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Concept: Campylobacter

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Campylobacter jejuni is an important cause of human foodborne gastroenteritis; strategies to prevent infection are hampered by a poor understanding of the complex interactions between host and pathogen. Previous work showed that C. jejuni could bind human histo-blood group antigens (BgAgs) in vitro and that BgAgs could inhibit the binding of C. jejuni to human intestinal mucosa ex vivo. Here, the major flagella subunit protein (FlaA) and the major outer membrane protein (MOMP) were identified as BgAg-binding adhesins in C. jejuni NCTC11168. Significantly, the MOMP was shown to be O-glycosylated at Thr(268); previously only flagellin proteins were known to be O-glycosylated in C. jejuni. Substitution of MOMP Thr(268) led to significantly reduced binding to BgAgs. The O-glycan moiety was characterized as Gal(β1-3)-GalNAc(β1-4)-GalNAc(β1-4)-GalNAcα1-Thr(268); modelling suggested that O-glycosylation has a notable effect on the conformation of MOMP and this modulates BgAg-binding capacity. Glycosylation of MOMP at Thr(268) promoted cell-to-cell binding, biofilm formation and adhesion to Caco-2 cells, and was required for the optimal colonization of chickens by C. jejuni, confirming the significance of this O-glycosylation in pathogenesis.

Concepts: Immune system, Protein, Bacteria, Glucose, Proteobacteria, Gastroenteritis, Campylobacter, Campylobacter jejuni

53

In August 2016, a local public health agency (LPHA) notified the Colorado Department of Public Health and Environment (CDPHE) of two culture-confirmed cases of Campylobacter infection among persons who consumed raw (unpasteurized) milk from the same herdshare dairy. In Colorado, the sale of raw milk is illegal; however, herdshare programs, in which a member can purchase a share of a herd of cows or goats, are legal and are not regulated by state or local authorities. In coordination with LPHAs, CDPHE conducted an outbreak investigation that identified 12 confirmed and five probable cases of Campylobacter jejuni infection. Pulsed-field gel electrophoresis (PFGE) patterns for the 10 cases with available isolates were identical using the enzyme Sma. In addition, two milk samples (one from the dairy and one obtained from an ill shareholder) also tested positive for the outbreak strain. Five C. jejuni isolates sent to CDC for antimicrobial susceptibility testing were resistant to ciprofloxacin, tetracycline, and nalidixic acid (1). Although shareholders were notified of the outbreak and cautioned against drinking the milk on multiple occasions, milk distribution was not discontinued. Although its distribution is legal through herdshare programs, drinking raw milk is inherently risky (2). The role of public health in implementing control measures associated with a product that is known to be unsafe remains undefined.

Concepts: Epidemiology, Milk, Proteobacteria, Pasteurization, Campylobacter, Campylobacter jejuni, Raw milk, Campylobacteriosis

42

This paper introduces a novel method for sampling pathogens in natural environments. It uses fabric boot socks worn over walkers' shoes allowing collection of composite samples over large areas. Wide area sampling is better suited to studies focussing upon human exposure to pathogens (e.g. recreational walking). This sampling method is implemented using a Citizen Science approach: groups of three walkers wearing boot socks undertook one of six routes, 40 times over 16 months in the North West (NW) and East Anglian (EA) regions of England. To validate this methodology we report the successful implementation of this Citizen Science approach, the observation that Campylobacter was detected on 47% of boot socks, and the observation that multiple boot socks from individual walks produced consistent results. Findings indicate elevated Campylobacter presence in the livestock dominated NW in comparison to EA (55.8% vs 38.6%). Seasonal variation in Campylobacter presence was found between regions, with indications of winter peaks in both regions, but a spring peak in NW. Campylobacter presence on boot socks was negatively associated with ambient temperature (p=0.011) and positively associated with precipitation (p<0.001), results which are consistent with our understanding of Campylobacter survival and the probability of material adhering to boot socks. C. jejuni was the predominant species found, with C. coli largely restricted to the livestock dominated NW. Source attribution analysis indicated that the potential source of C. jejuni was predominantly sheep in NW and wild birds in EA but did not vary between peak and non-peak periods of human incidence.Importance There is debate in the literature on the pathways through which pathogens transfer from the environment to humans. We report on the success of a novel method for sampling human-pathogen interactions using boot socks and citizen science techniques, which enable us to sample human-pathogen interactions that may occur through visits to natural environments. This contrasts with traditional environmental sampling, which is based upon spot sampling techniques and does not sample human-pathogen interactions. Our methods are of practical value to scientists trying to understand transmission of pathogens from the environment to people. Our findings provide insight into the risk of Campylobacter from recreational visits and an understanding of how these risks vary seasonally and the factors behind these patterns. We highlight the Campylobacter species predominantly encountered and the potential sources of the C. jejuni.

