Chapter 3 : Chromogenic and Accelerated Cultural Methods

MyBook is a cheap paperback edition of the original book and will be sold at uniform, low price.

Ebook: Choose a downloadable PDF or ePub file. Chapter is a downloadable PDF file. File must be downloaded within 48 hours of purchase

Buy this Chapter
Digital (?) $15.00

Preview this chapter:
Zoom in

Chromogenic and Accelerated Cultural Methods, Page 1 of 2

| /docserver/preview/fulltext/10.1128/9781555817121/9781555815424_Chap03-1.gif /docserver/preview/fulltext/10.1128/9781555817121/9781555815424_Chap03-2.gif


Analysis of food samples by cultural and enrichment techniques remains an integral part of the examination of food for the presence or enumeration of foodborne pathogens. Enrichment methods have afforded the ability to detect as few as one cell per 500 g of food. In addition, enrichment techniques aid in the recovery of injured bacteria, which are often present in foods in a stressed condition because some foods may lack the optimal nutrients essential for bacterial growth, the food may present an environment not conducive to growth of the microorganisms, or the bacteria may be damaged during processing of the food. In this regard, the enrichment process provides an optimal growth medium and a period of time that can be effectively used by injured/stressed microorganisms to perform repair of cellular damage, allowing for resuscitation of the bacterial cells. Additionally, enrichment processes provide an opportunity for target bacterial cells to proliferate, while the growth of competing background microflora is truncated. This chapter describes many differential and selective media, which have been developed in response to the presence of bacterial pathogens capable of causing illness in humans, by employing unique phenotypic characteristics of these microorganisms to develop detection platforms based on cultural approaches. The selective media includes enterohemorrhagic (EHEC) media, media, media, media, media, and enumeration media. Several groups have investigated the use of a single broth to simultaneously enrich the concentrations of several bacterial species, followed by downstream detection.

Citation: Goodridge L, Bisha B. 2011. Chromogenic and Accelerated Cultural Methods, p 47-61. In Hoorfar J (ed), Rapid Detection, Characterization, and Enumeration of Foodborne Pathogens. ASM Press, Washington, DC. doi: 10.1128/9781555817121.ch3
Highlighted Text: Show | Hide
Loading full text...

Full text loading...


