While the relevance of Bacillus (B.) cereus as a major cause of gastroenteritis is undisputed, the role of the closely related B. thuringiensis in foodborne disease is unclear. B. thuringiensis strains frequently harbor enterotoxin genes. However, the organism has only very rarely been associated with foodborne outbreaks, possibly due to the fact that during outbreak investigations, B. cereus is routinely not differentiated from B. thuringiensis. A recent EFSA scientific opinion stresses the urgent need for further data allowing for improved risk assessment, in particular as B. thuringiensis is a commonly used biopesticide. Therefore, the aim of this study was to gain further insights into the hazardous potential of B. thuringiensis. To this end, 39 B. thuringiensis isolates obtained from commercially used biopesticides, various food sources, as well as from foodborne outbreaks were characterized by panC typing, panC-based SplitsTree analysis, toxin gene profiling, FTIR spectroscopic analysis, a cytotoxicity assay screening for enterotoxic activity, and a sphingomyelinase assay. The majority of the tested B. thuringiensis isolates exhibited low (23%, n = 9) or mid level enterotoxicity (74%, n = 29), and produced either no (59%, n = 23) or low levels (33%, n = 13) of sphingomyelinase, which is reported to act synergistically with enterotoxins Nhe and Hbl. One strain isolated from rosemary was however classified as highly enterotoxic surpassing the cytotoxic activity of the high-level reference strain by a factor of 1.5. This strain also produced vast amounts of sphingomyelinase. Combining all results obtained in this study into a fingerprint pattern, several enterotoxic biopesticide strains were indistinguishable from those of isolates from foods or collected in association with outbreaks. Our study shows that many B. thuringiensis biopesticide strains exhibit mid-level cytotoxicity in a Vero cell assay and that some of these strains cannot be differentiated from isolates collected from foods or in association with outbreaks. Thus, we demonstrate that the use of B. thuringiensis strains as biopesticides can represent a food safety risk, underpinning the importance of assessing the hazardous potential of each strain and formulation used.
To determine antimicrobial drug resistance mechanisms of Shigella spp., we analyzed 344 isolates collected in Switzerland during 2004–2014. Overall, 78.5% of isolates were multidrug resistant; 10.5% were ciprofloxacin resistant; and 2% harbored mph(A), a plasmid-mediated gene that confers reduced susceptibility to azithromycin, a last-resort antimicrobial agent for shigellosis.
Mashed potato powder as well as powdered infant formula (PIF) are frequently contaminated with Bacillus cereus sensu lato (B. cereus s.l.), mainly with its spores. These products have also been implicated in foodborne illnesses. Here, we characterized B. cereus s.l. isolates originating from powdered products based on sporulation assays, toxin gene profiling, and panC typing combined with a SplitsTree analysis. Furthermore, cytotoxicity assays with B. cytotoxicus isolates were performed. 78% of PIF tested positive for B. cereus s.l., whereas 92% of all mashed potato powders were positive. In total, 43 isolates were further characterized. The nhe and cytK2 genes were most frequently detected. Moreover, a cereulide-producer was detected from PIF. Most isolates were assigned to panC group III, but members of group II, IV, V, and VII could also be found. Nine B. cytotoxicus were isolated out of nine mashed potato powders. All panC group VII isolates were positive for cytK1. Cytotoxicity assays of these nine isolates revealed one highly cytotoxic strain, while all other isolates exhibited no detectable cytotoxicity, underpinning that cytotoxicity of a certain B. cereus group strain cannot be deduced from the sole presence or absence of toxin genes.
Heated food is known to be often contaminated with B. cereus, leading to cases of diarrhoeal or emetic diseases. Battalion kitchens or army catering facilities present a food safety risk, as temperature abuse and long storage time can result in serious public health problems affecting a high number of served people. In contrast to civil catering facilities, no microbiological monitoring systems are currently implemented in Swiss military kitchens. In this study toxin gene profiles and cytotoxicity levels of 21 isolates of B. cereus originating from six different food categories were determined. Nearly all isolates (95%) harbored the nhe gene, whereas no hbl could be detected. Seven isolates displayed the cytK2 gene and one cereulide-producer was isolated out of vegetables. While most isolates displayed low cytotoxicity, highly cytotoxic strains were detected, with three isolates even exceeding the cytotoxicity level of the reference strain for high-level toxin production, underpinning that cytotoxicity cannot be deduced only from presence or absence of toxin genes. These findings further underline the importance of rapid cooling of foods or maintenance over 65°C before serving. This is especially important in mass catering facilities, such as military kitchens, in which food is often prepared a long time in advance.
In contrast to Bacillus cereus, the role of Bacillus thuringiensis in foodborne illness has been controversially discussed. As B. thuringiensis–based biopesticides containing a mixture of crystal toxins and viable spores are widely used, a current European Food Safety Authority opinion underlines the need for additional data to enable risk assessment. However, it is currently poorly understood if B. thuringiensis is able to multiply in food, which is crucial to sound risk assessment. Therefore, the aim of this study was to investigate growth of selected B. thuringiensis strains from food and insecticides in a ratatouille food model. To this end, the growth parameters of three B. thuringiensis strains were determined: insecticide strain ABTS-351 (CH_119, B. thuringiensis serovar kurstaki), insecticide strain ABTS-1857 (CH_121, B. thuringiensis serovar aizawai), and CH_48 (wild-type B. thuringiensis isolated from rosemary), producing extremely high levels of enterotoxins. After an initial drop in colony counts, we observed a statistically significant growth for the tested B. thuringiensis strains between 6 and 24 h at 22, 30, and 37°C, conditions mimicking prolonged holding times. We were also able to show that the enterotoxin overproducer CH_48 can grow up to 108 CFU/g in the ratatouille matrix within 24 h at 37°C. The two midlevel enterotoxin formers ABTS-351 (CH_119) and ABTS-1857 (CH_121) isolated from biopesticides exhibited growth between 6 and 24 h, with one of the strains growing to 107 CFU/g. To our knowledge, this is the first study providing evidence of B. thuringiensis growth in a food model with intact competitive flora. Our findings suggest strain-specific variation and stress the complexity of assessing the risk related to B. thuringiensis in food, indicating that some strains can represent a risk to consumer health when vegetable-based foods are stored under conditions of prolonged temperature abuse. HIGHLIGHTS
scite is a Brooklyn-based organization that helps researchers better discover and understand research articles through Smart Citations–citations that display the context of the citation and describe whether the article provides supporting or contrasting evidence. scite is used by students and researchers from around the world and is funded in part by the National Science Foundation and the National Institute on Drug Abuse of the National Institutes of Health.
hi@scite.ai
10624 S. Eastern Ave., Ste. A-614
Henderson, NV 89052, USA
Copyright © 2024 scite LLC. All rights reserved.
Made with 💙 for researchers
Part of the Research Solutions Family.