Gram negative bacterial species are identified by comparison to an online database.

Test 2 ID 32E (bioMérieux SA; Marcy-l’Etoile, France) [30] consists of 32 miniaturised enzyme assays with positive or negative scores these assays can be measured TGF-beta inhibitor either manually or automatically and Gram negative bacterial species are identified by comparison to an online database. Test 3 API Zym (bioMérieux SA; Marcy-l’Etoile, France) [31] consists of 20 cupules with 19 enzyme assays and one control. The assays produce a coloured response which is scored in intensity between 0 and 5. Test 4 Biotyping [1] is a series of biochemical tests for identifying bacteria. Tests are carried out for: indole production (Ind), motility at 36°C (Mot), acid production from PCI-32765 cell line i-inositol (Ino), malonate utilization (Malo) ornithine-Moellers (Orn), acid production from dulcitol (Dul), Methyl Red test (MR), Voges-Proskauer (VP) test, gas production (Gas), and nitrite AS1842856 mw metabolism (Nit). Details of all tests are given in [1]. The results of each test were represented by a separate dataset containing only the strains that have results for that test. The Test 1, Test 2, Test 3 and Test 4 datasets contained 91, 92, 65 and 76

strains respectively. There are 98 strains in total, 48 of these have data for all four tests. Further, 31 only have data for three out of four tests, and 14 for only two out of four tests. It should be noted that although there was a considerable overlap between the datasets, each dataset was considered separately. Each

strain was identified Benzatropine by its isolate number retrieved from the Cronobacter MLST database [13] as well as source, geographical location and date of isolation. These attributes were removed for the purpose of clustering but were used to label the data afterwards. The result of each enzyme assay was represented categorically. In the case of Tests 1, 2 and 4 this was 0 or 1 for a negative or positive result respectively. A positive result being one which shows activity for the enzyme in the sample. Test 3 had categories ranging from 0 to 5. 0 is indicative of no reaction, and categories 1-5 indicate a range of positive responses, with 5 being the strongest response. Thus, each strain from each dataset was represented by a vector of attributes with each attribute containing the result of one of the enzyme assays in the corresponding test. Features used The enzyme assays used in this study were not designed to discriminate between species or genotypes of Cronobacter. In all four tests there were assays where all (or almost all) strains were reported as producing the same result, either positive or negative. Attributes where all strains produce the same result, either positive or negative, for Tests 1, 2 and 4 or where all strains occupy one category in the case of Test 3 were removed from the list of features used for clustering. The features from each test used to perform clustering are listed in Table 7.