1. Aragon-Alegro, L. C.,, D. C. Aragon,, E. Z. Martinez,, M. Landgraf,, B. D. Gombossy de Melo Franco, and, M. T. Destro. 2008. Performance of a chromogenic medium for the isolation of Listeria monocytogenes in food. Food Control 19:483486.
2. Becker, B.,, S. Schuler,, M. Lohneis,, A. Sabrowski,, G. D. Curtis, and, W. H. Holzapfel. 2006. Comparison of two chromogenic media for the detection of Listeria monocytogenes with the plating media recommended by EN/DIN 11290-1. Int. J. Food Microbiol. 109:127131.
3. Cassar, R.,, and P. Cuschieri. 2003. Comparison of Salmonella chromogenic medium with DCLS agar for isolation of Salmonella species from stool specimens. J. Clin. Microbiol. 41:32293232.
4. Centers for Disease Control and Prevention. 1998. Outbreak of Vibrio parahaemolyticus infections associated with eating raw oysters—Pacific Northwest, 1997. MMWR Morb. Mortal. Wkly. Rep. 47:457462.
5. Centers for Disease Control and Prevention. 1999. Outbreak of Vibrio parahaemolyticus infection associated with eating raw oysters and clams harvested from Long Island Sound—Connecticut, New Jersey, and New York, 1998. MMWR Morb. Mortal. Wkly. Rep. 48:4851.
6. Chain, V. S.,, and D. Y. C. Fung. 1991. Comparison of Redigel, Petrifilm, spiral plate system, Isogrid, and aerobic plate count for determining the numbers of aerobic bacteria in selected foods. J. Food Prot. 54:208211.
7. Church, D. L.,, D. Emshey,, H. Semeniuk,, T. Lloyd, and, J. D. Pitout. 2007. Evaluation of BBL CHROMagar O157 versus sorbitol-MacConkey medium for routine detection of Escherichia coli O157 in a centralized regional clinical microbiology laboratory. J. Clin. Microbiol. 45:30983100.
8. Cooke, V. M.,, R. J. Miles,, R. G. Price, and, A. C. Richardson. 1999. A novel chromogenic ester agar medium for detection of salmonellae. Appl. Environ. Microbiol. 65:807812.
9. Daniels, N. A.,, L. MacKinnon,, R. Bishop,, S. Altekruse,, B. Ray,, R. M. Hammond,, S. Thompson,, S. Wilson,, N. H. Bean,, P. M. Griffin, and, L. Slutsker. 2000a. Vibrio parahaemolyticus infections in the United States, 1973–1998. J. Infect. Dis. 181:16611666.
10. Daniels, N. A.,, B. Ray,, A. Easton,, N. Marano,, E. Kahn,, A. L. McShan II,, L. Del Rosario,, T. Baldwin,, M. A. Kingsley,, N. D. Puhr,, J. G. Wells, and, F. J. Angulo. 2000b. Emergence of a new Vibrio parahaemolyticus serotype in raw oysters. JAMA 284:15411545.
11. de Boer, E.,, and A. E. Heuvelink. 2000. Methods for the detection and isolation of Shiga-toxin producing Escherichia coli. J. Appl. Microbiol. Symp. Suppl. 88:133S143S.
12. DePaola, A.,, J. Ulaszek,, C. A. Kaysner,, B. J. Tenge,, J. L. Nordstrom,, J. Wells,, N. Puhr, and, S. M. Gendel. 2003. Molecular, serological, and virulence characteristics of Vibrio parahaemolyticus isolated from environmental, food, and clinical sources in North America and Asia. Appl. Environ. Microbiol. 69:39994005.
13. Duan, J.,, and Y. C. Su. 2005. Comparison of a chromogenic medium with thiosulfate–citrate–bile salts–sucrose agar for detecting Vibrio parahaemolyticus. J. Food Sci. 70:125128.
14. El Marrakchi, A.,, N. Boumhandi, and, A. Hamama. 2005. Performance of a new chromogenic plating medium for isolation of Listeria monocytogenes from marine environments. Lett. Appl. Microbiol. 40:8791.
15. Food and Drug Administration. 2007. Bacteriological Analytical Manual, chapt. 5. Salmonella. (Online.) http://www.fda.gov/Food/ScienceResearch/LaboratoryMethods/BacteriologicalAnalyticalManualBAM/ucm070149.htm. Accessed 17 September 2010.
16. Funatsu, T.,, T. Taniyama,, T. Tajima,, H. Tadakuma, and, H. Namiki. 2002. Rapid and sensitive detection method of a bacterium by using a GFP reporter phage. Microbiol. Immunol. 46:365369.
17. Gaillot, O.,, P. di Camillo,, P. Berche,, R. Courcol, and, C. Savage. 1999. Comparison of CHROMagar Salmonella medium and Hektoen enteric agar for isolation of salmonellae from stool samples. J. Clin. Microbiol. 37:762765.
18. Gasanov, U.,, D. Hughes, and, P. M. Hansbro. 2005. Methods for the isolation and identification of Listeria spp. and Listeria monocytogenes: a review. FEMS Microbiol. Rev. 29:851875.
19. Greenwood, M.,, C. Willis,, P. Doswell,, G. Allen, and, K. Pathak. 2005. Evaluation of chromogenic media for the detection of Listeria species in food. J. Appl. Microbiol. 99:13401345.
