Growth and Laboratory Maintenance of <i>Campylobacter jejuni</i>

Davis, Lindsay; DiRita, Victor J.

doi:10.1002/9780471729259.mc08a01s10

Cited by 56 publications

(45 citation statements)

References 8 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Campylobacter spp. are known to grow best in vitro at an oxygen content of 5% (62,63), and Treponema phagedenis has been found to grow best in the authors' laboratory at approximately 2% oxygen (A. Krull and P. Plummer, unpublished data). While these bacteria may not be the cause of DD, they certainly could play a role in deteriorating skin integrity and setting up an ideal environment for more pathogenic bacteria to thrive.…”

Section: Discussionmentioning

confidence: 97%

Deep Sequencing Analysis Reveals Temporal Microbiota Changes Associated with Development of Bovine Digital Dermatitis

et al. 2014

View full text Add to dashboard Cite

bBovine digital dermatitis (DD) is a leading cause of lameness in dairy cattle throughout the world. Despite 35 years of research, the definitive etiologic agent associated with the disease process is still unknown. Previous studies have demonstrated that multiple bacterial species are associated with lesions, with spirochetes being the most reliably identified organism. This study details the deep sequencing-based metagenomic evaluation of 48 staged DD biopsy specimens collected during a 3-year longitudinal study of disease progression. Over 175 million sequences were evaluated by utilizing both shotgun and 16S metagenomic techniques. Based on the shotgun sequencing results, there was no evidence of a fungal or DNA viral etiology. The bacterial microbiota of biopsy specimens progresses through a systematic series of changes that correlate with the novel morphological lesion scoring system developed as part of this project. This scoring system was validated, as the microbiota of each stage was statistically significantly different from those of other stages (P < 0.001). The microbiota of control biopsy specimens were the most diverse and became less diverse as lesions developed. Although Treponema spp. predominated in the advanced lesions, they were in relatively low abundance in the newly described early lesions that are associated with the initiation of the disease process. The consortium of Treponema spp. identified at the onset of disease changes considerably as the lesions progress through the morphological stages identified. The results of this study support the hypothesis that DD is a polybacterial disease process and provide unique insights into the temporal changes in bacterial populations throughout lesion development.

show abstract

Section: Discussionmentioning

confidence: 97%

Deep Sequencing Analysis Reveals Temporal Microbiota Changes Associated with Development of Bovine Digital Dermatitis

et al. 2014

View full text Add to dashboard Cite

show abstract

“…Preparation of the growth curve inocula were performed by statically culturing NCTC 11,168, CjCm and CjK at 42°C in the broth phase of biphasic MH broth and agar (2% w/v) in a microaerophilic environment, as described previously (Davis and DiRita, 2008). Growth curves were performed in octuplet using microplates in a Bioscreen C plate reader (Growth Curves USA, Piscataway, NJ), measuring OD 600 every 2 h for 48 h. To limit aggregation, which may impact optical density values, microplates were shaken for 30 s prior to each OD 600 reading.…”

Section: Campylobacter Jejuni Motility and Growth Curvesmentioning

confidence: 99%

Intestinal colonization and acute immune response in commercial turkeys following inoculation with Campylobacter jejuni constructs encoding antibiotic-resistance markers

