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Escherichia coli

Escherichia coli (E. coli) is a bacterium found in the faeces of all warm-blooded animals, including man. In Europe and elsewhere it is used as an indicator of the sanitary quality of bivalve shellfish.

The level of E. coli in bivalve shellfish shows how much faecal pollution (human sewage or animal waste) they have been exposed to in the harvesting area. This in turn determines what, if any, treatment shellfish require before they are eaten. EU food law stipulates monitoring, classification and resulting treatments.

There are many different methods for counting/assessing the level of E. coli in food and environmental samples, but not all are suitable for bivalve shellfish. Under EU Regulations a reference method for E. coli in bivalve shellfish is stipulated for official control testing: ISO TS 16649-3 - Microbiology of food and animal feedings stuffs - Horizontal method for the enumeration of β-glucuronidase-positive Escherichia coli - Part 3: Most probable number technique using5-bromo-4-chloro-3-indolyl-β-D glucuronide. This reference method has been extensively tested in bivalve shellfish and provides a reliable indicator of the amount of faecal contamination that shellfish have been exposed to.

This reference method contains a specific recovery step, where E. coli cells damaged by exposure to seawater are able to recover. Acid production in minerals-modified glutamate broth is used in the first stage of the method, followed by confirmation of the presence of E. coli through β-glucoronidase production at 44°C using a chromogenic medium. The result is expressed as a "most probable number" (MPN) of E. coli per 100g shellfish. The MPN technique estimates microbial population sizes based on the pattern of positive and negative results following incubation of replicated diluted sample, rather than counting individual bacterial cells.

For bivalve shellfish the test is applied in a "5 by 3" format: five replicates are tested at each of three (or more) dilutions. This increases the method's precision, over the common "3 by 3" application.

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Last Modified: 26 September 2011