20. Hitchins, A. D. 2003. Listeria monocytogenes, p. 10.01–10.13. In Food and Drug Administration (ed.), Bacteriological Analytical Manual, 8th ed. AOAC International, Washington, DC.
21. Hoorfar, J.,, and D. Baggesen. 1998. Importance of pre-enrichment media for isolation of Salmonella spp. from swine and poultry. FEMS Microbiol. Lett. 169:125130.
22. Jenkins, C.,, J. Evans,, H. Chart,, G. A. Willshaw, and, G. Frankel. 2008. Escherichia coli serogroup O26—a new look at an old adversary. J. Appl. Microbiol. 104:1425.
23. Jenkins, C.,, M. C. Pearce,, A. W. Smith,, H. I. Knight,, D. J. Shaw,, T. Cheasty,, G. Foster,, G. J. Gunn,, G. Dougan,, H. R. Smith, and, G. Frankel. 2003. Detection of Escherichia coli serogroups O26, O103, O111, and O145 from bovine faeces using immunomagnetic separation and PCR/DNA probe techniques. Lett. Appl. Microbiol. 37:207212.
24. Jiang, J.,, C. Larkin,, M. Steele,, C. Poppe and, J. A. Odumeru. 1998. Evaluation of universal preenrichment broth for the recovery of foodborne pathogens from milk and cheese. J. Dairy Sci. 81:27982803.
25. Kanki, M.,, K. Seto,, J. Sakata,, T. Harada, and, Y. Kumeda. 2009. Simultaneous enrichment of Shiga toxin-producing Escherichia coli O157 and O26 and Salmonella in food samples using universal preenrichment broth. J. Food Prot. 72:20652070.
26. Kim, H.,, and A. K. Bhunia. 2008. SEL, a selective enrichment broth for simultaneous growth of Salmonella enterica, Escherichia coli O157:H7, and Listeria monocytogenes. Appl. Environ. Microbiol. 74:48534866.
27. Klein, E. J.,, J. R. Stapp,, M. A. Neill,, J. M. Besser,, M. T. Osterholm, and, P. I. Tarr. 2004. Shiga toxin antigen detection should not replace sorbitol MacConkey agar screening of stool specimens. J. Clin. Microbiol. 42:44164417.
28. Kobayashi, T.,, S. Enomoto,, R. Sakazaki, and, S. Kuwahara. 1963. A new selective medium for pathogenic vibrios: T.C.B.S. agar (Modified Nakanishi’s Agar). Jpn. J. Bacteriol. 18:387391.
29. Leimeister-Wächter, M.,, E. Domann, and, T. Chakraborty. 1991. Detection of a gene encoding a phosphatidylinositol-specific phospholipase C that is co-ordinately expressed with listeriolysin in Listeria monocytogenes. Mol. Microbiol. 5:361366.
30. Line, J. E. 2001. Development of a selective differential agar for the isolation and enumeration of Campylobacter spp. J. Food Prot. 64:17111715.
31. Manafi, M. 2000. New developments in chromogenic and fluorogenic culture media. Int. J. Food Microbiol. 60:205218.
32. Manafi, M.,, and B. Kremsmaier. 2001. Comparative evaluation of different chromogenic/fluorogenic media for detecting Escherichia coli O157:H7 in food. Int. J. Food Microbiol. 71:257262.
33. March, S. B.,, and S. Ratnam. 1986. Sorbitol-MacConkey medium for detection of Escherichia coli O157:H7 associated with hemorrhagic colitis. J. Clin. Microbiol. 23:869872.
34. Mead, P. S.,, L. Slutsker,, V. Dietz,, L. F. McCaig,, J. S. Bresee,, C. Shapiro,, P. M. Griffin, and, R. V. Tauxe. 1999. Food-related illness and death in the United States. Emerg. Infect. Dis. 5:607625.
35. Miller, R. G.,, and E. T. Mallinson. 2000. Improved detection of Salmonella of nontyphoid and typhoid salmonellae with balanced agar formulations. J. Food Prot. 63:14431446.
36. Murray, P.,, E. J. Baron,, J. H. Jorgensen,, M. A. Pfaller, and, R. H. Yolken (ed.). 2003. Manual of Clinical Microbiology, 8th ed. ASM Press, Washington, DC.
37. Nakanishi, Y. 1963. An isolation agar medium for cholerae and enteropathogenic halophilic vibrios. Modern Media 9:246.
38. National Advisory Committee on Microbiological Criteria for Foods. 2007. Analytical utility of Campylobacter methodologies. J. Food Prot. 70:241250.
39. Notermans, S. H.,, J. Dufrenne,, M. LeimeisterWachter,, E. Domann, and, T. Chakraborty. 1991. Phosphatidylinositol-specific phospholipase C activity as a marker to distinguish between pathogenic and nonpathogenic Listeria species. Appl. Environ. Microbiol. 57:26662670.
40. Novicki, T. J.,, J. A. Daly,, S. L. Mottice, and, K. C. Carroll. 2000. Comparison of sorbitol MacConkey agar and a two-step method which utilizes enzyme-linked immunosorbent assay toxin testing and a chromogenic agar to detect and isolate enterohemorrhagic Escherichia coli. J. Clin. Microbiol. 38:547551.
