Enterotoxigenic profiles and submerged and interface biofilms in Bacillus cereus group isolates from foods.

Biofilm formation by Bacillus cereus strains is now recognized as a systematic contamination mechanism in foods; the aim of this study was to evaluate the production of submerged and interface biofilms in strains of B. cereus group in different materials, the effect of dextrose, motility, the presence of genes related to biofilms and the enterotoxigenic profile of the strains. We determine biofilm production by safranin assay, motility on semi-solid medium, toxin gene profiling and genes related to biofilm production by PCR in B. cereus group isolated from food. In this study, we observe strains used a higher production of biofilms in PVC; in the BHI broth, no submerged biofilms were found compared to phenol red broth and phenol red broth supplemented with dextrose; no strains with the ces gene were found, the enterotoxin profile was the most common the profile that includes genes for the three enterotoxins. We observed a different distribution of tasA and sipW with the origin of isolation of the strain, being more frequent in the strains isolated from eggshell. The production and type of biofilms are differential according to the type of material and culture medium used.

Genetic diversity of enterotoxigenic Bacillus cereus strains in coriander in southwestern Mexico.

Background: Coriander, like other leafy green vegetables, is available all year round and is commonly consumed raw in Mexico as in other countries in the preparation of street or homemade food. Bacillus cereus (B. cereus) is a microorganism that can reach coriander because it is usually found in the soil and in some regions the vegetables are irrigated with polluted water. Therefore, the aim of this study was to determinate the presence of B. cereus in coriander used for human consumption in southwestern Mexico and determine the toxigenic profile, biofilm production, genes associated with the production of biofilms, sporulation rates, enzymatic profile, psychotropic properties, and genetic diversity of B. cereus. Methods: Fresh coriander samples were collected from several vegetable retailers in different markets, microbiological analysis was performed. Molecular identification, genes related to the production of biofilm, and toxin gene profiling of B. cereus isolates were determined by PCR. The biofilm formation was measured by performing a crystal violet assay. The genetic diversity of B. cereus strains was determined by PCR of repetitive elements using oligonucleotide (GTG) 5. Results: We found a frequency of B. cereus in vegetables was 20% (13/65). In this study, no strains with genes for the HBL toxin were found. In the case of genes related to biofilms, the frequency was low for sipW [5.8%, (1/17)] and tasA [11.7%, (2/17)]. B. cereus strains produce a low amount of biofilm with sporulation rates around 80%. As for genetic diversity, we observed that strains isolated from the same market, but different vegetable retailers are grouped into clusters. In the coriander marketed in southwestern Mexico, were found B. cereus strains with genes associated with the production of diarrheal toxins. Together, these results show actual information about the state of art of B. cereus strains circulating in the southwestern of Mexico.

Biofilm Production by Enterotoxigenic Strains of Bacillus cereus in Different Materials and under Different Environmental Conditions.

Foodborne illnesses, such as infections or food poisoning, can be caused by bacterial biofilms present in food matrices or machinery. The production of biofilms by several strains of Bacillus cereus on different materials under different culture conditions was determined, as well as the relationship of biofilms with motility, in addition to the enterotoxigenic profile and candidate genes that participate in the production of biofilms. Biofilm production of B. cereus strains was determined on five materials: glass, polystyrene, polyethylene, polyvinylchloride (PVC), PVC/glass; in three culture media: Phenol red broth, tryptic soy broth, and brain heart infusion broth; in two different temperatures (37 °C and 25 °C), and in two different oxygen conditions (oxygen and CO2 tension). Furthermore, the strains were molecularly characterized by end-point polymerase chain reaction. Motility was determined on semi-solid agar. The B. cereus strains in this study were mainly characterized as enterotoxigenic strains; statistically significant differences were found in the PVC material and biofilm production. Motility was positively associated with the production of biofilm in glass/PVC. The sipW and tasA genes were found in two strains. The results of this study are important in the food industry because the strains carry at least one enterotoxin gene and produce biofilms on different materials.

