On the Trail of Campylobacter

By: Jvo Siegrist, AnalytiX Volume 8 Issue 1

Detection, identification, differentiation and cultivation of Campylobacter

Product Manager, Microbiology ivo.siegrist@sial.com   

The Campylobacter is one of the leading causes of human gastroenteritis. Common Campylobacter species C. jejuni, C. coli, and C. lari are responsible for most cases of campylobacteriosis1,2. However, other species, like C. fetus, which causes spontaneous abortions, have also been associated with human illness2–6.
Campylobacter are Gram-negative, spiral-shaped, microaerophilic and motile bacteria with uni- or bi-polar fl agella (see Figures 1, 2 and 3).

 

Figure 1.Scanning electron microscope image: shows the characteristic spiral, or corkscrew, shape of C. jejuni cells.

Figure 1.Scanning electron microscope image: shows the characteristic spiral, or corkscrew, shape of C. jejuni cells.
Photo by De Wood; digital colorization by Chris Pooley, USDA/ARS

Figure 2. Scanning electron micrograph of the single polar flagellum and corkscrew shape of C. jejuni.

Figure 2. Scanning electron micrograph of the single polar flagellum and corkscrew shape of C. jejuni. These morphologic characteristics contribute to the characteristic darting motility of C jejuni in the viscous mucous layer of the intestinal lumen.
Sean F. Altekruse National Cancer Institute, Rockville

Figure 3. Electron microscope image of Campylobacters by Jochen Reetz, Bundesinstitut für Risikobewertung (BfR), Germany

Figure 3. Electron microscope image of Campylobacters by Jochen Reetz, Bundesinstitut für Risikobewertung (BfR), Germany

The size of a cell is roughly 0.2–0.8 x 0.5–5 µm and, interestingly, in culture they can undergo morphological change from spiral to spherical shape. Most species are catalase- and oxidase-positive7, with the exception of the catalase-negative C. sputorum, C. concisus, C. mucosalis and C. helveticus. The metabolism of Campylobacter is chemoorganotrophic, with amino acids and intermediates of the citric acid cycle serving as energy sources; typical carbohydrates cannot be used. Campylobacter reduce nitrate to nitrite, obtaining oxygen for their metabolism by this pathway. These distinctive metabolic reactions can be used for the differentiation and identification of Campylobacter species (Table 1). Commercially available tests from Sigma-Aldrich appear in Tables 2 and 3.

 

Table 1. Table of differentiating characteristics of Campylobacter species and subspecies

Table 1. Table of differentiating characteristics of Campylobacter species and subspecies

Table 2 Sigma-Aldrich tests for identification and differentiation of Campylobacter

Campylobacter test Cat. no.
Catalase Test 88597
Hippurate Disks 40405
Hydrogen Sulfide Test Strips 06728
Indoxyl Strips 04739
Nitrate Reagent A 38497
Nitrate Reagent B 39441
Nitrate Reagent Disks 08086
Oxidase Test 70439
Oxidase Strips 40560
Oxidase Reagent acc. Gaby-Hadley A 07345
Oxidase Reagent acc. Gaby-Hadley B 07817
Oxidase Reagent acc. Gordon-McLeod 18502


Table 3 Sigma-Aldrich Gram staining kit and single solutions

Gram stain Cat. no.
Gram staining kit 77730
Gram‘s crystal violet solution 94448
Gram‘s decolorizer solution 75482
Gram’s fuchsin solution 87794
Gram‘s iodine solution 90107
Gram‘s safranin solution 94635

 

Campylobacter are generally very fastidious microorganisms and grow only on complex media that have been amended with diverse essential amino acids and supplements, such as pyruvate, a-ketoglutarate, hemin, formate and other essential ions. For selective isolation of Campylobacter, the growth media can be supplemented with antibiotics like cefoperazone, vancomycin, trimethoprim, amphotericin, cycloheximide, rifampicin, cefsulodin and polymyxin B sulfate. Sigma- Aldrich offers a broad range of specifi c agars and broths for the detection, identifi cation, differentiation, enumeration and cultivation of Campylobacter (Table 4).

Table 4 Media for Campylobacter (detection, differentiation, and identification )

Nonselective media Cat. no.
Tryptic Soy Broth, Vegitone 41298
Columbia Agar 27688
Tryptic Soy Agar, Vegitone 14432
Tryptic Soy Broth 22092
CASO Agar 22095
Tryptic Soy Broth No. 2 51228
Tryptic Soy Agar 22091
Tryptic Soy Agar Plates (Diameter 55 mm) 57994
CASO Broth 22098
Nonselective nedia with differential system Cat. no.
Blood Agar Base No. 2 B1676
Hippurate Broth 53275
OF Test Nutrient Agar 75315
Selective media with differential system Cat. no.
MacConkey Agar No. 1 70143
Campylobacter selective media Cat. no.
Bolton Broth Base 67454
Karmali Campylobacter Agar (Base) 17152
Brucella Broth Base B3051
Campylobacter Selective Agar (Base) 21378
Park and Sanders Enrichment Broth (Base) 17189
Blood-free Campylobacter Selective Agar Base B2426

 

For more details about our products for analytical microbiology, please visit our website www.sigmaaldrich.com/microbiology

Scientific classification of Campylobacter:

Kingdom: Bacteria
Phylum: Proteobacteria
Class: Epsilon Proteobacteria
Order: Campylobacterales
Family: Campylobacteraceae
Genus: Campylobacter

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Materials

     

References

  1. Campylobacter jejuni Current Status and Future Trends. Nachamkin, I., Blaser, M. J., Tompkins, L. S., eds., American Society for Microbiology: Washington, DC (1992).
  2. Tauxe, R. V., Hargrett-Bean, N., Patton, C. M., Wachsmuth, I. K., Campylobacter isolates in the United States, 1982 – 86. Morbid. Mortal. Weekly Rep. (1988), 37, 1–13.
  3. Butzler, J. P., Campylobacter Infection in Man and Animals. CRC Press: Boca Raton, FL (1984).
  4. Linton, D., Owen, R. J., Stanley, J., Rapid identification by PCR of the genus Campylobacter and of five Campylobacter species enteropathogenic for man and animals. Res. Microbiol. (996), 147, 707–18.
  5. Patton, D. M., Shaffer, N., Edmonds, P., Barrett, T. J., Lampert, M. A., Baker, C., Perlman, D. M., Brenner, D., Human disease associated with Campylobacter upsaliensis (catalase-negative or weakly positive Campylobacter species) in the United States. J. Clin. Microbiol. (1989), 27, 66 –73.
  6. Tee, W., Anderson, B. N., Ross, B. C., Dwyer, B., Atypical campylobacters associated with gastroenteritis. J. Clin. Microbiol. (1987), 25, 1248 – 54.
  7. Ryan, K. J. and Ray C. G., Eds., Sherris Medical Microbiology, 4th ed., McGraw-Hill: New York, NY (2004), 378 – 80.

 

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