are among the most prevalent disorders of the nervous system. Because of the accessibility of the peripheral nervous system (PNS) to direct physiological and pathological study, neuropathies have traditionally played a unique role in developing our understanding of basic mechanism of nervous system injury and repair. At present they are providing new insight into the mechanisms of immune injury to the nervous system. A rapidly growing catalogue of PNS disorders are now suspected to be immune-mediated, and in the best understood of these disorders, the molecular and cellular targets of immune attack are known, and the pathophysiology follows directly from the specific immune injury. This review summarizes the immunologically relevant features of the PNS, then considers selected immune-mediated neuropathies, focusing on pathogenetic mechanisms. Finally, the PNS is providing a testing ground for new immunotherapies and approaches to protection and regeneration, including the use of trophic factors. The current status of treatment and implications for future approaches is reviewed.
Sixteen cases of Guillain-Barré syndrome occurred in the third week of a diarrhoea epidemic caused by water pollution in EL-Sult, Jordan. Of 30 000 people exposed to polluted water, 5000 developed diarrhoea, 74 typhoid, and 30 infectious hepatitis. Thirteen of the 16 patients with Guillain-Barré syndrome had been mildly affected by diarrhoea 8-24 days before the onset of peripheral neuropathy. Paralysis progressed rapidly, reaching a peak in one to five days, and recovery began three to seven days after the start of the most severe symptoms. All but four patients had recovered completely after one year. Rapid progress of paralysis and a delayed interval between maximum weakness and start of recovery were both associated with poor prognosis.
In early January, 1976, an outbreak of gastroenteritis caused by contamination of the water supply system occurred in Salt, Jordan. This outbreak was followed by the appearance of peripheral neuropathy in 19 cases diagnosed as Guillain-Barré Syndrome. Clinically, gastroenteritis was suggestive of Shigellosis and one of nine patients with Guillain-Barré Syndrome, grew Shigella boydii. Epidemiology of the 19 cases with peripheral neuropathy is detailed. Seventeen patients had definite gastroenteritis symptoms prior to their neuropathy. The median incubation period was seven days.
We report cases of 2 patients with pure motor Guillain-Barré syndrome of explosive onset who required mechanical ventilation for more than 2 months. Their electrophysiologic findings and poor clinical recoveries suggested severe axonal degeneration involving the motor nerves. Enzyme-linked immunosorbent assay and thin-layer chromatogram-immunostaining showed the sera of both patients had high IgG antibody titer against GD1a ganglioside. Their titers decreased with the clinical course of the illness. GD1a as well as GM1, appears to be the target pathogenic antigen in motor axon disorders. Elevated IgG anti-GD1a antibody titer may prove useful for predicting severe GBS.
Guillain-Barré syndrome (GBS), as we recognize it today, was first described by Landry in 1859 [32] and for years was known as Landry’s ascending paralysis. In 1916, Guillain, Barré and Strohl [23], using the newly developed technique of lumbar puncture, described albuminocytological dissociation, i.e., increased albumin with very few or no cells, in the cerebrospinal fluid of patients with Landry’s ascending paralysis and helped to distinguish the disorder from other paralytic disorders especially poliomyelitis. Over the next several years, Landry’s ascending paralysis with new diagnostic features in the cerebrospinal fluid (CSF) became kown as Guillain-Barré syndrome.
HLA typing for class II alleles was performed on 97 patients with Guillain-Barré syndrome or Miller Fisher syndrome and compared with 100 controls. There was a significant association between HLA-DQB1*03 and preceding Campylobacter jejuni infection (Pc = 0.05).
An O antigenic polysaccharide was liberated from the lipopolysaccharide of high M(r) from Campylobacter jejuni serotype O:19 by acetic acid hydrolysis of the ketosidic linkage to lipid A. The structure of the polysaccharide was established in several one- and two-dimensional 1H and 13C NMR experiments, fast atom bombardment mass spectrometry and methylation linkage analysis of the permethylated glycan and its degradation products. It is concluded that the glycan is a derivative of hyaluronic acid in which the beta-D-glucuronic acid residues in the alternating sequence [-4)-beta-D-GlcA-(1-->3)-beta-D-GlcNAc-(1]n are present as amides of 2-amino-2-deoxyglycerol. Parallel experiments were performed on O antigens liberated from lipopolysaccharides of high M(r) from bacterial isolates that had been obtained from two patients who subsequently developed the Guillain-Barré syndrome. Within the limits of structural analysis by NMR spectroscopy and methylation linkage analysis, both these O antigens were identical to that from the serostrain.
The localization, mode of action, and roles of complement in the Guillain-Barre syndrome have been controversial. We used high-resolution immunocytochemistry to localize complement activation products in early stages of the acute inflammatory demyelinating polyneuropathy (AIDP) pattern of Guillain-Barre syndrome. Three AIDP subjects who were autopsied had had symptoms for 3 to 9 days at the time of death. Immunocytochemistry was performed on etched, epoxy resin-embedded sections, and the next thin section was compared by electron microscopy (thick/thin sections). Many fibers had a rim of the complement activation marker C3d and the terminal complement complex neoantigen C5b-9 along the outer surface of the Schwann cells. Ultrastructural analysis of these C3d-positive fibers showed mild vesicular changes of the outermost myelin lamellae. Vesicular degeneration was seen before the invasion of macrophages into the myelin, and was the predominant change in the subject with symptoms for 3 days. C3d staining was not found on myelin membranes. The results suggest that at least some forms of AIDP are complement mediated. We speculate that complement is activated by antibody bound to epitopes on the outer surface of the Schwann cell and that the resulting complement activation initiates the vesiculation of myelin.