Prevalencia de cepas del grupo de Bacillus cereus productoras de biopelicula en helados comercializados en México / Prevalence of strains of the Bacillus cereus group biofilm producers in ice cream in Mexico

El helado es un vehículo para la transmisión de patógenos como Bacillus cereus. Por lo cual, se determinó la frecuencia de cepas del grupo Bacillus cereus en helados, perfil enterotoxigénico, psicrofilia y producción de biopelícula. Un total de 230 muestras de seis marcas de helado de producción y distribución nacional fueron colectadas en México. El análisis microbiológico incluyó aislamiento en agar manitol yema de huevo. Las cepas se identificaron molecularmente a partir de la amplificación del gen de la topoisomerasa (gyrB) y el perfil enterotoxigénico por la amplificación de regiones conservadas de los operones nheABC y hblABD y del gen cytK. Además, se determinó la producción de biopelícula en vidrio y policloruro de vinilo. La frecuencia de contaminación por cepas del grupo B. cereus fue de 3,6%, se encontró una cepa positiva para nheABC y cinco para cytK, el 87,5% de las cepas generó biopelícula en vidrio y todas en policloruro de vinilo, dos cepas fueron psicrofilicas. En conclusión, en el helado distribuido en México, se encontró una baja contaminación por cepas del grupo B. cereus con alta producción de biopelícula; sin embargo, no se debe subestimar el potencial enterotoxigénico de estas cepas

Ice cream is a medium for microbial growth due to its nutritional value and neutral pH. Therefore, the frequency of strains of the Bacillus cereus group in ice cream was determined, the enterotoxigenic profile, psychrophilic strains and biofilm production. A total of 230 samples of six brands of ice cream produced and distributed nationwide were collected in Mexico. The microbiological analysis was a cold pre-enrichment and isolation of the microorganism in egg yolk agar. The strains were identified molecularly from the amplification of the topoisomerase gene (gyrB) and the enterotoxigenic profile by the amplification of conserved regions of the nheABC and hblABD operons and of the cytK gene. In addition, the production of biofilm in glass and polyvinyl chloride and psychophilia was determined. The frequency of contamination by strains of the B. cereus group was 3.6%, a positive strain was found for nheABC and five for cytK, 87.5% of the strains generated biofilm in glass and all in polyvinyl chloride, two strains were psychrophilic. In the ice cream distributed in Mexico, a low contamination by strain of the B. cereus group was found, however, the enterotoxigenic potential of the strains should not be underestimated

Methicillin-Resistant Staphylococcus aureus (MRSA) in Artisanal Cheeses in México.

Milk and dairy foods have frequently been implicated in staphylococcal food poisoning, and contaminated raw milk is often involved. The aim of the study was to determine the occurrence of methicillin-resistant Staphylococcus aureus (MRSA) in raw cow milk cheese produced in Mexico. A total of 78 unpasteurized cow milk cheese samples were screened for S. aureus. The isolates were identified as S. aureus based on morphology, Gram stain, catalase test, coagulase test, and mannitol salt agar fermentation. Isolates were subjected to biotyping, the methicillin resistance was analyzed using the disk diffusion, and the Staphylococcus enterotoxin A (SEA) production was examined by a dot-blot analysis. From a total of 78 samples of unpasteurized cheeses analyzed in this study, 44 cheeses were positive for S. aureus; however, a differential contamination between the different types of cheeses was observed, with high risk of contamination in adobero cheese (12, 95% CI 1.75 to 94.20; p=0.002). In this study, the frequency of methicillin-resistant Staphylococcus aureus (MRSA) was 18.1% (8/44) and of enterotoxin A producers was 18.1% (8/44). When classified by biotypes, MRSA only belongs to the human ecovar biotype (2/8, 25%) and the D biotype (4/8, 50%). S. aureus producers of enterotoxin A were distributed in specific nonhost biotypes.