Plant protection products based on Bacillus thuringiensis have been used to fight agricultural pests for decades and are the world's most frequently applied biopesticide. However, there is growing concern that B. thuringiensis residues in food may occasionally cause diarrheal illness in humans. This has recently sparked a plethora of research activities and vivid discussions across the scientific community, competent authorities, and the public. To support this discussion, we provide a structured overview of the current knowledge on the role of B. thuringiensis as a causative agent of foodborne infections in humans and pinpoint research gaps that need to be addressed for improved risk assessment. We review (i) recent taxonomic changes in the B. cereus group; (ii) the role of B. thuringiensis in transforming agrosystems; and (iii) key considerations for assessing the hazard potential of B. thuringiensis strains detected in foods. We conclude that (i) the taxonomy of the B. cereus group is collapsing, (ii) B. thuringiensis based biopesticides play a key role in realizing the UN's sustainable development goals, and (iii) risk assessment needs to move from taxonomy-driven considerations to strain-specific identification of virulence and pathogenicity traits We also provide an overview of relevant risk-related data for commonly used biopesticide strains.

The present study investigated the microbiological safety of the increasingly popular plant‐based milk alternatives. No (10/27) or only very low microbial counts (17/27) were detected in the tested products. These were mainly identified as spore formers via MALDI‐ToF‐MS. Three products contained Bacillus cereus group isolates, which were able to form considerable amounts of enterotoxins and exhibited cytotoxicity towards CaCo‐2 cells. Preliminary tests showed good growth of B. cereus , Listeria monocytogenes , and Salmonella enterica in all tested products (maximum bacterial counts: 5 × 10 ¹² cfu/mL). These experiments also revealed strain‐, time‐, and temperature‐, but especially product‐specific enterotoxin production of B. cereus . In propagation and persistence tests according to DIN EN ISO 20976‐1:2019‐09, rapid bacterial proliferation was also detected in all products. B. cereus generally showed lower bacterial counts (10 ⁶ –10 ⁷ cfu/mL) compared to L. monocytogenes and S. enterica (10 ⁸ –10 ⁹ cfu/mL), but was detectable in a larger number of products over the test period of 6 weeks. pH values decreased (20/27) over time and visual and/or olfactory alterations (24/27) were observed. The present study provides information on the occurrence, growth and persistence of pathogenic bacteria in plant‐based drinking milk alternatives. It also points out that the accompanying changes in pH, odor, and appearance are not necessarily recognizable to the consumer. Practical Application: The present study contributes to the understanding of the microbial risk related to plant‐based drinking milk alternatives. It is crucial that the manufacturer ensures that particularly spore formers have been effectively eliminated from the products. Among them, especially toxin‐producing bacteria can pose a risk to the consumer, as these products promote proliferation and persistence of the bacteria.

The genes hblC, hblD and hblA encode the components Hbl L2, L1 and B of the pore forming enterotoxin haemolysin BL of Bacillus cereus. Two variants of the operon existand the more common one additionally contains hblB downstream of hblCDA. Up to now, it was completely unclear whether the corresponding protein, Hbl B’, is widely expressed among B. cereus strains and if it has a distinct function. In the present study, it was shown that the hblB gene is indeed expressed and the Hbl B’ protein is secreted by nearly all analysed B. cereus strains. For the latter, a detection system was developed based on monoclonal antibody 11A5. Further, a distinct reduction of cytotoxic and haemolytic activity was observed when recombinant (r)Hbl B’ was applied simultaneously with L2, L1 and B. This effect was due to direct interaction of rHbl B’ with L1. _D⁻⁶B. cereusAltogether, we present the first simple tool for the detection of Hbl B’ in B. cereus culture supernatants. Moreover, an important regulatory function of Hbl B’ in the mechanism of Hbl was determined, which is best described as an additional control of complex formation, balancing the amounts of Hbl B-L1 complexes and the corresponding free subunits.

The iron-binding glycoprotein lactoferrin is well known for its wide range of antibacterial effects. However, the aim of this study was to show that its antibacterial activity is not generally applicable to a bacterial species as a whole. In disk diffusion assays performed with 112 isolates from 13 bacterial species (including the foodborne pathogens Bacillus cereus and Staphylococcus aureus), a lactoferrin-based food supplement showed no inhibition of growth on 24%, moderate inhibition on 31%, and strong inhibition on 45% of all tested isolates. Minimal inhibitory concentrations against B. cereus and Bacillus thuringiensis strain-specifically ranged from 0.31 mg/mL to no impairment at all. Further 11 commercially available lactoferrin-based food supplements and purified bovine lactoferrin showed strain- as well as product-specific growth inhibition. In comparison to bovine lactoferrin, human lactoferrin showed no inhibitory effects. In summary, purified lactoferrin and lactoferrin-based food supplements inhibit bacterial growth in a dose-, strain-, and product-dependent manner. Thus, a general antimicrobial effect of lactoferrin against a specific bacterial species cannot be assumed.

Cow’s milk contains A1- and A2-β-caseins. The breakdown of A1-β-casein produces β-casomorphin-7 (BCM-7), a peptide with opioid-like properties that is associated with health aspects. In addition, A1- and A2-β-casein have different technological properties. The aim of the present study was to investigate whether cheese produced from the milk of homozygous A1A1 and A2A2 cows varies in terms of its physicochemical parameters and BCM-7 concentration. These parameters were analyzed during initial cheese processing, six weeks of ripening and 84 days of storage, including additional microbiological analyses during the storage period. The pH values of the A1A1 cheeses were higher than those of the A2A2 cheeses from the beginning of production until the starter culture bacteria were added. The yellowness values of the A1A1 cheeses were lower until the salt bath treatment. Water activity, lightness, hardness, fat, protein, NaCl and dry matter content, as well as color and microbiological parameters, were not affected by the β-casein genotype. BCM-7 concentrations were higher in the A1A1 cheeses after pressing and during ripening. We found mainly comparable quality characteristics and slightly different BCM-7 levels in the A1A1 and A2A2 cheeses. From this point of view, both varieties are equally suitable for cheese production.

One of the multiple factors determining the onset of the diarrhoeal disease caused by enteropathogenic Bacillus cereus is the ability of the bacteria to actively move towards the site of infection. This ability depends on flagella, but it also varies widely between different strains. To gain more insights into these strain-specific variations, polyclonal rabbit antisera as well as monoclonal antibodies (mAbs) were generated in this study, which detected recombinant and natural B. cereus flagellin proteins in Western blots as well as in enzyme immunoassays (EIAs). Based on mAb 1A11 and HRP-labelled rabbit serum, a highly specific sandwich EIA was developed. Overall, it could be shown that strain-specific swimming motility correlates with the presence of flagella/flagellin titres obtained in EIAs. Interestingly, mAb 1A11, recognizing an epitope in the N-terminal region of the flagellin protein, proved to inhibit bacterial swimming motility, while the rabbit serum rather decreased growth of selected B. cereus strains. Altogether, powerful tools enabling the in-depth characterization of the strain-specific variations in B. cereus swimming motility were developed.