Sylte

Johnson

Meyer

et al. 2019

Veterinary Immunology and Immunopathology

View full text Add to dashboard Cite

Consumption of contaminated poultry products is one of the main sources of human campylobacteriosis, of which Campylobacter jejuni subsp. jejuni (C. jejuni) is responsible for approximately 90% of the cases. At slaughter, the ceca of commercial chickens and turkeys are the main anatomical site where C. jejuni asymptomatically colonizes. We have previously colonized commercial turkey poults with different isolates of C. jejuni and evaluated different media to best enumerate Campylobacter from intestinal samples, but the hostresponse is unknown in turkeys. Enumeration of Campylobacter(colony forming units (cfu)/gram of intestinal contents) can be challenging, and can be confounded if animals are colonized with multiple species of Campylobacter. In order to precisely enumerate the C. jejuni isolate used to experimentally colonize turkeys, constructs of C. jejuni (NCTC 11,168) were tagged with different antibiotic resistance markers at the CmeF locus (chloramphenicol (CjCm) or kanamycin (CjK)). We sought to examine the kinetics of intestinal colonization using the antibiotic resistant constructs, and characterize the immune response in cecal tissue of turkeys. In vitroanalysis of the tagged antibiotic-resistant constructs demonstrated no changes in motility, morphology, or adherence and invasion of INT-407 cells compared to the parent isolate NCTC 11,168. Two animal experiments were completed to evaluate intestinal colonization by the constructs. In experiment 1, three-week old poults were colonized after oral gavage for 14 days, and CjCm and CjK cfu were recovered from cecal, but not ileal contents. In experiment 2, nine-week old poults were orally inoculated with CjCm, and the abundance of CjCm cfu/g of cecal contents significantly decreased beyond 14 days after inoculation. Significant lesions were detected in CjCm colonized poults at day 2 post-colonization. Using immunohistochemistry, Campylobacter antigen was detected in between cecal villi by day 7 of CjCm colonized poults. Quantitative RT-PCR of CjCm-colonized cecal tissue demonstrated significant downregulation of IL-1β, IL-10 and IL-13 mRNA, and significant up-regulation of IL-6, and IFNγ mRNA on day 2, and for some on day 7 post-colonization. All differentially expressed genes were similar to mock-infected poults by day 14. These data suggest that C. jejuni induced a brief inflammatory response in the cecum of poults that quickly resolved. Results from this study provide valuable insight into host-response and persistent colonization of the turkey cecum. These findings will help to develop and test strategies to promote food safety in commercial turkeys.

show abstract

“…Various methods of culture preparation including cell separation and concentration by filtration or centrifugation can be done to accelerate the enrichment process. 12…”

Section: Culture-based Methodsmentioning

confidence: 99%

Hyperspectral microscope imaging methods for multiplex detection of Campylobacter

Park

Eady

Oakley

et al. 2019

J. Spectral Imaging

View full text Add to dashboard Cite

Campylobacter is an emerging zoonotic bacterial threat in the poultry industry. The current methods for the isolation and detection of Campylobacter are culture-based techniques with several selective agars designed to isolate Campylobacter colonies, which is time-consuming, labour intensive and has low sensitivity. Several immunological and molecular techniques such as enzyme-linked immunosorbent assay (ELISA) and Latex agglutination are commercially available for the detection and identification of Campylobacter. However, these methods demand more advanced instruments as well as specially trained experts. A hyperspectral microscope imaging (HMI) technique with the fluorescence in situ hybridisation (FISH) technique has the potential for multiplex foodborne pathogen detection. Using Alexa488 and Cy3 fluorophores, the HMI (450–800 nm) technique was able to identify Campylobacter jejuni stains with high sensitivity and specificity. In addition, HMI was able to classify six bacteria using scattering intensity from their spectra without a FISH fluorophore. Overall classification accuracy of quadratic discriminant analysis (QDA) method for six bacteria including Bifidobacter longum, Campylobacter jejuni, Clostridium perfringens, Enterobacter cloacae, Lactobacillus salivarius and Shigella flexneri using the HMI technique without fluorescent markers was approximately 88.6 % with pixel-wise classification.

show abstract

Growth and Laboratory Maintenance of Campylobacter jejuni

Cited by 56 publications

References 8 publications

Deep Sequencing Analysis Reveals Temporal Microbiota Changes Associated with Development of Bovine Digital Dermatitis

Deep Sequencing Analysis Reveals Temporal Microbiota Changes Associated with Development of Bovine Digital Dermatitis

Intestinal colonization and acute immune response in commercial turkeys following inoculation with Campylobacter jejuni constructs encoding antibiotic-resistance markers

Hyperspectral microscope imaging methods for multiplex detection of Campylobacter

Contact Info

Product

Resources

About