Concepts: Sample, Human, Sample size, Sampling, Environment, Natural environment, Nature, Campylobacter

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In the United Kingdom, outbreaks of Campylobacter infection are increasingly attributed to undercooked chicken livers, yet many recipes, including those of top chefs, advocate short cooking times and serving livers pink. During 2015, we studied preferences of chefs and the public in the United Kingdom and investigated the link between liver rareness and survival of Campylobacter. We used photographs to assess chefs' ability to identify chicken livers meeting safe cooking guidelines. To investigate the microbiological safety of livers chefs preferred to serve, we modeled Campylobacter survival in infected chicken livers cooked to various temperatures. Most chefs correctly identified safely cooked livers but overestimated the public’s preference for rareness and thus preferred to serve them more rare. We estimated that 19%-52% of livers served commercially in the United Kingdom fail to reach 70°C and that predicted Campylobacter survival rates are 48%-98%. These findings indicate that cooking trends are linked to increasing Campylobacter infections.

Concepts: Microbiology, Infection, Pathogen, United Kingdom, Chicken, Preference, Campylobacter, Campylobacteriosis

28

Campylobacter jejuni was non-motile at 20°C but highly motile at ≥37°C, similarly to Helicobacter pylori but unlike Vibrio cholerae O1/O139. C. jejuni is a bacterium with one of the highest swimming speeds (>100 μm/s), especially at 42°C. Straight and spiral bacterial shapes shared the same motility. C. jejuni possessed a unique structure in the flagellate polar region, characterized by a cup-like structure (beneath the inner membrane), a funnel shape (opening onto the pole surface), and less-dense space (cytoplasm). Other Campylobacter species (coli, fetus, and lari) exhibited similar features of motility and flagellate polar structures, albeit with slight differences especially for C. fetus, which had a flagellum only at one pole and a cup-like structure composed of two membranes.

Concepts: Bacteria, Eukaryote, Flagellum, Helicobacter pylori, Proteobacteria, Campylobacter, Campylobacter jejuni, Vibrio

28

Recent advancements in the periodontal research field are consistent with a new model of pathogenesis according to which periodontitis is initiated by a synergistic and dysbiotic microbial community rather than by select ‘periopathogens’, such as the ‘red complex’. In this polymicrobial synergy, different members or specific gene combinations within the community fulfill distinct roles that converge to shape and stabilize a disease-provoking microbiota. One of the core requirements for a potentially pathogenic community to arise involves the capacity of certain species, termed ‘keystone pathogens’, to modulate the host response in ways that impair immune surveillance and tip the balance from homeostasis to dysbiosis. Keystone pathogens also elevate the virulence of the entire microbial community through interactive communication with accessory pathogens. Other important core functions for pathogenicity require the expression of diverse molecules (e.g. appropriate adhesins, cognate receptors, proteolytic enzymes and proinflammatory surface structures/ligands), which in combination act as community virulence factors to nutritionally sustain a heterotypic, compatible and proinflammatory microbial community that elicits a non-resolving and tissue-destructive host response. On the basis of the fundamental concepts underlying this model of periodontal pathogenesis, that is, polymicrobial synergy and dysbiosis, we term it the PSD model.

Concepts: Immune system, Bacteria, Microbiology, Protease, Periodontitis, Periodontal disease, Campylobacter, Holism

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Campylobacter jejuni is the most common bacterial cause of human gastroenteritis and often precedes development of Guillain-Barré syndrome (GBS), a life-threatening paralytic disease. The incorporation of the carbohydrate sialic acid into C. jejuni lipooligosaccharides (LOS) is associated with increased severity of gastroenteritis and with induction of GBS; however, the underlying mechanisms remain completely unknown. In this study, we demonstrate that sialic acids in C. jejuni endotoxin enhance the rapid production of IFN-β and TNF-α by human dendritic cells (DCs). Using neutralizing Abs and receptors it was shown that these DC-derived cytokines promote the proliferation of human mucosal B cells in a T cell-independent manner. The production of both IFN-β and TNF-α by DCs in response to LOS requires CD14, and the amplified response of DCs to sialylated C. jejuni LOS is CD14 dependent. Together, these results indicate that sialylation of C. jejuni LOS increases DC activation and promotes subsequent B cell responses through CD14-driven production of IFN-β and TNF-α. This enhanced DC/B cell response may explain the increased pathogenicity of sialylated C. jejuni and may be key to the initiation of B cell-mediated autoimmunity in GBS.

