Molecular characterization of an unauthorized genetically modified Bacillus subtilis production strain identified in a vitamin B 2 feed additive

Abstract

HighlightsGenetically modified Bacillus subtilis identified in a vitamin B2 product.Whole genome sequencing runs are performed for characterization of the isolated strain.Complex modifications of the genome are identified.Four putative recombinant plasmids are characterized.Real-time PCR methods are developed and available for testing vitamin B2 products.

EFSA statement on the risk posed to humans by a vitamin B2 produced by a genetically modified strain of Bacillus subtilis used as a feed additive

“…Additional information regarding the analysis linked to notification 371917 from the laboratory that performed the analysis and a scientific publication regarding the methods used to analyse the samples (Paracchini et al, 2017) were also made available to EFSA. The documents are related to Rapid Alert System for Food and Feed (RASFF) notification number 371917, from 2 October 2018 and reference 2018.2755, and follow-up notifications received until 7 January 2019.…”

“…The samples were analysed for the detection of recombinant DNA of B. subtilis with two different methods which are described in two publications Paracchini et al (2017) and Barbau-Piednoir et al (2015), respectively. Both methods were developed to detect the presence of a non-authorised genetically modified strain of B. subtilis found in 2014 by the French competent authorities in samples of vitamin B 2 produced by another company, and for which a RASFF notification was raised at that time (RASFF 2014(RASFF .1249).…”

“…Both methods were developed to detect the presence of a non-authorised genetically modified strain of B. subtilis found in 2014 by the French competent authorities in samples of vitamin B 2 produced by another company, and for which a RASFF notification was raised at that time (RASFF 2014(RASFF .1249). This strain was characterised by whole genome sequence analysis by the German authorities and the JRC (Paracchini et al, 2017). Those detection methods are based on realtime PCR (limit of detection 95% = 10 copies) using primers which target the following fragments:…”

“…The only information available is that the strain CGMCC13326 shares the insertion of the chloramphenicol resistance gene in the chromosome and the riboflavin operon in a pSM19035derived vector with the strain characterised and sequenced by Paracchini et al (2017). The only information available is that the strain CGMCC13326 shares the insertion of the chloramphenicol resistance gene in the chromosome and the riboflavin operon in a pSM19035derived vector with the strain characterised and sequenced by Paracchini et al (2017).…”

“…The European Commission specifically referred to the analytical reports linked to notifications 371917 (vitamin B 2 samples) and follow-up 38 for notification 371917 (ref 379444, referring to feed 2 ). Additional information regarding the analysis linked to notification 371917 from the laboratory that performed the analysis and a scientific publication regarding the methods used to analyse the samples (Paracchini et al, 2017) were also made available to EFSA. EFSA also considered the publication from Barbau-Piednoir et al (2015) that regards one of the analytical methods used.…”