41. Oyarzabal, O. A.,, K. S. Macklin,, J. M. Barbaree, and, R. S. Miller. 2005. Evaluation of agar plates for direct enumeration of Campylobacter spp. from poultry carcass rinses. Appl. Environ. Microbiol. 71:33513354.
42. Perry, J. D.,, M. Ford,, J. Taylor,, A. L. Jones,, R. Freeman, and, F. K. Gould. 1999. ABC medium, a new chromogenic agar for selective isolation of Salmonella spp. J. Clin. Microbiol. 37:766768.
43. Pignato, S.,, G. Giammanco, and, G. Giammanco. 1995. Rambach agar and SM- I D medium sensitivity for presumptive identification of Salmonella subspecies I-VI. J. Med. Microbiol. 43:6871.
44. Pontello, M.,, S. Russolo,, F. Carozzi, and, U. Bottiroli. 1987. Evaluation of a new, rapid method (MUCAP test) for the presumptive identification of Salmonella on primary isolation media. Abstr. 5th Int. Symp. Rapid Methods and Automation Microbiol. Immunol., Florence, Italy, p. 339.
45. Poupart, M. C.,, M. Mounier,, F. Denis,, J. Sirot,, C. Couturier,, A. M. Freydiere, and, Y. Gille. 1991. Detection of salmonellae by using Rambach agar and by a C8 esterase spot test. J. Clin. Microbiol. 29:23572359.
46. Rambach, A. 1990. New plate medium for facilitated differentiation of Salmonella spp. from Proteus spp. and other enteric bacteria. Appl. Environ. Microbiol. 56:301303.
47. Reissbrodt, R. 2004. New chromogenic plating media for detection and enumeration of pathogenic Listeria spp.—an overview. Int. J. Food Microbiol. 95:19.
48. Schönenbrücher, V.,, E. T. Mallinson, and, M. Bulte. 2008. A comparison of standard cultural methods for the detection of foodborne Salmonella species including three new chromogenic plating media. Int. J. Food Microbiol. 123:6166.
49. Sharma, V. K.,, and S. A. Carlson. 2000. Simultaneous detection of Salmonella strains and Escherichia coli O157:H7 with fluorogenic PCR and single-enrichment-broth culture. Appl. Environ. Microbiol. 66:54725476.
50. Silliker, J. H.,, and D. A. Gabis. 1973. ICMSF methods studies. I. Comparison of analytical schemes for detection of Salmonella in dried foods. Can. J. Microbiol. 19:475479.
51. Skirrow, M. B. 1977. Campylobacter enteritis: a new disease. Br. Med. J. 2:911.
52. Sperber, W. A.,, M. A. Moorman, and, T. A. Freier. 2001. Cultural methods for the enrichment and isolation of microorganisms, p. 45–51. In F. P. Downes and, K. Ito (ed.), Microbiological Examination of Foods. American Public Health Association, Washington, DC.
53. Su, Y. C.,, J. Duan, and, W. H. Wu. 2005. Selectivity and specificity of a chromogenic medium for detecting Vibrio parahaemolyticus. J. Food Prot. 68:14541456.
54. Taylor, W. I.,, and J. H. Silliker. 1961. Isolation of salmonellae from food samples. IV. Comparison of methods of enrichment. Appl. Microbiol. 9:484486.
55. van Dijk, S.,, M. J. Bruins, and, G. J. H. M. Ruijs. 2009. Evaluation and implementation of a chromogenic agar medium for Salmonella detection in stool in routine laboratory diagnostics. J. Clin. Microbiol. 47:456458.
56. Willis, C.,, T. Baalham,, M. Greenwood, and, F. Presland. 2006. Evaluation of a new chromogenic agar for the detection of Listeria in food. J. Appl. Microbiol. 101:711717.
57. Wong, H. C. 2003. Detecting and molecular typing of Vibrio parahaemolyticus. J. Food Drug Anal. 11:100107.
58. Yu, Y. G.,, H. Wu,, Y. Y. Liu,, S. L. Li,, X. Q. Yang, and, X. L. Xiao. 2010. A multipathogen selective enrichment broth for simultaneous growth of Salmonella enterica serovar Enteritidis, Staphylococcus aureus, and Listeria monocytogenes. Can. J. Microbiol. 56:585597.
59. Zadik, P. M.,, P. A. Chapman, and, C. A. Siddons. 1993. Use of tellurite for the selection of verocytotoxigenic Escherichia coli O157. J. Med. Microbiol. 39:155158.


Generic image for table

Examples of representative accelerated cultural methods for pre-enrichment/enrichment, plating, and confirmation of select foodborne pathogens

Citation: Goodridge L, Bisha B. 2011. Chromogenic and Accelerated Cultural Methods, p 47-61. In Hoorfar J (ed), Rapid Detection, Characterization, and Enumeration of Foodborne Pathogens. ASM Press, Washington, DC. doi: 10.1128/9781555817121.ch3

This is a required field
Please enter a valid email address
Please check the format of the address you have entered.
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error