Global overview of the risk linked to the Bacillus cereus group in the egg product industry: identification of food safety and food spoilage markers.

AIMS: To evaluate the food safety and spoilage risks associated with psychrotrophic Bacillus cereus group bacteria for the egg product industry and to search for relevant risk markers. METHODS AND RESULTS: A collection of 68 psychrotrophic B. cereus group isolates, coming from pasteurized liquid whole egg products, was analysed through a principal component analysis (PCA) regarding their spoilage and food safety risk potentials. The principal component analysis showed a clear differentiation between two groups within the collection, one half of the isolates representing a safety risk and the other half a spoilage risk. CONCLUSIONS: Relevant risk markers were highlighted by PCA, that is (i) for the food safety risk, the presence of the specific 16S rDNA-1m genetic signature and the ability to grow at 43°C on solid medium and (ii) for the spoilage risk, the presence of the cspA genetic signature. SIGNIFICANCE AND IMPACT OF THE STUDY: This work represents a first step in the development of new diagnostic technologies for the assessment of the microbiological quality of foods likely to be contaminated with psychrotrophic B. cereus group bacteria.

Biodiversity of psychrotrophic bacteria of the Bacillus cereus group collected on farm and in egg product industry.

The aim of the present study was (i) to type, by genotypic and phenotypic methods, a collection of psychrotrophic bacteria belonging to the Bacillus cereus group collected in a farm and in 6 egg breaking industries during a period covering a warm and a cold season, and (ii) to characterize the egg product spoilage (growth in liquid whole egg) and the sanitary risk potential (cytotoxic activity on Caco-2 cells and adhesion on stainless steel) of each isolate of the collection. The investigation of specific psychrotrophic and mesophilic signatures together with the study of ability to grow at 6 °C and/or at 43 °C on optimal agar medium allowed highlighting twelve profiles, the major one corresponding to the species Bacillus weihenstephanensis (46.2% of the collection). The diversity of the profiles depended on the season and on the origin of the isolates. In terms of food spoilage, all the isolates were able to grow at the same level in liquid whole egg and in optimal medium, even at low temperature. Under the same conditions, the cytotoxic activity depended on the isolate, the medium and the temperature. At 10 °C, no isolate was cytotoxic at 10 °C in liquid whole egg and only one, belonging to the Bacillus weihenstephansensis species, in the optimal medium. All the isolates were able to adhere on stainless steel at various levels, from 2.6±0.2 log cfu/cm(2) to 4.9±0.1 log cfu/cm(2). A large majority (80.8%) was strongly adhering and could lead to the formation of biofilms in industrial equipments.

A single amino acid, outside the AlcR zinc binuclear cluster, is involved in DNA binding and in transcriptional regulation of the alc genes in Aspergillus nidulans.

In Aspergillus nidulans, the transcriptional activator AlcR mediates specific induction of a number of alc genes. The AlcR DNA-binding domain is a zinc binuclear cluster that differs from the other members of the Zn2Cys6 family in several respects. Of these, the most remarkable is its ability to bind in vitro as a monomer to single sites, whereas only repeated sites (direct or inverted) are necessary and functional in vivo. Deletion of the first five amino acids (following the N-terminal methionine) upstream of the AlcR zinc cluster or mutation of a single residue, Arg-6, impairs the AlcR in vitro binding mainly to symmetrical sites. In vivo, the same mutations result in the inability of A. nidulans to grow on ethanol. The alc- phenotype results from a drastic decrease in activation of its own transcription and, in addition, that of the two structural genes, alcA and aldA, required for ethanol oxidation. This defect seems to be correlated to the inability of the Arg-6 AlcR mutant protein to bind to AlcR palindrome targets, which are essential in the three alc promoters. AlcR shows a unique pattern of binding and of transactivation among the Zn2Cys6 family.

Incorporation of 12(S)-hydroxyeicosatetraenoic acid into phospholipids and active diacylglycerols in rat liver epithelial cells: effects on DNA synthesis.

12(S)-Hydroxyeicosatetraenoic acid (12-HETE), the 12-lipoxygenase-derived metabolite of arachidonic acid, was incorporated into membrane phospholipids (PL) in various cells. PL are precursors of diacylglycerol (DAG), a protein kinase C (PKC) activator involved in cell-growth signaling. We studied 12-HETE incorporation into PL in non-transformed (NT-) and spontaneously transformed (T-) rat liver epithelial cells (RLEC), and its consequence on DNA synthesis. NT- and T-RLEC incorporated 12-HETE predominantly into phosphatidylcholine (PC). 12-HETE was incorporated at a greater rate, and with a higher phosphatidylethanolamine (PE)/PC ratio in T-cells. Preincubation of RLEC with 12-HETE at > or = 0.5 microM partially inhibited basal DNA synthesis in NT- and T-RLEC. Preincubation of NT-RLEC with 12-HETE (2.5 microM) also decreased bradykinin-stimulated DNA synthesis. Unstimulated RLEC produced 1-acyl-2-(12-HETE)DAG which was increased in NT-RLEC by bradykinin stimulation. Finally, 1-stearoyl-2-(12-HETE)DAG was as potent an in vitro PKC activator as 1-stearoyl-2-arachidonyl-DAG. These data demonstrate that 12-HETE incorporation into PL resulted in the production of active 12-HETE-containing DAG, together with reduced DNA synthesis.

Incorporation of arachidonic and eicosapentaenoic acids into phospholipids of non-transformed and spontaneously-transformed rat liver epithelial cells: effects on DNA-synthesis.

We compared the incorporation of arachidonic acid (AA) and eicosapentaneoic acid (EPA) into phospholipids of non-transformed (NT-) and spontaneously-transformed (T-) rat liver epithelial cells (RLEC), and their consequences on DNA-synthesis. In NT-cells, both radioactive fatty acids were preferentially incorporated into phosphatidylcholine (PC). In T-cells, in contrast, AA was predominantly incorporated into phosphatidylethanolamine (PE), whereas EPA remained preferentially incorporated into PC. After pulse labelling, we observed in both cell types a progressive decrease in AA- and EPA-labelled PC associated with an increase in AA- and EPA-labelled PE. Preincubation of NT-cells with increasing concentrations of AA or EPA (0.1 microM to 20 microM) resulted in a concentration-dependent DNA-synthesis stimulation with a stronger effect of AA compared with EPA. In T-cells, the same treatment had no effect on DNA-synthesis.