High titers of anti-GD1a antibodies have been found in patients with Guillain-Barre syndrome or motor neuropathy. To determine the possible diagnostic relevance of these antibodies, we measured serum anti-GD1a IgG and IgM antibodies by enzyme-linked immunosorbent assay in 195 patients with different motor syndromes and in 335 control subjects. Moderately high antibody titers (1/1,280-1/5,120) were occasionally found in patients with chronic inflammatory demyelinating polyneuropathy (5%), multifocal motor neuropathy (18%), lower motor neuron disease (3.8%), or amyotrophic lateral sclerosis (1.8%) and in immunological control subjects (1.2%), while titers of 1/20,480 or higher were only found in 2 patients with Guillain-Barre syndrome (IgG in both) and 2 with motor neuropathy and IgM lambda monoclonal gammopathy improving with immunotherapy. In both motor neuropathy patients and the Guillain-Barre syndrome patient who were retested during recovery, anti-GD1a titers decreased concomitantly with clinical improvement. High anti-GD1a antibody titers may be found in several motor syndromes but only markedly increased anti-GD1a titers are strictly associated with potentially treatable dysimmune neuropathies.
Three of six rabbits immunized with purified GD1b developed ataxic sensory neuropathy. They laid on the floor with their limbs splayed out, and their movements were awkward; but muscle power, tonus, and superficial sensation appeared to be intact. Sciatic nerve motor conduction studies were normal. Axonal degeneration was present in the dorsal column of the spinal cord, in the dorsal roots, and in the sciatic nerve. Some of the nerve cell bodies in the dorsal root ganglia had degenerated and disappeared. No demyelinative lesions or mononuclear cell infiltrations were seen in those regions. No pathological changes were present in the other three immunized rabbits that showed no clinical symptoms. Control rabbits inoculated only with adjuvants showed neither clinical symptoms nor pathological changes. Anti-GD1b antibody was raised in the sera from all six rabbits immunized with GD1b. The monoclonal anti-GD1b antibody GGR12 immunostained about one-half the rabbit primary sensory neurons. Sensitization with GD1b, therefore, may cause ataxic sensory neuropathy in rabbits due to antibody-mediated damage to the primary sensory neurons.
To develop and characterise an animal model of paralytic neuropathy after Campylobacter jejuni infection. Campylobacter infection precedes development of many cases of Guillain-Barré syndrome and is particularly associated with cases having prominent axonal degeneration. Understanding the pathogenesis of Guillain-Barré syndrome after C jejuni infection has been slowed by the lack of animal models.
A spontaneous paralytic neuropathy is described that developed in chickens from the farms of four patients with Guillain-Barré syndrome. The production of paralytic neuropathy in chickens experimentally fed Campylobacter jejuni isolated from one of these patients is reported. The sciatic nerves of the spontaneously paralysed chickens were examined pathologically in teased fibres, in plastic embedded sections, and by electron microscopy. Two large groups of chickens were then fed cultures of a C jejuni (Penner type O:19) isolated from one of these patients.
The chickens with spontaneous paralysis had pathologically noninflammatory neuropathy. Pathology in the sciatic nerves ranged from no detectable changes to severe Wallerian-like degeneration. In the experimentally inoculated groups, an average of 33% of the chickens became paralysed. The median time after inoculation to paralysis was 12 days. The lesions found in the first few days of paralysis included nodal lengthening and paranodal demyelination. In those animals that survived for several days after onset of weakness, the pathology was dominated by extensive Wallerian-like degeneration. Animals that survived for weeks with no clinically apparent neuropathy had paranodal remyelination in some teased nerve fibres, reflecting earlier paranodal demyelination.
Experimental inoculation with C jejuni may provide a new model for understanding some forms of Guillain-Barré syndrome.
The flagellins of Campylobacter spp. differ antigenically. In variants of C. coli strain VC167, two antigenic flagellin types determined by sero-specific antibodies have been described (termed T1 and T2). Post-translational modification has been suggested to be responsible for T1 and T2 epitopes, and, using mild periodate treatment and biotin hydrazide labelling, flagellin from both VC167-T1 and T2 were shown to be glycosylated. Glycosylation was also shown to be present on other Campylobacter flagellins. The ability to label all Campylobacter flagellins examined with the lectin LFA demonstrated the presence of a terminal sialic acid moiety. Furthermore, mild periodate treatment of the flagellins of VC167 eliminated reactivity with T1 and T2 specific antibodies LAH1 and LAH2, respectively, and LFA could also compete with LAH1 and LAH2 antibodies for binding to their respective flagellins. These data implicate terminal sialic acid as part of the LAH strain-specific epitopes. However, using mutants in genes affecting LAH serorecognition of flagellin it was demonstrated that sialic acid alone is not the LAH epitope. Rather, the epitope(s) is complex, probably involving multiple glycosyl and/or amino acid residues.