Concepts: Immune system, Protein, Sialic acid, Toll-like receptor, Humoral immunity, Gastroenteritis, Campylobacter, Campylobacter jejuni

26

The antimicrobial and antioxidant activities of some cultivars of the nopal cactus have not been determined. In this study, 8 cultivars of nopal cacti from Mexico were assayed for phenolic content, antioxidant activities, and antimicrobial activities against Campylobacter Jejuni, Vibrio cholera, and Clostridium Perfringens. Plant material was washed, dried, and macerated in methanol. Minimum bactericidal concentrations (MBCs) were determined using the broth microdilution method. Antioxidant activities were quantitatively determined using spectrophotometric methods. The MCBs of the nopal cacti ranged from 1.1 to 12.5 mg/mL for c. jejuni, 4.4 to 30 mg/mL for V. cholera, and 0.8 to 16 mg/mL for C. perfringens in the cultivars Cardon Blanco, Real de Catorce, and Jalpa, respectively. High quantities of total phenols and total flavonoids were found in the Jalpa cacti (3.80 mg of gallic acid equivalent GAE/g dry weight [DW] and 36.64 mg of quercetin equivalents [QE]/g DW, respectively). 2,2-Diphenyl-1-picrylhydrazyl (DPPH) radical scavenging activities (RSA) were correlated to bioactive compound contents. The Villanueva cacti had the highest %RSA at 42.31%, and the lowest activity was recorded in Copena V1 at 19.98%. In conclusion, we found that some of the 8 cactus pear cultivars studied may be used for their antioxidant compounds or antimicrobials to control or prevent the contamination of foods.

Concepts: Microbiology, Opuntia, Cactus, Opuntia ficus-indica, Gastroenteritis, Clostridium perfringens, Campylobacter, Campylobacter jejuni

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Despite education efforts, consumers often practice unsafe food handling and storage behaviors. Little is known about how these unsafe practices contribute to contamination of the home kitchen with foodborne pathogens. In addition, only a limited number of studies have examined the role of the kitchen as a reservoir for pathogens. The purpose of this study was to characterize microbial contamination and foodborne pathogens found in home kitchens and determine whether contamination was significantly associated with unsafe or unsanitary conditions observed in the kitchen. Swab samples were collected from food contact and preparation surfaces in homes (n = 100) in Philadelphia, PA. The samples were tested for coliforms, fecal coliforms, Escherichia coli , Staphylococcus aureus , Salmonella, Campylobacter, and Listeria. Fecal coliforms were found in 44% of homes (most often in samples from kitchen sinks, sponges, and dishcloths), and E. coli was found in 15% of homes (mostly in samples from kitchen sinks). Nearly half (45%) of the homes tested positive for a foodborne pathogen, and 12% had multiple pathogens present in the kitchen. S. aureus was isolated from 39% of homes, most often from countertops and refrigerator door handles. Listeria spp., including L. monocytogenes and L. innocua , were present in 15% of homes, most often in samples from refrigerator meat drawers. C. jejuni was isolated from 3% of homes. Contamination with Listeria was significantly associated with higher refrigerator temperatures. The contamination of surfaces with fecal coliforms and S. aureus was significantly associated with a lack of cleaning materials: dish soap and paper or cloth towels in the kitchen, and any type of towel in the nearest bathroom. The contamination of a sponge or dishcloth with either fecal coliforms or S. aureus was predictive of other surfaces in the kitchen having the same contamination, indicating that sponges and dishcloths are both reservoirs and vectors for bacteria in the kitchen.

Concepts: Bacteria, Microbiology, Pneumonia, Staphylococcus aureus, Antibiotic resistance, Escherichia coli, Gastroenteritis, Campylobacter

23

Antibiotic-resistant Campylobacter jejuni strains are rapidly emerging worldwide. Here, we aimed to determine the antibiotic-resistance patterns and genetic structure of C. jejuni from stool samples of symptomatic patients in Dalmatia, the largest Croatian county.

Concepts: Bacteria, Escherichia coli, Proteobacteria, Gastroenteritis, Campylobacter, Campylobacter jejuni