Evaluation of the ID 32E for the identification of Gram-negative glucose-fermenting and glucose-non-fermenting bacilli

O'Hara, C M; Miller, J. M.

doi:10.1111/j.1469-0691.1999.tb00141.x

Cited by 7 publications

(7 citation statements)

References 7 publications

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“…Specifically, each sample was transferred into buffered peptone water (Oxoid) and incubated at 25 °C and 37 °C for 24 h. Subsequently, the samples were streaked onto MacConkey agar n. 3 (Oxoid) plates and incubated at 25 °C and 37 °C for 24 h. All isolated strains were primarily identified on the basis of their colonial morphology, lactose metabolism, pigment production, and standard biochemical tests. The isolates were then confirmed using the Analytical Profile Index system (bioMérieux), and the identification to the species level was considered successful when reading provided at least “Very Good id.” (%id > 99.0 and T > 0.5) [38].…”

Section: Methodsmentioning

confidence: 99%

Gastrointestinal investigation of parasites and Enterobacteriaceae in loggerhead sea turtles from Italian coasts

et al. 2019

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BackgroundCaretta caretta is the most abundant sea turtle species in the Mediterranean, and studies on this species have vastly expanded during recent years, including those investigating gut bacterial and parasitic communities. Members of these communities have been reported with variable prevalence and pathogenicity, mainly depending on their host and environment (e.g. lifespan, distribution, habitat, diet, health status and stressors). Indeed, many species commonly inhabiting the sea turtle gastrointestinal tract exhibit an opportunistic behaviour.This study aimed to provide baseline data on enterobacterial and parasitic composition, through bacteriological culture-based methods and the FLOTAC parasitological technique, in cloacal and faecal samples of 30 live Caretta caretta, examined upon their arrival at the Marine Turtle Research Centre (Portici, Italy).ResultsEnterobacteriaceae were isolated in 18/23 cloacal samples (78.3%), with Citrobacter and Morganella as the most common genera, followed by Proteus, Enterobacter, Providencia, and Hafnia. Parasitic elements were detected in 11/30 faecal samples (36.7%), with Enodiotrema, Rhytidodes, and Eimeria as most common genera, followed by Pachypsolus and Cymatocarpus. Additionally, Angiodyctium is reported for the first time in this host. The majority (47.8%) of sea turtles hosted exclusively Enterobacteriaceae, whereas 30.4% hosted both parasites and Enterobacteriaceae; the remaining 21.8% hosted neither of the agents.ConclusionsBacteria and parasites evaluated in the present study are common in Mediterranean loggerhead sea turtles, with slight differences between the western and eastern basin. Although naturally present in the gastrointestinal system of free-living sea turtles, their relationship with these hosts might range from mutualism to parasitism. Indeed, members of the gut community might express their pathogenic potential in immune-compromised animals, such as those in rehabilitation facilities. Therefore, it is advisable to include in the standard work-up of rescued sea turtles a screening procedure for such opportunistic agents, in order to better evaluate the animal’s health status and achieve timely intervention with appropriate treatment, thus improving rehabilitation. Furthermore, data collected from free-living sea turtles represent a starting point for investigating wild populations. However, further studies are needed to clarify the differences between sea turtle’s normal gut microbiome and pathobiome.

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Section: Methodsmentioning

confidence: 99%

Gastrointestinal investigation of parasites and Enterobacteriaceae in loggerhead sea turtles from Italian coasts

et al. 2019

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“…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 either manually or automatically and Gram negative bacterial species are identified by comparison to an online database.…”

Section: Methodsmentioning

confidence: 99%

In Silico identification of pathogenic strains of Cronobacter from Biochemical data reveals association of inositol fermentation with pathogenicity

et al. 2011

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BackgroundCronobacter, formerly known as Enterobacter sakazakii, is a food-borne pathogen known to cause neonatal meningitis, septicaemia and death. Current diagnostic tests for identification of Cronobacter do not differentiate between species, necessitating time consuming 16S rDNA gene sequencing or multilocus sequence typing (MLST). The organism is ubiquitous, being found in the environment and in a wide range of foods, although there is variation in pathogenicity between Cronobacter isolates and between species. Therefore to be able to differentiate between the pathogenic and non-pathogenic strains is of interest to the food industry and regulators.ResultsHere we report the use of Expectation Maximization clustering to categorise 98 strains of Cronobacter as pathogenic or non-pathogenic based on biochemical test results from standard diagnostic test kits. Pathogenicity of a strain was postulated on the basis of either pathogenic symptoms associated with strain source or corresponding MLST sequence types, allowing the clusters to be labelled as containing either pathogenic or non-pathogenic strains. The resulting clusters gave good differentiation of strains into pathogenic and non-pathogenic groups, corresponding well to isolate source and MLST sequence type. The results also revealed a potential association between pathogenicity and inositol fermentation. An investigation of the genomes of Cronobacter sakazakii and C. turicensis revealed the gene for inositol monophosphatase is associated with putative virulence factors in pathogenic strains of Cronobacter.ConclusionsWe demonstrated a computational approach allowing existing diagnostic kits to be used to identify pathogenic strains of Cronobacter. The resulting clusters correlated well with MLST sequence types and revealed new information about the pathogenicity of Cronobacter species.

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“…Although phenotypic variants (5,6,15,20,21,23) can cause identification problems for some E. coli strains, nu- merous commercial systems or previous study on ID 32E systems (17,23) have been shown to identify E. coli with a fairly high degree of accuracy (11, 13, 14, 16, 19-22, 25-29, 32, 33, 36, 37). One strain serotyped as O74:H52 was found to be indole negative without affecting its identification as E. coli.…”

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confidence: 99%

“…A new well-based identification system, ID 32E, was recently developed by bioMérieux Vitek for the identification of Enterobacteriaceae and other nonfastidious gram-negative bacteria, after an incubation period of 22 Ϯ 2 h at 37 Ϯ 1°C (17,23). This system is an upgrading of the API 20E gallery and consists of 32 individual conic test wells (13 enzymatic, 3 biochemical, and 16 sugar utilization tests) in polystyrene trays (Table 1).…”

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confidence: 99%

Particular Biochemical Profiles for Enterohemorrhagic Escherichia coli O157:H7 Isolates on the ID 32E System

et al. 2001

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The ability of the ID 32E system to identify and discriminate 74 Escherichia coli O157 isolates among 106 E. coli non-O157 isolates was evaluated. The results showed atypical biochemical reactions but accurate identification at the species level and no unique biochemical profile numbers for E. coli O157, although these numbers were distinct from those of other serotypes.Verocytotoxigenic Escherichia coli (VTEC), a new public health problem worldwide, is represented by more than 100 serotypes that produce verocytotoxins, with two main serotypes, O157:H7 and O157:H(Ϫ). These last two serotypes, classified as enterohemorrhagic E. coli (EHEC), are implicated in hemorrhagic colitis, hemolytic-uremic syndrome, and thrombotic thrombocytopenic purpura. EHEC strains have been described as important and emergent food-borne pathogens (4, 18, 31). Other non-O157 VTEC serotypes, not deprived of pathogenicity, have been increasingly implicated in sporadic enteric diseases and outbreaks (18,31).Detection of E. coli O157:H7 is based on its recovery from samples and the presence of its virulence-associated factors (verocytotoxins) or the detection of its O157 antigens (3,30,31). Selective media for isolating O157:H7 strains rely on the fact that most of these strains display characteristic biochemical reactions (31): no ␤-glucuronidase activity and no D-sorbitol fermentation within 24 h at 37°C, except for some German or U.S. strains (10, 12). However, these last two biochemical features are not commonly used in routine clinical and food laboratories because of the simplicity of well-established commercial identification systems. The use of these systems on presumptive E. coli strains offers not only the advantage of confirming strains at the genus and/or species level but also the possibility to detect the presence of O157 EHEC by identifying profiles that are unique to these strains. For the Microscan dried conventional gram-negative identification panel (DadeMicroScan International, West Sacramento, Calif.), 90% of the tested E. coli O157:H7 strains shared two common biochemical profiles (1), whereas in the case of the API 20E (bioMérieux Vitek, Inc., Hazelwood, Mo.), 92% of the tested strains displayed the same profile (8).A new well-based identification system, ID 32E, was recently developed by bioMérieux Vitek for the identification of Enterobacteriaceae and other nonfastidious gram-negative bacteria, after an incubation period of 22 Ϯ 2 h at 37 Ϯ 1°C (17, 23). This system is an upgrading of the API 20E gallery and consists of 32 individual conic test wells (13 enzymatic, 3 biochemical, and 16 sugar utilization tests) in polystyrene trays (Table 1).A total of 180 serotyped E. coli strains were tested with the ID 32E system. These included 55 O157:H7 strains, 19 O157: H(Ϫ) strains confirmed by PCR on the rfb O157 locus (3), and 106 reference strains and laboratory isolates of non-O157:H7 or O157H(Ϫ) E. coli harboring (76 strains) or not harboring (30 strains) verocytotoxin genes, originating from patients, animals, and food...

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Evaluation of the ID 32E for the identification of Gram-negative glucose-fermenting and glucose-non-fermenting bacilli

Cited by 7 publications

References 7 publications

Gastrointestinal investigation of parasites and Enterobacteriaceae in loggerhead sea turtles from Italian coasts

Gastrointestinal investigation of parasites and Enterobacteriaceae in loggerhead sea turtles from Italian coasts

In Silico identification of pathogenic strains of Cronobacter from Biochemical data reveals association of inositol fermentation with pathogenicity

Particular Biochemical Profiles for Enterohemorrhagic Escherichia coli O157:H7 Isolates on the ID 32E System

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